经表面修饰的β-TCP对BMSCs细胞增殖及分化作用的研究

Surface Modified β-Tricalcium Phosphate Ceramic on Proliferation and Differentiation of Bone Marrow Stromal Cells

目的研究三种材料,材料一:-磷酸三钙支架(-TCP);材料二:明胶/低结晶度羟基磷灰石涂层--磷酸三钙支架(gel/lc-HA--TCP);材料三:明胶/高结晶度羟基磷灰石涂层--磷酸三钙(gel/hc-HA--TCP)。三种材料的细胞毒性及兔骨髓基质细胞(BMSCs)与材料共培养增殖的情况。方法取兔股骨骨髓腔细胞,进行贴壁培养BMSCs。在成骨诱导液中诱导BMSCs。将诱导后的BMSCs与三种支架材料在培养板内共培养1、3、5、7、9d。采用形态学观察、MTT法及ALP检测试剂盒等方法检测材料的BMSCs细胞毒性及BMSCs细胞在材料表面的增殖和分化能力。结果光镜及电镜下观察各组无显著差异。细胞毒性在0-1级。与细胞共培养,-TCP组在第7、9天与阴性对照组差异有统计学意义。ALP检测,-TCP组在第7、9天与阴性对照组差异有统计学意义(P<0.05),gel/lc-HA--TCP在第9天与阴性对照组差异有统计学意义(P<0.05)。结论新型明胶/高结晶度羟基磷灰石涂层--磷酸三钙支架和明胶/低结晶度羟基磷灰石涂层--磷酸三钙支架与BMSCs生物相容性好,适合作为支架材料负载BMSCs构建组织工程骨。

Objective To research three materials ,the first material： β-TCP scaffold ;the second material：β-TCP scaffold coated with gel/low crystalline hydroxyapatite （gel/lc-HA-β-TCP）;the Third material： β-TCP scaffold coated with gel/high crystalline hydroxyapatite （gel/hc-HA-β-TCP）.To study three materials cytotoxicity and on Proliferation of Bone Marrow Stromal Cells（BMSCs）. Method BMSCs were harvested from the medullary cavity of femurs and obtained by using adhesive culture method .BMSCs were induced with osteogenic medium .Then the induced BMSCs were cultured in vitro and implanted on three materials .The proliferation and differentiation of BMSCs were measured at ld, 3d, 5d, 7d and 9d through morphology, MTT detection and ALP kit. Results The BMSCs had uniformity appearance on three materials under the inverted microscope and the scanning electron microscope. And the toxicity ranges from 0 to 1.Proliferative effect of scaffolds on MSCs： On the 7th and 9th day, the proliferative effect of scaffolds on BMSCs in the β-TCP group was lower than that in the control group, and there were significant differences （P〈 0.05） . Activity of alkaline phosphatase in BMSCs： The activity of alkaline phosphatase in β-TCP group was obviously lower than that in the control group at days 7 and 9. And the gel/Ic-HA-β-TCP group was obviously lower than that in the control group at 9th day, and there were significant differences （P〈 0.05）. Conclusion β-TCP scaffold coated with gel/high crystalline hydroxyapatite and - TCP scaffold coated with gel/low crystalline hydroxyapatite have a good biocompatibility with BMSCs, and they were suitable scaffolds for BMSCs to fabricate tissue engineered bone.