Sekiya K, Nishimura M, Suehiro F, Nishimura H, Hamada T, Kato Y
Department of Prosthetic Dentistry, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
Int J Artif Organs. 2008 Aug;31(8):708-15. doi: 10.1177/039139880803100804.
This study investigates concanavalin A (ConA) as a novel factor that may enhance osteogenesis of mesenchymal stem cells (MSCs) in vitro. Various factors, such as cytokine bone morphogenetic protein-2 (BMP-2), have been studied for their possible promotion of MSC osteogenesis in vivo and in vitro. However, the factor that might be safer, more effective, and less expensive than these has not been determined. We therefore cultured human MSCs in osteogenic medium in the presence or absence of ConA, and used calcium assays to compare the effects of ConA and BMP-2 on MSC calcification. We also used enzyme-linked immunosorbent assay (ELISA) and quantitative polymerase chain reaction (PCR) to evaluate the expression levels of bone-specific markers. ConA and BMP-2 enhanced calcification with comparable effectiveness. The combination of ConA and BMP-2 further enhanced calcification slightly but significantly. ConA also increased osteocalcin and BMP-2 protein levels in MSC culture medium. Furthermore, ConA increased osteocalcin, RUNX2, BMP-2, BMP-4, and BMP-6 mRNA expression levels. However, the gene expression pattern of ConA-stimulated MSCs was different from that of MSCs stimulated by BMP-2. Together, these results suggest that ConA and BMP-2 enhance MSC osteogenesis via different pathways. ConA-induced bone formation in MSC cultures may be useful in regenerative medicine or tissue engineering in clinical studies, as well as in basic research on bone formation.
本研究调查了伴刀豆球蛋白A(ConA)作为一种可能在体外增强间充质干细胞(MSC)成骨作用的新因子。人们已经对各种因子,如细胞因子骨形态发生蛋白-2(BMP-2),在体内和体外促进MSC成骨的可能性进行了研究。然而,尚未确定比这些因子更安全、更有效且更便宜的因子。因此,我们在有或没有ConA的情况下,将人MSC在成骨培养基中培养,并使用钙测定法比较ConA和BMP-2对MSC钙化的影响。我们还使用酶联免疫吸附测定(ELISA)和定量聚合酶链反应(PCR)来评估骨特异性标志物的表达水平。ConA和BMP-2以相当的效果增强了钙化。ConA和BMP-2的组合进一步轻微但显著地增强了钙化。ConA还增加了MSC培养基中骨钙素和BMP-2的蛋白水平。此外,ConA增加了骨钙素、RUNX2、BMP-2、BMP-4和BMP-6的mRNA表达水平。然而,ConA刺激的MSC的基因表达模式与BMP-2刺激的MSC不同。总之,这些结果表明ConA和BMP-2通过不同途径增强MSC成骨。ConA诱导的MSC培养物中的骨形成在临床研究的再生医学或组织工程以及骨形成的基础研究中可能有用。