Liu Jinzhong, Zhao Liang, Ni Ling, Qiao Chunyan, Li Daowei, Sun Hongchen, Zhang Zongtao
Department of Oral Pathology, School of Stomatology, Jilin University Changchun 130021, China.
State Key laboratory Inorganic Synthesis and Preparative Chemistry, Jinlin University Changchun 130012, China.
Am J Transl Res. 2015 Sep 15;7(9):1588-601. eCollection 2015.
The reconstruction of large bone defects has been the focus in bone tissue engineering research. By acting as synthetic frameworks for cell growth and tissue formation, biomaterials can play a critical role in bone tissue engineering. Among various biomaterials, calcium phosphate based materials include hydroxyapatite (HA), α-tricalcium phosphate (α-TCP), and β-tricalcium phosphate (β-TCP) are widely used as scaffold materials in bone tissue engineering. However, little is known about the effect of α-TCP alone on the osteogenic differentiation of the BMSCs. To this end, we synthesized α-TCP using a novel co-precipitation method. The synthetic α-TCP was then incubated with rat BMSCs under osteogenic inductive medium culture conditions, followed by the analysis of the mRNA levels of various osteogenesis-related genes, including ALP, Rux2, COL-I, and SP7, using a quantitative RT-PCR method. Following incubation of BMSCs with 20 μg/ml α-TCP, cells reached confluency after 7 days. Additionally, the MTT analysis showed that α-TCP at concentration of 10-20 μg/ml had good biocompatibility with BMSCs, showing no significant inhibition of rat BMSCs proliferation. Furthermore, the synthetic α-TCP (20 μg/ml), when incubated with rat BMSCs in the osteogenic culture medium, increased the mRNA levels of various osteogenesis-related genes, including ALP, Rux2, COL-I, and SP7. Finally, treatment of synthetic α-TCP (20 μg/ml) potentiated calcium nodule formations after incubation with rat BMSCs in osteogenic culture medium for 21 days, as compared with non-treated control. Taken together, the results in the present study suggested that α-TCP alone likely promotes rat BMSCs osteogenic differentiation through up-regulating ALP, Col-I, Runx2, and SP7 gene expression.
大骨缺损的修复一直是骨组织工程研究的重点。生物材料作为细胞生长和组织形成的合成支架,在骨组织工程中起着关键作用。在各种生物材料中,基于磷酸钙的材料包括羟基磷灰石(HA)、α-磷酸三钙(α-TCP)和β-磷酸三钙(β-TCP),被广泛用作骨组织工程中的支架材料。然而,关于单独的α-TCP对骨髓间充质干细胞(BMSCs)成骨分化的影响知之甚少。为此,我们采用一种新型共沉淀法合成了α-TCP。然后将合成的α-TCP在成骨诱导培养基培养条件下与大鼠BMSCs孵育,随后使用定量逆转录聚合酶链反应(RT-PCR)方法分析各种成骨相关基因的mRNA水平,包括碱性磷酸酶(ALP)、Rux2、I型胶原(COL-I)和SP7。用20μg/ml的α-TCP孵育BMSCs后,细胞在7天后达到汇合。此外,MTT分析表明,浓度为10-20μg/ml的α-TCP与BMSCs具有良好的生物相容性,对大鼠BMSCs的增殖没有明显抑制作用。此外,合成的α-TCP(20μg/ml)在成骨培养基中与大鼠BMSCs孵育时,会增加各种成骨相关基因的mRNA水平,包括ALP、Rux2、COL-I和SP7。最后,与未处理的对照组相比,合成的α-TCP(20μg/ml)在成骨培养基中与大鼠BMSCs孵育21天后,促进了钙结节的形成。综上所述,本研究结果表明,单独的α-TCP可能通过上调ALP、Col-I、Runx2和SP7基因表达来促进大鼠BMSCs的成骨分化。