Department of Orthopedics, Southern Medical University Affiliated Fengxian Hospital, Shanghai, China;Department of Orthopedics, The People's Hospital of Three Gorges University, The First People's Hospital of Yichang, Yichang, Hubei, China.
Department of Orthopedics, Southern Medical University Affiliated Fengxian Hospital, Shanghai, China;The Third School of Clinical Medicine, Southern Medical University, Guangzhou, China.
Acta Orthop Traumatol Turc. 2021 Mar;55(2):94-101. doi: 10.5152/j.aott.2021.18480.
This study aims to determine expression profiles of relevant genes in the early stages of post-traumatic heterotopic ossification (HO) in a rat model of Achilles tenotomy.
A total of 80 male Sprague-Dawley rats were randomly assigned to two groups: the HO group and the control group. Tenotomy was performed in the Achilles tendon of the rats in the HO group, and no intervention was conducted in the control group. On the 3rd, 5th, 8th, and 14th days after the operation, 8 rats were taken from each group at each time point, and the Achilles tendon and surrounding tissue specimens were collected. Gene expressions of TGF-β, BMP, GDF, IL, and MMP families as well as TNF-α, HIF-1α chordin, gremlin, noggin, and NODAL were analyzed by qRT-PCR. The relevant genes that were highly expressed at different time points were screened, and immunohistochemical staining was then used to verify their expression. At the 10th week, HO formation was explored by radiographic and histological examination in the remaining 8 rats of each group.
Both the radiographic and histological analyses indicated that all the rats developed HO in the HO group (100%), whereas no HO occurred in the control group. Surrounding tissues obtained from the HO group showed significantly higher gene expressions of TGF-β1, BMP-1, IL1β, HIF-1α, and MMP-2 but lower expressions of BMP-4, GDF-8, and TNF-α compared with the control group. In addition, immunohistochemical staining confirmed the higher protein expression levels of relevant genes in the HO group.
TGF-β1, BMP-1, IL-1β, HIF-1α, MMP-2, BMP4, GDF-8, and TNF-α may be associated with the formation of traumatic HO; and BMP4, GDF-8, and TNF-α may play a protective role in the early stage of HO. In this study, we investigated the expression levels of the related cytokines in the early stages of traumatic HO in the Achilles tendon tenotomy rat model to better understand the pathogenesis of HO.
本研究旨在确定创伤后异位骨化（HO）早期大鼠模型中相关基因的表达谱。
将 80 只雄性 Sprague-Dawley 大鼠随机分为两组：HO 组和对照组。HO 组大鼠行跟腱切断术，对照组大鼠不做任何干预。术后第 3、5、8、14 天，每组各时间点取 8 只大鼠，采集跟腱及周围组织标本。采用 qRT-PCR 检测 TGF-β、BMP、GDF、IL、MMP 家族以及 TNF-α、HIF-1α、chordin、gremlin、noggin 和 NODAL 等基因的表达情况。筛选出不同时间点高表达的相关基因，并用免疫组织化学染色验证其表达。每组剩余 8 只大鼠在第 10 周进行影像学和组织学检查，以探索 HO 的形成情况。
影像学和组织学分析均显示 HO 组所有大鼠（100%）均发生 HO，而对照组大鼠无一发生 HO。HO 组周围组织中 TGF-β1、BMP-1、IL1β、HIF-1α、MMP-2 基因表达明显升高，BMP-4、GDF-8、TNF-α 基因表达明显降低，与对照组相比差异有统计学意义。此外，免疫组织化学染色也证实了 HO 组相关基因的蛋白表达水平较高。
TGF-β1、BMP-1、IL-1β、HIF-1α、MMP-2、BMP4、GDF-8、TNF-α 可能与创伤性 HO 的形成有关；BMP4、GDF-8、TNF-α 可能在 HO 早期发挥保护作用。本研究通过建立大鼠跟腱切断术模型，探讨创伤后早期跟腱 HO 相关细胞因子的表达水平，以期更好地了解 HO 的发病机制。
