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      Role of Cultured Skin Fibroblasts in Aesthetic and Plastic Surgery

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          Abstract

          The skin is the largest tissue of man with several functions. Cosmetics/cosmeceuticals agents always are needed to be evaluated for their detrimental effects on the skin. Nowadays, the therapeutic potential of stem cells has and fibroblasts have increased the hope for a successful cell therapy in aesthetic medicine. Stem cells are unspecialized cells capable of renewing themselves through cell division without limit as long as the person is still alive. Each new cell has the potential either to remain a stem cell or become another type of cell. 1 In a healthy individual, skin integrity is supported by epidermal stem cells that self-renew and generate daughter cells undergoing terminal differentiation. In addition to accumulation of senescence markers in aged skin, epidermal stem cells are maintained at normal levels throughout life. So, skin ageing is provided by an impair in stem cell mobilization or a reduction in the number of stem cells being capable to respond to proliferative signals. The self-renewal and multi-lineage differentiation of skin stem cells make these cells attractive for ageing process studies but also for regenerative medicine, tissue repair, gene therapy and cell-based therapy with autologous adult stem cells. 2 Healing in skin wound demonstrates an extraordinary mechanism of cascading cellular functions. Migration of fibroblasts along with the fibrin network and start of reepithelialization from the wound edges, angiogenesis and neovascularization get activated by capillary sprouting. 3 Sakrak et al. (2012) noted that cultured fibroblasts, particularly with a dermal support, contribute to the wound healing process; reduce the contraction of the wound; and support collagen synthesis and neovascularization. 4 Many potential mechanisms exist to impair healing. One contributory mechanism may be inhibition of fibroblast proliferation and induction of a stress-induced premature senescence phenotype by the continuing inflammation found in chronic wounds. They showed that recognition of the role of fibroblast senescence in wound chronicity may allow for identification of those wounds that will respond positively to these products. 5 Adipose-derived stem cells (ADSC) were shown to have relative advantages in accessibility and abundance compared to other kinds of stem cells in treatment of many dermatologic diseases. Subcutaneous injection of ADSC could significantly increase the collagen synthesis in hairless mice, and dermal thickness, collagen density and fibroblast number, angiogenesis, procollagen type I protein and mRNA expression also increased. They recommended that ADSC therapy may be useful in ageing skin as ADSC have antioxidant, whitening and wound-healing effects in the skin by secretion of growth factors and by activating fibroblasts. 6 The ADSC and its secretory factors were demonstrated to be effective for UVB-induced wrinkles too, and the antiwrinkle effect was mainly mediated by reducing UVB-induced apoptosis and stimulating collagen synthesis of HDF. 7 Cell therapy for facial anti-aging in clinical experience was introduced including cultured gingival fibroblasts injection lasting for at least one year, making it a good option for patients. 8 Park et al. (2008) demonstrated that ADSCs and their secretory factors showed promise for application in cosmetic dermatology, especially in the treatment of skin aging too. 9 In aging, loss of collagen and elastin are the important visible processes. Schmidt (2011) reported that the US Food and Drug Administration (FDA) approved laViv (azficel-T), a first-in-class personalized cell therapy to remove fine wrinkles or nasolabial folds around the nose and mouth. 10 Autologous fibroblasts grown in culture (azficel-T) were shown to correct the appearance of aging and wrinkles by replacing lost dermal constituents showing that autologous cell therapy can mark the beginning of a new phase in aesthetic therapy. 11 Eca et al. (2012) showed that injection of skin fibroblasts cultivated in medium supplemented with human serum was a viable method and had no side effects. Four injections at 15-day intervals containing 6.4×106 fibroblasts/mL could significantly improve periorbital skin flaccidity. 12 In a study by Khodadadi et al. (2010) on 10 patients with stable vitiligo, it was shown that intra-epidermal injection of dissociated autologous epidermal cells could improve the patches in 80% of subjects. 13 Dermal fibroblasts are responsible for synthesizing and organizing the dermis with three layers of (i) epidermis containing keratinocytes, melanocytes, and Langherans cells; (ii) dermis that is populated with fibroblasts, vessels and dendritic cells; and (iii) subcutaneous tissue. A basement membrane separates epidermis from dermis by composed of collagens and laminins, synthesized by fibroblasts and keratinocytes. Type I collagen is the most protein in the dermis produced by fibroblasts, synthesizing other collagens (III, V, VII), elastin, proteoglycans, and fibronectin too. The half-life of type I collagen in human skin was shown to be greater than 1 year. 14 Fibroblasts have a crucial role in wound healing. They produce matrix metalloproteinases and plasmin too. Its synthesis is increased in remodeling of an injured area after a wound 15 , 16 and also its production has an increasing trend in fibrotic diseases 17 , 18 while has a decreasing trend during aging and after a sun exposure. 19 For decades, skin organ culture has successfully been used to study skin ex vivo . 20 Normal human dermal fibroblast cultures can be divided into three phases of (i) primary cultures established by enzymatic digestion of the dermis, or by outgrowth of fibroblasts from explanted tissue pieces; (ii) secondary cultures as actively proliferating cells, provided from passage and expansion of primary cultures and (iii) terminal cultures eventually reaching a state of replicative senescence to be aging at the cellular level. 21 . 22 Cryogenic preservation of low passage-number fibroblasts is useful to maintain reserves of cells for further therapeutic measures. Therefore, it is easy to rapidly build a bank of fibroblasts from a limited number of skin samples. Culture of skin fibroblasts in collagen gels was first described by Bell et al. 23 The collagen gel serves as a stimulus to modulate fibroblast behavior. 24 In monolayer, fibroblasts are flat, spindle-shaped, and organized in parallel arrays, but in a three-dimensional collagen gel, they are elongated and have several dendrites. Fibroblasts in collagen gels usually proliferate slower than in monolayer cultures. 25 They are cultured in Dulbecco’s MEM supplemented with non-essential amino acids and 10% fetal bovine serum (DMEM-FBS). Cell growth is undertaken at 37◦C in 5% CO2 and 95% air. Fibroblasts are subcultured using trypsin/EDTA as required, and are used at passage 3 to 5. 26 Therefore, it seems that autologous fibroblasts are good sources to correct the appearance of aging and wrinkles by replacing lost dermal constituents and have a crucial role in healing showing that the autologous cell therapy can be a new phase in aesthetic therapy with no side effects for the patient. CONFLICT OF INTEREST The authors declare no conflict of interest.

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          Most cited references21

          • Record: found
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          The myofibroblast in wound healing and fibrocontractive diseases.

          G Gabbiani (2003)
          The demonstration that fibroblastic cells acquire contractile features during the healing of an open wound, thus modulating into myofibroblasts, has open a new perspective in the understanding of mechanisms leading to wound closure and fibrocontractive diseases. Myofibroblasts synthesize extracellular matrix components such as collagen types I and III and during normal wound healing disappear by apoptosis when epithelialization occurs. The transition from fibroblasts to myofibroblasts is influenced by mechanical stress, TGF-beta and cellular fibronectin (ED-A splice variant). These factors also play important roles in the development of fibrocontractive changes, such as those observed in liver cirrhosis, renal fibrosis, and stroma reaction to epithelial tumours. Copyright 2003 John Wiley & Sons, Ltd.
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            Effect of collagen turnover on the accumulation of advanced glycation end products.

            Collagen molecules in articular cartilage have an exceptionally long lifetime, which makes them susceptible to the accumulation of advanced glycation end products (AGEs). In fact, in comparison to other collagen-rich tissues, articular cartilage contains relatively high amounts of the AGE pentosidine. To test the hypothesis that this higher AGE accumulation is primarily the result of the slow turnover of cartilage collagen, AGE levels in cartilage and skin collagen were compared with the degree of racemization of aspartic acid (% d-Asp, a measure of the residence time of a protein). AGE (N(epsilon)-(carboxymethyl)lysine, N(epsilon)-(carboxyethyl)lysine, and pentosidine) and % d-Asp concentrations increased linearly with age in both cartilage and skin collagen (p < 0.0001). The rate of increase in AGEs was greater in cartilage collagen than in skin collagen (p < 0.0001). % d-Asp was also higher in cartilage collagen than in skin collagen (p < 0.0001), indicating that cartilage collagen has a longer residence time in the tissue, and thus a slower turnover, than skin collagen. In both types of collagen, AGE concentrations increased linearly with % d-Asp (p < 0.0005). Interestingly, the slopes of the curves of AGEs versus % d-Asp, i.e. the rates of accumulation of AGEs corrected for turnover, were identical for cartilage and skin collagen. The present study thus provides the first experimental evidence that protein turnover is a major determinant in AGE accumulation in different collagen types. From the age-related increases in % d-Asp the half-life of cartilage collagen was calculated to be 117 years and that of skin collagen 15 years, thereby providing the first reasonable estimates of the half-lives of these collagens.
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              UV-light-induced signal cascades and skin aging.

              L Rittié (2002)
              UV irradiation acts as a broad activator of cell surface growth factor and cytokine receptors. This ligand-independent receptor activation induces multiple downstream signaling pathways that regulate expression of multiple genes. These signaling pathways converge to stimulate transcription factor AP-1. Among genes whose expression is regulated by AP-1 are several matrix-metalloproteinase (MMP) family members and type I procollagen. UV-enhanced matrix degradation is accompanied with decreased collagen production mediated not only by activation of AP-1, but also by inhibition of transforming growth factor (TGF)-beta signaling. Several alterations to skin connective tissue that occur during aging are mediated by mechanisms that are similar to those that occur in response to UV irradiation. Thus, skin aging is associated with increased AP-1 activity, increased MMP expression, impaired TGF-beta signaling, enhanced collagen degradation, and decreased collagen synthesis. Knowledge gained from examining molecular responses of human skin to UV irradiation provides not only a framework for understanding mechanisms involved in skin aging, but also may help in development of new clinical strategies to impede chronological and UV-induced skin aging.
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                Author and article information

                Journal
                World J Plast Surg
                World J Plast Surg
                WJPS
                World Journal of Plastic Surgery
                Iranian Society for Plastic Surgeons (Tehran, Iran )
                2228-7914
                2252-0724
                January 2013
                : 2
                : 1
                : 2-5
                Affiliations
                [1 ]Stem Cell Research and Transgenic Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
                [2 ]Student Research Committee, Tehran University of Medical Sciences, Tehran, Iran
                Author notes
                [* ]Corresponding Author: Navid Manafi, Student Research Committee, Tehran University of Medical Sciences, Tehran, Iran. Tel: +98-2188797882, E-mail: dramanafi@ 123456yahoo.com
                Article
                wjps-2-002
                4238328
                25489497
                1efa9a51-5605-47f4-8668-cb6f0bcb9627

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, ( http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 6 September 2012
                : 1 November 2012
                Categories
                Editorial

                skin,fibroblast,aesthetic surgery
                skin, fibroblast, aesthetic surgery

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