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丝素纤维增强透明质酸水凝胶在软骨组织工程中的应用

Silk Fiber-Reinforced Hyaluronic Acid-Based Hydrogel for Cartilage Tissue Engineering.

机构信息

Department of Anatomy, Christian-Albrechts-University Kiel, 24118 Kiel, Germany.

Department of Oral and Maxillofacial Surgery, Division of Regenerative Orofacial Medicine, University Medical Center Hamburg-Eppendorf, 20251 Hamburg, Germany.

出版信息

Int J Mol Sci. 2021 Mar 31;22(7):3635. doi: 10.3390/ijms22073635.

DOI:10.3390/ijms22073635
PMID:33807323
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8036422/
Abstract

A continuing challenge in cartilage tissue engineering for cartilage regeneration is the creation of a suitable synthetic microenvironment for chondrocytes and tissue regeneration. The aim of this study was to develop a highly tunable hybrid scaffold based on a silk fibroin matrix (SM) and a hyaluronic acid (HA) hydrogel. Human articular chondrocytes were embedded in a porous 3-dimensional SM, before infiltration with tyramine modified HA hydrogel. Scaffolds were cultured in chondropermissive medium with and without TGF-β1. Cell viability and cell distribution were assessed using CellTiter-Blue assay and Live/Dead staining. Chondrogenic marker expression was detected using qPCR. Biosynthesis of matrix compounds was analyzed by dimethylmethylene blue assay and immuno-histology. Differences in biomaterial stiffness and stress relaxation were characterized using a one-step unconfined compression test. Cell morphology was investigated by scanning electron microscopy. Hybrid scaffold revealed superior chondro-inductive and biomechanical properties compared to sole SM. The presence of HA and TGF-β1 increased chondrogenic marker gene expression and matrix deposition. Hybrid scaffolds offer cytocompatible and highly tunable properties as cell-carrier systems, as well as favorable biomechanical properties.

摘要

软骨组织工程中用于软骨再生的一个持续挑战是为软骨细胞和组织再生创造合适的合成微环境。本研究旨在开发一种基于丝素蛋白基质（SM）和透明质酸（HA）水凝胶的高度可调的混合支架。将人关节软骨细胞嵌入多孔 3D SM 中，然后用酪氨酸改性的 HA 水凝胶渗透。支架在软骨形成培养基中培养，有无 TGF-β1。使用 CellTiter-Blue 测定法和 Live/Dead 染色评估细胞活力和细胞分布。使用 qPCR 检测软骨形成标志物的表达。通过二甲亚甲基蓝测定法和免疫组织化学分析分析基质化合物的生物合成。使用一步法无约束压缩试验表征生物材料的刚度和应力松弛差异。通过扫描电子显微镜研究细胞形态。与单纯的 SM 相比，混合支架显示出更好的软骨诱导和生物力学性能。HA 和 TGF-β1 的存在增加了软骨形成标志物基因的表达和基质沉积。混合支架作为细胞载体系统具有细胞相容性和高度可调的特性，以及良好的生物力学性能。

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