摘要
目的构建纤维蛋白-Kartogenin(KGN)释放体系,探索其对肩袖损伤修复小鼠模型腱-骨愈合的作用及机制。方法使用纤维蛋白封闭剂负载KGN,使用高效液相色谱法测定体外释放效率。采用随机数字表法将小鼠分为KGN组和对照组,每组36只。构建肩袖损伤修复小鼠模型,每只小鼠均接受单侧冈上肌腱分离和经骨修复,对照组在腱-骨修复部位应用纤维蛋白空载体系,KGN组在腱-骨修复部位应用纤维蛋白-KGN释放体系。采用qRT-PCR检测两组小鼠肩关节中肌腱重塑和成熟相关分子腱调蛋白(TNMD)、血小板衍生生长因子-AA(PDGFAA)、软骨形成基因Y染色体性别决定区盒转录因子9(SOX9)、成骨基因SP7转录因子(SP7)mRNA表达水平;采用免疫组化染色法检测小鼠肩关节TNMD、PDGFAA、SOX9、SP7蛋白表达情况;采用HE染色及阿尔新蓝染色染色观察纤维软骨面积,采用天狼星红染色观察胶原纤维密度。结果纤维蛋白-KGN释放体系中的KGN可释放5 d,释放量随时间递减,以前3天为主,占释放体系中KGN总量的34.29%。与对照组比较,KGN组小鼠肩关节中TNMD、PDGFAA、SOX9、SP7 mRNA相对表达水平均上调(均P<0.01),相应蛋白的表达增加,纤维软骨面积减小,而胶原纤维密度增加。结论KGN可以促进肩袖损伤修复小鼠模型腱-骨愈合,可能是通过促进骨生成和肌腱的重塑与成熟实现的。
Objective To construct Kartogenin(KGN)-fibrin release system and to explore its effect and mechanism on tendon-bone healing of mouse model with rotator cuff tear repair.Methods KGN was loaded with fibrin sealant,and the release efficiency in vitro was determined by high performance liquid chromatography.Mice were divided into KGN group and control group with 36 mice in each group by random number table method.Mouse model with rotator cuff tear repair was constructed,and each mouse received unilateral supraspinatus tendon separation and bone repair.The control group was treated with fibrin no-load system at the tendone-bone repair site,while the KGN group was treated with KGN-fibrin release system at the tendone-bone repair site.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expressions of tendon remodeling and maturation-associated tenomodulin(TNMD),platelet-derived growth factor-AA(PDGFAA),sex-determing region of Y chromosome box transcription factor 9(SOX9),and SP7 transcription factor(SP7)mRNA in the shoulder of mice in two groups.Immunohistochemical staining was used to detect the levels of TNMD,PDGFAA,SOX9 and SP7 protein in shoulder joints of mice.HE and Alcian blue were used to observe fibrocartilage area,and Sirius red staining was used to observe density of collagen fibers.Results KGN in KGN-fibrin release system could be released for 5 days,and the release amount decreased with time,mainly in the previous 3 days,accounting for 34.29%of the total KGN in the released system.Compared with the control group,the relative mRNA expressions of TNMD,PDGFAA,SOX9,SP7 and the levels of corresponding proteins in tendon-bone tissue of mice in KGN group were up-regulated(all P<0.01).The area of fibrocartilage in KGN group was significantly smaller than that in the control group,while the density of collagen fibers was significantly increased.Conclusion KGN can promote tendon-bone healing of mouse model with rotator cuff tear repair,which may be achieved by promoting osteogenesis and tendon remodeling and maturation.
作者
徐天波
刘德国
张颉鸿
郑宇翔
侯振海
XU Tianbo;LIU Deguo;ZHANG Jiehong;ZHENG Yuxiang;HOU Zhenhai(The Third Department of Orthopaedics,the 903 Hospital of the Joint Logistics Support Force of the Chinese people's Liberation Army,Hangzhou 310013,China)
出处
《浙江医学》
CAS
2024年第2期151-155,I0006,共6页
Zhejiang Medical Journal
基金
浙江省医药卫生科技计划项目(2022RC238)。
