Study Of Human Gingival Fibroblasts Induction Into Skeletal Muscle-like Cells

Objective:Oral gingival tissue is used as the periodental support tissue which corvedon the tooth surface and formed dento-gingival junction. Gingival tissue ismainly composed of fibroblasts. Previous studies have proved that gingivalfibroblasts possess the properties of stem cells which are strongly potential torepair, immigration and differentiation from different subpopulation cells.Basic physiological functions of oral cavity including mastication,pronunciation and facial motion such as smile are performed by skeletalmuscle contraction. However, it is so weak that skeletal muscle cellsregenerate when they has been damaged. The reconstruction of the lostedskeletal muscle tissue is renovated by fibroblasts and finially replaced by scartissue. So it is becoming urgent to look for new sources of myoblasts. Iffibroblasts could be differentiated into skeletal muscle cells, a sort of newmyoblast seed cells could be found to form new muscle tissues, so that toreduce the scar formation during the repairation of muscle cells damaged. Thisstudy was to reveal if human gingival fibroblasts possess the properties ofmulti-lineage differentiation as stem cells by inducing into skeletalmuscle-like cells.Methods:1Cultured and identified of human gingival fibroblasts.Normal human gingival tissue was extracted from tooth impacted therapy ofa patient (female,28years old) in Department of Oral&Maxillofacial Surgery,Hospital of Stomatology, Hebei Medical University.Gingival fibroblast cells were cultured in DMEM-low glucose contained20%FBS by using traditional method of primary culture in vitro. The thirdgeneration cells were identified by immunocytochemical stain againstvimentin and α-sarcometric actin&myosin antibodies respectively.2Induced and identified of human gingival fibroblasts Given concentrations of5-Aza, Myod，IGF-1were added to the3passagesof human gingival fibroblasts in DMEM/F12contained20%FBS. Themorphology was observed by phase-contrastmicroscopy. The cells inducedwere identified by immunocytochemical stain against α-sarcometricactin&myosin and vimentin antibody repectively. They were also identified byMasson stain. Electron Scanning Microscope (SEM) was used respectively toobserve the cells that the human gingival fibroblasts induced.Result:1Cultured and identified of human gingival fibroblastsSpindle cells with round or ellipse nucleus containing1~3nucleolus wereobserved in primary culture. Immunocytochemical stain against vimentinantibody was positive in the cytoplasms of human gingival fibroblasts.2Induction of human gingival fibroblastsAfter38-day induced, the cell morphology changed from long fusiform tocolumnar and long cylindrical feature was found by Phase-ContrastMicroscopy and SEM.Immunocytochemical stain against mixed α-sarcometric actin&myosinantibody was positive in the cytoplasms of the skeletal muscle-like cells afterinduced, which showed the immunocytological feature of skeletal muscle cells.Masson stain showed positive in the cytoplasm of skeletal muscle-like cells.The results of SEM showed that the skeletal muscle-like cells grewcolumnar with round nucleus containing1-2nucleolus, and the skeletalmuscle-like cells structures were formed.Conclusion： Human gingival fibroblasts could be induced anddifferentiated to skeletal muscle-like cells.