Comparison of various reagents for preparing a decellularized porcine cartilage scaffold

American Journal of Translational Research

Li Shi¹  Zucheng Luo²  Shi Li³  Wenting Su⁴  Yujie Bian⁵ 

[1] Department of Dermatology, Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine, Wenzhou 325000, Zhejiang, P. R. China;Department of Orthopedics, The Second Affiliated Hospital and Yuying Childrens Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, P. R. China;Department of Plastic and Reconstructive Surgery, Shanghai Ninth Peoples Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, P. R. China;The Second School of Medicine, Wenzhou Medical University, Wenzhou 325000, Zhejiang, P. R. China;Zhejiang Provincial Key Laboratory of Orthopedics, Wenzhou 325000, Zhejiang, P. R. China

关键词: Cartilage;    decellularization;    detergent;    acellular scaffolds;    tissue engineering;   

DOI  :  

学科分类：医学（综合）

来源: e-Century Publishing Corporation

【 摘 要 】

Cartilage lesion repair is difficult due to the limited self-repair capability of cartilage and its lack of vascularization. Our previous study established a sandwich model for engineering cartilage with acellular cartilage sheets (ACSs) and chondrocytes. However, there is still debate over which agent achieves the optimal decellularization of cartilage sheets. In addition, changes in the extracellular matrix after decellularization are worth studying. We aimed to determine the optimal decellularization reagents and decellularization time for preparing cartilage sheets. This study compared the effects of 2 extraction chemicals [t-octylphenoxypolyethoxyethanol (Triton X-100) and sodium dodecyl sulfate (SDS)] on cartilage sheets. The sheets were soaked in various concentrations (0.1-2%) of the extraction solutions for various time periods (24-72 h). After the decellularization process with the various treatments, we examined the cell removal and preservation of the matrix components and microstructure to determine which method was the most efficient while inducing minimal damage to the perichondrium. Both protocols achieved decellularization within an acceptable time. DNA analysis showed that the reagent removed nearly all of the DNA from the cartilage sheets. The growth factor contents in the Triton X-100 samples were higher than those in the SDS samples, quantified by enzyme-linked immunosorbent assay (ELISA). Furthermore, Triton X-100 decreased the glycosaminoglycan (GAG) and increased the chondromodulin-I contents compared with SDS. The results of a Cell Counting Kit-8 (CCK-8) assay revealed that the ACSs were not cytotoxic. In conclusion, our results demonstrate that cartilage sheets decellularized by 1% SDS for 24 h or by 2% Triton X-100 for 48 h may be suitable candidate scaffolds for cartilage tissue engineering.

【 授权许可】

CC BY-NC