Implantation of a nerve protector embedded with human GMSC-derived Schwann-like cells accelerates regeneration of crush-injured rat sciatic nerves.
Implantation of a nerve protector embedded with human GMSC-derived Schwann-like cells accelerates regeneration of crush-injured rat sciatic nerves.
复制标题
DOI：
10.1186/s13287-022-02947-4
复制
发表时间：
2022-06-20
影响因子：
7.5
通讯作者：
中科院分区：
文献类型：
--
作者：
周围神经损伤（PNI）仍然是最大的临床挑战之一，因为他们相当大的长期残疾的潜力。出生后的神经嵴来源的多能干细胞，包括牙龈来源的间充质干细胞（GMSC），代表了一个有前途的种子细胞来源的组织工程和再生治疗的各种疾病，包括PNI。在这里，我们产生了GMSC再填充的神经保护器，并在大鼠坐骨神经挤压损伤模型中评估其治疗效果。 将GMSC与甲基丙烯酸化胶原蛋白混合并培养48小时，使GMSC转化为许旺样细胞（GiSC）。通过对雪旺细胞标志物表达的荧光研究来验证GiSC的表型。将包封在甲基丙烯酸化3D-胶原凝胶中的GMSC与THP-1源性巨噬细胞共培养，并通过ELISA测定上清液中抗炎细胞因子IL-10或炎性细胞因子TNF-α和IL-1β的分泌。此外，将混合在甲基丙烯酸化胶原中的GMSC填充到由脱细胞小肠粘膜下细胞外基质（SIS-ECM）制成的神经保护器中并培养24小时，从而产生用GiSC再填充的功能化神经保护器。我们植入神经保护器包裹大鼠坐骨神经损伤部位，并在术后4周进行功能和组织学评估。 包封在甲基丙烯酸化3D-胶原水凝胶中的GMSC直接转化为Schwann样细胞（GiSC），其特征在于S-100β、p75 NTR、BDNF和GDNF的表达。在体外，包封在3D-胶原水凝胶中的GMSC与巨噬细胞的共培养显著增加了IL-10的分泌，IL-10是促再生（M2）巨噬细胞的抗炎细胞因子特征，但强烈减少了LPS刺激的TNF-1α和IL-1β的分泌，TNF-1α和IL-1β是促炎（M1）巨噬细胞的两种细胞因子特征。此外，我们的研究结果表明，植入功能化的神经保护器，再填充GiSC显着加速挤压损伤的大鼠坐骨神经的功能恢复和轴突再生，伴随着促再生（M2）巨噬细胞的浸润增加，而促炎（M1）巨噬细胞的浸润减少。总的来说，这些研究结果表明，从GMSC转化的Schwann样细胞通过其双重功能，神经营养作用和促炎（M1）/促再生（M2）巨噬细胞的免疫调节，代表了PNI再生治疗的支持细胞的有希望的来源。在线版本包含补充材料，可通过10.1186/s13287-022-02947-4获得。
Peripheral nerve injuries (PNIs) remain one of the great clinical challenges because of their considerable long-term disability potential. Postnatal neural crest-derived multipotent stem cells, including gingiva-derived mesenchymal stem cells (GMSCs), represent a promising source of seed cells for tissue engineering and regenerative therapy of various disorders, including PNIs. Here, we generated GMSC-repopulated nerve protectors and evaluated their therapeutic effects in a crush injury model of rat sciatic nerves. GMSCs were mixed in methacrylated collagen and cultured for 48 h, allowing the conversion of GMSCs into Schwann-like cells (GiSCs). The phenotype of GiSCs was verified by fluorescence studies on the expression of Schwann cell markers. GMSCs encapsulated in the methacrylated 3D-collagen hydrogel were co-cultured with THP-1-derived macrophages, and the secretion of anti-inflammatory cytokine IL-10 or inflammatory cytokines TNF-α and IL-1β in the supernatant was determined by ELISA. In addition, GMSCs mixed in the methacrylated collagen were filled into a nerve protector made from the decellularized small intestine submucosal extracellular matrix (SIS-ECM) and cultured for 24 h, allowing the generation of functionalized nerve protectors repopulated with GiSCs. We implanted the nerve protector to wrap the injury site of rat sciatic nerves and performed functional and histological assessments 4 weeks post-surgery. GMSCs encapsulated in the methacrylated 3D-collagen hydrogel were directly converted into Schwann-like cells (GiSCs) characterized by the expression of S-100β, p75NTR, BDNF, and GDNF. In vitro, co-culture of GMSCs encapsulated in the 3D-collagen hydrogel with macrophages remarkably increased the secretion of IL-10, an anti-inflammatory cytokine characteristic of pro-regenerative (M2) macrophages, but robustly reduced LPS-stimulated secretion of TNF-1α and IL-1β, two cytokines characteristic of pro-inflammatory (M1) macrophages. In addition, our results indicate that implantation of functionalized nerve protectors repopulated with GiSCs significantly accelerated functional recovery and axonal regeneration of crush-injured rat sciatic nerves accompanied by increased infiltration of pro-regenerative (M2) macrophages while a decreased infiltration of pro-inflammatory (M1) macrophages. Collectively, these findings suggest that Schwann-like cells converted from GMSCs represent a promising source of supportive cells for regenerative therapy of PNI through their dual functions, neurotrophic effects, and immunomodulation of pro-inflammatory (M1)/pro-regenerative (M2) macrophages. The online version contains supplementary material available at 10.1186/s13287-022-02947-4.
登录
查看更多内容
影响因子：
5.6
作者：
Matsuoka H;Ebina K;Tanaka H;Hirao M;Iwahashi T;Noguchi T;Suzuki K;Nishimoto S;Murase T;Yoshikawa H
通讯作者：
Yoshikawa H
影响因子：
5.2
作者：
Boddupally, Keerthi;Wang, Guangfang;Kobielak, Agnieszka
通讯作者：
Kobielak, Agnieszka
影响因子：
21.3
作者：
Fernandes, KJL;McKenzie, IA;Miller, FD
通讯作者：
Miller, FD
影响因子：
6
作者：
Hopf A;Schaefer DJ;Kalbermatten DF;Guzman R;Madduri S
通讯作者：
Madduri S
影响因子：
6
作者：
Ganz, Javier;Arie, Ina;Offen, Daniel
通讯作者：
Offen, Daniel