Sánchez Angulo P, Meseguer Olmo L, Meseguer Henarejos A B, Cragnolini F
Cirugía Ortopédica y Traumatología, Hospital General Universitario Reina Sofía, Murcia, España.
Rev Esp Cir Ortop Traumatol. 2012 May-Jun;56(3):179-87. doi: 10.1016/j.recot.2011.11.003. Epub 2011 Dec 22.
The purpose of this work is to evaluate the in vitro behaviour and the capacity to induce osteoblastic differentiation of a 55S vitro-ceramic (Vc) on a population of adult rabbit mesenchymal stem-cells (MSCs).
The material was obtained using the sol-gel method. The cells were obtained from rabbit bone-marrow aspirate and seeded over the Vc, and over plastic (control). The MSCs were cultivated in two culture media; one a standard DMEM (growth medium), and the other an osteoblastic phenotype inducer, composed of DMEM complemented with dexamethasone, ß-glycerophosphate and ascorbic acid-2-phosphate (osteogenic medium). The morphology of the cells that grew was assessed using a scanning electronic microscope. The tetrazolium salt reduction test was used for evaluating the cell growth. For cell differentiation, osteocalcin production and loss of CD90 bone surface antigen, characteristic of MSCs, were quantified.
During the culture time the MSCs adhered, proliferated and formed a mineralised extracellular matrix over the Vc. An osteoblastic phenotype finally being shown, producing osteocalcin and decreasing the expression of the CD90 antigen, regardless of the culture medium used.
Based on these results we can state that Vc 55S behaved like a material capable of supporting adhesion and growth of MSCs and, in turn, inducing the differentiation of the MSCs to osteoblastic cell lines, thus showing osteoconduction and osteoinduction properties.
本研究旨在评估一种55S体外陶瓷(Vc)在成年兔间充质干细胞(MSCs)群体上的体外行为以及诱导成骨细胞分化的能力。
材料采用溶胶 - 凝胶法制备。细胞取自兔骨髓抽吸物,接种于Vc及塑料(对照)表面。MSCs在两种培养基中培养;一种是标准的DMEM(生长培养基),另一种是成骨细胞表型诱导剂,由添加地塞米松、β - 甘油磷酸酯和抗坏血酸 - 2 - 磷酸酯的DMEM组成(成骨培养基)。使用扫描电子显微镜评估生长细胞的形态。采用四唑盐还原试验评估细胞生长。对于细胞分化,定量检测骨钙素的产生以及MSCs特征性的CD90骨表面抗原的丢失情况。
在培养期间,MSCs在Vc表面黏附、增殖并形成矿化的细胞外基质。最终呈现出成骨细胞表型,产生骨钙素并降低CD90抗原的表达,且与所使用的培养基无关。
基于这些结果,我们可以表明55S Vc表现得像一种能够支持MSCs黏附与生长,进而诱导MSCs分化为成骨细胞系的材料,从而显示出骨传导和骨诱导特性。