The Cytocompatibility Study Of Carbon Fibre-reinforced Polyetheretherketone Hydroxyaapatite Composites In Vitro

ObjectiveTo evaluate the cytotocicity of the carbon fibre-reinforced polyeterethebrketone hydroxyapatite composites on osteoblasts and the effect of behaviors by co-culturing w-ith the osteoblasts in vitro.Methods1. A modified method for isolating and culturing pure osteoblasts from SD rats calvaria in vitro. Osteoblasts obtained from newly SD rat calvaria by means of enzymatical digestion serveral times, were cultured and subcultured to investigate their biological features. Inverted phase contrast microscope (IPM)was used to observe the cell-ular morphology The osteoblasts were identified by cell morphology, histochemical al-kaline phosphatase(ALP) staining and calcified nodules staining. The growth cure was recorded.2.Cytotocicity of carbon fibre-reinforced polyeterethebrketone hydroxyapatite composites on osteoblasts. Osteoblasts were cultured by leaching liquros of composites materials which were prepared according to IS010993-1.Cell relative growth(RGR) measured by CCK-8 kits was used as a quantitative assessment for cytotoxicity of the biomaterials.3.Effect of carbon fibre-reinforced polyeterethebrketone hydroxyapatite compoites on the behaviors of osteoblasts. The various composite samples were seeded with osteo blastic cells and miantained for a period of 1-14days. Adhesion in 24h were assessed. The influence on cell cycle of osteoblasts were detected by flow cytometry (FCM) for 1,3,and 7days. The gene expression of ALP, COL-I and osteocalcin(OCN) of different composites were detected by fluorescent real-time quantitative PCR at di-fferent culture time. Scanning electron microscopy(SEM) revealed the co-cultured cells could proliferate and grew normally in composites materials. Results1. Primary osteoblasts were round-like and then became triangular after 1-3day culture.Osteoblasts began attachto the dish bottom at 2h, Histochemical staining of alkaline phosphatase demonstrated 95%purity of cultured osteoblsts. The mineralized nodes were studied with alizarin red-S staining.2. The classification of cytotoxicity of carbon fibre-reinforced polyeterethe-brketone hydroxyapatite composites material was grade 0 to 1.3. No difference was found in the trend of cell proliferation and adhesion between differnet the differtent composites. Expressions of ALP mRNA,COL-Im RNA were increased with rising of HA in PEEK-HA-CF composites. Expressions of BGP mRNA were no significance among different groups(P>0.05). There were a great of bone-lik e nodules in high HA group (20%wt)under electron micros cope.Conclusions1. The results showed isolated osteoblasts were pure and retained their unique property in culture. This method of primary osteoblast culture is feasibe and provides pure osteblasts for the following tests.2. The biological responses of PEEK-HA-CF composites carried out in vitro v-erified cytocompibility and bioactive nature of the compostie materials,70PEEK-20H-A-10CF group have the most biological activiety.PEEK-HA-CF composites may be us-ed for orthopedic implanted application.