Supokawej Aungkura, Korchunjit Wasamon, Wongtawan Tuempong
Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Nakhon Pathom 73170, Thailand.
Laboratory of Cellular Biomedicine, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand.
J Equine Sci. 2022 Jul;33(2):19-26. doi: 10.1294/jes.33.19. Epub 2022 Jul 6.
The Wingless and Int-1 (WNT) and bone morphogenic protein/growth differentiation factor (BMP/GDF) signalling pathways contribute significantly to the development of the musculoskeletal system. The mechanism by which they contribute is as follows: BMP/GDF signalling usually promotes tendon differentiation, whereas WNT signalling inhibits it. We hypothesised that inhibiting WNT and subsequently stimulating BMP signalling may enhance the tenogenic differentiation of stem cells. The objective of this study was to determine whether a combination of WNT inhibitor (KY02111) and BMP12/GDF7 protein could enhance the differentiation of bone marrow-derived equine mesenchymal stromal cells (BM-eMSCs) into tenocytes. Cells were cultured in five treatments: control, BMP12, and three different combinations of BMP12 and KY02111. The results indicated that a 1-day treatment with KY02111 followed by a 13-day treatment with BMP12 resulted in the highest tenogenic differentiation score in this experiment. The effect of KY02111 is dependent on the incubation time, with 1 day being better than 3 or 5 days. This combination increased tenogenic gene marker expression, including SCX, TNMD, DCN, and TNC, as well as COL1 protein expression. In conclusion, we propose that a combination of BMP12 and KY02111 can enhance the in vitro tenogenic differentiation of BM-eMSCs more than BMP12 alone. The findings of this study might be useful for improving tendon differentiation protocols for stem cell transplantation and application to tendon regeneration.
无翅型MMTV整合位点家族（WNT）信号通路以及骨形态发生蛋白/生长分化因子（BMP/GDF）信号通路对肌肉骨骼系统的发育起着重要作用。它们发挥作用的机制如下：BMP/GDF信号通路通常促进肌腱分化，而WNT信号通路则抑制肌腱分化。我们推测，抑制WNT信号通路并随后激活BMP信号通路可能会增强干细胞的成腱分化。本研究的目的是确定WNT抑制剂（KY02111）与BMP12/GDF7蛋白联合使用是否能够增强马骨髓间充质干细胞（BM-eMSCs）向肌腱细胞的分化。将细胞分为五种处理组进行培养：对照组、BMP12组以及BMP12与KY02111的三种不同组合。结果表明，在本实验中，先用KY02111处理1天，随后用BMP12处理13天，可获得最高的成腱分化评分。KY02111的作用效果取决于孵育时间，1天优于3天或5天。这种联合处理增加了成腱基因标志物的表达，包括SCX、TNMD、DCN和TNC，以及COL1蛋白的表达。总之，我们认为，与单独使用BMP12相比，BMP12与KY02111联合使用能够增强BM-eMSCs在体外的成腱分化。本研究结果可能有助于改进干细胞移植的肌腱分化方案，并应用于肌腱再生。
