Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, 600 Yishan Road, Shanghai, 200233, China.
Calcif Tissue Int. 2019 Mar;104(3):320-330. doi: 10.1007/s00223-018-0501-6. Epub 2019 Jan 11.
Distraction osteogenesis (DO) is used to treat specific disorders associated with growth abnormalities and/or loss of bone stock secondary to trauma or disease. However, a high rate of complications and discomfort hamper its further application in clinical practice. Here, we investigated the effects of all-trans retinoic acid (ATRA) on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs) and bone consolidation in a rat DO model. Different doses of ATRA were used to treat rBMSCs. Cell viability and osteogenic differentiation were assessed using CCK-8 and alkaline phosphatase staining, respectively. The mRNA expression of osteogenic differentiation-genes (including ALP, Runx2, OCN, OPN, OSX, and BMP2) and angiogenic genes (including VEGF, HIF-1, FLK-2, ANG-2, and ANG-4) were determined by quantitative real-time PCR analysis. Further, we locally injected ATRA or PBS into the gap in the rat DO model every 3 days until termination. X-rays, micro-computed tomography (Micro-CT), mechanical testing, and immunohistochemistry stains were used to evaluate the quality of the regenerates. ATRA promoted osteogenic differentiation of rBMSCs. Moreover, ATRA elevated the mRNA expression levels of osteogenic differentiation-genes and angiogenic genes. In the rat model, new bone properties of bone volume/total tissue volume and mechanical strength were significantly higher in the ATRA-treatment group. Micro-CT examination showed more mineralized bone after the ATRA-treatment, and immunohistochemistry demonstrated more new bone formation after ATRA-treatment than that in the PBS group. In conclusion, as a readily available and very cost effective bio-source, ATRA may be a novel therapeutic method to enhance bone consolidation in the clinical setting.
牵引成骨术（DO）用于治疗与生长异常和/或创伤或疾病引起的骨量丢失相关的特定疾病。然而，高并发症率和不适感阻碍了其在临床实践中的进一步应用。在这里，我们研究了全反式视黄酸（ATRA）对大鼠骨髓间充质干细胞（rBMSCs）成骨分化和大鼠 DO 模型中骨整合的影响。使用不同剂量的 ATRA 处理 rBMSCs。通过 CCK-8 和碱性磷酸酶染色分别评估细胞活力和成骨分化。通过定量实时 PCR 分析测定成骨分化基因（包括 ALP、Runx2、OCN、OPN、OSX 和 BMP2）和血管生成基因（包括 VEGF、HIF-1、FLK-2、ANG-2 和 ANG-4）的 mRNA 表达。进一步，我们将 ATRA 或 PBS 局部注射到大鼠 DO 模型中的间隙中，每 3 天一次，直到实验结束。X 射线、微计算机断层扫描（Micro-CT）、机械测试和免疫组织化学染色用于评估再生体的质量。ATRA 促进 rBMSCs 的成骨分化。此外，ATRA 提高了成骨分化基因和血管生成基因的 mRNA 表达水平。在大鼠模型中，ATRA 治疗组的骨体积/总体积和机械强度的新骨特性显著更高。Micro-CT 检查显示 ATRA 治疗后有更多的矿化骨，免疫组织化学显示 ATRA 治疗后比 PBS 组有更多的新骨形成。总之，作为一种易得且非常经济有效的生物来源，ATRA 可能是一种增强临床骨整合的新治疗方法。
