Bioprinting a resilient and transparent cornea stroma equivalent: harnessing dual crosslinking strategy with decellularized cornea matrix and silk fibroin hybrid.
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引用次数: 0
Abstract
Bioprinting a resilient yet optically transparent corneal tissue substitute remains a challenge. In this study we introduce an innovative methodology aimed at bolstering the mechanical and optical attributes of silk fibroin (SF) hydrogels, pivotal for the progression of cornea tissue engineering. We devised a unique eosin Y-based photoinitiator system to instigate di-tyrosine linkages within highly concentrated pristine SF solutions under green light exposure. This pioneering technique resulted in SF hydrogels fortified by dityrosine covalent bonds, preserving exceptional transparency and soft elastomeric qualities devoid of spontaneous transitions to stiff, opaque beta-sheet conformations. Furthermore, we synergistically combined SF with decellularized cornea matrix (DCM) hydrogel, leveraging photo-polymerization under green light followed by thermal gelation to establish resilient and stable gel formation. The ensuing dual crosslinked hybrid hydrogels exhibited superior mechanical and thermal resilience in comparison to dual crosslinked DCM hydrogels. The inclusion of SF in DCM further augmented the hydrogel's elasticity and shear recovery, positioning it as an optimal bioink for cornea bioprinting endeavors. During the extrusion printing process, photocrosslinking of the bioink superficially fortified SF and DCM polymer chains via di-tyrosine linkages, furnishing initial stability and mechanical fortitude. Subsequent post-printing thermal gelation further reinforced collagen chains through self-assembly. Notably, the bioprinted cornea constructs, housing human limbal mesenchymal stem cells, manifested transparency, structural integrity, and optimal functionality, underscored by the expression of keratocyte proteoglycans. In summation, our engineered 3D constructs exhibit promising potential forin vivoapplications in cornea tissue engineering, marking a significant stride forward in the field's advancement.
生物打印具有弹性且光学透明的角膜组织替代物仍然是一项挑战。在本研究中,我们介绍了一种创新方法,旨在增强丝纤维蛋白(SF)水凝胶的机械和光学属性,这对角膜组织工程的进展至关重要。我们设计了一种独特的基于曙红 Y 的光引发剂系统,可在绿光照射下在高浓度原始 SF 溶液中引发二酪氨酸连接。这项开创性的技术使 SF 水凝胶得到了双酪氨酸共价键的强化,保持了优异的透明度和柔软的弹性,不会自发转变为僵硬、不透明的β片构象。此外,我们还将 SF 与脱细胞角膜基质(DCM)水凝胶协同结合,利用绿光下的光聚合反应,然后通过热凝胶化来建立弹性和稳定的凝胶形成。与双交联 DCM 水凝胶相比,这种双交联混合水凝胶具有更优越的机械和热回弹性。在 DCM 中加入 SF 进一步增强了水凝胶的弹性和剪切恢复能力,使其成为角膜生物打印的最佳生物墨水。在挤压打印过程中,生物墨水的光交联通过二酪氨酸连接使 SF 和 DCM 聚合物链表面强化,从而提供了初始稳定性和机械强度。随后的印刷后热凝胶技术通过自组装进一步强化了胶原蛋白链。值得注意的是,生物打印的角膜构建体容纳了人类角膜缘间充质干细胞(hLMSCs),表现出透明度、结构完整性和最佳功能性,角膜细胞蛋白多糖的表达更凸显了这一点。总之,我们的工程三维构建物在角膜组织工程的体内应用中展现出了巨大的潜力,标志着该领域向前迈出了一大步。
期刊介绍:
Biofabrication is dedicated to advancing cutting-edge research on the utilization of cells, proteins, biological materials, and biomaterials as fundamental components for the construction of biological systems and/or therapeutic products. Additionally, it proudly serves as the official journal of the International Society for Biofabrication (ISBF).