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Purpose of a preliminary study the role of nano-hydroxyapatite suspension of human periodontal ligament fibroblasts, and provide the basis for nano-hydroxyapatite used in the treatment of periodontal disease. 2,-based research group future studies of drug-loaded nano-and mesoporous materials related apoptosis of fibroblasts to prepare. Method 1. Culture method using tissue blocks were identified and passaged in human periodontal ligament fibroblasts, the to first 4-6 fibroblast experiment. Laboratory preparation of nano-hydroxyapatite, sodium carboxymethyl cellulose, sodium hexametaphosphate as a dispersing agent, contrast observed the effect of two kinds of dispersants on the dispersion of nano-hydroxyapatite and simple cells culture, compared to cell growth, to select the appropriate dispersant. 3 using the dispersing agent to be dispersed on the nano hydroxyapatite, l% concentration of nano hydroxyapatite suspension was prepared, were diluted to 0.5%, 0.25%, 0.1%, 0.05%, 0.025%, and a set of separate 0.1% concentration of nano-hydroxyapatite group, a separate 0.1% concentration of sodium carboxymethyl cellulose group, 0.005% mesoporous silicon group, 0.01% calcium hydroxide group and the control group on human periodontal ligament fibroblasts cultured 24h, 48h, 72h after the groups cell proliferation using the MTT assay measured, evaluation groups into human periodontal ligament fibroblast proliferation. 4. Selected concentration of 0.5% cell suspension and the control group continued for 48h, the the groups cultured cells using flow cytometry cell cycle distribution, and analysis of cells in each group on human periodontal ligament fibroblasts cycle. Results 1. Success cultured human periodontal ligament fibroblasts, determined by the anti-vimentin, anti-cytokeratin: anti-vimentin-positive, anti-keratin negative. Laboratory were prepared by chemical precipitation of nano-hydroxyapatite meet this experiment required particle size detection resulting nano-hydroxyapatite particle size in the 20-40 nm. By comparing six partial sodium phosphate and sodium carboxymethyl cellulose dispersion effect of nano-hydroxyapatite, sodium hexametaphosphate better dispersion of nano-hydroxyapatite, but cell cultures can be drawn using six partial sodium dispersed nano-hydroxyapatite culture has led to a large number of human periodontal ligament fibroblasts death, cell growth and sodium carboxymethyl cellulose dispersed nano-hydroxyapatite group. So choose configure nano hydroxyapatite suspension, sodium carboxymethyl cellulose as a dispersing agent for subsequent experiments. 3.MTT cell proliferation assay grouping of different concentrations and multiple control group, found that 0.5% concentration of nano-hydroxyapatite suspension group is more obvious than the other group effect on cell proliferation. Nano-hydroxyapatite suspension group of cell proliferation in the same concentration (0.1%), is superior to nano-hydroxyapatite group. Using flow cytometry results showed that nano-hydroxyapatite suspension group can reduce the number of cells in the G0/G1 phase, increasing the number of cells in S phase, lead to periodontal membrane into fiber cells in the cell proliferation cycle soon complete the G0/G1, period of transition to the s. Conclusion 1. Nano-hydroxyapatite suspension on human periodontal ligament fibroblasts promote proliferation. Allow osmolality in cell culture, the more obvious role in promoting the higher the concentration of cell proliferation, and nano-hydroxyapatite on human periodontal ligament fibroblast proliferation promoting the most obvious role in the first 24h. Nano-hydroxyapatite suspensions role in human periodontal ligament fibroblasts, can shorten cell in Go/G1, time to make a relatively large number of cells into the S phase of mitosis, accelerate the proliferation rate of HPDLCs. Through this study, further affirmed nHA used in periodontal soft tissue reconstruction may provide a reference for nHA used in the treatment of periodontal disease.
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