Abstract：
Despite significant advancements in tissue engineering and regenerative medicine during the last two decades, the fabrication of proper scaffolds with appropriate cells can still be considered a critical achievement in this field. Hypoxia is a major stumbling block to chronic wound healing, which restrains tissue engineering plans because a lack of oxygen may cause cell death. This study evaluated the cocultured human keratinocytes and human adipose-derived mesenchymal stem cells (AMSCs) on a multilayer oxygen-releasing electrospun scaffold based on PU/PCL.Sodium percarbonate (SPC)-gelatin/PU. The scaffold was characterized using Fourier transform infrared (FTIR) and scanning electron microscopy (SEM) methods. Flow cytometry confirmed mesenchymal stem cells, and then the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay and DAPI staining were used to assess the in vitro biocompatibility of the scaffold. The experimental results showed that the multilayer electrospun scaffold containing 2.5% SPC could efficiently produce oxygen. Furthermore, according to cell viability results, this structure makes a suitable substrate for the coculture of keratinocytes and AMSCs. Gene expression analysis of various markers such as Involucrin, Cytokeratin 10, and Cytokeratin 14 after 14 days confirmed that keratinocytes and AMSCs coculture on PU/PCL.SPC-gelatin/PU electrospun scaffold promotes dermal differentiation and epithelial proliferation compared to keratinocytes single-cell culture. Therefore, our study supports using oxygen-releasing scaffolds as a potential strategy to hasten skin tissue regeneration. Based on the results, this structure is suggested as a promising candidate for cell-based skin tissue engineering. Given that the developed oxygen-generating polymeric electrospun scaffolds could be used as part of a future strategy for skin tissue engineering, the PU/PCL.SPC-gelatin/PU hybrid electrospun multilayer scaffold in combination with keratinocyte/AMSC coculture is proposed as an effective substrate for skin tissue engineering and regenerative medicine platforms.
摘要：
尽管在过去的二十年中，组织工程和再生医学取得了重大进展，用适当的细胞制造适当的支架仍然可以被认为是该领域的一项重大成就。缺氧是慢性伤口愈合的主要绊脚石，这限制了组织工程计划，因为缺氧可能导致细胞死亡。本研究评估了基于PU/PCL的多层释氧电纺支架上共培养的人角质形成细胞和人脂肪间充质干细胞（AMSCs）。过碳酸钠(SPC)-明胶/PU。使用傅里叶变换红外（FTIR）和扫描电子显微镜（SEM）方法对支架进行表征。流式细胞术证实间充质干细胞，然后用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)测定和DAPI染色来评估支架的体外生物相容性。实验结果表明，含有2.5%SPC的多层静电纺丝支架可以有效地产生氧气。此外，根据细胞活力的结果，这种结构为角质形成细胞和AMSCs的共培养提供了合适的基质。各种标记如Involucrin的基因表达分析,细胞角蛋白10和细胞角蛋白14在14天后证实角质形成细胞和AMSC在PU/PCL上共培养。与角质形成细胞单细胞培养相比，SPC-明胶/PU电纺支架促进真皮分化和上皮增殖。因此,我们的研究支持使用氧释放支架作为加速皮肤组织再生的潜在策略.根据结果,这种结构被认为是基于细胞的皮肤组织工程的有希望的候选者。鉴于开发的产氧聚合物电纺支架可用作未来皮肤组织工程战略的一部分，PU/PCL。SPC-明胶/PU混合电纺多层支架结合角质形成细胞/AMSC共培养被建议作为皮肤组织工程和再生医学平台的有效基质。
