Abstract
Purpose :
Hydrogels derived from decellularized tissues are widely considered as promising biomaterials for tissue regeneration. However, their rapid biodegradation may disrupt in vitro cultivation and in vivo reconstruction. The aim of our study was to retard the biodegradation of a hydrogel derived from porcine decellularized lacrimal glands (dLG-HG) by crosslinking with genipin, a natural crosslinker, and to increase the mechanical stability without affecting the secretory function as well as viability of lacrimal gland (LG) associated cells.
Methods :
dLG-HG was prepared according to an established protocol and crosslinked with 0.01, 0.1, 0.5, 1, or 2, 5 mM genipin (n=6). Proliferation of LG epithelial cells (EpC), mesenchymal stem cells (MSC), and endothelial cells (EC) on unmodified dLG-HG was assessed at day 7 with a viability assay comparative to genipin crosslinked dLG-HG (G-HG). The secretory capacity of EpC was analyzed by β-hexosaminidase assay. EpC-dependent biodegradation of G-HG was quantified by the area of hydrogel remaining in culture and compared with unmodified dLG-HG over a 7-day period. Rheological characterisation (gelation kinetics and strength) was performed using an oscillating rheometer.
Results :
Proliferation of EpC, MSC, and EC on G-HG was not significantly affected by genipin concentrations of ≤ 0.5 mM (p > 0.05). Moreover, genipin concentrations of ≤ 0.5 mM showed no negative effect on EpC secretion compared to unmodified dLG-HG (p > 0.05). After 7 days cultivation of EpC on G-HG, cell-mediated degradation was significantly delayed by ≥ 0.1 mM genipin compared to unmodified dLG-HG (> 99.6 % ± 0.6 versus 51.9 ± 24.7.0 %; p ≤ 0.05). The shear modulus of dLG-HG was improved with increasing genipin concentration (p < 0.0001).
Conclusions :
Crosslinking of dLG-HG with genipin ≥ 0.1 mM was able to prevent cell-dependent degradation and mechanical strength significantly, without negatively affecting viability of LG associated cells and secretory capacity of EpC. G-HG may be suitable as a biomaterial for tissue engineering of the lacrimal gland.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.