Department of Orthopaedic Surgery, Shanghai Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 200233, P.R. China.
Mol Med Rep. 2013 Apr;7(4):1129-36. doi: 10.3892/mmr.2013.1341. Epub 2013 Feb 26.
The objective of this study was to examine the proliferation and differentiation behaviors of different compositions of strontium-containing (from 0-12 mol%) borate glasses with mesenchymal stem cells (MSCs). The Cell Counting Kit-8 (CCK-8) assay revealed that after three days of culturing, the 6Sr group had the highest cell growth rate. Analysis of cell morphology revealed that cells proliferated well near the particles of the samples in all the groups on day 3. On day 7, cells in the 6Sr group demonstrated a higher proliferation rate than other 4 groups under the microscope. When performing the Live-Dead staining experiment, the 6Sr group had the least number of dead cells. Total DNA qualification indicated that the 6Sr group had a statistically higher concentration compared with the remaining groups. It was found that on day 7, compared with the 0Sr group, the core binding factor α1 (Cbfa1) mRNA expression level was significantly higher in the 6Sr, 9Sr and 12Sr groups. On day 14, compared with the 0Sr group, the bone sialoprotein (BSP) mRNA level was significantly higher in the 6Sr group. Additionally, on day 21, the 6Sr and 9Sr groups demonstrated higher osteocalcin (OCN) mRNA expression levels compared with the 0Sr group. In the alkaline phosphatase (ALP) activity test, on day 21, the 6Sr group presented a higher activity than the 0Sr group. Further, the number of mineralized nodules per unit in MSCs was measured by Alizarin Red S staining. The results showed that the 6Sr and 9Sr groups had the greatest number of mineralized nodules. Therefore, it could be concluded that borate glasses containing strontium oxide of 0, 3, 6, 9 and 12 mol% demonstrate a significant level of proliferation when interacting with MSCs. The borate glass containing 6 mol% strontium oxide had the greatest level of proliferation when cultured with MSCs. The borate glass containing 6 and 9 mol% strontium oxide facilitated an improved bone formation ability compared with the remaining two compositions.
本研究旨在探讨不同组成(含 0-12mol%氧化锶)的硼酸盐玻璃与间充质干细胞(MSCs)相互作用时的增殖和分化行为。细胞计数试剂盒(CCK-8)检测结果显示,培养 3 天后,6Sr 组细胞生长速度最快。细胞形态分析结果显示,培养第 3 天,各组样本颗粒附近的细胞均生长良好。培养第 7 天,在显微镜下观察,6Sr 组细胞的增殖速度高于其他 4 组。进行活/死染色实验时,6Sr 组的死细胞数量最少。总 DNA 定量分析表明,6Sr 组的浓度明显高于其他组。结果发现,与 0Sr 组相比,培养第 7 天,6Sr、9Sr 和 12Sr 组的核心结合因子α1(Cbfa1)mRNA 表达水平显著升高。培养第 14 天,6Sr 组的骨涎蛋白(BSP)mRNA 水平显著高于 0Sr 组。此外,培养第 21 天,6Sr 和 9Sr 组的骨钙素(OCN)mRNA 表达水平均显著高于 0Sr 组。碱性磷酸酶(ALP)活性试验中,培养第 21 天,6Sr 组的活性明显高于 0Sr 组。此外,通过茜素红 S 染色测量 MSCs 中每单位的矿化结节数。结果表明,6Sr 和 9Sr 组的矿化结节数最多。综上所述,氧化锶含量为 0、3、6、9 和 12mol%的硼酸盐玻璃与 MSCs 相互作用时具有显著的增殖水平。培养 MSCs 时,含 6mol%氧化锶的硼酸盐玻璃的增殖水平最高。与其余两种成分相比,含 6 和 9mol%氧化锶的硼酸盐玻璃促进了骨形成能力的提高。