Interleukin-10 engineered human umbilical cord mesenchymal stem cells for superior treatment of inflammatory bowel disease

doi:10.12307/2025.512

Abstract

Abstract: BACKGROUND: Mesenchymal stem cells have been widely used in the treatment of various diseases due to their wide range of sources, their ease of proliferation in vitro and their ability to secrete a range of immunomodulatory factors to suppress inflammation and promote tissue repair and regeneration. However, in the treatment of many diseases, the therapeutic effect is limited. The effort to engineer and modify mesenchymal stem cells for specific disease pathogenesis or intervention targets is an important development for cell therapy in the future.
OBJECTIVE: Interleukin-10 is a typical anti-inflammatory cytokine that helps to modulate the immune response and induces macrophage polarization towards an anti-inflammatory phenotype. This study investigated the therapeutic effect of interleukin-10 engineered human umbilical cord mesenchymal stem cells on inflammatory bowel disease.
METHODS: Human umbilical cord mesenchymal stem cells stably overexpressing interleukin-10 were established by electro-transfection, and screened for clinical-grade cells based on the cell therapy product criteria. C57BL/6J mice were given 5% aqueous dextran sulfate sodium ad libitum to establish a model of acute colitis. Empty plasmid-transfected human umbilical cord mesenchymal stem cells or interleukin-10-human umbilical cord mesenchymal stem cells (1×106 cells/each mouse) were injected on day 1 before modeling (tail vein injection) and day 4 (intraperitoneal injection) after modeling, respectively. On day 6 after modeling, colon tissue sections were taken for hematoxylin-eosin staining to assess histological changes. Immunofluorescence staining was performed to detect the expression of proliferating cell nuclear antigen and CD31.
RESULTS AND CONCLUSION: The engineered human umbilical cord mesenchymal stem cells stably overexpressing interleukin-10 were constructed, and met the quality standard of clinical-grade human umbilical cord mesenchymal stem cells. Human umbilical cord mesenchymal stem cells could repair acute colitis in mice. The therapeutic effect of interleukin-10-human umbilical cord mesenchymal stem cells was more efficacious, which more significantly suppressed body weight loss (P < 0.05), colon shortening (P < 0.05), and damage of colonic tissues (P < 0.05) in acute colitis mice. In interleukin-10-human umbilical cord mesenchymal stem cell treatment group, there were significantly more proliferating cell nuclear antigen-positive cells and CD31-positive cells in the colon sections than in the human umbilical cord mesenchymal stem cell treatment group, suggesting that interleukin-10-overexpressing human umbilical cord mesenchymal stem cells contributed to the repair of colon tissue by significantly promoting the proliferation of intestinal tissues and angiogenesis.

Key words: interleukin-10, human umbilical cord mesenchymal stem cell, engineered mesenchymal stem cell, inflammatory bowel disease, regeneration and repair, cell proliferation, intestinal regeneration

CLC Number:

Feng Yirui, Gao Tianyun, Wang Yaping, Huang Yahong, Wang Bin. Interleukin-10 engineered human umbilical cord mesenchymal stem cells for superior treatment of inflammatory bowel disease[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(23): 4878-4887.

Figures/Tables 3

2.2 IL10-UCMSCs优效治疗葡聚糖硫酸钠盐诱导的小鼠急性结肠炎模型葡聚糖硫酸钠盐造模成功的小鼠主要表现为腹泻、粪便黏稠、肉眼血便以及体质量减轻。造模组明显的体质量减轻与疾病活动指数评分升高证明5% 葡聚糖硫酸钠盐自由饮水成功诱导了C57BL/6J小鼠急性结肠炎。UCMSCs组与IL10-UCMSCs组都有效抑制了小鼠的体质量下降，其中IL10-UCMSCs组表现出更为明显的抑制效果，见图8A。结肠炎会使结肠长度缩短，PBS组小鼠的结肠最短，UCMSCs组和IL10-UCMSCs组小鼠的结肠长度都明显恢复，但二者相比之下，IL10-UCMSCs的缓解更为突出，见图8C，D。疾病活动指数评分进一步说明IL10-UCMSCs治疗相较于UCMSCs表现更为优效，见图8B。 2.3 IL10-UCMSCs治疗更显著地减少肠道损伤并促进结肠组织再生修复结肠组织病理切片苏木精-伊红染色结果显示，葡聚糖硫酸钠盐可致小鼠结肠中免疫细胞大量浸润、杯状细胞缺失和隐窝破坏，而这些症状通过UCMSCs和IL10-UCMSCs治疗得到显著恢复，并且IL10-UCMSCs治疗组小鼠结肠的破坏程度更轻，见图9A-E。增殖细胞核抗原是一种DNA聚合酶的辅助蛋白，与细胞DNA合成关系密切，在细胞增殖的启动上起重要作用，可作为细胞增殖状态的指标。用免疫荧光染色结肠切片中的增殖细胞核抗原阳性细胞，发现UCMSCs和IL10-UCMSCs都促进了肠道损伤后的组织修复，而IL10-UCMSCs表现出更为明显的促增殖作用，见图10A-E。 CD31是血管内皮分化标志物，通过免疫荧光染色发现UCMSCs组与IL10-UCMSCs组相比于造模组，结肠组织中CD31阳性的内皮细胞数量显著增加以及平均血管面积增加，表明UCMSCs和IL10-UCMSCs促进了结肠部位的血管生成，其中IL10-UCMSCs的促血管生成作用更为明显，见图11A-E。"

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