School of Dentistry, Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Shandong University, Jinan, Shandong 250012, P.R. China.
Department of Stomatology, Qingdao Municipal Hospital, Qingdao, Shandong 266011, P.R. China.
Int J Mol Med. 2018 Apr;41(4):2297-2305. doi: 10.3892/ijmm.2018.3437. Epub 2018 Jan 29.
Stem cell‑based therapies are promising strategies to stimulate bone regeneration. Carbon monoxide releasing molecule‑3 (CORM‑3) exhibits multiple regulatory effects in a number of cells by releasing carbon monoxide (CO). The present study aimed to investigate the influence of CORM‑3 on the osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). BMSCs were divided into five groups: A CORM‑3‑osteogenic group, in which cells were pretreated with CORM‑3 and subjected to osteogenic differentiation induction using osteogenic medium; an osteogenic group, in which cells were cultured in osteogenic medium; a degassed CORM‑3‑osteogenic group, in which cells were pretreated with degassed CORM‑3 and subjected to osteogenic differentiation induction; a CORM‑3 group, in which cells were cultured in control medium containing CORM‑3; and a control group, in which cells were cultured in control medium alone. The osteo‑specific mRNA and protein expression of runt‑related transcription factor 2 (Runx2), osteocalcin (OCN) and osteopontin (OPN) were assessed using reverse transcription‑quantitative polymerase chain reaction and western blot analysis. Alkaline phosphatase (ALP) activity was also examined and mineralization was detected using alizarin red staining. Levels of Runx2, OCN and OPN mRNA and protein in the CORM‑3‑osteogenic group were significantly increased compared with the osteogenic group (P<0.05), with the exception of OCN protein levels on day 3. The mRNA and protein expression of Runx2, OCN and OPN in the degassed CORM‑3‑osteogenic and osteogenic groups were similar. In addition, the mRNA and protein expression of Runx2, OCN and OPN in the CORM‑3 and control group were similar. ALP activity in the CORM‑3‑osteogenic group was increased from day 3 and remained significantly higher compared with all other groups on days 3, 5 and 7 (P<0.05). Additionally, the results indicated that the optical density value of alizarin red staining in the CORM‑3‑osteogenic group was significantly increased compared with the other groups (P<0.05). Therefore, the present study demonstrated that CORM‑3 may promote the osteogenic differentiation of BMSCs by releasing CO.
基于干细胞的疗法是一种很有前途的策略，可刺激骨再生。一氧化碳释放分子 3（CORM-3）通过释放一氧化碳（CO）在许多细胞中表现出多种调节作用。本研究旨在探讨 CORM-3 对大鼠骨髓间充质干细胞（BMSCs）成骨分化的影响。BMSCs 分为五组：CORM-3-成骨组，细胞用 CORM-3 预处理，然后用成骨培养基进行成骨诱导分化；成骨组，细胞在成骨培养基中培养；脱气 CORM-3-成骨组，细胞用脱气 CORM-3 预处理，然后用成骨诱导分化；CORM-3 组，细胞在含 CORM-3 的对照培养基中培养；对照组，细胞单独在对照培养基中培养。采用逆转录定量聚合酶链反应和 Western blot 分析检测 runt 相关转录因子 2（Runx2）、骨钙素（OCN）和骨桥蛋白（OPN）的成骨特异性 mRNA 和蛋白表达。还检测碱性磷酸酶（ALP）活性，并通过茜素红染色检测矿化。与成骨组相比，CORM-3-成骨组的 Runx2、OCN 和 OPN 的 mRNA 和蛋白水平显著升高（P<0.05），但第 3 天的 OCN 蛋白水平除外。脱气 CORM-3-成骨组和成骨组的 Runx2、OCN 和 OPN 的 mRNA 和蛋白表达相似。此外，CORM-3 组和对照组的 Runx2、OCN 和 OPN 的 mRNA 和蛋白表达相似。CORM-3-成骨组的 ALP 活性从第 3 天开始增加，并且在第 3、5 和 7 天与所有其他组相比显著升高（P<0.05）。此外，结果表明，CORM-3-成骨组的茜素红染色光密度值明显高于其他组（P<0.05）。因此，本研究表明，CORM-3 通过释放 CO 可能促进 BMSCs 的成骨分化。
