Guo Fang, Jin Jianhua, Lin Wenjing, Gorostidi Mikel, Yang Jie, Liu Li, Chen Xinyan
Department of Gynecology, Wenzhou People's Hospital, Wenzhou, China.
Department of Obstetrics and Gynecology, Donostia University Hospital, San Sebastián, Basque Country, Spain.
Gland Surg. 2024 Dec 31;13(12):2372-2382. doi: 10.21037/gs-24-474. Epub 2024 Dec 19.
Infertility is a special reproductive health defect. For women, congenital uterine malformations, extensive adhesions in the uterine cavity, and hysterectomy are associated with infertility. Uterine transplantation is technically feasible, but its clinical application and development are limited by donor shortages and immune rejection. Thus, uterine tissue engineering research has promising prospects. This study sought to explore the ideal perfusion strategy and evaluation process for successfully preparing natural uterine decellularized scaffolds using decellularized perfusion technology to provide a good platform for uterine tissue engineering research.
Female Sprague-Dawley rats were selected. Eluents, including TritonX-100 supplemented with sodium dodecyl sulfate, were perfused into the uterus through the uterine artery after physical freezing, thawing, and enzymatic hydrolysis. After decellularization, each scaffold was evaluated by general observation, methylene blue staining, hematoxylin and eosin staining, immunohistochemical staining, quantitative analysis of genomic DNA, collagen detection and identification, cytokine content determination, transmission electron microscopy (TEM), and scanning electron microscopy (SEM).
After perfusion, a transparent uterine scaffold was established, and the histological examination and TEM showed that it contained no cell residue. The DNA content was shown to be less than 5% that of the normal uterus. Methylene blue staining and SEM showed that the vascular network and spatial structure were intact. Immunohistochemical staining and collagen quantification showed that the extracellular matrix components of the scaffold were completely preserved. In addition, the enzyme-linked immunosorbent assay results showed that the cytokines, including epidermal growth factor, basic fibroblast growth factor, and transforming growth factor beta, had been retained in the decellularized scaffold, and still showed some biological activity.
A decellularized uterine scaffold was successfully established, and its physical and chemical properties were preserved; consequently, it may be used as an alternative platform for uterine tissue engineering research.
不孕症是一种特殊的生殖健康缺陷。对于女性而言，先天性子宫畸形、宫腔广泛粘连以及子宫切除术均与不孕有关。子宫移植在技术上是可行的，但其临床应用和发展受到供体短缺和免疫排斥的限制。因此，子宫组织工程研究具有广阔的前景。本研究旨在探索理想的灌注策略和评估流程，以利用去细胞灌注技术成功制备天然子宫去细胞支架，为子宫组织工程研究提供良好的平台。
选用雌性斯普拉格 - 道利大鼠。在进行物理冷冻、解冻和酶解后，将含有十二烷基硫酸钠的吐温 -100等洗脱液通过子宫动脉灌注到子宫内。去细胞处理后，通过大体观察、亚甲蓝染色、苏木精 - 伊红染色、免疫组织化学染色、基因组DNA定量分析、胶原蛋白检测与鉴定、细胞因子含量测定、透射电子显微镜（TEM）和扫描电子显微镜（SEM）对每个支架进行评估。
灌注后建立了透明的子宫支架，组织学检查和TEM显示其不含细胞残留。DNA含量显示低于正常子宫的5%。亚甲蓝染色和SEM表明血管网络和空间结构完整。免疫组织化学染色和胶原蛋白定量显示支架的细胞外基质成分得到完全保留。此外，酶联免疫吸附测定结果表明，包括表皮生长因子、碱性成纤维细胞生长因子和转化生长因子β在内的细胞因子保留在去细胞支架中，并且仍表现出一定的生物活性。
成功建立了去细胞子宫支架，其物理和化学性质得以保留；因此，它可作为子宫组织工程研究的替代平台。
