人脐带间充质干细胞联合富含血小板血浆促进大鼠跟腱损伤的修复

王位; 付宇翀; 周梅; 唐红; 张吉强; 唐康来

doi:10.16016/j.1000-5404.201607181

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人脐带间充质干细胞联合富含血小板血浆促进大鼠跟腱损伤的修复

基础医学 | 更新时间：2020-08-05

- 人脐带间充质干细胞联合富含血小板血浆促进大鼠跟腱损伤的修复
- Synergistic effect of human umbilical cord mesenchymal stem cells and platelet-rich plasma on promoting healing of Achilles tendon injury in rats
- 第三军医大学学报 2017年39卷第2期页码：185-191
- 作者机构：
  
  1.400038 重庆, 第三军医大学: 西南医院骨科, 全军矫形外科中心
  2.400038 重庆, 第三军医大学: 基础医学部神经生物学教研室
- 作者简介：
  
  唐康来, E-mail: tangkanglai@hotmail.com Tang Kanglai, E-mail: tangkanglai@hotmail.com
- 基金信息：
  
  全军医学科研“十二五”计划(创新专项重点 13CXZ008)
- DOI：10.16016/j.1000-5404.201607181
  中图分类号： R329.2;R686.1;R977.8
- 纸质出版日期：2017-1-30，
  
  收稿日期：2016-7-26，
  
  录用日期：2016-9-4
扫描看全文
王位, 付宇翀, 周梅, 等. 人脐带间充质干细胞联合富含血小板血浆促进大鼠跟腱损伤的修复[J]. 第三军医大学学报, 2017,39(2):185-191.

Wei Wang, Yuchong Fu, Mei Zhou, et al. Synergistic effect of human umbilical cord mesenchymal stem cells and platelet-rich plasma on promoting healing of Achilles tendon injury in rats[J]. J Third Mil Med Univ, 2017,39(2):185-191.
王位, 付宇翀, 周梅, 等. 人脐带间充质干细胞联合富含血小板血浆促进大鼠跟腱损伤的修复[J]. 第三军医大学学报, 2017,39(2):185-191. DOI： 10.16016/j.1000-5404.201607181.

Wei Wang, Yuchong Fu, Mei Zhou, et al. Synergistic effect of human umbilical cord mesenchymal stem cells and platelet-rich plasma on promoting healing of Achilles tendon injury in rats[J]. J Third Mil Med Univ, 2017,39(2):185-191. DOI： 10.16016/j.1000-5404.201607181.

摘要

目的

探讨不同浓度富含血小板血浆(platelet-rich plasma，PRP)在体外对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells，hUC-MSCs)增殖、分化的影响，评估hUC-MSCs联合PRP对大鼠跟腱损伤修复的影响。

方法

PRP活化后成为PRP-clot releasate(PRCR)。取P3代hUC-MSCs于培养皿中培养24 h后，分别加入含0%(对照组)、10%、20%、40% PRCR的DMEM/F12培养基，于第1、2、3、4天测定hUC-MSCs的增殖速度。P3代hUC-MSCs按相同分组培养，于第1、3、5天收集细胞并提取蛋白。Western blot检测hUC-MSCs中成肌腱分化标志性基因肌腱蛋白C(tenascin c

TNC)、SCX(scleraxis)、Ⅰ型胶原的表达。选取48只8周龄雄性SD大鼠，Ⅰ型胶原酶溶液50 μL注射于大鼠右侧跟腱制备跟腱损伤模型，3 d后于损伤部位再次分别注射50 μL生理盐水(对照)、hUC-MSCs悬液、PRP或PRP+hUC-MSCs混悬液，术后2、4周安乐死大鼠，Western blot检测跟腱中TNC、SCX及Ⅰ型胶原的表达，跟腱切片HE染色行组织学评估。

结果

PRCR在体外对hUC-MSCs增殖均有促进作用，20% PRCR组促增殖作用最显著(

＜0.05)；20% PRCR组较对照组在体外可以显著地促进hUC-MSCs成肌腱分化标志性基因的表达(

＜0.05)；术后2、4周，PRP+hUC-MSCs组跟腱中成肌腱分化相关基因在蛋白水平表达更高，跟腱切片HE染色显示PRP+hUC-MSCs组纤维排列更规整，炎细胞更少。

结论

适宜浓度PRP在体外显著促进了hUC-MSCs的增殖、成肌腱分化；PRP联合hUC-MSCs能有效促进大鼠受损跟腱的修复。

Abstract

Objective

To determine the effect of different concentrations of platelet-rich plasma (PRP) on the proliferation and differentiation of human umbilical cord mesenchymal stem cells (HUC-MSCs)

in vitro

and evaluate the effect of hUC-MSCs combined with PRP in the treatment of collagenase-induced Achilles tendon injury in rats.

Methods

PRP was activated to prepare into PRP-clot releasate (PRCR). hUC-MSCs at passage 3 were incubated in control medium or medium supplemented with 10%

20% or 40% PRCR

and the proliferation of hUC-MSCs on days 1

3 and 4 was evaluated. Tenogenesis related genes (scleraxis

tenascin-C

and collagen typeⅠ) were assessed by Western blotting on days 1

3 and 5. In 2 and 4 weeks after hUC-MSCs

PRP

hUC-MSCs+PRP or saline (as control) injection into the injured Achilles tendon

the tendon tissue was harvested and evaluated by histology and Western blotting.

Results

PRCR promoted the proliferation of hUC-MSCs

in vitro

and 20% PRCR prominently increased the number of hUC-MSCs (

＜0.05) as well as the protein levels of collagen typeⅠ

tenascin-C and scleraxis (

＜0.05). Combination of hUC-MSCs and PRP resulted in better healing effect of the injured tendon by improving histological characteristics and increasing expression of tenogenesis related genes in 2 and 4 weeks after operation.

Conclusion

Appropriate concentration of PRP promotes the proliferation and tenogenic differentiation of hUC-MSCs. Combination of hUC-MSCs and PRP has synergistic effect on promoting healing of the injured Achilles tendon.

关键词

人脐带间充质干细胞富含血小板血浆大鼠模型跟腱损伤

Keywords

human umbilical cord mesenchymal stem cellsplatelet-rich plasmarat modelAchilles tendon injury

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