Abstract
Gelatin methacryloyl (GelMA) stands out for its biocompatibility, tunability, and functionality, being often selected as a scaffolding material. However, the biological modulations induced by its photocrosslinking process on mesenchymal stem cells as well as stress mitigation measures remain insufficiently explored. By using GelMA of Good Manufacturing Practice (GMP) grade, this study aimed (a) to achieve a comprehensive understanding of the biological effects of photocrosslinking process with a specific focus on oxidative stress and (b) to develop a strategy to mitigate the adverse effects by employing conditioned medium (CM) by dental pulp stem cells (DPSCs). Following photocrosslinking, pathways related to oxidative phosphorylation and DNA repair were enriched in the presence of DPSC-CM carrying various antioxidants such as peroxiredoxin (PRDX) 1-6 and superoxide dismutase type 1 (SOD1), while the control samples exhibited enrichment in inflammatory signaling pathways. Incorporating DPSC-CM into the hydrogel notably reduced the degree of cellular oxidation caused by photocrosslinking and stress responses, resulting in improved cell viability, growth, motility, and osteogenic differentiation, as well as fewer apoptotic and senescent cells compared to those without DPSC-CM. The deteriorated biocompatibility of freshly crosslinked GelMA hydrogel was confirmed by the disrupted vasculature of chorioallantoic membranes in chicken embryos after implantation, which was prevented by DPSC-CM. In conclusion, this study demonstrates the robust antioxidative effects of DPSC-CM, mitigating the negative effect of GelMA photocrosslinking processes.