Department of Urology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, No.639, Zhizaoju Road in Huangpu District, Shanghai, 200011, China.
Stem Cell Res Ther. 2020 Dec 11;11(1):535. doi: 10.1186/s13287-020-02052-4.
Tissue engineering is a promising alternative for urethral reconstruction, and adipose-derived stem cells (ADSCs) are widely used as seeding cells. Hypoxia preconditioning can significantly enhance the therapeutic effects of ADSCs. The low oxygen tension of postoperative wound healing is inevitable and may facilitate the nutritional function of ADSCs. This study aimed to investigate if hypoxia-preconditioned ADSCs, compared to normoxia-preconditioned ADSCs, combined with scaffold could better promote urethral reconstruction and exploring the underlying mechanism.
In vitro, paracrine cytokines and secretomes that were secreted by hypoxia- or normoxia-preconditioned ADSCs were added to cultures of human umbilical vein endothelial cells (HUVECs) to measure their functions. In vivo, hypoxia- or normoxia-preconditioned ADSCs were seeded on a porous nanofibrous scaffold for urethral repair on a defect model in rabbits.
The in vitro results showed that hypoxia could enhance the secretion of VEGFA by ADSCs, and hypoxia-preconditioned ADSCs could enhance the viability, proliferation, migration, angiogenesis, and glycolysis of HUVECs (p < 0.05). After silencing VEGFA, angiogenesis and glycolysis were significantly inhibited (p < 0.05). The in vivo results showed that compared to normoxia-preconditioned ADSCs, hypoxia-preconditioned ADSCs combined with scaffolds led to a larger urethral lumen diameter, preserved urethral morphology, and enhanced angiogenesis (p < 0.05).
Hypoxia preconditioning of ADSCs combined with scaffold could better promote urethral reconstruction by upregulating angiogenesis and glycolysis. Hypoxia-preconditioned ADSCs combined with novel scaffold may provide a promising alternative treatment for urethral reconstruction.
组织工程是一种有前途的尿道重建替代方法，脂肪来源的干细胞（ADSCs）被广泛用作种子细胞。缺氧预处理可以显著增强 ADSCs 的治疗效果。术后伤口愈合的低氧张力是不可避免的，并且可能促进 ADSCs 的营养功能。本研究旨在探讨与常氧预处理 ADSCs 相比，缺氧预处理 ADSCs 与支架结合是否能更好地促进尿道重建，并探讨其潜在机制。
在体外，将缺氧或常氧预处理的 ADSCs 分泌的旁分泌细胞因子和 secretomes 添加到人脐静脉内皮细胞（HUVECs）的培养物中，以测量其功能。在体内，将缺氧或常氧预处理的 ADSCs 接种到多孔纳米纤维支架上，用于修复兔缺陷模型中的尿道。
体外结果表明，缺氧可以增强 ADSCs 分泌 VEGFA，缺氧预处理 ADSCs 可以增强 HUVECs 的活力、增殖、迁移、血管生成和糖酵解（p<0.05）。沉默 VEGFA 后，血管生成和糖酵解明显受到抑制（p<0.05）。体内结果表明，与常氧预处理 ADSCs 相比，缺氧预处理 ADSCs 与支架结合导致尿道管腔直径更大，保持尿道形态，并增强血管生成（p<0.05）。
支架结合缺氧预处理 ADSCs 可以通过上调血管生成和糖酵解来更好地促进尿道重建。支架结合新型缺氧预处理 ADSCs 可能为尿道重建提供一种有前途的替代治疗方法。
