The Study Of Rabbit Bone Marrow Stromal CellsAdhesion To Cementum Slices In Vitro

Cementum is similar with bone in both structure and characteristic, but not in ability to regenerate because of the lack of regeneration cells, such as periodontal cells. Bone marrow stromal cells(BMSCs) are easy to obtain and culture in vivro, and they have potentials to differentiate to various kinds of cells. What people are interest in is if BMSCs can be used as seed cells in cementum regeneration. The goal of this project is to detect BMSCs ashesion and proliferation on cementum. This may help people find a new way in periodontal disease therapy.Methods1. Bone marrow was aspirated from New Zealand white rabbit and culture-expanded in vitro by the method of total marrow. Subsequently, passages of BMSCs were cultured in DMEM media containing dexamethasone(10-8 mol/L), beta-sodium glycerophosphate(10mmol/L and ascorbic acid(50mg/L). Cells were observed continually by inverted phase contrast microscope, Alkaline phosphatase(ALP) and Calcifying nodule were stained by Gomori Calcium-Cobalt and Von Kossa methods.2. Central incisor were extracted from same rabbit and were shaped to cementum slices(3mm X 2.5mm X 0.5mm). BMSCs(passage 3-4) were seeded on cementum and coverslip(use as control). Each sample were seeded with 1 X 106 cells, then culture in incubator at 37C, 5%CO23. HE staining and scanning electron microscope(SEM) were performanced to check BMSCs ashesion and proliferation on cementum slices at different time point.Results1. BMSCs show fibroblast-like morphology, most cells were like circle and fusifoum shape. ALP and Calcifying nodule staining are both show positive results2. HE staining show BMSCs has good adhesion on cementum slice and they can grow confluent. After 4 weeks, new tissue formation were observed on cementum slice.3. SEM results show BMSCs grow as well as normal cultured cells after seed on cementum slice, calcium crystal deposition were observed aroundcells. After 4 weeks, mineral layer were observed. Conclusion1. BMSCs cultured in vitro can be induced to show osteoblast characteristics: show strong ability about mineralization2. BMSCs seeded on cementum slice has good adhesion, proliferation and defferentiation3. BMSCs has potential to be used as seed cell in cementum regeneration...