Phosphorylated Chitooligosaccharides(PCOS)/Basic Fibroblast Growth Factor(bFGF) Exert Regulatory Effects On Proliferation And Mineralization In Human Alveolar Osteoblasts

Objectives: Basic fibroblasts play an important role in the regeneration of alveolar bone.The study investigated the effects of phosphorylated chitooligosaccharides / basic fibroblast growth factor(PCOS/b FGF)on the proliferation and mineralization of alveolar osteoblasts(HAOBs)in healthy adults and the expression of osteogenic-related gene,which can provide reference for periodontal tissue regeneration and repair.Methods: 1 Adult healthy HAO Bs were cultured using a modified pre-digested tissue block method in vitro.Morphological observations were performed by electron microscope.Characteristic of mineralization was performed using alkaline phosphatase(ALP)assay and cytochrome-alizarin red-calcium nodular staining.2 MTT assay was used to detect the proliferation of HAOBs after the administration of PCOS,b FGF and PCOS/b FGF.3 The induction and no-induction of HAOBs mineralization was detected by ALP staining,Sirius red staining,and alizarin red calcium nodule staining,the degree of mineralization of HAOBs was determined in groups of PCOS?b FGF and.different proportions of PCOS/b FGF.The expression levels of mineralization-related genes in the induced HAOBs(ALP,COL1a1,RUNX2,OSX,OCN)were detected by RT-q PCR.Results: 1 HAOBs were shuttle or triangular in shape at the 6th day in shape.90% of cells were positive expression visualizing by ALP after induction of mineralization.The results of alizarin red staining showed cel s produce calcified nodules.2 MTT results showed that: When b FGF was selected as 20ng/m L,the proliferation ratios of HAOB streated with PCOS/b FGF were significantly different compared with blank cell group at 48 h and 72 h(P<0.01).But there was no significant difference in PCOS/b FGF treated cell groups at different ratios.3 10 days after osteogenic mineralization inducted in HAO Bs,the b FGF-containing experimental group stained lighter than the induction group alone,and the induced b FGF group showed the lightest staining;and the expression level of ALP and COL1a1 was lowest in the b FGF group(P<0.05).Conclusion: 1 HAOBs can be successfully cultured in vitro using modified pre-digestion method;mineralization of HAOBs in vitro requires the effect of an osteogenic inducer.2 Basic fibroblast growth factor can promote the proliferation of HAOBs,but inhibit mineralization of HAOBs induced by the inducer solution.3 Phosphorylated chitooligosaccharides had no significant effect on the proliferation of HAOBs.phosphorylated chitooligosaccharides combined with basic fibroblast growth factor could reduce the mineralization of HAOBs after induction,presumably because phosphorylated chitooligosaccharides provide the phosphorus to promote HAOBs mineralization.However,there was no significant difference in PCOS/b FGF treated cell groups at different ratios.