肿瘤细胞治疗研究
英文原题：0.1% Peracetic Acid Treatment Is the Optimal Process for Ovarian Decellularized Extracellular Matrix.
0.1% PAA 是 OV-dECM 的最佳灭菌工艺，具有更好的灭菌能力和生物相容性。影响声明：本研究确定 0.1% 过氧乙酸是卵巢脱细胞细胞外基质（OV-dECM）的最佳灭菌方法，解决了人工卵巢（AO）开发的关键障碍。
本研究旨在确定卵巢脱细胞细胞外基质（OV-dECM）的最佳灭菌流程。研究制备 OV-dECM，并评估四种灭菌方法：紫外线、1 mg/L CuCl₂ 加 0.5% H₂O₂、70% 乙醇和 0.1% 过乙酸（PAA）。研究评估了灭菌效率以及体外和体内生物相容性。苏木精-伊红染色、4′,6-二脒基-2-苯基吲哚（DAPI）染色和 DNA 定量结果表明 OV-dECM 制备成功。0.1% PAA 和 70% 乙醇的灭菌效果较好。经 70% 乙醇或 0.1% PAA 灭菌的 OV-dECM 均可与人脐带间充质干细胞相容；两组在第 7 天和第 10 天的差异分别为 p=0.187 和 p=0.293（均 p>0.05），并能维持细胞活率（CCK-8，细胞计数试剂盒-8）和细胞形态（F-actin 染色）。两种灭菌组的 CD31（新生血管标志物）融合指数均良好，组间差异不显著（p=0.288；p>0.05）；而 0.1% PAA 组的 α-平滑肌肌动蛋白（另一种新生血管标志物）高于 70% 乙醇组（p=0.012；p<0.05）。CD68、CD86 和 CD206（巨噬细胞表型标志物）评估显示，小鼠植入 OV-dECM 后，0.1% PAA 组较 70% 乙醇组更可能促进巨噬细胞由 M1 向 M2 转化。综上，0.1% PAA 是 OV-dECM 的最佳灭菌流程，灭菌能力和生物相容性更好。 影响说明：本研究确定 0.1% 过乙酸是卵巢脱细胞细胞外基质（OV-dECM）的最佳灭菌方法，解决了人工卵巢（AO）开发中的关键障碍。该方法可确保 OV-dECM 无菌，同时保留生物相容性，并促进巨噬细胞由 M1 向 M2 极化以减轻炎症。这项工作推动 AO 转化应用，为癌症/白血病患者的生育力保存带来希望，也为再生医学中 OV-dECM 灭菌标准化提供指导。
This research aimed to find the optimal sterilization process for ovarian decellularized extracellular matrix (OV-dECM). OV-dECM was prepared. Four different sterilization processes, such as ultraviolet ray, 1 mg/L CuCl 2 + 0.5% H 2 O 2 , 70% ethanol, and 0.1% peracetic acid (PAA), were evaluated. The sterilization efficiency and the in vitro and in vivo biocompatibility were assessed. Hematoxylin and eosin, 4',6-diamidino-2-phenylindole staining, and DNA quantitation indicated the successful production of OV-dECM. 0.1% PAA and 70% ethanol achieved better sterilization. The sterilized OV-dECM in the 70% ethanol and 0.1% PAA groups could integrate with human umbilical cord mesenchymal stem cell (between them: 7 days: p = 0.187; 10 days: p = 0.293; both p > 0.05) and keep the cell viability (CCK-8, cell counting kit-8) and cell morphology (F-actin staining). The sterilized OV-dECM in both 70% ethanol and 0.1% PAA groups had good CD31 (a neovascularization maker) fusion indexes (between them: p = 0.288; p > 0.05), while -smooth muscle actin (another neovascularization maker) in the 0.1% PAA group was higher than that in the 70% ethanol group (between them: p = 0.012; p < 0.05). The assessment of CD68, CD86, and CD206 (macrophage phenotype makers) demonstrated that the 0.1% PAA group had better chance to promote the M1-M2 transformation of macrophage than 70% ethanol after the implantation of OV-dECM in mice. In conclusion, 0.1% PAA is the optimal sterilization process for OV-dECM with better sterilization capacity and biocompatibility.Impact StatementThis study identifies 0.1% peracetic acid as the optimal sterilization method for ovarian decellularized extracellular matrix (OV-dECM), resolving a critical barrier to artificial ovary (AO) development. It ensures OV-dECM sterility while preserving biocompatibility, promoting macrophage M1-to-M2 polarization to reduce inflammation. This advances AO translational potential, offering hope for fertility preservation in patients with cancer/leukemia and guiding standardized OV-dECM sterilization in regenerative medicine.
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