Instituto de Bioingeniería, Universidad Miguel Hernández, Avda. Universidad s/n, 03202 Elche, Alicante, Spain.
Mater Sci Eng C Mater Biol Appl. 2013 Oct;33(7):4009-20. doi: 10.1016/j.msec.2013.05.043. Epub 2013 May 25.
Bioceramic samples with osteogenic properties, suitable for use in the regeneration of hard tissue, were synthesized. The materials consisting of α-tricalcium phosphate (αTCP) and also αTCP doped with either 1.5 wt.% or 3.0 wt.% of dicalcium silicate (C2S) in the system Dicalcium Silicate-Tricalcium Phosphate (C2S-TCP) were obtained by solid state reaction. All materials were composed of a single phase, αTCP in the case of a pure material, or solid solution of C2S in αTCP (αTCPss) for the doped αTCP. Viability, proliferation and in vitro osteoinductive capacity were investigated by seeding, adult mesenchymal stem cells of human origin (ahMSCs) which were CD73(+), CD90(+), CD105(+), CD34(-) and CD45(-) onto the 3 substrates for 30 days. Results show a non-cytotoxic effect after applying an indirect apoptosis test (Annexin V/7-AAD staining), so ahMSCs adhered, spread, proliferated and produced extracellular matrix (Heparan-sulfate proteoglycan (HS) and osteopontin (OP)) on all the ceramics studied. Finally, the cells lost the cluster differentiation marker expression CD73, CD90 y CD105 characteristic of ahMSCs and they showed an osteoblastic phenotype (Alkaline phosphatase activity (ALP), Osteocalcin production (OC), Collagen type I expression (Col-I), and production of mineralization nodules on the extracellular matrix). These observations were more evident in the αTCP ceramic doped with 1.5 wt.% C2S, indicating osteoblastic differentiation as a result of the increased concentration of solid solution of C2S in αTCP (αTCPss). Overall, these results suggest that the ceramics studied are cytocompatible and they are able to induce osteoblastic differentiation of undifferentiated ahMSCs.
合成了具有成骨性能的生物陶瓷样品，适用于硬组织再生。通过固态反应获得了由 α-磷酸三钙（αTCP）组成的材料，以及在硅酸二钙（C2S）-磷酸三钙（C2S-TCP）体系中掺杂 1.5wt.%或 3.0wt.%C2S 的 αTCP 材料。纯材料中所有材料均由单相 αTCP 组成，掺杂αTCP 中为 C2S 在 αTCP 中的固溶体（αTCPss）。通过接种人源骨髓间充质干细胞（ahMSCs），研究了这些材料的细胞活力、增殖和体外成骨能力，ahMSCs 为 CD73(+)、CD90(+)、CD105(+)、CD34(-)和 CD45(-)。将 ahMSCs 接种到 3 种底物上培养 30 天后，结果表明通过间接凋亡试验（Annexin V/7-AAD 染色）无细胞毒性作用，因此 ahMSCs 能够黏附、伸展、增殖并产生细胞外基质（硫酸乙酰肝素蛋白聚糖（HS）和骨桥蛋白（OP））。最后，细胞失去簇分化标志物 CD73、CD90 和 CD105 的表达，表现出成骨细胞表型（碱性磷酸酶活性（ALP）、骨钙素生成（OC）、I 型胶原表达（Col-I）和细胞外基质中矿化结节的生成）。在掺杂 1.5wt.%C2S 的 αTCP 陶瓷中观察到这些现象更为明显，表明由于 αTCP 中 C2S 固溶体浓度的增加，促进了成骨细胞分化。总体而言，这些结果表明研究的陶瓷具有细胞相容性，并能够诱导未分化 ahMSCs 向成骨细胞分化。
