Qiu Gengtao, Wang Ping, Li Guangjun, Shi Zhanjun, Weir Michael D, Sun Jinyu, Song Yang, Wang Jixing, Xu Huakun H, Zhao Liang
Department of Orthopaedic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.
Department of Orthopaedic Surgery, Shunde First People Hospital, Shunde, 528300, Guangdong, China.
Mol Biotechnol. 2016 Nov;58(11):748-756. doi: 10.1007/s12033-016-9974-6.
Calcium phosphate cements (CPCs) are a new generation of bone repair materials with good biocompatibility for various stem cells. The minipig is a recommended large animal model for bone engineering research. This study aimed to evaluate the feasibility of utilizing CPC scaffolds for the adhesion, proliferation, and osteogenic differentiation of minipig's bone marrow mesenchymal stem cells (pBMSCs). Passage 3 pBMSCs were seeded on the CPC scaffold and cultured with osteogenic culture medium (osteogenic group) or normal medium (control group). The density of viable cells increased in both groups, and pBMSCs firmly attached and spread well on the CPC scaffold. The alkaline phosphatase (ALP) activity in the osteogenic group had significantly increased on day 7 (D7) and peaked on D14. qRT-PCR revealed that mRNA levels of ALP and three osteogenic marker genes were significantly higher on D4, D7, and D14 in the osteogenic group. Alizarin Red S staining showed a significantly higher degree of bone mineralization from D7, D14 to D21 in the osteogenic group. These results indicated that pBMSCs can attach, proliferate well on CPC scaffold, and be successfully induced to differentiate into osteogenic cells. Our findings may be helpful for bone tissue engineering and the studies of bone regeneration.
磷酸钙骨水泥（CPCs）是新一代对各种干细胞具有良好生物相容性的骨修复材料。小型猪是骨工程研究推荐使用的大型动物模型。本研究旨在评估利用CPC支架促进小型猪骨髓间充质干细胞（pBMSCs）黏附、增殖和成骨分化的可行性。将第3代pBMSCs接种于CPC支架上，分别用成骨培养基培养（成骨组）或正常培养基培养（对照组）。两组活细胞密度均增加，且pBMSCs在CPC支架上牢固黏附并铺展良好。成骨组碱性磷酸酶（ALP）活性在第7天（D7）显著升高，并在D14达到峰值。qRT-PCR显示，成骨组在D4、D7和D14时ALP及三个成骨标记基因的mRNA水平显著更高。茜素红S染色显示，成骨组从D7、D14到D21骨矿化程度显著更高。这些结果表明，pBMSCs能够在CPC支架上黏附、良好增殖，并成功诱导分化为成骨细胞。我们的研究结果可能有助于骨组织工程和骨再生研究。
