| Background:Periodontics result in destruction of periodontal tissues,including cementum,bone,and periodontal ligament,with eventual tooth loss if left untreated. Periodontal treatments are effective at promote the regeneration of periodontal structures. The healing of a periodontal wound associated with the number of periodontal cell,it has capacity of regeneration and differentiate in the periodontal. This cell may progress and differentiate into fibroblasts,osteoblasts,cementoblasts,etc. For the periodontic,the source of normal cell is limited in the periodontal tissues,especially in the older people. The stromal cell system associated with the soft connective tissue of marrow and bone surfaces contains multipotential stem cells that can differentiate into fibroblastic,adipocytic,ostegenic,and other cell lines under appropriate conditions. The capacity and superiorities of bone marrow stromal cells being seeded cells in the GTR. The recent studyindicated that the periodontal bone defect of furcation can be fully repairedwith BMSC transplantation.Objective:To evaluate the effects of alone or combination of human recombinant basic fibroblast growth factor(rhbFGF),recombinant human transforming growth factor- & 1 (rhTGF- ?1) and recombinant human bone morphogenetic protein-2(rhBMP-2) on the proliferation and differentiate of rat bone marrow stromal cells(BMSC) in vitro. Methods:Rat BMSC was cultured in vitro,rhbFGF(lug/L),rhTGF- & l(5ug/L) and rhBMP-2(200ug/L) alone or in combination,were added to medium,cells were examined by phase contrast microscope,cells growth were measured with cell counting,population doubling time of cell was calculated,cells proliferate were detected by [3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide,MTTJ.The cell differentiation was measured by the activity of alcaline phosphatase (ALP). BMSC cultured with conditional medium were tested with Von Kossa staining. Results:The result demonstrated that the cell population of groups was promoted on the 3th day,the cells proliferation of control group,t,f,f+t and f+t+b group were stationary on the 4th day,cells of b and f+b groups have strong proliferative ability,the cell of f+b group has the shortest population doubling time about 32 hours,proliferation of cells was stimulated by rhBMP-2(200ug/L) group and rhbFGF(lug/L)+rhBMP-2(200ug/L) as compared with control group in cell counting and MTT (P<0.05). Alkalinephosphatase activity was stimulated by rhBMP-2. t,f,f+t and f+t+b groupsinhibited alkaline phosphatase activity (P<0.05).Conclusion:1. rhbFGF(lug/L)+ rhBMP-2(200ug/L) group could stimulate BMSC to proliferate,rhbFGF(lug/L)+ rhBMP-2(200ug/L) can be used for the best culture condition of BMSC proliferation in vitro.2. Alkaline phosphatase activity and cell proliferation was stimulated by rhBMP-2.3. The alkaline phosphatase activity was inhibited by rhbFGF and rhTGF-pl alone or combination.4. The cell proliferation and the alkaline phosphatase was inhibited by rhBMP-2 and rhTGF-pl combination.5. Combined treatment with rhbFGF,rhTGF-pl and rhBMP-2 inhibited the alkaline phosphatase activity and had no effect on proliferation.6. rhTGF-pl increased the transcriptional and polypeptide levels of osteopontin with dexamethasone,p-glycerophosphate and ascorbic acid. Cadherins increased the intercellular adherins. |
