In vitro enhancement of chondrogenic and epithelial genes differentiation of human adipose mesenchymal stem cells in decellularized xenograft tracheal scaffold: Implications for tracheal disease management

Tracheal diseases and chronic respiratory conditions often require complex surgical interventions. Traditional treatment methods have limitations, prompting the need for innovative solutions. This study assesses the effectiveness of human adipose mesenchymal stem cell (ADMSC)-seeded decellularized goat trachea for tracheal tissue engineering. By investigating chondrogenic and epithelial gene expression, this study aims to develop a viable alternative for tracheal reconstruction and improve treatment strategies and patient outcomes for chronic respiratory diseases. Human adipose mesenchymal stem cells (ADMSCs) were isolated and cultured from adipose tissue samples obtained from consenting patients at Dr. Soetomo General Academic Hospital, Surabaya, Indonesia. Tracheal segments from Capra hircus L. goats were decellularized using a combination of 0.5% SDS and 3% H2O2 solution. ADMSCs were seeded onto decellularized tracheal scaffolds and cultured for 14, 21, and 28 days. Gene expression analysis was performed using quantitative PCR, with statistical analysis using the Kruskal-Wallis test. AGC gene expression peaked at day 14, showing a fivefold increase compared to days 21 and 28. SOX9 expression was highest at day 21, declining by day 28. Tubb4a and CK18 peaked at day 14, while E-Cadherin peaked at day 21. Despite these variations, no significant differences were observed between groups (p > 0.05). The ideal duration for seeding h-ADMSCs onto decellularized goat tracheal scaffolds is 14 and 21 days. Further exploration of decellularization methods, cell sources, densities, and seeding techniques is necessary to optimize epithelial and chondrocyte attachment and proliferation on the goat tracheal scaffold.