Phenotypic study of human gingival fibroblasts in a medium enriched with platelet lysate.
ABSTRACT The modulation abilities of gingival fibroblasts open new therapeutic strategies for the treatment of vascular diseases (e.g., aneurism) and irradiation burns. Culture media are classically supplemented with animal sera to provide nutriments. Unfortunately, because of their potential for interspecies transmission of microorganisms, these media are not used for cells destined for human transplantation. This preliminary phenotypic study aims to test a serum-free (SF) culture medium for human gingival fibroblasts (hGF) supplemented with human platelet lysates (PLs) for rapid cell expansion.
An SF medium was first elaborated to compete with hGF proliferation in a reference medium containing 10% fetal bovine serum (BSmedium). Adhesion, proliferation, and doubling kinetics were run in the presence of PLs (SF+PL). Cytoskeletal proteins were analyzed and chromosomal abnormalities were evaluated by karyotype analyses. The SF+PL influence on secretion of molecules implied in tissue remodeling (i.e., matrix metalloproteinases [MMPs], their tissue inhibitors [TIMPs], and several growth factors) was studied.
SF+PL increased the proliferation rate 1.5-fold in a week compared to BSmedium. Cytoskeleton protein expression was similar in BSmedium and in SF+PL. Chromosomal abnormalities were rare in SF+PL. MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, TIMP-1, and the growth factors interleukin-1β and -4 and transforming growth factor-β1 secretions were stable during the experiment. TIMP-2 and interleukin-6 were slightly decreased in SF+PL compared to BSmedium.
While waiting confirmation from a proteomic approach, this SF culture medium could allow a secured faster hGF proliferation adapted for human cell transplant therapy.
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ABSTRACT: Les cellules souches mésenchymateuses (CSMs) représentent une population hétérogène de cellules ressemblant à des fibroblastes et qui sont présentent dans différentes localisations, notamment la moelle osseuse, le tissue adipeux, les tissus extra-fœtaux, la gencive et le derme. Les propriétés des CSMs (multipotence, prolifération in vitro et immunotolérance) sont intéressantes en vue de leur utilisation en réparation tissulaire et en médecine régénératrice. Aujourd’hui, différents travaux et études cliniques soulignent l’intérêt des CSMs pour aider la réparation cutanée, soulignant que leurs effets bénéfiques sont essentiellement dus aux nombreux facteurs que ces cellules sécrètent. Les CSMs sont aussi utilisées pour développer des substituts cutanés. Dans cette étude, nous discuterons d’abord les différentes sources de CSMs disponibles. Nous présenterons ensuite des résultats montrant que des CSMs issues de la moelle osseuse et préparées en accord avec les bonnes pratiques de production généralement définies, sont capables, lorsqu’elles sont utilisées pour construire un derme équivalent, de promouvoir de façon tout à fait satisfaisante, la croissance et la différenciation épidermique. Ces résultats démontrent que les CSMs issues de la moelle osseuse représentent une alternative intéressante à l’utilisation des fibroblastes dermiques. De plus, les CSMs contribuent au développement de l’épiderme et peuvent ainsi faciliter la réparation des plaies.Pathologie Biologie 04/2014; DOI:10.1016/j.patbio.2014.01.002 · 1.07 Impact Factor
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ABSTRACT: Human gingival stem cells (HGSC) can be easily isolated and manipulated in culture to investigate their multipotency. Osteogenic differentiation of bone marrow derived mesenchymal stem cells (BMMSC) has been well documented. HGSC derive from neural crests, however, and their differentiation capacity has not been fully established. The aim of the present report was to investigate whether HGSC can be induced to differentiate to osteoblasts and chondrocytes. HGSC were cultured either in a classical monolayer culture or in 3-dimensional floating micromass pellet cultures in specific differentiation media. HGSC differentiation to osteogenic and chondrogenic lineages was determined by protein and gene expression analyses, and also by specific staining of cells and tissue pellets. HGSC cultured in osteogenic differentiation medium showed induction of Runx2, alkaline phosphatase and osterix expression, and subsequently formed mineralized nodules consistent with osteogenic differentiation. Interestingly, HGSC micromass cultures maintained in chondrogenic differentiation medium showed SOX9-dependent differentiation to both chondrocyte and synoviocyte lineages. Chondrocytes at different stages of differentiation were identified by gene expression profiles and by histochemical and immunohistochemical staining. In 3-week-old cultures, peripheral cells in the micromass cultures organized in layers of cuboidal cells with villous structures facing the medium. These cells were strongly positive for cadherin-11, a marker of synoviocytes. In summary, the findings indicate that HGSC have the capacity to differentiate to osteogenic, chondrogenic and synoviocyte lineages. Therefore, HGSC could serve as an alternative source for stem cell therapies in regenerative medicine for patients with cartilage and joint destructions, such as observed in rheumatoid arthritis.Stem Cells and Development 07/2014; DOI:10.1089/scd.2013.0547 · 4.20 Impact Factor
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ABSTRACT: Skin aging shows an imbalance between synthesis and degradation of the extracellular matrix. The overproduction of degradative enzymes (MMPs) during the chronology-and photo-induced aging leads to a degradation of the elastic and collagen networks. In a model of collagen and elastin destruction, we showed that the gingival fibroblast was able to preserve these macromolecules by inhibiting the overproduction of metalloproteinases by overproduction of TIMP-1 and modulation of the inflammatory cytokines activity. The objective of this study is to evaluate the effect of the gingival fibroblasts on human skin. The results in vitro and ex vivo show that the gingival fibroblast protects the skin collagen and elastic network by the inhibition of MMPs which leads to an overproduction of the TIMP-1. Moreover, the gingival fibroblast modulates the activity of some enzymes responsible for the inflammation; they inhibit the IL-1β and stimulate the production of TGF-β1. In vivo studies with a duration of six months and 50 women with pronounced wrinkles show that the culture supernatant of gingival fibroblasts diluted to 5% leads to a statistically significant decrease in the number and length of wrinkles.Journal of Cosmetics, Dermatological Sciences and Applications 01/2011; 01(01). DOI:10.4236/jcdsa.2011.11002