Autologous dermal fibroblasts for the correction of age related skin changes SPRS therapy (r) Results of 2 year clinical trials and postmarketing clinical studies
One of the innovative methods to correct the age related skin changes is the use of cultured fibroblasts derived from the patient's own skin. The technology called SPRS therapy(r) (Service for Personal Regeneration of Skin) has been officially approved in 2011 in Russia (developed by the public biotech company Human Stem Cells Institute (HSCI) Moscow) The results of two year clinical trials with the application of SPRS therapy to correct the agerelated skin defects demonstrated, that after the intradermal transplantation (injections) cultivated autoDF (autologous dermal fibroblasts) fully integrated into the dermis and their biosynthetic activity maintained at least for 12 months. As a result, the increase of collagen content and skin hydration was observed as well as the increased thickness of the dermis. The abovementioned changes resulted in the increase of skin tension, elasticity, and thickness and in the decrease of an amount of wrinkles and their depth. The results of post marketing studies conducted by HSCI after the first year of the application of technology in clinics confirmed the safety and clinical efficacy of SPRS therapy for correction of skin aging changes. On the aesthetic medicine market, SPRStherapy was introduced by HSCI in January 2011. Today the SPRS therapy service is primarily offered through the leading dermatology and cosmetics clinics of Moscow and cities across Russia and CIS >40) Currently, the total number of patients that received the procedure is more than 750, with more than 60% of those that have returned for the second and/ or additional skin treatments (included in other areas besides the face) SPRS therapy complex, developed for correction of patient's skin changes and aging prophylaxis, includes also SPRS diagnostics service (the evaluation of morpho functional properties of patient's skin fibroblasts called Skin Passport(r) which allows to build an individual SPRS therapy program as well to provide recommendations on other instrumental and cosmetological procedures for the patient. An individual cryogenic bank of skin fibroblasts is created for each patient allowing to repeat the procedure on a regular basis without additional skin sampling. IntroductIon Aging skin, as well as the organism aging in general, represents a complex biological process which is influenced by many factors including genetic, epigenetic, and environmental ones. It has been demonstrated that processes developing in the skin during aging are based on fundamental changes connected with the main cellular population of the derma and fibroblasts, namely, with their amount, morphology, proliferative potential, and functional activity1 4. In the skin, fibroblasts are responsible for the whole range of various and sophisticated This article has been presented at "Regenerative Medicine 2.0: Redefining the Practice" on June 23 25, 2016. Produced by Regenera Global, Inc. in partnership with the Grand Bahama Port Authority, Freeport, Grand Bahama 2 V.L. Zorin, A.I. Zorina, P.B. Kopnin, S.Z. Kantserov, A.A. Isaev remained. Later (2003 2008) the company Fibrocell Science, previously known as Isolagen, (USA) conducted multiple randomized, blinded, placebocontrolled clinical studies for the treatment of facial wrinkles. And in July 2011, this technology was used in the USA as technology LAVIVTM (azticel T)6. Since July 2010, the use of the technology of intradermal administration of autologous dermal fibroblasts has been officially approved in Russia7. The technology is termed the SPRS (Service for Personal Regeneration of Skin) therapy (OJSC "Human Stem Cells Institute" SPRS therapy represents a set of personalized diagnostics and treatment procedures which help to improve skin appearance based on the innovative technology of applying autoDF to correct skin damage due to aging and other structural changes. the clInIcAl results of the sPrs therAPy The Human Stem Cells Institute in cooperation with the Central Research Institute of Dental and Maxillofacial Surgery (Moscow) and with participation of a number of Moscow cosmetic clinics conducted two year clinical trials (non randomized, prospective, open label, single group study with comparison to baseline facial skin status) with the application of autoDF to correct the age related skin defects in order to obtain the data on duration of clinical effect, the persistence of biosynthetic activity of transplanted fibroblasts, and mechanisms of qualitative and quantitative changes occurring in human skin after injections of fibroblasts. Patients. The study enrolled 17 healthy subjects (4 men, 13 women) at the age of 45 65 years (the mean age 53 years) with mild to moderate signs of aging facial changes (wrinkles, reduced skin turgor) who did not receive other treatments to correct aging facial changes in the study period (were eligible for inclusion) Ethics. The clinical studies were carried out in accordance with the medical technology approved by Russian Healthcare Regulation Authority (Roszdravnadzor)7 and by the decision of the Ethics Committee and Academic Council of the Central Research Institute of Dental and Maxillofacial Surgery #4/276 from 14.04.2010) All patients signed the informed consent. SPRS product (autoDF) represents a suspension of fibroblasts which are derived from skin (4 mm) taken from behind the patient's ear (an area which functions. They not only maintain homeostasis of the dermal intercellular matrix supporting its remodeling and renewing (due to degradation of "used" dermal components and synthesis of the new ones collagen, elastin, and the main substance) but also play a significant role in maintaining the physiological status of the other skin layers representing a key chain in the skin metabolism1. When age increases, the amount of dermal fibroblasts is decreased, as well as their ability to synthesize biomolecules, and the balance between synthesis and degradation of components of the intercellular dermal matrix is destroyed. Thus, the study of Varani et al2 has shown that in old people the total amount of dermal fibroblasts is averagely less by 35% as compared with young people. According to the data of Fisher et al4, the collagen production in the skin of old people is decreased by 75% on average. The natural consequences of such a decrease include thinning of the skin, the loss of the skin's flexibility and elasticity, and wrinkle formation. Thus, the age related skin changes are accompanied by decreasing the dermal population of fibroblasts as well as their proliferative and biosynthetic abilities, which results in deterioration of quantitative and qualitative characteristics of the dermal intercellular matrix. Currently, in the practice of aesthetic medicine, a large arsenal of methods is used such as peels, fractional photothermolysis, radio wave therapy, dermabrasion, etc. with the main purpose of the application being stimulation of the functional activity of fibroblasts. A special place in this series is the method relating to regenerative medicine the use of cultured fibroblasts derived from the skin of the patients themselves to correct their age related changes. The peculiarity of this technology is that it allows you to replenish age diminished with the population of resident fibroblasts of young and functionally active cells, grown in a special laboratory environment that allows for the unique biological mechanisms effectively to correct age related skin defects. As early as in 1995 (Boss Jr. et al) it was shown that intradermal administration of autologous dermal fibroblasts (autoDF) promoted the effective correction of aging skin changes5. Cultured dermal fibroblasts actively synthesized collagen and other components of extracellular matrix in vitro, and after dermal transplantation, such synthetic activity Autologous dermal fibroblasts for the correction of age related skin changes (SPRS therapy(r) 3 groups, without signs of mitosis; newly synthesized elements of the intercellular matrix, newly synthesized collagen fibers, increased dermal volume and hydration were observed8,9. In injection sites, cultured autoDF remained at least for 12 months. Hereby, the prolonged synthetic activity was observed which was manifested as a synthesis of young (argyrophil) collagen fibers shown as thin crimped black threads within fibroblast groups. It resulted in a progressive increase of dermal thickness which was shown as changes of the mean distance between cutaneous appendages (Figure 1) The immunohistochemical skin analysis after SPRS therapy showed that transplanted cells did not proliferate and have no pathological differentiation (i.e. to myofibroblasts) which was demonstrated with antibodies to Ki 67 and smooth muscle actin (a SMA) which, in its turn, neutralized the minimal theoretical risk of excessive proliferation of transplanted cells or fibrous tissue formation. All preparations did not reveal an excessive number of phagocytes in derma, their number in injection sites and surrounding tissues did not significantly differ8,9. There were positive dynamic skin changes after application of autoDF on the histological level which is fully consistent with the clinical picture. Improvement of facial skin increased firmness, reduced relief, improved facial color and contours was observed by patients within 10 14 days after the course of cell therapy. The effect was progressive. Thus, if 88% of patients evaluated the clinical is exposed to harmful UV rays to the least extent) in concentration 15kh106 cells/ml of saline solution for injections. chArActerIzAtIon of Autodf Immunophenotype. The immunophenotypic analysis of dermal fibroblasts cultures showed high level of collagen (type I, III) elastin and vimentin expression, revealed markers (CD73+ CD90+ and CD105+ which confirmed mesenchymal origin of used cells and lack of hemopoietic cell (CD34, CD45 and epithelial cell (cytokeratins 14, 15, 16, 19) mar