Kajiyama Sohtaro, Ujiie Yuko, Nishikawa Sumio, Inoue Kohji, Shirakawa Satoshi, Hanada Nobuhiro, Liddell Robert, Davies John E, Gomi Kasuhiro
School of Dental Medicine, Tsurumi University, Tsurumi-ku, Yokohama, Kanagawa, Japan.
Research Center of Electron Microscopy, Tsurumi University, School of Dental Medicine, 2-1-3, Tsurumi, Tsurumi-ku, Yokohama, 230-8501, Japan.
J Biomed Mater Res A. 2015 Aug;103(8):2807-14. doi: 10.1002/jbm.a.35396. Epub 2015 Feb 11.
We investigated the possibility of employing human umbilical perivascular cells (HUCPVCs) within the context of finding an alternative source of mesenchymal stromal cells (MSC) for bone tissue engineering. Since it has previously been reported that conditioned medium (CM) from osteogenic bone marrow (BM) MSCs can potentiate osteogenic differentiation in a secondary cell population, we also employed BM-MSCs to generate CM to stimulate osteogenesis in the HUCPVCs. The BM-MSCs were a commercially available immortalized human cell line. In vitro assays showed negligible levels of osteogenic gene expression in HUCPVCs compared to BM-MSC, but alkaline phosphatase was detected when HUCPVC were cultured in osteogenic medium in the presence of CM from BM-MSC. An in vivo assay employing a rat calvarial osteotomy defect, together with a collagen sponge scaffold, showed that HUCPVCs provided statistically significant bony repair compared to controls. BM-MSC loaded scaffolds were not statistically different from either controls or HUCPVCs. The addition of BM-MSC CM to HUCPVCs also produced no statistically significant difference to the bone formed by HUCPVCs alone. Our results demonstrate that the in vitro assays employed did not predict in vivo outcomes, and that the BM-MSC cell line employed, or CM from such cells, provided no osteogenic advantage over the use of HUCPVCs alone.
我们在寻找用于骨组织工程的间充质基质细胞（MSC）替代来源的背景下，研究了使用人脐周血管细胞（HUCPVCs）的可能性。由于先前有报道称，成骨骨髓（BM）间充质干细胞的条件培养基（CM）可增强次级细胞群体中的成骨分化，我们还利用BM-MSCs来制备CM，以刺激HUCPVCs的成骨作用。BM-MSCs是一种市售的永生化人类细胞系。体外试验表明，与BM-MSC相比，HUCPVCs中的成骨基因表达水平可忽略不计，但当HUCPVC在含有BM-MSC的CM的成骨培养基中培养时，可检测到碱性磷酸酶。一项使用大鼠颅骨切开术缺损并结合胶原海绵支架的体内试验表明，与对照组相比，HUCPVCs能提供具有统计学意义的骨修复。负载BM-MSC的支架与对照组或HUCPVCs相比，在统计学上没有差异。向HUCPVCs中添加BM-MSC CM与单独使用HUCPVCs形成的骨相比，也没有产生统计学上的显著差异。我们的结果表明，所采用的体外试验无法预测体内结果，并且所使用的BM-MSC细胞系或此类细胞的CM，与单独使用HUCPVCs相比，没有成骨优势。
