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预退火丝弹性蛋白样共重组体中的软骨再生:在体外培养平台中,将成熟软骨细胞包埋于可注射水凝胶中。

Cartilage Regeneration in Preannealed Silk Elastin-Like Co-Recombinamers Injectable Hydrogel Embedded with Mature Chondrocytes in an Ex Vivo Culture Platform.

机构信息

Technical Proteins Nanobiotechnology S.L. , Paseo Belén 9A , 47001 Valladolid , Spain.

LifeTec Group B.V. , 5611 ZS Eindhoven , The Netherlands.

出版信息

Biomacromolecules. 2018 Nov 12;19(11):4333-4347. doi: 10.1021/acs.biomac.8b01211. Epub 2018 Oct 22.

Abstract

Tissue engineering for cartilage repair requires biomaterials that show rapid gelation and adequate mechanical properties. Although the use of hydrogel is the most promising biomaterial, it often lacks in rigidity and anchorage of cells when they are surrounded by synovial fluid while they are subjected to heavy loads. We developed and produced the Silk Elastin-Like co-Recombinamer (SELR), which contains both the physical interaction from elastin motifs and from silk motifs. In the first part of this work, we set up and optimized a preannealing treatment based on the evolution of silk motifs into β-sheet structures in order to fulfill the required mechanical properties of hydrogels for cartilage repair. The new preannealed SELRs (pA(EIS)-(IR)) were characterized with the combination of several experimental techniques (CD, TEM, SEM, and rheology) to provide a deep insight into the material features. Finally, the regeneration properties of the pA(EIS)-(IR) hydrogel embedded with chondrocytes were evaluated. After 4 weeks of culturing in a standardized and representative ex vivo model, the biochemical and histological analysis revealed the production of glycosaminglycans and collagen. Moreover, the immunohistochemistry showed the absence of fibro-cartilage and the presence of hyaline cartilage. Hence, we conclude that the pA(EIS)-(IR) hydrogel presents improved mechanical properties while conserving the injectability, which leads to successful regeneration of hyaline cartilage in an ex vivo model.

摘要

组织工程学用于软骨修复需要具有快速凝胶化和足够机械性能的生物材料。虽然水凝胶的使用是最有前途的生物材料,但当它们被滑膜液包围并承受重载时,往往缺乏刚性和细胞的附着。我们开发并生产了丝弹性蛋白样共重组体(SELR),它包含弹性蛋白基序和丝基序的物理相互作用。在这项工作的第一部分,我们建立并优化了一种预退火处理,该处理基于丝基序向β-折叠结构的演变,以满足软骨修复用水凝胶所需的机械性能。通过几种实验技术(CD、TEM、SEM 和流变学)的结合对新的预退火 SELR(pA(EIS)-(IR))进行了表征,以深入了解材料特性。最后,评估了包埋软骨细胞的 pA(EIS)-(IR)水凝胶的再生特性。在标准化和代表性的体外模型中培养 4 周后,生化和组织学分析显示了糖胺聚糖和胶原蛋白的产生。此外,免疫组织化学显示没有纤维软骨,只有透明软骨。因此,我们得出结论,pA(EIS)-(IR)水凝胶在保持可注射性的同时提高了机械性能,从而在体外模型中成功地再生了透明软骨。

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