| Objective:Fabricate Collagen/Hyaluronic acid composites coating on implant surface via Layer-by-Layer technique. Evalucate the biological properties of the CA/HA coating by culture mouse preosteoblasts and human gingival fibroblasts (HGF) respectively on the surface in vitro.Method:Divide Titanium discs into three groups according to the treatment methods, a CA/HA coating Ti group (A), an acid etched Ti group (B) and a machine-smooth Ti group (C). The Ti discs'formation and properties of three groups were investigated by SEM observation, contact angles measurement and XPS detection. Then, mouse preosteoblasts and human gingival fibroblasts were respectively cultured to evalute the biological properties of different Ti discs by different treatment methods. Observe cell attachment and spreading on Ti discs at cell culture 1h,3h and 24h by fluorescence microscope and take fluorescence images. Use a shape factor to discribe cell morphology. According to the results of measuring ALP activity, OC release, total protein and collagen type I release to compare the situation of cell proliferation and differentiation on different Ti discs.Results:The characterization of the surface morphology observed by SEM and the chemical component of Ti surface which detected by XPS showed that Ca/HA coating could be fabricated on Ti discs surface successfully via Layer-by-Layer technique.By 24h, the cells cultured on Group A were more elongated and exhibited numerous cellular processes than that on Group B and Group C, and he shape factor of Group A was the lowest among the three different surfaces at any time point.The DNA content of proliferated preosteoblasts of Group A is significantly higher than that of other groups at 7d. In the case of HGF, the DNA content of Group C is the highest at 4d. But at 7d and 14d, the DNA content followed the order of Group A> Group B> Group C, and the difference was significantly.The ALP activity and the amount of OC of Group A were significantly higher than that of Group B and Group C. Only at 4d, the production of collagen type I in SM-Ti group was the highest. At 7d and 14d, the Col level in Group A was significantly higher than that in the other two groups.The expression levels of Akp-2 mRNA, OC mRNA and Col mRNA in Group A were significantly higher than those in the other two groups at all three time points.Conclusion:The Ca/HA coating could be fabricated on Ti discs surface successfully via Layer-by-Layer technique. The results of cell culture showed the Ti discs of Group A could accelerate the proliferation and differentiation of mouse preosteoblasts and human gingival fibroblasts compared with the other two groups. It's means that the CA/HA coating could be favorable to both osseintegration and soft tissue integration.
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