In Vitro Assessment of Adipose-Derived Mesenchymal Stem Cell attachment to Injured Cartilage in Agarose Hydrogel Biomaterial

Abstract

Articular cartilage (AC) injuries are a prevalent problem leading to osteoarthritis (OA), which significantly impacts the quality of life of millions worldwide. Tissue engineering using mesenchymal stem cells (MSCs) holds potential as a promising therapeutic approach. This study aimed to investigate the attachment of adipose-derived MSCs (A-MSCs) to cartilage with four types of injuries: scraping, load, 3 mm biopsy punch, and scalpel cut. Injured explants and A-MSCs were placed inside 12 culture wells embedded in 2% agarose, one explant per well, hydrogel biomaterial to mimic native cartilage in vitro. The hypothesis was that A-MSCs would attach to the injured sites. Bovine cartilage explants (N=3) were subjected to the four injury types, totalling 56 explants, and the study was conducted over seven days, with attachment assessment on days 5 and 7 using confocal laser scanning microscopy (CLSM). The results revealed a decreased cartilage viability in both load and scraping injuries, with higher Propidium Iodide (PI)-labelled levels observed on CLSM. Additionally, the number of A-MSCs attachment was higher on these injuries compared to the 3mm biopsy punch and scalpel cut. Additionally, day 7 viability percentage was lower compared to day 5 across all injury samples. These findings suggest that injured areas show increased A-MSC attachment, indicating the potential of A-MSCs for promoting tissue repair. This study has implications for tissue engineering in developing a model to study A-MSCs behaviour to injured cartilage explants. Future recommendations include increasing the culture time to promote chondrogenesis and utilising a human model for a more clinically relevant approach. Understanding the attachment behaviour of A-MSCs to injured cartilage enhances the knowledge base in cartilage tissue engineering and contributes to future therapeutic advancements.