The effect of hyaluronan on osteoblast proliferation and differentiation in rat calvarial-derived cell cultures
The effect of hyaluronan on osteoblast proliferation and differentiation in rat calvarial-derived cell cultures
复制标题
DOI：
10.1002/jbm.a.10535
复制
发表时间：
2003-09-15
影响因子：
4.9
通讯作者：
Kumta, SM
中科院分区：
文献类型：
--
作者：
Huang, L;Cheng, YY;Kumta, SM
透明质酸（Hyaluronan，HA）是细胞外基质的重要组成部分。它与其他大分子相互作用，并在组织形态发生、细胞迁移、分化和粘附中起主导作用。HA的细胞信号传导功能通过CD-44受体介导，并取决于聚合物的分子量。我们假设，在最佳浓度下单独使用适当分子量的HA可以诱导成骨细胞分化和骨形成。将来自2日龄新生大鼠的酶消化的颅骨来源的间充质细胞与三种不同分子量（2300、900和60 kDa）的HA一起培养。在第一次接种细胞时，我们向培养基中加入0.5、1.0和2.0 mg/mL HA（每种分子量）。在培养7至20天后，通过测量胸苷掺入、碱性磷酸酶活性和骨钙素基因表达来评价细胞增殖和分化。茜素红染色法观察HA对骨形成的影响。结果表明，低分子量HA（60 kDa）能显著刺激细胞生长，并呈剂量依赖性增加骨钙素mRNA表达，但对碱性磷酸酶活性和骨矿化无明显影响。另一方面，高重量HA（900和2300 kDa）显着增加所有检查的参数，特别是碱性磷酸酶活性，在剂量依赖性的方式和刺激细胞矿化的126%和119%的控制，分别为1.0 mg/mL的剂量。我们的研究结果表明，HA具有分子量特异性和剂量特异性的作用模式，由于其对成骨细胞的刺激作用，可能会增强骨移植材料和替代品的成骨和骨诱导特性。(C)2003 Wiley Periodicals，Inc.
Hyaluronan (or hyaluronic acid, HA) is an essential component of extracellular matrices. It interacts with other macromolecules and plays a predominant role in tissue morphogenesis, cell migration, differentiation, and adhesion. The cell signaling functions of HA are mediated through the CD-44 receptor and are dependent upon the molecular weight of the polymer. We hypothesized that an HA of appropriate molecular weight alone in optimal concentration may induce osteoblast differentiation and bone formation. Enzyme-digested calvarial-derived mesenchymal cells from 2-day-old newborn rats were cultured with the addition of HA of three different molecular weights (2300, 900, and 60 kDa). We added, 0.5, 1.0, and 2.0 mg/mL HA for each molecular weight to the medium at the first plating of cells. After 7 to 20 days in culture, cell proliferation and differentiation were evaluated by measuring thymidine incorporation, alkaline phosphatase activity, and osteocalcin gene expression. The effects of HA on bone formation were examined by using Alizarin red staining for mineralization. The results showed that low molecular weight HA (60 kDa) significantly stimulated cell growth, increased osteocalcin mRNA expression in a dose-dependent manner, but showed no apparent effects on alkaline phosphatase activity and bone mineralization. On the other hand, high-weight HA (900 and 2300 kDa) significantly increased all the parameters examined, particularly alkaline phosphatase activity, in a dose-dependent manner and stimulated cell mineralization to 126% and 119% of the controls, respectively, in the 1.0 mg/mL dose. Our findings suggest that HA has a molecular weight-specific and dose-specific mode of action that may enhance the osteogenic and osteoinductive properties of bone graft materials and substitutes due to its stimulatory effects on osteoblasts. (C) 2003 Wiley Periodicals, Inc.