羟基磷灰石/二氧化锆复合材料颗粒对骨髓间充质干细胞增殖及成骨分化的影响

目的 采用分子生物学方法评价羟基磷灰石/二氧化锆(HA/ZrO2)复合材料颗粒对兔骨髓间充质干细胞(MSCs)增殖及成骨分化的影响.方法 按不同比例将纳米HA粉体与纳米ZrO2粉体混合在高温下烧结制备HA/ZrO2复合材料颗粒,分离和培养兔MSCs.MTT法检测复合材料颗粒悬液对兔MSCs增殖促进作用,ALP法测定碱性磷酸酶活性,RT-PCR检测细胞Ⅰ型胶原(collagen Ⅰ),骨钙蛋白(osteocalcin),骨桥蛋白(osteopontin)mRNA表达.结果 HA和含有HA的复合材料颗粒均对细胞增殖有一定促进作用.Vonkossa染色显示复合材料和纯HA颗粒可使细胞阳染程度降低.细胞在HA和含有HA的复合材料颗粒的成骨诱导培养液中的ALP活性均显著高于常规培养基质及纯ZrO2组(P＜0.05).RT-PCR结果显示复合材料颗粒能够促进collagen Ⅰ和osteocalcin基因的表达.结论 HA/ZrO2复合材料颗粒可促进骨髓间充质干细胞增殖及成骨分化.

Abstract：

Objective To evaluate the effect of the HA/ZrO2 composite particle on proliferation and osteogenesis of rabbit mesenchymal stem cells (MSCs) by using molecular biology methods.Methods The HA/ZrO2 composite particles were sintered at high temperature using the powder of HA and ZrO2 with different proportions.MSCs were isolated from rabbits and cultured.The effect of the composite particles on promoting cell proliferation of rabbit MSCs was detected using MTT method.Alkaline phosphatase activities were measured with ALP method.RT-PCR method was applied to measure the expression of collagen Ⅰ , osteocalcin and osteopontin mRNA.Results Pure HA pauicles and composite particles which contain HA promoted MSCs proliferation.Vonkossa staining showed that both pure HA particks and composite particles decreased the degree of positive stain of MSCs.ALP test showed that cell activity in the culture medium that contained pure HA particles or composite particles was significantly higher than that in the conventional culture medium and that contained pure ZrO2 particles (P ＜ 0.05＝.RT-PCR results showed that composite particles promoted the expression of collagen Ⅰ and osteocalcin gene.Conclusion The HA/ZrO2 composite particles promote proliferation and osteogenesis of MSCs.

The effect of the HA/ZrO2 composite particle on proliferation and osteogenesis of MSCs

Objective To evaluate the effect of the HA/ZrO2 composite particle on proliferation and osteogenesis of rabbit mesenchymal stem cells (MSCs) by using molecular biology methods.Methods The HA/ZrO2 composite particles were sintered at high temperature using the powder of HA and ZrO2 with different proportions.MSCs were isolated from rabbits and cultured.The effect of the composite particles on promoting cell proliferation of rabbit MSCs was detected using MTT method.Alkaline phosphatase activities were measured with ALP method.RT-PCR method was applied to measure the expression of collagen Ⅰ , osteocalcin and osteopontin mRNA.Results Pure HA pauicles and composite particles which contain HA promoted MSCs proliferation.Vonkossa staining showed that both pure HA particks and composite particles decreased the degree of positive stain of MSCs.ALP test showed that cell activity in the culture medium that contained pure HA particles or composite particles was significantly higher than that in the conventional culture medium and that contained pure ZrO2 particles (P ＜ 0.05＝.RT-PCR results showed that composite particles promoted the expression of collagen Ⅰ and osteocalcin gene.Conclusion The HA/ZrO2 composite particles promote proliferation and osteogenesis of MSCs.