Wu Jiani, Jiang Yingzi, Wang Guanyu, Wang Liliao, Bao Jie, Wang Jun
Department of Rehabilitation Medicine, Wuxi Ninth People's Hospital, Wuxi Jiangsu, 214000, P. R. China.
School of Physical Education, Soochow University, Suzhou Jiangsu, 215000, P. R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2024 Nov 15;38(11):1391-1398. doi: 10.7507/1002-1892.202405090.
To investigate the anti-adhesive effect and underlying mechanism of dynamic and static stress stimulation on the early healing process of rat Achilles tendon injury.
Achilles tendon tissues of 15 male Sprague Dawley (SD) rats aged 4-6 weeks were isolated and cultured by enzyme digestion method. Rat Achilles tendon cells were treated with tumor necrosis factor α to construct the Achilles tendon injury cell model, and dynamic stress stimulation (dynamic group) and static stress stimulation (static group) were applied respectively, while the control group was not treated. Live/dead cell double staining was used to detect cell activity, ELISA assay was used to detect the expression of α smooth muscle actin (α-SMA), and real-time fluorescence quantitative PCR was used to detect the mRNA expression of collagen type Ⅰ (COL1A1), collagen type Ⅲ (COL3A1), and Scleraxis (SCX). Thirty male SD rats aged 4-6 weeks underwent Achilles tendon suture and were randomly divided into dynamic group (treated by dynamic stress stimulation), static group (treated by static stress stimulation), and control group (untreated), with 10 rats in each group. HE staining and scoring were performed to evaluate the healing of Achilles tendon at 8 days after operation. COL1A1 and COL3A1 protein expressions were detected by immunohistochemical staining, α-SMA and SCX protein expressions were detected by Western blot, and maximum tendon breaking force and tendon stiffness were detected by biomechanical stretching test.
cell experiment, when compared to the static group, the number of living cells in the dynamic group was higher, the expression of α-SMA protein was decreased, the relative expression of COL3A1 mRNA was decreased, and the relative expression of SCX mRNA was increased, and the differences were all significant ( <0.05). In the animal experiment, when compared to the static group, the tendon healing in the dynamic group was better, the HE staining score was lower, the expression of COL1A1 protein was increased, the expression of COL3A1 protein was decreased, the relative expression of SCX protein was increased, the relative expression of α-SMA protein was decreased, and the tendon stiffness was increased, the differences were all significant ( <0.05).
Compared with static stress stimulation, the dynamic stress stimulation improves the fibrosis of the scar tissue of the rat Achilles tendon, promote the recovery of the biomechanical property of the Achilles tendon, and has obvious anti-adhesion effect.
探讨动态和静态应力刺激对大鼠跟腱损伤早期愈合过程的抗粘连作用及潜在机制。
取15只4 - 6周龄雄性Sprague Dawley（SD）大鼠的跟腱组织，采用酶消化法进行分离培养。用肿瘤坏死因子α处理大鼠跟腱细胞构建跟腱损伤细胞模型，分别施加动态应力刺激（动态组）和静态应力刺激（静态组），对照组不做处理。采用活/死细胞双染色检测细胞活性，酶联免疫吸附测定法检测α平滑肌肌动蛋白（α-SMA）的表达，实时荧光定量PCR检测Ⅰ型胶原（COL1A1）、Ⅲ型胶原（COL3A1）和硬骨素（SCX）的mRNA表达。30只4 - 6周龄雄性SD大鼠行跟腱缝合术，随机分为动态组（给予动态应力刺激）、静态组（给予静态应力刺激）和对照组（不做处理），每组10只。术后8天进行苏木精-伊红（HE）染色及评分，评估跟腱愈合情况。采用免疫组织化学染色检测COL1A1和COL3A1蛋白表达，蛋白质免疫印迹法检测α-SMA和SCX蛋白表达，通过生物力学拉伸试验检测最大肌腱断裂力和肌腱刚度。
细胞实验中，与静态组相比，动态组活细胞数量增多，α-SMA蛋白表达降低，COL3A1 mRNA相对表达量降低，SCX mRNA相对表达量升高，差异均有统计学意义（<0.05）。动物实验中，与静态组相比，动态组跟腱愈合较好，HE染色评分较低，COL1A1蛋白表达升高，COL3A1蛋白表达降低，SCX蛋白相对表达量升高，α-SMA蛋白相对表达量降低，肌腱刚度增加，差异均有统计学意义（<0.05）。
与静态应力刺激相比，动态应力刺激可改善大鼠跟腱瘢痕组织的纤维化，促进跟腱生物力学性能的恢复，具有明显的抗粘连作用。
