Within-group analysis revealed a significant increase in ROM and a significant decrease in medial and lateral gastrocnemius (LG) stiffness for the foam roller group after the intervention (p<0.05). In addition, further analysis of the preintervention data revealed a significant negative correlation between ankle dorsiflexion ROM and AT stiffness (r=−0.378 and p=0.007). These results suggest that self-myofascial release using a foam roller on the calf is an effective method for decreasing the stiffness of the gastrocnemius and increasing ankle dorsiflexion ROM.Increased muscle stiffness can contribute to reduced range of motion (ROM) and impaired function. Reduced ankle dorsiflexion ROM has been associated with increased injury risk in the ankle. Self-myofascial release (SMR) has been widely used in clinical and sports settings, but the effects of SMR on gastrocnemius and Achilles tendon (AT) stiffness are unclear. Therefore, we investigated the effects of self-myofascial release using a foam roller (FR) on the stiffness of the gastrocnemius–AT complex and ankle dorsiflexion ROM. Fifty healthy, untrained, and non-sedentary participants (age=22.5±2.6years) were randomly divided into an intervention group (FR group) and a control group. The subjects in the intervention group received a single foam roller intervention (three sets of 1min), while the subjects in the control group performed a 5-min sedentary rest. Stiffness of the gastrocnemius–AT complex was evaluated using MyotonPRO and the ankle dorsiflexion ROM was assessed using the weight-bearing lunge test. For the foam roller and control groups, the between-group analysis revealed a statistically significant difference in gastrocnemius stiffness and ankle dorsiflexion ROM after intervention ( Stiffness is one of the mechanical properties of muscle that can affect exercise capacity and has been correlated with muscle strain injury risk, especially in high-intensity activities . The musIn recent years, self-myofascial release (SMR) has been widely used in clinical and sports settings. SMR is a self-treatment method involving the application of compressive forces to soft tissue. It claims to mimic the effects of manual therapy techniques and aims to address soft tissue dysfunction . The foain vitro biomechanical testing; however, they can reflect the relative stiffness of soft tissues  the acute effect of self-myofascial release using the foam roller on the stiffness of the gastrocnemius and AT; (2) the acute effect of a foam roller intervention on passive ankle dorsiflexion range of motion; and (3) the associations of passive ankle dorsiflexion ROM with gastrocnemius and AT stiffness.2; (3) no known cardiovascular, pulmonary, or neurological diseases; and (4) no skin lesions in the lower limbs. Participants were excluded if they met the following criteria: (1) acute or chronic musculoskeletal system diseases or neuromuscular diseases; (2) any gastrocnemius or Achilles tendon dysfunction; (3) a history of musculoskeletal injuries to the ankle joint  within the previous 6months; (4) a history of surgery in the lower extremities; or (5) inability to complete the whole experiment. Fifty participants were randomly divided into an intervention group  and a control group .Fifty healthy, untrained, and non-sedentary participants  were recruited for the present study. The inclusion criteria were as follows: (1) age range: 18–35years; (2) Body mass index (BMI) range: 16–28kg/mAbout 1week before the testing sessions, participants attended a standardized familiarization session including the testing procedure and the foam roller intervention to minimize learning effects. All tests were performed in the dominant limb in a room with a temperature of 25°C. The demographic characteristics  and the dominant limb were recorded when participants arrived at the testing area. The dominant limb was defined by the side the participant used when asked to kick a ball . To furtIn this study, FR exercises were conducted on the calf of the subject’s dominant limb with a GRID Foam Roller . According to a previous study, the GRID is composed of a hollow inner core enclosed in a 15mm thick layer of ethylene-vinyl acetate foam, with a total Young’s modulus of 103kPa . For theThe ankle dorsiflexion ROM of the dominant limb was measured using the weight-bearing lunge test . The weiA MyotonPRO  was used to quantify the stiffness of the medial gastrocnemius (MG), lateral gastrocnemius (LG), and AT in the present study. In our previous studies, we found a good intra- and inter-rater reliability (ICC>0.84) for assessing the MG, LG, and AT stiffness using the MyotonPRO . The basThe MG measurement site was located at 30% of the length between the popliteal fossa and lateral malleolus, where cross-sectional areas of the gastrocnemius are almost maximum . LG stif2). A paired t-test was used to examine the differences pre- to postintervention for ROM, gastrocnemius stiffness, and AT stiffness in the control group and intervention group. The differences in gastrocnemius stiffness, AT stiffness, and ankle dorsiflexion ROM between the foam roller and control groups were compared using an independent sample t-test. Pearson correlation analysis (r) was used to analyze the correlation among gastrocnemius stiffness, Achilles tendon stiffness, and ROM. In addition, the effect size was also calculated using Cohen’s d . Descriptive data and all stiffness data are presented as the mean±SD. The Shapiro-Wilk test was used to assess the normal distribution of all data. Homogeneity of variances was evaluated using Levene’s test. BMI was calculated by the following formula: BMI=weight (kg)/height . After the intervention, the LG and MG stiffness in the foam roller group was significantly lower than that in the control group (p<0.05), while there was no statistical difference in AT stiffness between the foam roller and control group (p>0.05). As shown in p>0.05).p<0.05). There was a significant increase in ankle dorsiflexion ROM in the FR group after the intervention . However, for muscle, no significant correlation was apparent between MG stiffness and ankle dorsiflexion ROM (p>0.05). Similarly, there was no significant correlation between LG stiffness and ankle dorsiflexion ROM (p>0.05). After the FR intervention, no significant correlation was apparent between the ankle dorsiflexion ROM and LG, MG, AT in the FR group .The present study investigated gastrocnemius stiffness, AT stiffness, and ankle dorsiflexion ROM before and after an intervention in foam roller and control groups. Accordingly, our results showed a significant decrease in gastrocnemius stiffness and a significant increase in ankle dorsiflexion ROM only in the foam roller group, with no significant change in AT stiffness. In addition, there was no correlation between ankle dorsiflexion ROM and LG and MG stiffness, but a significant negative correlation was observed between ankle dorsiflexion ROM and AT stiffness.The results of this study showed that gastrocnemius stiffness significantly decreases after the foam roller intervention, whereas there was no significant change in the Achilles tendon stiffness. A recent study investigated stiffness variation in the hamstring muscles after performing a foam roller protocol using shear wave elastography, and the authors found a significant decrease in average hamstring muscle stiffness 5min after FR . Reiner The decrease in muscle stiffness is one of the proposed mechanisms for acute morphological responses after SMR . Foam roIn the present study, our findings revealed an increase in ankle dorsiflexion ROM with 11.44% in the foam roller group from pre- to postintervention. There are many theories that attempt to explain the increased ROM after foam roller interventions. Some scholars have suggested that SMR can positively affect fascial sliding properties by eliminating fascial restriction or loosening crosslinks . Kiyono In this study, we found no significant correlation between ankle dorsiflexion ROM and LG or MG stiffness, but ankle dorsiflexion ROM was negatively correlated with AT stiffness. To the best of our knowledge, this is the first study to explore the relationship between ankle dorsiflexion ROM and AT stiffness. We found a negative correlation between ankle dorsiflexion ROM and AT stiffness, but not muscle stiffness. However, the reason for this is still unclear. The possible explanation may due to different the elastic properties of muscle and tendon. Furthermore, ankle dorsiflexion ROM may be associated with various factors . Apart fSome limitations to this study should be recognized. First, we evaluated the stiffness variations in specific regions of the gastrocnemius and Achilles tendon, but this is not representative of changes in other regions. Second, we did not record the variation in stiffness and ankle dorsiflexion ROM with time after the foam roller intervention. In addition, we did not compare the effects of foam roller intervention with other common interventions in clinical practice, such as static stretching. In this study, we measured the stiffness of gastrocnemius and Achilles tendon in the relaxed position of the ankle. However, the resting ankle angle varies from person to person and may have some influence on these results. Finally, we performed only one foam roller intervention protocol and did not explore the effects of different FR intervention intensities on the gastrocnemius-Achilles tendon complex stiffness and ankle dorsiflexion ROM. Therefore, future studies should investigate the effects of different FR durations and frequencies on soft tissue stiffness and ankle dorsiflexion ROM or other functional outcomes.We conclude that a single foam roller intervention on the calf can increase the ankle dorsiflexion ROM and reduce the stiffness of the gastrocnemius. The present study suggests a negative correlation between ankle dorsiflexion ROM and AT stiffness in the healthy population.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.This study was approved by the Human Subjects Ethics Committee of Luoyang Orthopedic Hospital of Henan Province (2019-001-01). Experimental procedures conformed to the Declaration of Helsinki principles and national guidelines. The patients/participants provided their written informed consent to participate in this study.T-TC designed the study and drafted the manuscript. Z-JZ and XQ-W helped to conceive the study. ZL and Z-JZ helped to perform statistical analysis. T-TC, ZL, and Y-CZ participated in the data collection. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Salvelinus alpinus erythrinus from Arctic Canada. The genome sequences were 16,652 bp in length; the genome organization and GC content (45.6%) are consistent with those of charr mitochondrial genomes published previously. The difference between the two genomes studied is low, 0.02%. Our results indicate the phylogenetic closeness of S. alpinus erythrinus and Salvelinus sp. 4 from Lake Nachikinskoe, Kamchatka and their origin from a common ancestor.The complete mitochondrial genomes were sequenced in two individuals of charr  Salvelinus alpinus Linnaeus, 1758  is widely distributed in the northern Holarctic  based on their zoogeographic patterns and morphological characteristics  from Arctic Canada and compared them with genomes of charrs from East Asia to North America to assess the phylogeographic patterns of charr.The Arctic charr S. alpinus erythrinus from Jayko River, Victoria Island, Nunavut, Canada . The fish specimens are stored in the collection of the Genetics Laboratory, NSCMB FEB RAS, Vladivostok, Russia (www.imb.dvo.ru) under accession numbers JAY11.J2 and JAY11.J3. Total genomic DNA was isolated from fin fragments using standard phenol–chloroform extraction and ethanol precipitation methods (Sambrook and Russell Table S1) were designed with the MitoPrimer_V1 . The mitogenomes of S. alpinus from northeastern Québec, Canada  from the Scandinavian Peninsula were included for comparison. Nucleotide content, codon position, the average number of nucleotide substitutions per site between groups , were calculated using MEGA X  genes, 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, a light-strand replication origin (OL), and a control region (CR). The overall base composition was 28.0% A, 26.4% T, 17.0% G, and 28.6% C, and the GC content of the mitogenomes was 45.6%. This is consistent with previous results for charr mitogenomes  and S. alpinus erythrinus (MW664922); the total sequence divergence was 0.000180 ± 0.000097. There were 193 single-nucleotide substitutions  between mitogenome sequences of S. alpinus erythrinus and S. alpinus alpinus, and a length difference due to an insertion (GA) in the non-coding region (OL). Of these, 145 substitutions were found in overall PCGs, and 31 substitutions in CR. A similar distribution of 190 single-nucleotide substitutions was observed when comparing mitogenomes of S. alpinus erythrinus and S. alpinus oquassa (NC_000861): 147 substitutions in PCGs and 29 substitutions in CR. Protein coding genes had a different degree of variability, but variability of the NADH dehydrogenase subunit genes was highest for the mitogenomes compared .The mitogenome of Salvelinus, Parahucho, and Salmo indicates a close affinity of S. alpinus erythrinus to a congeneric species, S. taranetzi (and closely related taxa) (Dxy) between them was 0.00278 ± 0.00033, which is in close agreement with the values of intraspecific divergence previously reported for S. taranetzi  is lowest, 0.0011 ± 0.0002. The S. alpinus erythrinus mitogenomes studied here showed similar sequence divergence (0.0110 ± 0.0007 on average) from the GenBank mitogenomes of S. alpinus alpinus and S. alpinus oquassa (NC_000861). These values correspond to the level of intraspecific variability in the genus ) and North East Asia belong to the same species that was originally described as S. taranetzi Kaganovsky, 1955 from Lake Achchen (Chukotka).The comparison of mitogenomes now obtained with 27 mitogenomes of related groups available in GenBank including genera ed taxa) . The difClick here for additional data file.


In this study, we have analysed the effects of a silane coupling agent on the volume fraction of zirconia for digital light processing (DLP)-based additive manufacturing processes. Zirconia suspension was prepared by the incorporation of silane-modified zirconia particles  or untreated zirconia particles (control group). Furthermore, the control and experimental group were subdivided into three groups based on the volume fraction  of zirconia particles. The disk-shaped zirconia samples were 3D  printed using the DLP technique and their physical and mechanical properties were evaluated. The addition of a silane coupling agent to the zirconia samples was found to have influence of about 6% on the hardness and biaxial flexural strength. Moreover, the decrease in minute air gaps inside the zirconia layers significantly increased the material density . Thus, from this study, it was established that the silane-modified zirconia particles had a positive effect on the physical properties of the zirconia parts. Various materials have been used for prostheses in dental clinics, such as titanium, cobalt–chromium, zirconia, and glass ceramics ,4,5,6,7.Additive manufacturing includes spot scanning-based stereolithography, which uses a stereolithography apparatus (SLA), digital light processing (DLP), and Polyjet printing based on mask image projection . Of thesMethyltrimethoxysilane (MTMS), an organosilane with one methyl group and three hydrolyzable methoxy substituents, has been successfully used to impart hydrophobicity and oleophobicity to a variety of substrates. As previously reported, MTMS controls the hydrolysis conditions, so that the hydrolysis and coating processes are usually in simple ambient aqueous conditions, which represents an obvious advantage and is easily compatible with existing manufacturing processes .In this study, the zirconia suspensions with a volume fraction of 52–56% without added silane were set as the control group , whereasCommercially available zirconia powder  with an average particle size of 310 nm was used. Three monomers , i.e., acrylic resin-based IBA (isobornyl acrylate), HDDA , and PNPGDA (propoxylated neopentyl glycol diacrylate), were utilised to manufacture a photocurable binder. Other additives were a photoinitiator , a dispersant to improve the dispersibility of zirconia powder binder , and a silane coupling agent . MTMS mixed solution was prepared by stirring at a mass ratio of ethanol, distilled water, and MTMS (90:8:2) for 1 h. Then, 100 g of zirconia was mixed with the MTMS mixed solution. After adding the particles, it was ball milled for 24 h and then dried overnight at room temperature. The physical properties of the silane-modified zirconia particles were evaluated using FT-IR spectroscopy . The silane-modified zirconia powders were used to prepare suspensions of zirconia volume fractions of 52, 54, and 56 vol%. A planetary centrifugal mixer  was used to mix the suspension homogeneously. The groups were mainly divided into Z (control group) and ZS ; further, it was sub-divided into three control groups and three experimental groups based on zirconia volume fraction. The zirconia suspension compositions are summarised in A DLP machine was used for light-curing the zirconia suspension  ; the wavThe additively manufactured zirconia samples were degreased and sintered using a sintering machine . The degreasing of each experimental group proceeded as follows. The temperature was gradually increased by 0.5 °C/min until it reached 200 °C and maintained for 1 h. Then, it was increased to 300 °C at 0.5 °C/min and again held for 1 h. Finally, the temperature was incrementally increased to 500 °C at 0.5 °C/min and maintained for 1 h. After the degreasing process, the sintering process proceeded as follows. The temperature was gradually increased to 1450 °C at 10 °C/min and held for 2 h. Then, the samples were naturally cooled  26]. Th. Th26]. n = 10) was measured using a vernier calliper (ASTM C 326: 2009). The diameter and height of samples were measured before and after sintering according to the volume fraction of zirconia. The following formula is used:3) of zirconia before and after sintering, respectively.The linear shrinkage of fabricated zirconia samples (1 is the dry mass (g), m2 is the underwater mass (g), m3 is water mass (g), and ρ(Liq) is the density of liquid infiltration solution (density of water at room temperature = 1.0 g/m3); relative density (%) is calculated by  × 100.The relative density of zirconia samples was measured according to the volume fraction with reference to ISO 18754: 2007. After grinding the surface to reduce the influence of surface roughness, relative density is calculated using the following formula:n = 10). The sample surface was finely ground using diamond pastes with 3 µm and 1 µm particles . The ground samples were pressed five times for 12 s with a force of 1 kgf (9.8 N) using a Micro-Vickers hardness tester  to determine the micro-hardness at each instance.To measure the hardness of the final sintered zirconia samples , these were fixed to multiple diamond plates and ground under constant pressure using grits in the order #400, #800, #1200, and #2400 . The load was increased at a cross-head speed of 1 mm/min until the samples were completely fractured. Calculations are performed using the following formulae:To observe the microstructure of zirconia samples, their surfaces were ground using diamond pastes with 3 µm and 1 µm particles . Then, they were cleaned with alcohol and distilled water in an ultrasonic cleaner  for 20 min. To observe the cross-section, the samples were cleaned with alcohol and distilled water for 20 min. The platinum coating was applied to sample surfaces using a sputter coater . After coating, the zirconia sample layers were observed using a scanning electron microscope .p < 0.05.The sample size was evaluated using G-Power 3.1 software  . The exp−1 and 2900 cm−1, respectively. The bending vibration of –CH is 1405 cm−1. At 1270 cm−1 the stretching vibration of C–O appears, while the stretching vibration of –Si–O– attached to the –CH is at 1052 cm−1, the stretching vibration of –O–O– appears at 890 cm−1. From the results, it was confirmed that the zirconia particles were treated with the silane coupling agent, and a zirconia slurry was prepared using the treated powder.FT-IR spectroscopy was performed to determine the organic characterization, i.e., the effect of silanization of zirconia particles. As shown in The linear shrinkage of additively manufactured zirconia samples was measured according to the zirconia volume fraction by comparing them before and after sintering .In the previous experiment, the shrinkage range of zirconia powder samples sintered at 1200–1400 °C was 19–20%; consequently, the average linear shrinkage of zirconia ceramic was found to be 18–25% . The linThe relative density values of additively manufactured zirconia samples in control groups and experimental groups are shown in p < 0.05). As for the experimental groups, the difference between ZS54 and ZS56 was insignificant, whereas that between these two and ZS52 was significant. Based on the addition of silane coupling agent, the difference between Z52 and ZS52 and that between Z54 and ZS54 did not significantly differ; however, a significant difference between Z56 and ZS56 (p > 0.05) was observed. Regardless of whether the silane coupling agent was added to the samples, the relative density increased as the volume fraction of zirconia increased. However, when a high-volume fraction of zirconia was added, density was further increased with the addition of a silane coupling agent. The silane coupling agent was presumed to enable the homogeneous dispersion of zirconia particles in the suspension. As a result, density was evenly distributed during particle growth [The difference between Z52, Z54, and Z56 was significant . All zirconia samples exhibited significant variations in Vickers average hardness (p < 0.05). Group ZS56, which had a zirconia volume fraction of 56 vol% and contained a silane coupling agent, exhibited the highest hardness value; it was followed by Z56, ZS54, ZS52, Z54, and Z52. The hardness values among these groups significantly differed . Similar to the results of Vickers hardness measurement, the following results are attributed to the commixture of photocuring polymer and zirconia promoted by the addition of a silane coupling agent, thereby increasing dispersibility [The biaxial flexural strengths between Z52 and ZS52, between Z54 and ZS54, and between Z56 and ZS56 significantly differed according to the added silane coupling agent  for DLP additive manufacturing technology has been demonstrated. It was established that the density of the printed zirconia samples significantly increased with an increase in the zirconia volume fraction . Interestingly, the experimental group (ZS56) with the addition of silane coupling agent to the suspension of 56 vol% exhibits improved strength and hardness (5–6%) compared to those without silane coupling agent (control group). However, it is essential to focus future research activities on enhancing the dispersibility of zirconia suspension using the silane coupling agent for DLP printing. In future experiments, we plan to evaluate the dispersion stability of the zirconia suspension by varying conditions, such as pH control, solution composition, and hydrolysis time, for the encapsulation of the zirconia particles with the silane coupling agent.


A critical overview of current approaches to the development of starch-containing packaging, integrating the principles of green chemistry (GC), green technology (GT) and green nanotechnology (GN) with those of green packaging (GP) to produce materials important for both us and the planet is given. First, as a relationship between GP and GC, the benefits of natural bioactive compounds are analyzed and the state-of-the-art is updated in terms of the starch packaging incorporating green chemicals that normally help us to maintain health, are environmentally friendly and are obtained via GC. Newer approaches are identified, such as the incorporation of vitamins or minerals into films and coatings. Second, the relationship between GP and GT is assessed by analyzing the influence on starch films of green physical treatments such as UV, electron beam or gamma irradiation, and plasma; emerging research areas are proposed, such as the use of cold atmospheric plasma for the production of films. Thirdly, the approaches on how GN can be used successfully to improve the mechanical properties and bioactivity of packaging are summarized; current trends are identified, such as a green synthesis of bionanocomposites containing phytosynthesized metal nanoparticles. Last but not least, bioinspiration ideas for the design of the future green packaging containing starch are presented. Politicians, organizations, companies and consumers around the world are increasingly aware that traditional food packaging, especially plastic goods, causes tremendous damage to the environment, water supplies and the entire ecosystem ,2,3,4. TThe term “green” was developed in the 1980s as a business concept to use environmental issues as competitive advantages  defined Green economic examples in food packaging are: limiting the waste creation; maximizing sustainable materials ; use of renewable energy for packaging production . The bigAt the marketing level, GP can be obtained by using adequate logistic techniques , adequaGP can be implemented by applying green management and green politics, directly and indirectly promoted by governments, institutions or companies through adequate laws, regulations, or taxation, and by institutional rules and strategies at company level . The ecoTraditionally, food-packaging materials have been chosen to allow transport and to protect food from environmental interactions . HoweverWhen a packaging material such as a film or a coating is food-derived and is eaten with the food, it is named “edible”; sometimes these two terms—edible films or coatings—are used as synonymous, but they are applied differently ,29. EdibGiven that we are daily attacked by more and more aggressive microorganisms and viruses, a new challenge for the designers of food packages appears: is it possible to support human health through intelligent materials? The same question is current for the specialists in environment. How these goals could be achieved? We aim to provide a first answer to these questions, to show new points of view and to aggregate new ideas to obtain intelligent food packaging to help maintain human and environment health. In this context, sustainable starch-based primary packaging materials can be a reliable solution. Considering the starch-based films and coatings as reference, we propose to start “connecting-the-dots” between the concept of GP and two other green concepts—GC and GT—with a healthy integrative point of view; present work provides an overview of the recent studies regarding:-Bioactive compounds  and their role for the immune system;-GC methods to incorporate bioactive compounds into the starch matrix and their role for the human health;-GT, with emphasis on physical treatments with low environmental impact  on the starch-based packaging with or without incorporated bioactive ingredients, together with a Green Nanotechnology (GN) approach in developing nanomaterials combined with starch for food packaging.Ideas for designing future starch-based packaging are given, too, based on novel materials biocompatible with starch and on new concepts.Among the biopolymers that can be used as food GP materials , starch Starch meets many of the expected features for an ideal candidate material for green packaging ,41, beinThe initial properties of starch can be modified/improved by chemical, physical, biochemical, and genetic actions and/or by their combination . StarcheThe most commonly used methods for producing starch-based films are extrusion and casting; they are described in Starch shows shape memory. Shape-memory materials are advanced biopolymeric materials which undergo a phase transition between an initial temporary phase (leading to temporary shape) and a permanent phase (leading to permanent shape) , when exCurrent research in the field of starch-based films is oriented in the following directions: improving the green production technology ; applyinConsidering starch-based packaging, a challenging issue is to make it a smart packaging material by identifying the adequate reason and technological solution. In the following section we focus on plant-derived bioactive compounds  which coPlant-derived bioactive compounds such as vitamins, polyphenols, essential oils, minerals, amino acids and lipids are all important for human health. Vitamins A, B, C, D and E are beneficial to the immune system. Vitamins A, C and E are responsible for maintaining the epithelial  and mucoB-Complex vitamins improve the immune response through their antioxidant effect and protect the body from diseases by stimulating the immunity . VitaminVitamin C has beneficial effects on the cellular functions of the innate and adaptive immune system. It also possesses an immunomodulatory effect, and it is very effective antioxidant due to its capacity to donate electrons. Therefore, it protects some biomolecules  against oxidants provided by the normal cellular metabolism or oxidants which come from hypoxia and/or exposure to toxins and pollutants (for example smoking). It stimulates the migration of neutrophils to the site of infection and increases phagocytosis. Vitamin C protects the host cell from excessive damage by increasing neutrophil apoptosis, macrophage elimination and decreasing neutrophil necrosis. Increasing the function of the immune cells by vitamin C leads to prevention and treatment of respiratory and systemic infections . Vitamin+T lymphocytes [Vitamin D modulates innate antimicrobial and adaptive (acquired) immune responses. Vitamin D deficiency increases susceptibility to infections . The vitphocytes . The diephocytes , fish, mphocytes .Vitamin E increases humoral and cellular immunity and phagocytic functions. Vitamin E has a greater effect in infectious diseases involving phagocytosis but is less effective in cases of cell-mediated immune defense. Studies show that a daily intake of 200 mg of vitamin E increases antibody responses to various vaccines in healthy subjects. Daily supplementation may rise the efficiency of the immune response to a specific antigen. Vitamin E also increases resistance to viral infections in the elderly people. Vitamin E is exclusively obtained from diet, and the richest sources of it are the vegetable oils , nuts, seeds (sunflower), almonds, hazelnuts, vegetables with green leaves , cereals, peanut butter, kiwi, mango and tomatoes .Polyphenols are secondary plant metabolites, which have aromatic rings and hydroxyl groups in their chemical structure, and are found in various parts of plants, like leaves, fruits, seeds, wood and root . The claPolyphenols are traditionally extracted from plants by infusion, decoction, heat reflux, maceration, percolation or Soxhlet extraction, but these methods require a large amount of solvents and a very long extraction time. New technologies for the extraction of polyphenols are based on the use of microwaves, ultrasound, pulsed electric field and enzyme-assisted methods , and thePolyphenols have different beneficial effects for humans, including immunomodulatory , antioxiApiaceae (Umbelliferae), Asteraceae (Compositae), Cupressaceae, Hypericaceae, Lamiaceae, Lauraceae, Fabaceae, Liliaceae, Myrtaceae, Pinaceae, Piperaceae, Rosaceae, Rutaceae, Santalaceae, Zingiberaceae and Zygophyllaceae families [Essential oils (EOs) are secondary plant metabolites that contain mixtures of lipophilic and volatile compounds with an aromatic odor. EOs have a complex composition, in which terpenes (mono- and sesquiterpenes) and their oxygenated derivatives predominate ,86. Theyfamilies . Essentifamilies . EOs arefamilies , Soxhletfamilies , and colfamilies . Qualitafamilies . Origanum vulgare L. volatile oil showed antimicrobial and antioxidant effects. The antioxidant effect of volatile oregano oil is due to rosmarinic acid [Escherichia coli, Salmonella typhimurium, Bacillus cereus, Pseudomonas aeruginosa and Staphylococcus aureus [S. aureus and E. coli [Laurus nobilis and Myrtus communis volatile oils have shown antibacterial effects against S. aureus [Due to their antioxidant and antimicrobial activity, EOs are gaining increasing attention from the food industry , mostly nic acid . Cinnamonic acid . Thymol nic acid . Some res aureus . Volatil E. coli . Laurus . aureus . Due to . aureus .Minerals are inorganic compounds grouped in three classes, depending on the amount required by the body: macroelements , microelements  and ultramicroelements . Ca maintains the proper functioning of intracellular communication, the skeletal and muscular systems, it has a role in blood clotting, and participates in the activation of several enzymes . Along wAmino acids (AAs) are the building blocks of proteins, and they are essential nutrients in our diet, playing an absolute critical role in many cellular and organismal functions. For example, tyrosine (Tyr) helps iron assimilation , and theEuphausia superba) is rich in the long-chain n-3 PUFAs eicosapentaenoic acid  and docosahexaenoic acid , which have immunomodulatory and anti-inflammatory properties [2 and CO2 [Lipids are a category of hydrophobic or amphiphilic molecules including: fatty acids, mono-, di- and triglycerides, animal and vegetable oils and fats, fat-soluble vitamins, sterols, phospholipids, lecithins, sphingolipids, natural resins, waxes . They aroperties . Moreoveoperties . Taken t and CO2 . It has  and CO2 . Liposomes, lipid entities that mimic biomembranes, are used to build-up bionanomaterials with huge potential applications in the biomedical field and food industry . Thus, improved biocompatibility, antioxidant, antimicrobial and anticancer activities were achieved by incorporation of active phytoingredients, drugs and phytonanometals into biomimicking lipid bilayers ,119,120.There has been reported a trend of the incorporation of total or fractionated plant extracts into food packaging since these substances contain complex matrixes of bioactive ingredients  with synergistic actions in the context of properties like augmented antioxidant, antimicrobial, anticancer properties, and immunity boosting. Thus, an active plant extract-based food packaging is expected to facilitate the relocation of bioactive compounds from the packaging material to the food, preventing the oxidative damage of the foodstuff, and avoiding the food spoilage by reducing microbial contamination. The antimicrobial action of plant extracts is also attributed to the presence of phenolic compounds that have hydroxyl groups (bound to a benzene ring) and a system of delocalized electrons . FurtherGC was defined by the Environmental Protection Agency (EPA) as “the design, development, and implementation of chemical products and processes to reduce or eliminate the use and generation of substances hazardous to human health and the environment” . The GC Incorporating bioactive compounds into the starch-based films agrees with the GC aspects related to the economy, safe materials and minimizing the pollution. The literature research in the last few years on publications presenting starch-based films and coatings incorporating vitamins, polyphenols, essential oils, minerals, amino acids, peptides, proteins and enzymes, lipids and lipids-based nanostructures, together with plant extracts containing bioactive compounds, is presented below.Different techniques, such as casting and extrusion, can be used to produce starch-based edible films incorporating biocompounds. The extrusion process of edible film production does not use liquid solvents, such as water or alcohol; whereas for the dry process, heat is applied to the film-forming materials in order to increase the temperature needed to overcome the melting point of the film-forming starchy materials, thus causing their flow . ExtrusiThe casting process uses solvents for the dispersion of film-forming materials, followed by drying to remove the solvent and form a film structure. The film-forming dispersions should be applied to flat surfaces using a sprayer, spreader, or dipping roller, and dried to eliminate the solvent, forming a film structure ,130. CasThe research in incorporating vitamins into starch-based films and coatings is just beginning. The incorporation of the vitamins which possess antioxidant effect may be done during the manufacturing process because they protect against the film’s oxidation . The mosOther researchers evaluated the properties of rice starch film with ascorbic acid .Maranta arundinaceae L. can be used in packaging fruit stripes as a source of nutrients or as coating for sushi [Ascorbic acid film obtained by incorporating cranberry powder into the starch matrix of or sushi .S. aureus and E. coli [They are many researches on adding polyphenols into biopolymers-based films. The alginate films with polyphenols from tea showed antioxidant and anti-inflammatory effects . Polyphe E. coli . The addition of EOs to food packaging can increase the shelf life and quality of food by protecting the consumer from the harmful effects of oxidative stress and microorganisms. The introduction of EOs in food packaging may change the characteristics of the films used as packaging and this influences the acceptance of the product by the consumer. The frequent use of food packaging with essential oils can lead to hypersensitivity reactions .S. aureus, E. coli, L. monocytogenes) [Botryodiplodia theobromae Pat. and Colletotrichum gloeosporioides Penz. [Nisar et al. investigated the properties of citrus pectin films with clove bud EO. This study showed that the dispersion of clove oil in the films led to an increase in the water barrier and opacity of the films. The films with clove volatile oil exhibited antioxidant activity. Clove bud EO incorporated in citrus pectin films showed inhibitory effect against some foodborne pathogens (togenes) . In anotes Penz. . Eugenoles Penz. . Other res Penz. . Anotheres Penz. .EOs can be easily fabricated as microcapsules and nanoparticles, which increases their stability and solubility. Hence, EOs are considered as the most usable additives in future functional foods .Minerals form complex starch structures through coordinative bonds that are established between the hydroxyl groups of starch and the metal ion. During the heating of the mixture of starch and minerals, an increase in viscosity was observed; therefore the time required for the drying of the films is prolonged. Iron and manganese ions give a yellow-brown color to starch films. Manganese and zinc ions increase the ability of films to retain water. It has been observed that the incorporation of minerals into the starch matrix leads to a decrease in water vapor pressure and an increase in tensile strength (TS) .2) and titanium dioxide (TiO2) have antimicrobial activity, but they also protect against UV radiation. Zinc oxide and magnesium oxide are the most suitable and safest mineral compounds used for food packaging [The incorporation of metals, metal oxides or organic compounds into films can increase their antimicrobial activity. Magnesium oxide (MgO), zinc oxide (ZnO), silicon dioxide , aspartic acid (Asp) and arginine (Arg)) could modify the physicochemical properties of starch gels , while neutral amino acids (methionine (Met) and phenylalanine (Phe)) did not induce modifications .Proteins are polymers of AAs linked together by peptide bonds in a specific spatial architecture providing unique biological function. These valuable biopolymers are exploited in developing “green” starch-based packaging materials. The most commonly used proteins in food industry are those derived from milk, whey, eggs, soybean, wheat, corn, sunflower, peanut, cottonseed, rice, fish, silk, etc. ,109. Whey proteins–starch films embedded with rambutan peel extract and cinnamon oil exhibited antioxidant and antibacterial activities, and also improved mechanical and barrier properties . 2 and CO2 but do not resist water diffusion. Moreover, films made of proteins and carbohydrates are excellent barriers to O2, since the formation of ordered H-bonded networks between the two types of biopolymers [The films based on proteins provide a barrier for Opolymers . The addpolymers  to extenpolymers . Starch–proteins blends processed in the presence of plasticizers lead to obtaining biodegradable plastics with enhanced properties. Compared with animal proteins, plant protein sources are preferred due to their low cost, wide availability, and renewability .Some studies demonstrated that starch-protein interaction modified the film’s surface energy, enhancing surface hydrophobicity. Quéré stated that wettability of hydrophilic surfaces increased with increasing roughness . The incLeroy’s research team  preparedOne of the most commonly used proteins for food packaging film production is gelatin, an animal protein with excellent film-forming properties, obtained by hydrolysis of collagen, which is generated during animal slaughtering and processing. The main sources of gelatin are: fish, bovine, and porcine . In addiIn addition, protein-based films and coatings are biodegradable and compostable, and their degradation provides a source of nitrogen, which acts as a fertilizer. Moreover, protein hydrolysis results in the formation of peptides, which are short chains of AAs linked together by peptide bonds; these biomolecules are bioactive and possess health benefits  . An inteListeria innocua and/or Saccharomyces cerevisiae.Resa and coworkers  developeListeria monocytogenes and Clostridium perfringens. Recently, Meira et al.  disperseEnzymes represent a group of proteins which catalyze the chemical processes in living systems, which also have application in food packaging. Lysozyme, an important class of enzymes, present in many foods, such as milk and eggs, has a hydrophilic monopeptide chain and inhibit bacterial infections , by hydrolyzing the peptidoglycans which are the main bacterial cell wall components of Gram(+) bacteria, causing loss of intracellular content and, finally, bacterial death ,164. BhaThe packaging starch films are generally effective oxygen hinderers at intermediate to low humidity, but they are a poor water vapor barrier because of the starch hydrophilic properties . Lipids B. subtilis and E. coli more effective than chitosan alone [On the other hand, the addition of lipids into starch packaging materials improves the mechanical barrier and optical properties of the film due to the change in inner structure and film surface . Therefoan alone ,170. Another study  showed tSlavutsky and Bertuzzi showed that coating of lipid nanolayer (sunflower oil) with hydrophilic film (starch) caused an increase in TS and also low water diffusion coefficients since the structure and the hydrophobic nature of the lipids and oils restrict the migration of gas and vapors .Natural oils, especially vegetable oils, are currently used as green plasticizers for the compatibilization of polymer blends to obtain novel green materials for food packaging applications. Plant-derived oils are interesting due to their triglyceride structure, consisting of a glycerol backbone which is chemically bonded to different fatty acids through ester bonds. To give desired functionality to a vegetable oil, different chemical modifications are required: epoxidation, hydroxylation, maleinization, acrylation, etc. Thus, Gonzalez et al. reported that the addition of maleinized linseed oil to poly(lactic acid)/diatomaceous earth (PLA/DE) composites resulted in an increase in thermal stability, chain mobility, and Eb, together with a decrease in rigidity and TS . These fLipid-based nanostructures  are anoTagetes patula). Vegetable-based lipid nanoformulations are a current research topic in the design of active packaging materials. Exploiting natural resources in combination with soft nanotechnology gives rise to valuable products with improved biological performances. The research team of Lacatusu  develope2-eugenol liposomes with antioxidant activity and great potential as a food packaging material [In last few decades, liposomes have gained more attention in many fields , due to their fascinating properties: self-healing, ability to carry both hydrophilic and hydrophobic molecules, capacity to improve the bioactivities of the substances they carry, great resemblance to cell membranes. Liposomes are self-closed lipid vesicles containing one or more concentric phospholipid bilayers that include aqueous compartments. They are nontoxic, biodegradable, biocompatible, nonimmunogenic, and they can entrap hydrophobic (within lipid bilayers), hydrophilic (in the aqueous regions) and amphiphilic (at the aqueous–lipid interface) molecules ,178. Thematerial .Lactococcus lactis, entrapped in phosphatidylcholine liposomes, was incorporated to biopolymer-based films , resulting in active packaging materials that exhibit antimicrobial activity against L. monocytogenes [Liposomes have also been explored as antimicrobial incorporation systems for the development of new biocidal packaging materials . Thus, sytogenes ,182,183.Rosmarinus officinalis) extracts in edible and biodegradable cassava starch film (plasticized with glycerol), and the effects on the physicochemical properties of developed film, were firstly reported by Piñeros-Hernandez et al. [The incorporation of aqueous rosemary  leaves’ aqueous extract, a rich source of polyphenols, was inserted into chitosan/starch film formulations to provide edible films with remarkable antioxidant activity and improved mechanical properties (TS), with great potential in food preservation [Thyme (ervation .Vaccinium corymbosum L.) peels, and sorbitol into cassava starch, resulting in starch films with low water solubility and enhanced UV-light barrier properties [Furthermore, Luchese et al. incorporated dried and grinded blueberry (operties . This isRosa canina L.) extract, rich in antioxidants (such as ascorbic acid and carotenoids), was added to rye starch films and improved the mechanical and chemical properties of these films  [Rosehip (perties) . These sAnother study  reportedVigna unguiculata) for starch-based films’ preparation [Aristotelia chilensis) extract (rich in antioxidant biomolecules such as delphinidin derivatives) into cowpea starch films and demonstrated their applicability in the food industry for salmon packaging.The research group of Baek reported for the first time the use of cowpea (paration . They inNephelium lappaceum) peel ethanolic extract and cinnamon (Cinnamomum zeylanicum) oil. These starch-based films presented good antioxidant activity attributed to the presence of phenolic compounds such as corilagin, geraniin, and ellagic acid [Bacillus cereus, Staphylococcus aureus and Escherichia coli. It should be mentioned that corilagin  is ydrolysableble tannin (an ellagitannin) with a wide therapeutic spectrum, such as antioxidant, anti-inflammatory, antimicrobial, hepatoprotective, antihypertensive, antidiabetic, and antitumor activities [Chollakup et al.  reportedgic acid , arisingtivities . Moreovetivities ,192.Staphylococcus aureus, and also have antiviral activity against HIV (human immunodeficiency virus) and HSV-2 (herpes simplex virus 2) [In addition, tannins act as immunomodulatory agents in the battle against infectious diseases, being effective against the methicillin-resistant virus 2) .Eugenia uniflora L.  aqueous extract was incorporated in cassava starch (CS) and chitosan (CH) blend films, which resulted in effective antioxidant and antifungal food packaging material [material  with a UThis innovative food packaging concept based on vegetal extracts should be the center of attention of researchers since the incorporation of plant extracts into starch films is a promising method to prevent or reduce food quality deterioration, thus preserving and extending food shelf-life by providing improved physical, mechanical, structural and barrier properties .By taking into account all aspects discussed here, the main actions of starch-based primary packaging in the form of edible films and coatings incorporating the investigated bioactive compounds and the plant extracts containing these biomaterials can be grouped as:-Actions sustaining consumers’ health and safety: boosting the immune system, antimicrobial and antioxidant activity; -Actions on the packaged products: increasing their shelf life, improving of the mechanical and sensorial properties.An overview on this approach is given in Even though the chemical methods are the most widely spread starch modification methods , in the Physical methods used for starch treatments involve the application of various intensities of heat, pressure, and radiation . As mentNew research has also been focused on utilizing green physical methods to improve physical and chemical properties of the starch-based bioplastics . We highIrradiation is the process of radiation energy that is exposed to a certain type of the polymer or starch to achieve desirable changes . The mosGamma irradiation is the process of using an isotope from Cobalt-60 and results in a high energy photon . Gamma iCold plasma techniques are based on plasma and the term “plasma” has been used since 1928 to define the fourth out of five states of matter . A plasmAccording to , UV radiNawapat and Thawien  analyzedAt the treatment of a sago starch/PVA film with UV, TS increased with the rise in the radiation intensity; however, the TS started to decrease after reaching a maximum value, this behavior being due to the polymer chain cross-linking density. The TS value decreased when the concentration of starch increased; the cross-linking density increased with increased doses of UV radiation, which led to higher Eb . MonomerMoreover, UV radiation affects the transparency of the starch films; transparency decreases under UV-C (200–280 nm) treatment , this phThe microstructure of the starch films treated with different wavelength  does not show cracks on the surface for long time UV exposure (more than 6 h) .Due to starch’s hydrophilic structure, starchy films have poor barrier properties . The WVPEven though starch films are excellent barriers to oxygen , their mTo overcome the concerns caused by the limitation of the native starch, modifications have been made to improve its functions and properties . ElectroThe physicochemical properties of sago starch have been impacted by the EB irradiation. Amylose and amylopectin degradation resulted in a shift in physicochemical properties. When the irradiation dose was increased, peak viscosity, yellowness, redness, solubility, and free acidity were increased, whereas the gel pressure, swelling power and pH were reduced. The intrinsic viscosity was reduced by the increase in the EB irradiation .Zhou et al.  analyzedZhai, Yoshii, and Kume  reportedIrradiated corn starch was used for casting. Irradiation doses of 10, 20, 30 and 40 kGy resulted in a positive increase in the films’ TS. The water vapor permeability of the film decreased. The best properties of the films with the improved characteristics were shown at the irradiation doses of 30 kGy .During irradiation, the chemical reaction leads to the creation of an intact network structure in PVA/chitosan (CS) films. Radiation-induced crosslinked reaction increases the TS of PVA/CS films. However, under high energy irradiation, three forms of species are formed and may become entangled in polymers, ionic species, radicals, and peroxides. Postirradiation can be caused by both radicals and peroxides, and the various active centers can lead to various chemical transformations such as crosslinking and degradation .The gamma irradiation of cassava starch gave foams developing stable formulations at doses of 3 kGy, 6 kGy, 12 kGy and 25 kGy; the mechanical (compression resistance) and barrier (water solubility) properties increased compared to foams obtained with nonirradiated starch. With the increase in the irradiation dose, the absorbed amount of the water increased, whereas the mechanical and barrier tests did not follow the same trend .Regarding starch modification by plasma, attention must be paid when choosing from the multitude of the existing plasma types  to achieA multitude of review articles were published in recent years that address cold plasma treatments of starch ,243,244.It can easily be seen that the atmospheric pressure plasma method requires a less complicated set-up, with fewer components, and is an instant ready-to-use method. On top of the beforementioned advantages, atmospheric pressure cold plasma treatments are seen as innovative techniques for food processing . TherefoStarch presents some limitations attributed to its hydrophilic character, and like any biopolymer, starch’s mechanical properties are poor, and it is brittle . The nanParticulated 3D nanoreinforcements ;Fibrillated 2D nanoreinforcements ;Laminated 1D nanoreinforcements (nanoclays).2NPs, MgONPs), nanosilica (SiO2NPs), carbon nanotubes/fullerenes, etc. [The main nanoreinforcers used in packaging include: natural nanoparticles , nanoclays (montmorillonite), metallic nanoparticles , metal oxide nanoparticles, MONPs , and plasticized with sorbitol/glycerol (3:1) .L. monocytogenes and E.coli [A group of researchers incorporated ZnO nanoparticles in different films made from agar, carrageenan and carboxymethyl cellulose. They observed that nanoparticles were uniformly distributed into the matrix and the colors of the films were modified by ZnO nanoparticles. The incorporation of ZnO nanoparticles into the films led to increased hydrophobicity, UV barrier, Eb value, moisture content and thermal stability of the films, whilst the water vapor barrier and TS were lower. The films with ZnO nanoparticles showed antimicrobial effects against d E.coli .A significant decrease in the moisture sensitivity of the biopolymer matrix was achieved by Hietala et al. , by incoThrough a casting-evaporation method, Salaberria and coworkers  obtainedA simple casting method was employed by Gonzalez and Igarzabal to develop soy protein films reinforced with starch nanocrystals (SNC) at different concentrations. The presence of SNC as nanofillers was found to decrease the affinity of the films for water, and thus the water solubility and swelling of the protein film, as well as its WVP, have decreased ,262.Listeria innocua and Escherichia coli) and fungi (Aspergillus niger and Penicillium expansum). They observed that the silver ions’ release was slowed down in the nonpolar medium (involving oleic acid), suggesting the usage of this film in fat-rich products.Amazing features could be achieved by the insertion of MNP or MONPs as nanofillers. Metallic nanoparticles are promising in active packaging systems for innovative technology in food preservation based principally on mass transfer interactions between food and packaging materials . Cano’s Obtaining green MNPs opened a new era in nanotechnology. Vegetal extracts are a good choice for “green” synthesis of metallic nanoparticles, since plant extracts contain many bioactive compounds  that can bioreduce and stabilize the nanoparticles . We coul2NPs proved that only a small number of particles migrated from nanocomposites packaging to foods, being below the limits prescribed by the European Commission [Many research studies are needed to evaluate the nanomaterials’ cytotoxicity. Nanosized systems can migrate from package to food, resulting in serious health concerns. The migration rate increases if the nanoparticle size and polymer viscosity decrease, and if the nanoparticles’ concentration increases ,277. If mmission . Abreu emmission  preparedIn the last few years, ecofriendly products have attracted wide interest as safe and nontoxic food packaging materials, but further improvements are also needed to develop optimal formulations for bionanocomposites that comply with the regulations relating to the migration of nanomaterials from packaging to food ,279.Recently, bioinspiration became a new trend in green technology. As is known, the structure and optical properties are very important features of food packaging systems, so biomimetics offer new interesting aspects in this regard, since they have regular and self-assembled layered micro-/nanostructures which confer characteristic optical properties at the interaction with light , and als3) plates and the biomatrix  [A source of inspiration from nature is nacre (“mother-of-pearl”), a material fabricated by pearl oysters, being remarkable for its highly regular “brick-and-mortar” arrangement of the alternatively packed aragonite (CaCOroteins) ,281,282.roteins) ,284. MMTroteins) . Inspired from the amazing ability of mussels to adhere to various kinds of surfaces via marine mussel adhesive proteins (MAPs), the researchers designed biomimetic adhesive coatings from polydopamine (PDA) , which iBy combining the two biomimetic approaches, nacre and MAPs, researchers designed, by ecofriendly methods, starch-based biocomposites with improved mechanical properties . Thus, the first study regarding composite materials of starch and MMT modified with polydopamine, was reported by Zhou and Xu, in 2015 . In 2016“Lotus effect” is another remarkable biological model for bioinspiration in food packaging applications. Starting from the studies of Barthlott and Neinhuis in 1997 , the resIn this paper we review new research trends that present starch, a natural sustainable biopolymer, as a primary green packaging material to be used as films or coatings on food. Starch is used here as model to connect the concept of Green Packaging (GP), along with another two “green” concepts, namely the Green Chemistry (GC) and Green technology (GT), by incorporating bioactive compounds into the starchy matrix and by using physical treatments which are environmentally friendly on the films resulted or for the obtaining of packaging. In relation to GC, the focus is on the characterization and incorporation of the results into the starch-based films or coatings of the biocompounds involved in supporting health: vitamins, minerals, polyphenols, essential oils, amino acids, proteins and lipids, isolated from plants or in the form of plant extracts, rich in mixes of these bioactive compounds. Some compounds, such as vitamins or minerals, are not yet extensively used. By comparison, studies regarding the use of essential oils, polyphenols, proteins and lipids, and their effect on food coated or packed into starch-based films containing them are numerous. A large number of studies regarding the incorporation of plant extracts directly into the starch packaging, with very good results on the antimicrobial, antioxidant and/or mechanical properties of the films resulted and/or the food where the film was applied as primary packaging, are published. The development of starch-based primary films or coatings incorporating compounds with biological effect on health could be the next revolution in GP. As a way to connect GP to GT through the starch-based packaging, green physical techniques such as UV and ionizing radiation, together with atmospheric pressure cold plasma, are presented. Generally, radiation improves the mechanical properties of films, but this action is dependent on the radiation type and dose as well as the treatment time. Atmospheric pressure cold plasma induces various modifications directly into the starch granule, thus making this treatment method a promising technology to obtain films or coatings; however, investigations are only just beginning in this research field.Moving forward, we also propose to expand the concept of GP to another challenging field—that of Green Nanotechnology (GN)—by showing the actual status in developing green nanopackaging. The research is still in its initial stage and only a few publications on starch-based nanopackaging were found, with good results in improving the films’ mechanical properties and antimicrobial activity.In the end, our review presents a few proposals regarding using the nature as source of inspiration for future research, such as nacre-mimetics in combination with clay, mussel adhesive proteins or superhydrophobic materials to be combined with starch with the aim of improving its characteristics and developing a new generation of superpackaging.


The design of zirconia-based scaffolds using conventional techniques for bone-regeneration applications has been studied extensively. Similar to dental applications, the use of three-dimensional (3D) zirconia-based ceramics for bone tissue engineering (BTE) has recently attracted considerable attention because of their high mechanical strength and biocompatibility. However, techniques to fabricate zirconia-based scaffolds for bone regeneration are in a stage of infancy. Hence, the biological activities of zirconia-based ceramics for bone-regeneration applications have not been fully investigated, in contrast to the well-established calcium phosphate-based ceramics for bone-regeneration applications. This paper outlines recent research developments and challenges concerning numerous three-dimensional (3D) zirconia-based scaffolds and reviews the associated fundamental fabrication techniques, key 3D fabrication developments and practical encounters to identify the optimal 3D fabrication technique for obtaining 3D zirconia-based scaffolds suitable for real-world applications. This review mainly summarized the articles that focused on in vitro and in vivo studies along with the fundamental mechanical characterizations on the 3D zirconia-based scaffolds. Th. Th91]. In contrast, Zhang et al., fabricated a porous zirconia toughened HA composite scaffold for bone-tissue engineering using the DLP 3D-printing process with HA as the primary ratio of the scaffold; additionally, a small amount of zirconia was used as a structural stabiliser . The cer3) porous scaffold (CaSiO3/nano-zirconia scaffold) via the SLS technique for bone-recreation applications with improved mechanical properties [3/nano-zirconia (20%) scaffolds had the ability to mock bone features by forming a bone-like apatite layer and promoting superior assembly of MG-63 cells. The authors concluded that the porous scaffolds with the addition of 20% of nano-zirconia exhibited good biocompatibility and mechanical properties.SLS is a well-established additive-manufacturing technique that employs laser energy as a primary source to fuse powder particles into the desired 3D architecture . It has operties . The autElectrospinning is a unique method in which an electric field induced by a high direct-current voltage between a roller connection and a needle tip is used to spin bioceramic fibers with a constant diameter into an interconnected flexible bioceramic scaffold ,95,96. WThe interwoven PCL/zirconia composite nanofiber scaffolds fabricated by the author had high material stability and porosity, which are favourable for imitating the dynamic atmosphere of natural tissues. Compared with the pristine PCL scaffold, PCL/zirconia composite nanofiber scaffolds exhibited controlled degradation, swelling and superior bioactivity. Additionally, the PCL/zirconia composite nanofiber scaffolds were nontoxic, as indicated by a cell-viability test using the 3T3 mouse fibroblast cell line. This study further confirmed that electrospinning can be used to fabricate PCL/zirconia composite nanofiber scaffolds that are favourable for tissue-engineering.Additionally, Esfahani et al., conducted electrospinning of polyamide 6(PA6)/HA on the surface of the ZTA nanocomposites with improved bioactivity suitable for bone-repair applications . Even thIn contrast to the foregoing replica techniques, Feng et al., designed an acrylonitrile butadiene styrene (ABS) template with organised channel openings using a 3D Rapid Prototyper to construct zirconia scaffolds with different shapes  and porosities . The autSapkal et al., developed a biocomposite scaffold by infusing 10%, 20%, 30%, 40% and 50% zirconia with a β-TCP background via an indirect casting technique. The ABS was first printed using a conventional 3D printer, followed by the impregnation of a zirconia/β-TCP ceramic slurry and subsequent annealing to obtain a zirconia/β-TCP bioceramic scaffold. Osteosarcoma cell line (MG-63) results and compressive-testing results for a sample with a zirconia content of 30% indicated that it was suitable for bone-tissue engineering. Scaffold fabrication with proper porosities suitable for cortical and cancellous bone must be verified .In the aforementioned studies, the direct fabrication of flexible zirconia-containing composite scaffolds via electrospinning was demonstrated. However, a pioneering methodology combining the traditional sponge replica and modern electrospinning methods for fabricating an artificial small bone was reported by Lee et al., in 2011 . ZirconiRecently, Sakthiabirami et al., fabricated porous zirconia 3D scaffolds via FDM . The scaA summary of the zirconia-based scaffolds discussed in the previous sections that have been used for bone-regeneration applications over the past two decades is presented in 2 and its composite materials [Very recently, Weng et.al summarized the properties and construction of ZrOaterials . MoreoveThe non-degradability of zirconia-based scaffolds is a major bottleneck of zirconia-based in comparison with other calcium phosphate bioceramics-based bone-tissue engineering applications. Considerable research attention has been directed toward resolving this concern, and zirconia-based scaffolds have seen advancements over the decades owing to the repeated efforts of bone-tissue engineers. The successful long-term use of zirconia-based implants in dental applications has been validated via clinical trials with 90% survival rates. Even though zirconia-based ceramics are found to undergo fracture in the earlier stage, the absence of surplus parasitic reactions indicates that zirconia can cause no harm to the surrounding biological environments. Hence, zirconia-based scaffolds with 3D porous architectures can provide sufficient osteointegration; once, the bone-remodelling initiated earlier fracture will not be a major issue.In this paper, we comprehensively reviewed the evolution of 3D zirconia-based scaffolds over the years, detailing different fabrication techniques, e.g., the conventional foam replica technique and modern 3D-printing methods. As indicated by the pie chart in In the design aspects, pursuing the finest scaffold design for bone-repair applications has greatly increased; the recent evolution of 3D-printing techniques for designing 3D bioceramic scaffolds has encouraged bone-tissue engineers to construct 3D zirconia-based scaffolds with precise dimensions and interconnected porosity. Even though 3D printing is well established for calcium-based bioceramics aimed for bone-tissue engineering, few studies have been performed on 3D-printing techniques, including extrusion-based, DIW, DLP and SLS techniques, for fabricating zirconia-based scaffolds for bone-tissue engineering. This clearly indicates that 3D-printing techniques are still in a stage of infancy. Among the few studies reported thus far, a multi-extrusion technique-derived 3D zirconia-based scaffold exhibited considerable promise, with improved mechanical and biological properties. However, the surfactants, organic solvents and binders used for the 3D-printing process must be carefully selected, as they can be harmful to host cells and tissues. Given the limited number of reports on zirconia-based scaffolds fabricated via 3D-printing techniques, it may be challenging for future bone-tissue researchers to clearly understand the degradation, mechanical stability and bone-regeneration ability of 3D-printed zirconia scaffolds in load-bearing applications in human biological environments.To address the poor interfacial compatibility between the zirconia scaffolds and incoming cells, three main strategies have been employed by bone-tissue engineers: surface reformation, interface-phase introduction and crystal-structure variation . For insFinally, it is essential to discuss the clinical possibilities of zirconia-based scaffolds. The direct comparison of the strength of the zirconia-based scaffolds to the strength of the natural bone  lacks among the reviewed articles, which is very essential to understand the ability of zirconia-based scaffolds to satisfy the critical bone defects requirements. In most of the collected literature, the in vitro studies focused on validating the metabolic activity of osteogenic cells b. SurpriFew 3D scaffolding technologies have been commercially available, among which CAD/CAM is commonly used and the most dependable. After the successful fabrication of zirconia-based scaffolds for dental applications using the CAD/CAM technique. Apart from the in vitro and in vivo performance, 3D zirconia-based scaffolds fabricated using the CAD/CAM technique has been successfully employed for human mandibular reconstruction. Even though 3D-printed zirconia-based scaffolds were claimed to be the best possible alternatives to the conventional technologies, few in vivo studies support the clinical possibilities of 3D-printed zirconia-based scaffolds. Hence, it is essential to closely monitor the in-depth reproduction of natural mechanical and biological inducements and the rate of vascularisation of 3D-printed zirconia-based scaffolds in the in vivo environment. Multifaceted research should be targeted to systematically appreciate the perspective of 3D printing in bone-restoration applications.The primary goal of the scaffold is to get completely absorbed and replaced by body tissue. However, in large defects, scaffold or implanted artificial bone materials like HA or TCP are not absorbed completely and remained for a long time in vivo studies. And the healing period of each bone is different from several weeks to several months (ex. Femur 6 months). Until now researchers are not able to control the absorption rate of scaffold depending on new bone formation. Over the past 20 years, the zirconia-based scaffold has played a substantial role in critical-sized bone-defect applications, following the tremendous success of modern restorative dentistry. For the large sized loading area, a zirconia-based scaffold that has comparable strength to that of natural bone is needed to support bone formation during the long healing time. However, it remains challenging for orthopaedic experts and bone-tissue researchers to heal critical-sized bone deficiencies using zirconia-based scaffolds, owing to the immature fabrication techniques and unfavourable composite/coating materials for enhancing the bioactivity. Clinically feasible possibilities for promoting surface activation, along with appropriate 3D fabrication practices, are likely to be a focus area for future zirconia-based -scaffold design. The advancements in material engineering in association with biomechanical engineering could be critical in determining zirconia-based scaffolds as a successful biomaterial. Undoubtedly, with the expansion of advanced 3D-printing technology and unique coating strategies, the liability of zirconia-based scaffolds for critical-sized bone-replacement applications is not far away.


Kidney transplantation is the optimal treatment for patients with end-stage renal disease. Short- and long-term kidney graft survival is influenced by a number of donor and recipient factors. Predicting the success of kidney transplantation is important for optimizing kidney allocation.The aim of this study was to predict the risk of kidney graft failure across three temporal cohorts  based on donor and recipient characteristics. We analyzed a large data set comprising over 50,000 kidney transplants covering an approximate 20-year period.We applied machine learning–based classification algorithms to develop prediction models for the risk of graft failure for three different temporal cohorts. Deep learning–based autoencoders were applied for data dimensionality reduction, which improved the prediction performance. The influence of features on graft survival for each cohort was studied by investigating a new nonoverlapping patient stratification approach.Our models predicted graft survival with area under the curve scores of 82% within 1 year, 69% within 5 years, and 81% within 17 years. The feature importance analysis elucidated the varying influence of clinical features on graft survival across the three different temporal cohorts.In this study, we applied machine learning to develop risk prediction models for graft failure that demonstrated a high level of prediction performance. Acknowledging that these models performed better than those reported in the literature for existing risk prediction tools, future studies will focus on how best to incorporate these prediction models into clinical care algorithms to optimize the long-term health of kidney recipients. Kidneys are vital for the health of an individual, as they filter waste products from the blood and produce hormones and urine . PatientKidney transplantation ,6 and diPrediction modeling using machine learning (ML) algorithms has gained attention in recent years  for prednonoverlapping patient stratification approach, whereby a patient’s graft failure was recorded only in the temporal cohort when it actually happened, that is, a graft failure event in the preceding cohort was not included and (2) the traditional overlapping patient stratification approach that provides an accumulative count of graft outcomes until a specific time point. To develop the prediction models, we investigated multiple ML algorithms using both patient stratification approaches. Nonoverlapping temporal cohorts were considered to investigate the influence of clinical features on predicting graft survival over time, as the temporal partitioning of the data allowed for the establishment of feature influence across distinct temporal windows. We applied the feature importance method based on the mean decrease in impurity (Gini).In this study, the intent is to investigate kidney transplant allograft survival, that is, estimating the time-to-event and the evolving influence of clinical features leading to an event—within three temporal cohorts of 1 year, >1-5 years, and >5 years of a kidney transplant. We predicted the outcome of graft failure after kidney transplant based on the analysis of donor and recipient features. We applied ML methods to (1) predict the graft status over different temporal periods and (2) analyze the changing effect of donor-recipient–related predictors across different periods. To develop the prediction models, we analyzed a large data set of over 50,000 transplants covering approximately a 20-year period of kidney transplants in the United States. To generate the clinically meaningful temporal cohorts, we experimented with two patient stratification approaches: (1) a novel The contributions of this research are as follows: (1) ML-based prediction models that are trained on a large data set, offering improved prediction performance compared with previous studies ; (2) data dimensionality reduction based on a deep learning framework to handle the high-dimensional and complex kidney transplant data set; (3) a novel nonoverlapping patient stratification approach to provide fine-grained feature importance within a specific period while avoiding bias from preceding cohorts; (4) explaining the influence of the different clinical features, during different periods, toward the prediction performance of ML prediction models. This finding allows the selection of the most important features to predict graft outcomes within a specific temporal window; and (5) a comparison between the two stratification approaches with respect to the performance of the prediction models. The future practical outcome of this study is the provision of a data-driven decision support tool to assist nephrologists in the kidney allocation process by identifying the best donor and recipient pair that will lead to the highest likelihood of graft survival for a given recipient.Patients can receive a kidney from either deceased donors or living donors. The donor-recipient matching process becomes relatively more complex with deceased donors because of the need to account for additional clinical factors  . Given tData-driven methods are now being used for organ matching; these methods are used to establish clinical compatibility beyond the blood group and tissue type. Conventional data-driven prediction methods use statistical techniques such as Cox proportional hazard models and Kaplan-Meier estimates to perform time-to-event analyses . SignifiML-based data analysis to develop prediction models for predicting outcomes is usually performed using classification methods, whereas regression methods are used for time-to-event analysis. There are two prominent approaches to predict kidney allograft outcomes using ML-based classification methods. The first approach is to predict graft survival over time by dividing a longitudinal data set into different time cohorts based on the occurrence of a given adverse event or the last follow-up date from the date of transplant. Each time cohort has a binary target variable, that is, success or failure of the graft, which is used to train the classification model to predict graft survival ,16. The Due to the high dimensionality of existing data sets for organ transplantation, feature selection is applied to filter out redundant features. A stacked autoencoder, which is an unsupervised neural network, is an efficient dimensionality reduction technique with promising performance for deep representation of medical data  that recRight-censored data are a common problem for survival analysis, as it represents cases for which the adverse event is not available or recorded because of either the subject having been lost to follow-up or not experiencing the event during the study period. Multiple approaches have been adopted in previous studies to address this problem. The study on kidney transplants by Topuz et al  discardeThe influence of clinical features  on graft survival tends to vary over time , as showIn previous studies ,20-22,25Methods presents the study’s methodology; Results presents the results of the prediction; and Discussion discusses the significance of clinical features toward graft status prediction across different time cohorts and offers a conclusion and future research directions.This study is organized into five major sections: To predict graft survival over time and to analyze the influence of clinical features on graft survival, our data analytics methodology  compriseThis study used data from the Scientific Registry of Transplant Recipients (SRTR). The SRTR data system includes data on all donors, wait-listed candidates, and transplant recipients in the United States, submitted by the members of the Organ Procurement and Transplantation Network. The Health Resources and Services Administration and the US Department of Health and Human Services provided an overview of the activities of the Organ Procurement and Transplantation Network and SRTR contractors.The data set provided pretransplant clinical features and outcomes of 277,316 kidney transplants between 2000 and 2017. Survival was reported in terms of graft outcome and patient status. For the purposes of this study, graft failure was defined as (1) graft loss or (2) death with a functioning graft.We analyzed the data and used only complete cases , which comprised a total of 52,827 kidney transplants. Data preparation for learning the ML-based prediction models consisted of data cleaning, partitioning the data set into temporal cohorts, and addressing class imbalances.Data cleaning involved removing (1) all patient identifying features  , (2) posoverlapping and our novel nonoverlapping patient stratification approaches to partition the data set into three temporal cohorts representing graft status at 1 year, >1-5 years, and 5-17 years  during this period; cohort 2, which reported graft status from year0 to year5 and overlapped with cohort 1 such that it included the patients in cohort 1; and cohort 3, which reported graft status from year0 to year10+, thus overlapping with both earlier cohorts. As per the overlapping approach, a graft failure in the preceding cohort was also counted in the proceeding cohort.The overlapping patient stratification approach (used in previous graft status prediction studies) provides a cumulative analysis of graft outcomes up to a specific time point. In our study, the overlapping data stratification resulted in the following three cohorts: cohort 1, spanning from year, spanning year0 to year1, reporting graft outcomes in this period; cohort 2, spanning year1 to year5 and reporting graft outcomes only in this period, resulting in the exclusion of graft failures reported in cohort 1 and only reporting the graft outcomes of patients who survived cohort 1; and cohort 3, spanning year5 to year10+, reporting the graft outcomes of patients who survived cohort 2. In a nonoverlapping cohort approach, there was no looking back beyond the cohort’s starting point, as such a graft failure in the preceding cohort was not counted in the proceeding cohort.Our nonoverlapping patient stratification approach yielded three cohorts: cohort 1survivors in each cohort. First, we removed all the censored observations from the cohort being analyzed and labeled all the remaining instances as survived. For instance, the censored data that were removed for cohort 2 were all instances with a missing outcome by the end of cohort 2. The remaining instances were considered to have survived. The first phase of our approach reduced the number of censored observations, but the surviving observations were still relatively high compared with the graft failure cases in each cohort. In the second phase, we applied the approach of Topuz et al [When partitioning the data into cohorts, we accounted for the presence of censored data, that is, the lack of information about the occurrence of an adverse event for a surviving patient. There is no concrete method to determine survivors when confronted with censored data. We initially assumed that those patients who did not fail in a certain cohort could be presumed as survivors. However, this assumption led to two problems: (1) it included censored patients who might have experienced graft failure during the study, and (2) it led to a severe class imbalance between the graft failure and surviving patients. To overcome these problems, we took a two-phase heuristic approach to remove the censored observations to identify uz et al  to furthi in the equation is the ending year of our defined cohorts, that is, 1, 5, and 17. The equation first calculates the total number of graft failures that occurred after the end of the cohort. This fraction of instances was considered as confirmed survivors for the cohort under analysis. The second part of the equation deals with the second phase of the approach. It attempts to identify the potential survivors from the censored data by removing all the observations that did not have a graft failure within 8 years (2920 days) following a transplant. The first part of this equation illustrates the first phase of the proposed approach. The graft failure had a significantly lower number of instances compared with graft survival. Techniques such as Synthetic Minority Oversampling Technique (SMOTE) and random undersampling have been widely used in the literature [graft failure), we would need to generate 600% additional synthetic samples (at least for cohort 1), which would have led to overfitting. Therefore, we doubled our minority class to achieve a workable class balance to prevent the overfitting of the classification models. As cohort 2 of overlapped stratification and cohort 3 of nonoverlapped stratification had a class balance, they were not considered for oversampling.Our data set had two outcomes: the presence or absence of graft failure. There was a significant class imbalance whereby the terature ,29 for cterature , which iThis step involved both the removal and construction of features with the intent to reduce the dimensionality of the feature space.paired features. The following four donor and recipient features were generated: sex, age, CMV , and race. In addition, three types of HLA Antigen Mismatch features—ie, HLA Antigen Mismatch at the A Locus, HLA Antigen Mismatch at the B Locus, and HLA Antigen Mismatch at the DR Locus were also combined into a single HLA Antigen Mismatch feature.A set of paired features was constructed by combining the related features. Typically, graft predictions use discrete individual donor and recipient features. We examined the underlying correlation between the donor and recipient features and paired the highly related features to generate new We transformed the categorical features into multiple dummy features to make them compatible with the stacked autoencoders, which cannot process categorical features. In addition, it was also a necessary operation because of the functional constraint of the scikit-learn library .Finally, we used 86 transformed categorical features as inputs to the stacked autoencoders for feature reduction to subsequently train the ML prediction models. Continuous features were also initially considered as a part of the input vector to stacked autoencoders . Given that logistic regression (LR) has been widely used in prior studies to develop graft prediction models ,33, we tAll classification models were trained using a 10-fold stratified cross-validation training approach. The stratification ensured that outcome class ratio in each fold is maintained to avoid any sampling bias that may affect the classification results. We mainly used the scikit-learn library  to trainRandom forest (RF) was used as both a standalone classifier and a base learner for the adaptive boosting (AdaBoost) algorithm. It has been widely used to predict survival data ,27.The AdaBoost algorithm was applied to two weak learners, RF and LR. The study by Thongkam et al  used thiA backpropagation algorithm was used to train a neural network–based binary classifier. Generally, artificial neural networks (ANNs) perform well on survival data sets ,35.Classification models using support vector machines (SVMs) have been applied to predict survival data ,31,32. TThe nonoverlapped time cohorts were used to calculate the feature importance scores to understand the changing relevance of features over time. We calculated these scores by training an RF classifier on the complete data set. The scores were calculated using Gini. Feature influence scores were used to understand the effect of features over the three cohorts.Below, we present the prediction performance of the four ML classifiers using both overlapped and nonoverlapped cohorts. As LR has been extensively used to predict time-to-event in organ transplant studies ,29, it iThe AUC scores  show thaAlthough ANN and LR (the baseline model) showed no significant improvement across all three cohorts, the results confirmed the effectiveness of our deep learning architecture of stacked autoencoders for feature selection. For the subsequent prediction modeling analysis, we used the reduced feature set.The classifiers performed differently across the three cohorts—SVM offered the highest prediction performance for short-term predictions, that is, for cohort 1, whereas AdaBoost offered the highest performance for the remaining cohorts. The SD across the different folds was nominal, confirming the stability of the classifiers. . Therefore, the prediction model for cohort 2 should give a high prediction score for predicting graft failures from year0 to year5, but when applied to cohort 3, the cohort 2 prediction model would be unable to predict graft failure as it has not been trained on cohort 3 data. To further investigate the prediction efficacy of the ML-based classifiers, we evaluated the prediction performance of the best-performing classifier for all three cohorts by testing the prediction of graft failure events by a classifier trained for a specific cohort with data from other cohorts, that is, testing the classifier for cohort 2 with randomly selected data from cohorts 1 and 3. The underlying assumption is that the classifier should not produce good prediction results for data from other cohorts. As this evaluation considers survivors across progressive cohorts, we used the F1 score to measure prediction performance. A sound prediction model for cohort 2 will give a high graft failure prediction score for data from cohort 1 but a low prediction score for data from cohort 3, the rationale being that the overlapping cohort 2 classifier is trained on graft failure cases in both cohorts 1 and 2To determine if the prediction differences between the different models were statistically significant, we used the Wilcoxon signed-rank test to compare the scores between different models. Because the best scores in each cohort were usually produced by SVM and AdaBoost models, the Wilcoxon signed-rank test was conducted with each combination of these models with the other models.P values between the models were quite small and less than the threshold value of P=.05, confirming that the performance difference is statistically significant.The second objective of this research is to analyze the influence of clinical features on the prediction of graft survival over different periods. The intent was to understand the factors responsible for graft survival at different periods after transplant. The nonoverlapped cohorts  were used to ensure that there was no cascading influence of the features over time. For comparison purposes, we also examined the feature importance for overlapping cohorts. The feature importance scores represent the relative importance of the feature among all features, that is, the total of all the features’ importance scores add up to 100%; hence, if one feature gains a higher importance score, it will be at the expense of the importance score of other features.In general, the top 10% of the important features remained consistent in both the nonoverlapped and overlapped cohorts; however, we note that the nonoverlapped cohorts identified a larger group of important features. For instance, peak panel reactive antibody (Pkpra) and pre-emptive recipient status (Preemptive) had negligible importance in overlapping cohorts but were important during the 2-5 years and 6-17 years in the nonoverlapping cohorts. Below, we analyze the importance of features in each cohort and show the influence of features over time using nonoverlapping cohorts.According to the top features shown in Both HLAMMs and VINTAGE remained highly important in cohort 2. In addition, ESRDDXSIMP was noted as a highly important feature. Interestingly, we note that few features, such as donor age and recipient age, were rather insignificant in cohort 1 but were noted to be significant in both cohort 2 and further in cohort 3.ESRDDXSIMP showed a relative downward trend; however, it remained a highly significant feature. Unlike earlier cohorts, HLAMMs and VINTAGE were noted to not maintain their importance in the long term, whereas the recipient’s status of diabetes was noted to be the most important feature, along with recipient age and their functional status. Donor age was noted to maintain a medium importance score between 5% and 10%.Analysis of the values of categorical features provided novel insights into the influence of a feature. The cross-cohort prediction results  confirm We compared the prediction performance of our ML-based prediction models with comparable organ transplant studies that involved similar-sized observations and temporal windows. When comparing our results with prior studies, it is noted that although our cohort 2 prediction performance  is lower than that of Lin et al , it was A limitation of our research lies in the removal of censored instances. We removed all successful cases that were censored before 8 years following transplant. Although this type of approach has previously been used, including censored cases is a potential consideration for future analyses.Understanding the impact of donor and recipient factors that predict short- and long-term kidney transplant allograft survival is important for patients and providers. Kidney transplantation is the optimal form of kidney replacement therapy, but kidney allografts are a limited resource. In addition, the alternative to kidney transplantation  is considerably costlier.In this study, we present an ML-based framework to predict the status of kidney allografts, based on donor-recipient features, over a period of 17 years. We applied ML-based data analysis methods for feature engineering to reduce data dimensionality, develop prediction models for three distinct temporal cohorts, and investigate the changing relevance of clinical features across different temporal cohorts. We introduced the concept of nonoverlapped cohorts to analyze the changing relevance of features in three defined periods. In conclusion, our results emphasize that ML can be effective in predicting graft survival using donor and recipient factors that are routinely collected as part of patient care. As a next step, we plan to incorporate the prediction models into clinical care at the time of allocation; models that best predict short- and long-term kidney graft survival may be used as a pragmatic prognostic tool to aid clinicians in maximizing the best possible matching of donors and recipients while preserving existing allocation rules that are used to promote equity .