An Experimental Study On Osteoblast Proliferation Activity Of Coral Hydroxyapatite And Pure-phase Beta-tricalcium Phosphate

Objective:Through the cultivation of St2 cells, amplified by the mineralization induction into osteoblasts (OBs) differentiation, and induced the cells into bone cells identified, and then lines were cultured osteoblasts; will become bone cells and coral hydroxyl phosphorus transformed gray Stone (days Bo bone meal, CHA), pure-phaseβ-tricalcium phosphate (Cerasorb,β-TCP) composite in vitro culture, observation of CHA,β-TCP on osteoblast cell proliferation activity. From the cellular level evaluation CHA,β-TCP scaffold in both species and the proliferation of osteoblasts, thus scaffold for tissue engineering applications based on the choice of materials experimental basis.Method:1. St2 cells were cultured and passaged: St2 cell lines by culture, stable good experimental cell, application of low-sugar medium on St2 complete DMEM were passaged cultured cells, and cells were frozen as stocks.2. St2 cells into bone cells, identification and cultivation: Will grow well St2 cells into osteoblasts in normal culture medium, so that St2 cells into bone cells, and then the cells induced by Gomori Gomori method, alkaline phosphatase (ALP) staining, alizarin red ore of nodule staining, osteoblasts were identified osteoblast culture were amplified.3. Osteoblasts group inoculated CHA,β-TCP scaffolds were cultured. Combined with in vitro cell culture, osteoblasts were observed in the CHA and theβ-TCP on the proliferation, detected by direct counting the first 3,5,7 d osteoblasts and CHA,β-TCP composite cultured cells inoculation rate; then MTT method culturing 3,5,7 d complex when, CHA,β-TCP on osteoblast cell proliferation; detected with PNPP when the first 3,5,7 d, CHA,β-TCP on osteoblast ALP activity.Results:1. St2 cells induced to differentiate into osteogenic cells, osteoblast cells were identified:Gomori Gomori method, ALP staining cells were strongly positive reaction showed gray-black granules within the cytoplasm; alizarin red stained mineralized nodules mineralization nodus, dark red, nodular surrounding cytoplasm also was stained.2. Direct counting assay of osteoblasts and the CHA,β-TCP composite cultured cells inoculated with the situation:2.1 CHA,β-TCP with the surface of osteoblasts culture time, cells gradually increased, CHA, P-TCP have better biocompatibility.2.2 osteoblast proliferation rate in the CHA's surface faster than theβ-TCP.3. MTT assayβ-TCP, CHA on osteoblast cell proliferation and viability show:the surface of osteoblast proliferation activity in the CHA was higher thanβ-TCP.4. PNPP assay ALP activity in each group showed:OBs+CHA group of ALP activity was higher than OBs+β-TCP group (p<0.05).Conclusion:1. CHA on osteoblast proliferation rate than in theβ-TCP on fast, more suitable for bone tissue engineering scaffolds.2. Mineralization induced by osteoblast can get better withβ-TCP, CHA complex in vitro culture, and capable of rapid proliferation.3. Direct counting method using cell proliferation rate, the method can be directly reflected in the growth of cells in the surface condition, can be better evaluatedβ-TCP, CHA cell compatibility of two materials;4..β-TCP, CHA had good biocompatibility.