The development of functional therapies for corneal repair and regeneration is a pressing issue. Corneal stroma provides the principal functions of the cornea. However, because of the highly organized nature of the stromal matrix, the attempts to reproduce corneal stroma might follow a scar model. Here, we have developed a protocol for the efficient generation of a cell-laden and orthogonal-multilayer tissue-engineered (TE) corneal stroma, which is induced by the mechanical effects of compressed collagen (CC) or stretched compressed collagen (SCC). Within SCC, with applied compression and force extension, collagen microfibres and corneal stromal cells (CSCs) are arranged orderly, while collagen fibres and CSCs in CC are randomly arranged. Dehydrated SCC has higher tensile strength than dehydrated CC. Hydrated SCC has similar transparency with hydrated native corneal stroma. Compared with those cultured on tissue culture plates (TCP), down-regulation of the genes and proteins of cytoskeleton, activation, proliferation, collagen and TRPV4, up-regulation of proteoglycans, gap junction proteins and TRPA1 are in CSCs of CC and SCC. Moreover, SCC and CC grafts displayed biocompatibility and integration with host corneal tissue after rabbit intra-corneal stromal transplantation by wk 6 under slit lamp microscopy, in vivo confocal microscopy and histological examination. The SCC model facilitates the construction of physiological feature TE corneal stroma, which serves as a foundation for physiological TE construction of other tissues.

Statement of significance

The development of functional therapies for corneal repair and regeneration is a pressing issue. Corneal stroma provides the principal functions of the cornea. Here, we have developed a protocol for the efficient generation of a cell-laden and orthogonal-multilayer tissue-engineered (TE) corneal stroma, which is induced by the mechanical effects of compressed collagen (CC) or stretched compressed collagen (SCC). These models facilitate the construction of physiological feature TE corneal stroma, which serves as a foundation for physiological TE construction of other tissues and helps to reverse fibrosis pathologies in general.

中文翻译：

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机械胶原微环境和移植在兔模型中诱导细胞负载和正交多层组织工程的角膜基质。

用于角膜修复和再生的功能疗法的发展是一个紧迫的问题。角膜基质提供角膜的主要功能。但是，由于基质的高度组织性，重塑角膜基质的尝试可能遵循疤痕模型。在这里，我们已经开发出一种协议，可以有效生成细胞负载和正交多层组织工程化（TE）的角膜基质，该协议是由压缩胶原蛋白（CC）或拉伸压缩胶原蛋白（SCC）的机械作用诱导的。在SCC中，通过施加压缩和力扩展，胶原微纤维和角膜基质细胞（CSC）有序排列，而CC中的胶原纤维和CSC则随机排列。脱水SCC具有比脱水CC更高的拉伸强度。水合SCC与水合天然角膜基质具有相似的透明度。与组织培养板（TCP）上培养的细胞相比，细胞骨架的基因和蛋白质，激活，增殖，胶原和CC和SCC的CSC中存在TRPV4，蛋白聚糖，间隙连接蛋白和TRPA1的上调。此外，在裂隙灯显微镜，体内共聚焦显微镜和组织学检查下，第6周Wk 6兔角膜内基质移植后，SCC和CC移植物显示出生物相容性并与宿主角膜组织整合。SCC模型促进了生理特征TE角膜基质的构建，这为其他组织的生理TE构建奠定了基础。

重要声明

用于角膜修复和再生的功能疗法的发展是一个紧迫的问题。角膜基质提供角膜的主要功能。在这里，我们已经开发出一种协议，可以有效生成细胞负载和正交多层组织工程化（TE）的角膜基质，该协议是由压缩胶原蛋白（CC）或拉伸压缩胶原蛋白（SCC）的机械作用诱导的。这些模型促进了生理特征TE角膜基质的构建，这为其他组织的生理TE构建奠定了基础，并有助于总体上逆转纤维化病理。