The effect of crosslinking density on nasal chondrocytes' redifferentiation

In this manucript, we investigate the effect of different crosslinking density of the thiolated hyalonric acid hydrogel as the supporting matrix for monolayer expanded chondrocyte's chondrogenic redifferentiation.A wide ragne of gene expression level was found to be correlated with teh level of crosslinking, and the biological processes and KEGG pathways associated with this panel of genes are related to cell death, immune response, and regulation of various foundamental developmental processes. A combination of thiolate hyaluronic acid and thiolate gelatin (1:1, 10mg/mL) solution was mixed with PEDGA crosslinker solution (10mg/mL) in a solid content ratio of 1:2, 1:4, 1:6, 1:8. Tissue engineered constructs were made from nasal chondrocytes mixed with these four hydrogels with different crosslinking density. The tissue engineered constructs were cultured for 6 weeks in normal oxygen condition (21% oxygen, NRX). After 6 weeks of culture, the constructs were assessed vis histology, immunofluorescence, GAG/DNA assays, RNA sequencing.

本研究论文中，我们探究了巯基化透明质酸（thiolated hyaluronic acid）水凝胶作为单层扩增软骨细胞软骨向再分化支持基质时，不同交联密度对其产生的影响。研究发现，诸多基因的表达水平与交联密度显著相关，且该基因集所关联的生物学过程及KEGG通路（Kyoto Encyclopedia of Genes and Genomes pathway）均与细胞死亡、免疫应答以及多种基础发育过程的调控密切相关。我们将巯基化透明质酸与巯基化明胶按1:1的质量比（总浓度为10mg/mL）配制成混合溶液，再与PEDGA交联剂溶液（浓度为10mg/mL）按固含量比1:2、1:4、1:6、1:8进行混合，得到四种不同交联密度的水凝胶体系。将鼻腔软骨细胞与上述四种水凝胶混合，制备得到组织工程构建体。将该组织工程构建体置于常氧条件（21%氧浓度，NRX）下培养6周。培养6周后，通过组织学检测、免疫荧光分析、糖胺聚糖/DNA定量实验以及RNA测序对该构建体进行评估。