Particle size of hydroxyapatite granules calcified from red algae affects the osteogenic potential of human mesenchymal stem cells in vitro
Particle size of hydroxyapatite granules calcified from red algae affects the osteogenic potential of human mesenchymal stem cells in vitro
复制标题
DOI：
10.1159/000093062
复制
发表时间：
2006-01-01
影响因子：
2.7
通讯作者：
Turhani, Dritan
中科院分区：
文献类型：
--
作者：
Weissenboeck, Martina;Stein, Elisabeth;Turhani, Dritan
羟基磷灰石（HA）微粒作为可注射组织工程骨填充物的载体被认为是治疗颅颌面区小骨缺损的有希望的候选者。从红藻中钙化的透明质酸颗粒，大小不一，在体外评估了它们作为人间充质干细胞（hMSCs）载体的适用性。制备3组粒径为10-100、200500和600- 1000 μ m的HA颗粒，在成骨分化条件下将hMSCs在HA颗粒上播种培养3、6和9 d，观察细胞增殖（四氮唑盐，XTT）、碱性磷酸酶（ALP）特异性活性和总蛋白合成。通过检测骨特异性基因骨钙素、骨桥蛋白和i型胶原蛋白来评价细胞的成骨表型。XTT法显示，培养9天后，最小粒径的细胞增殖显著提高（p < 0.01）。关于alp特异性活性，在最小晶粒上生长的细胞中检测到明显更高的活性水平。不同粒径对骨钙素和骨桥蛋白的分泌无显著影响。最大粒径的细胞I型胶原产量显著高于其他两种粒径的细胞（p < 0.05）。这些结果表明，HA微粒的颗粒大小影响培养的hMSCs的成骨潜能，并得出颗粒大小对alp特异性活性和I型胶原生成有不同影响的结论。
Hydroxyapatite (HA) microparticles as a carrier in an injectable tissue-engineered bone filler are considered promising candidates for the treatment of small bone defects in the craniomaxillofacial region. HA granules calcified from red algae, varying in size, were evaluated in vitro for their suitability to be used as a carrier for human mesenchymal stem cells (hMSCs). Three groups of granules were produced in grain sizes of 10-100, 200500 and 600-1,000 mu m. After seeding and culturing hMSCs under osteogenic differentiation conditions onto HA particles for 3, 6 and 9 days, cellular proliferation (tetrazolium salt, XTT), alkaline phosphatase (ALP)-specific activity and total protein synthesis were investigated. The osteoblastic phenotype of the cells was evaluated by assaying the bone-specific genes osteocalcin, osteopontin and Collagen type I. XTT assay revealed significantly higher (p < 0.01) proliferation of cells grown on the smallest grain size after 9 days of culture. Regarding ALP-specific activity, significantly higher levels of activity were detected in cells grown on the smallest grain size. Different grain sizes had no significant effects on the secretion of osteocalcin and osteopontin. Collagen type I production was significantly higher (p < 0.05) in cells grown on the biggest grain size in comparison with the two other grain sizes. These results show that the particle size of HA microparticles affects the osteogenic potential of cultured hMSCs and lead to the conclusion that particle size has differential effects on ALP-specific activity and collagen type I production.