| Objective:This study investigated the effects of different porosity and polarization treatment for porous HA/BaTiO3biological piezoelectric ceramic (HABT)on adhesion, proliferation and differentiation of osteoblast cells in vitro,and lay the theoretical foundation for the research and development of new biological piezoelectric implant materials.Materials and methods:Firstly this study prepare dense and porosity of40%,50%,60%of the biological piezoelectric ceramic by ice-templating method; secondly conducted the toxicity experiment by MTT method on the rat fibroblast cells L929; thirdly co-culture of the different porosity porous biological piezoelectric ceramic with osteoblasts in vitro conditions, set dense piezoelectric ceramic group (A) for the control group, and set40%porosity group (B),50%porosity group (C),60%porosity group (D) for the experimental group, assessed the influence of different porosity porous piezoelectric ceramic on adhesion、proliferation and differentiation of osteoblast cells MG63in vitro; lastly poling was conducted at110℃in silicone oil at2.5kV/mm for20min using a DC power supply for the50%porosity HABT, co-culture of the polarized biological piezoelectric ceramic with osteoblasts in vitro conditions, set non-polar materials for the control group,the polarized material group for the experimental group, compared two groups of materials on osteoblasts cells MG63adhesion, proliferation and differentiation of function in vitro.Results:1) The MTT test showed the leaching liquor of the porous biological piezoelectric for the90%BaTiO3group,70%BaTiO3group and50%BaTiO3group was nontoxic to L929. 2) Biological piezoelectric ceramic cultured with MG63in vitro, after cultured3、7days, the proliferation results of MG63showed that the amount of cells in C Group> B the group> D group>A group, and there was no significant difference between the number of cells in Group B and Group D (P>0.05), the rest of the group difference was significant (P<0.05); observing cell culture in vitro on14th day by SEM, we found that the shape of cells was irregular in group A, the cells tiled on the surface of the material, formed a multi-layer structure, and the structure of the surface of the piezoelectric ceramic materials could barely be seen, the cells had pseudopods; and group B cells had irregular polygons, stretched well, a plurality of cell protrusions firmly attached between the surface of the particles of the material, and some cells into the pores formed the ingrown trend; group C and group D cells began to form pore growth, the pseudopodium of cell adhered to the surface of the bore above the bilateral particles like the drawbridge, forming cells bridge, some cells showed the growth of the vertical direction, and the more cells had grown into the material pores, a plurality of cell projections firmly attached to the top of the particles of the material the hole wall and linked together to form a mesh structure in the pores; after cultured3days, the ALP and BGP productions of group B、group C、group D were higher than group A, the difference was remarkable (P<0.05),after cultured7、14days, it showed the ALP、BGP content in group C> group D> group B> group A, of which the ALP content in group C, group D without significantly differences (P>0.05),and BGP content in group B, group D without significantly differences (P>0.05),and the remaining group differences twenty-two were significantly (P<0.05).3) The porous HABT and MG63cells co-cultured after3、7and14days under polarization conditions, detecting MG63cells’ proliferation and differentiation in vitro, the results show that there was no significant difference between the number of cells、the ALP and BGP content in the polarization group and the non-polarized group(P>0.05). Observing cell culture in vitro on7th day by SEM, we found that osteoblast cells mainly spreaded on the surface of the material irregularly, the cells was fusiform, multiple cells projection attached to the surface of the material between the particles, formed inwardly growing trend, and there was no difference between the control group and the polarized group on the number of cell morphology; on14th day, the cell morphology was similar with7th day, the number of adherent growing cells significantly increased, and there was no significant difference between the polarized group with the control group.Conclusion:1) The biological piezoelectric ceramics prepared by the ice template are nontoxic to fibroblast cells in vitro.2) the connected pore structure of porous biological piezoelectric ceramic can promote osteoblasts adhesion、proliferation and differentiation,50%porosity group was better than40%、60%porosity group for the role of promote MG63cells’ proliferation and differentiation.3) the porous HABT materials for the promotion of osteoblasts adhesion、 proliferation and differentiation was not obvious under the polarization conditions without applied loading. |
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