[1]许 青,孙香香,陈锦阳,等.人毛囊真皮鞘细胞移植治疗兔跟腱炎实验研究[J].陕西医学杂志,2024,(5):583-588.[doi:DOI:10.3969/j.issn.1000-7377.2024.05.002]
　XU Qing,SUN Xiangxiang,CHEN Jinyang,et al.Experimental study of human hair follicle dermal sheath cells transplantation in treatment of achilles tendonitis of rabbit[J].,2024,(5):583-588.[doi:DOI:10.3969/j.issn.1000-7377.2024.05.002]
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人毛囊真皮鞘细胞移植治疗兔跟腱炎实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年5期
页码:
583-588
栏目:
基础研究
出版日期:
2024-05-05

文章信息/Info

Title:
Experimental study of human hair follicle dermal sheath cells transplantation in treatment of achilles tendonitis of rabbit
作者:
许 青1; 孙香香1; 陈锦阳2; 庄 盼2; 刘军权2; 3
(1.陆军第七十一集团军医院检验科,江苏 徐州 221004; 2.浙江卫未生物医药科技有限公司,浙江 杭州 310053; 3.杭州金域医学检验所有限公司,浙江 杭州 310053)
Author(s):
XU Qing; SUN Xiangxiang; CHEN Jinyang; ZHUANG Pan; LIU Junquan
(Department of Clinical Laboratory,the 71st Group Military Hospital of PLA Army,Xuzhou 221004,China)
关键词:
跟腱炎;  人毛囊真皮鞘细胞;  移植;  胶原纤维;  炎症反应;  兔
Keywords:
Achilles tendinitis;  Human hair follicle dermal sheath cells;  Transplantation;  Collagenous fiber;  Inflammatory response;  Rabbit
分类号:
R -33
DOI:
DOI:10.3969/j.issn.1000-7377.2024.05.002
文献标志码:
A
摘要:
目的:探讨人毛囊真皮鞘细胞(hHF-DSCs)移植对兔跟腱炎的治疗效果。方法:采集人后枕部毛囊单位,并进行分离培养获得hHF-DSCs。选取30只雄性大白兔随机分为对照组、模型组和治疗组,每组10只。模型组及治疗组于兔左侧跟腱止点上1 cm处多点注射Ⅰ型胶原酶建立跟腱炎兔模型。建立跟腱炎模型成功后于第0、3、6、13天,治疗组在兔跟腱患侧注射0.5 ml的hHF-DSCs,模型组与对照组在兔跟腱相同部位注射相同体积0.9%氯化钠溶液。观察各组兔跟腱外观以及组织病理学染色结果。采用ELISA法检测跟腱组织羟脯氨酸(HYP)含量以及肌腱蛋白C(TNC)、基质金属蛋白酶-9(MMP-9)、白细胞介素(IL)-1β、IL-6、IL-10和肿瘤坏死因子-α(TNF-α)表达水平。采用免疫组化法检测跟腱组织Ⅰ、Ⅲ型胶原(collagen Ⅰ、Ⅲ)表达水平。采用牵拉法检测跟腱组织生物力学。结果:分离培养后的hHF-DSCs表面CD14、CD34、CD45和人类白细胞抗原-DR(HLA-DR)几乎不表达,CD49d、CD90、CD73和CD105表达率接近100%。与对照组比较,模型组步态改变明显,跟腱红肿、无弹性、无光泽。HE染色结果显示,模型组胶原纤维排列疏松,原核细胞较少,分布紊乱不均匀,表明跟腱炎兔模型构建成功。治疗组跟腱组织弹性和光泽度好于模型组,且治疗组HE染色显示胶原纤维排列整齐,无明显纤维坏死,炎症细胞较少,胶原纤维原核细胞数量正常。模型组HYP含量低于对照组和治疗组,对照组高于治疗组(均P<0.05)。模型组MMP-9水平高于对照组和治疗组,治疗组高于对照组(均P<0.05)。模型组TNC水平低于对照组和治疗组,治疗组低于对照组(均P<0.05)。与对照组比较,治疗组和模型组兔跟腱组织IL-1β、IL-6、IL-10及TNF-α表达水平升高(均P<0.05)。与模型组比较,治疗组IL-1β、IL-6、TNF-α表达水平降低(均P<0.05)。与模型组比较,治疗组collagen Ⅰ、Ⅲ升高(均P<0.05)。与对照组比较,模型组collagen Ⅰ、Ⅲ降低(均P<0.05)。模型组跟腱最大负荷低于对照组,治疗组高于模型组(均P<0.05)。结论:hHF-DSCs移植治疗兔跟腱炎,能够促进受损跟腱组织修复及胶原合成,降低胶原降解,减轻炎症反应。
Abstract:
Objective:To investigate the therapeutic effect of human hair follicle dermal sheath cells(hHF-DSCs)transplantation on achilles tendonitis of rabbit.Methods:The hHF-DSCs were isolated and cultured from the hair follicles of human posterior occipital.Thirty male white rabbits were randomly divided into control group,model group and treatment group,with 10 rabbits in each group.The model group and the treatment group were given multiple injection of type Ⅰ collagenase 1 cm above the left achilles tendon insertion point to establish the rabbit model of achilles tendinitis.On day 0,3,6 and 13 after the successful establishment of the achilles tendinitis model,0.5 ml of hHF-DSCs were injected into the affected side of the achilles tendon in the treatment group,and the same volume of 0.9% sodium chloride solution was injected into the same part of the achilles tendon in the model group and the control group.The appearance and histopathological staining of achilles tendon in each group were observed.The content of hydroxyproline(HYP)and the expression levels of tendinocyte C(TNC),matrix metalloproteinase-9(MMP-9),interleukin(IL)-1β,IL-6,IL-10 and tumor necrosis factor-α(TNF-α)were detected by ELISA.The expression levels of collagen Ⅰ and Ⅲ in achilles tendon tissues were detected by immunohistochemical method.The biomechanics of achilles tendon tissue was measured by stretch method.Results: CD14,CD34,CD45 and HLA-DR were almost not expressed on the surface of hHF-DSCs after isolation and culture,and the expression rates of CD49d,CD90,CD73 and CD105 were close to 100%.Compared with the control group,the gait of the model group was obviously changed,and the achilles tendon was red,swollen,inelastic and dull.The results of HE staining showed that the collagen fibers in the model group had loose arrangement,few prokaryotic cells and uneven distribution,indicating that the rabbit models of achilles tendinitis were successfully constructed.The elasticity and gloss of the achilles tendon tissue in the treatment group were better than those in the model group,and HE staining showed that the collagen fibers in the treatment group were arranged neatly,no obvious fiber necrosis,fewer inflammatory cells,and normal number of collagen fiber prokaryotic cells.HYP content in model group was lower than that in control group and treatment group,while that in control group was higher than that in treatment group(all P<0.05).The level of MMP-9 in model group was higher than that in control group and treatment group,and that in treatment group was higher than that in control group(all P<0.05).The TNC level of model group was lower than that of control group and treatment group,and that of treatment group was lower than that of control group(all P<0.05).Compared with the control group,the expression levels of IL-1β,IL-6,IL-10 and TNF-α in the achilles tendon tissues of rabbits in the treatment group and model group were increased(all P<0.05).Compared with model group,the expression levels of IL-1β,IL-6 and TNF-α in treatment group were decreased(all P<0.05).Compared with model group,collagen Ⅰ and Ⅲ in treatment group were increased(all P<0.05).Compared with control group,collagen Ⅰ and Ⅲ in model group were decreased(all P<0.05).The maximum load of achilles tendon in the model group was lower than that in the control group,and that in the treatment group was higher than that in the model group(all P<0.05).Conclusion:hHF-DSCs transplantation in the treatment of rabbit achilles tendinitis can promote the repair of damaged achilles tendon tissue and collagen synthesis,reduce collagen degradation and alleviate inflammatory response.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(82072512)
更新日期/Last Update: 2024-05-06