Umbilical cord-derived mesenchymal stem cells on scaffolds facilitate collagen degradation via upregulation of MMP-9 in rat uterine scars.
Umbilical cord-derived mesenchymal stem cells on scaffolds facilitate collagen degradation via upregulation of MMP-9 in rat uterine scars.
复制标题
支架上的脐带间充质干细胞通过上调大鼠子宫疤痕中的 MMP-9 促进胶原蛋白降解
DOI：
10.1186/s13287-017-0535-0
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发表时间：
2017-04-18
影响因子：
7.5
通讯作者：
Dai J
中科院分区：
文献类型：
--
作者：
Xu L;Ding L;Wang L;Cao Y;Zhu H;Lu J;Li X;Song T;Hu Y;Dai J
背景子宫严重损伤可能引发子宫疤痕形成，最终导致不孕或产科并发症。迄今为止，很少有方法能够充分解决子宫疤痕中胶原蛋白沉积的问题。脐带间充质干细胞（UC-MSC）在临床应用中显示出巨大的前景。本研究的目的是探讨支架/UC-MSCs构建体对子宫壁全层切除后大鼠子宫疤痕胶原蛋白降解和功能再生的影响。方法为了建立子宫疤痕大鼠模型，从每个子宫角切除长约1.0厘米、宽0.5厘米（子宫周长的三分之一）的子宫壁。 64只大鼠128只疤痕子宫角被随机分为4组，包括PBS组（n= 32子宫角）、支架组（n= 32子宫角）、UC-MSCs组（n= 32子宫角）和支架/UC-MSCs组（n= 32子宫角），以研究不同治疗的效果子宫疤痕的结构和功能。将PBS、可降解胶原纤维、UC-MSC或与凝胶状可降解胶原纤维混合的UC-MSC分别注射到每个子宫疤痕周围的四个预先标记的点中。在移植后第30天和第60天，对每组的一部分大鼠（n=≥8个子宫角）实施安乐死，并制备包含手术区域的子宫组织的连续切片。对 MMP-2、MMP-9、α-SMA 和 vWF 进行苏木精-伊红染色、Masson 三色染色和免疫组织化学染色。最后，每组的另一组大鼠（n = 16 子宫角）在移植后第 60 天与雄性大鼠交配，并在阴道塞存在后 18 天安乐死，以检查胎儿的数量、大小和重量以及植入部位。结果与其他三组相比，支架/UC-MSC 组表现出明显的胶原蛋白降解。移植后第60天,支架/UC-MSCs组MMP-9阳性细胞数(25.96±3.63)显着高于PBS组(8.19±1.61,P<0.01)、支架组(7.25±2.17,P<0.01)和UC-MSCs组(8.31±2.77,P<0.01)。支架/UC-MSCs组的妊娠率(10/16)也显着高于PBS组(2/16,P< 0.017)、支架组(1/16,P< 0.017)和UC-MSCs组(3/16,P< 0.017)。结论支架/UC-MSCs系统通过促进子宫疤痕中的胶原蛋白降解。移植的UC-MSCs分泌的MMP-9上调，促进子宫内膜、子宫肌层和子宫疤痕血管的再生。此外，支架/UC-MSCs治疗的子宫疤痕显示受孕能力几乎完全恢复。
BackgroundSevere injuries of the uterus may trigger uterine scar formation, ultimately leading to infertility or obstetrical complications. To date, few methods have adequately solved the problem of collagen deposition in uterine scars. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) have shown great promise in clinical applications. The objective of this study was to investigate the effect of a scaffold/UC-MSCs construct on collagen degradation and functional regeneration in rat uterine scars following full-thickness excision of uterine walls.MethodsIn order to establish a rat model of uterine scars, the uterine wall of approximately 1.0 cm in length and 0.5 cm in width (one-third of the uterine circumference) was excised from each uterine horn. A total of 128 scarred uterine horns from 64 rats were randomly assigned to four groups, including a PBS group (n= 32 uterine horns), scaffold group (n= 32 uterine horns), UC-MSCs group (n= 32 uterine horns) and scaffold/UC-MSCs group (n= 32 uterine horns) to investigate the effect of different treatments on the structure and function of uterine scars. PBS, degradable collagen fibres, UC-MSCs or UC-MSCs mixed with gelatinous degradable collagen fibres were injected into four pre-marked points surrounding each uterine scar, respectively. At days 30 and 60 post-transplantation, a subset of rats (n= 8 uterine horns) from each group was euthanized and serial sections of uterine tissues containing the operative region were prepared. Haematoxylin-eosin staining, Masson’s trichrome staining, and immunohistochemical staining for MMP-2, MMP-9, α-SMA and vWF were performed. Finally, another subset of rats (n= 16 uterine horns) from each group was mated with male rats at day 60 post-transplantation and euthanized 18 days after the presence of vaginal plugs to check numbers, sizes and weights of fetuses, as well as sites of implantation.ResultsThe scaffold/UC-MSCs group exhibited obvious collagen degradation compared with the other three groups. At day 60 post-transplantation, the number of MMP-9-positive cells in the scaffold/UC-MSCs group (25.96 ± 3.63) was significantly higher than that in the PBS group (8.19 ± 1.61,P< 0.01), the scaffold group (7.25 ± 2.17,P< 0.01) and the UC-MSCs group (8.31 ± 2.77,P< 0.01). The pregnancy rate in the scaffold/UC-MSCs group (10/16) was also significantly higher than that in the PBS group (2/16,P< 0.017), the scaffold group (1/16,P< 0.017) and the UC-MSCs group (3/16,P< 0.017).ConclusionsThe scaffold/UC-MSCs system facilitated collagen degradation in uterine scars via upregulation of MMP-9, which was secreted by transplanted UC-MSCs, and promoted regeneration of the endometrium, myometrium and blood vessels in uterine scars. Furthermore, the scaffold/UC-MSCs-treated uterine scars showed nearly complete restoration of receptive fertility.