Akahane M, Shimizu T, Kira T, Onishi T, Uchihara Y, Imamura T, Tanaka Y
Department of Public Health, Health Management and Policy, Nara Medical University Faculty of Medicine, Kashihara, Nara, Japan
Department of Orthopedic Surgery, Nara Medical University Faculty of Medicine, Kashihara, Nara, Japan.
Bone Joint Res. 2016 Nov;5(11):569-576. doi: 10.1302/2046-3758.511.BJR-2016-0013.R1.
To assess the structure and extracellular matrix molecule expression of osteogenic cell sheets created via culture in medium with both dexamethasone (Dex) and ascorbic acid phosphate (AscP) compared either Dex or AscP alone.
Osteogenic cell sheets were prepared by culturing rat bone marrow stromal cells in a minimal essential medium (MEM), MEM with AscP, MEM with Dex, and MEM with Dex and AscP (Dex/AscP). The cell number and messenger (m)RNA expression were assessed in vitro, and the appearance of the cell sheets was observed after mechanical retrieval using a scraper. β-tricalcium phosphate (β-TCP) was then wrapped with the cell sheets from the four different groups and subcutaneously implanted into rats.
After mechanical retrieval, the osteogenic cell sheets from the MEM, MEM with AscP, and MEM with Dex groups appeared to be fragmented or incomplete structures. The cell sheets cultured with Dex/AscP remained intact after mechanical retrieval, without any identifiable tears. Culture with Dex/AscP increased the mRNA and protein expression of extracellular matrix proteins and cell number compared with those of the other three groups. More bridging bone formation was observed after transplantation of the β-TCP scaffold wrapped with cell sheets cultured with Dex/AscP, than in the other groups.
These results suggest that culture with Dex/AscP improves the mechanical integrity of the osteogenic cell sheets, allowing retrieval of the confluent cells in a single cell sheet structure. This method may be beneficial when applied in cases of difficult tissue reconstruction, such as nonunion, bone defects, and osteonecrosis.Cite this article: M. Akahane, T. Shimizu, T. Kira, T. Onishi, Y. Uchihara, T. Imamura, Y. Tanaka. Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure. Bone Joint Res 2016;5:569-576. DOI: 10.1302/2046-3758.511.BJR-2016-0013.R1.
评估在同时含有地塞米松（Dex）和抗坏血酸磷酸酯（AscP）的培养基中培养所形成的成骨细胞片的结构及细胞外基质分子表达，并与单独使用Dex或AscP培养的情况进行比较。
通过在最低限度基本培养基（MEM）、含AscP的MEM、含Dex的MEM以及含Dex和AscP（Dex/AscP）的MEM中培养大鼠骨髓基质细胞来制备成骨细胞片。在体外评估细胞数量和信使核糖核酸（mRNA）表达，并在使用刮刀机械剥离后观察细胞片的外观。然后将β-磷酸三钙（β-TCP）用来自四个不同组的细胞片包裹，并皮下植入大鼠体内。
机械剥离后，来自MEM组、含AscP的MEM组和含Dex的MEM组的成骨细胞片呈现出破碎或不完整的结构。用Dex/AscP培养的细胞片在机械剥离后仍保持完整，没有任何可见的撕裂。与其他三组相比，用Dex/AscP培养可增加细胞外基质蛋白的mRNA和蛋白质表达以及细胞数量。在用Dex/AscP培养的细胞片包裹的β-TCP支架移植后，观察到的桥接骨形成比其他组更多。
这些结果表明，用Dex/AscP培养可改善成骨细胞片的机械完整性，使汇合的细胞能够以单细胞片结构剥离。当应用于诸如骨不连、骨缺损和骨坏死等困难的组织重建情况时，这种方法可能是有益的。引用本文：M. 赤羽根、T. 清水、T. 基拉、T. 大西、Y. 内原、T. 今村、Y. 田中。用地塞米松和抗坏血酸培养骨髓细胞可改善成骨细胞片结构。《骨与关节研究》2016；5：569 - 576。DOI：10.1302/2046 - 3758.511.BJR - 2016 - 0013.R1。
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