IADR Abstract Archives
Antioxidative Potential of Dental Pulp Stem Cell Conditioned Medium
Objectives: Dental pulp-derived mesenchymal stem cells (DPSCs) are known for their rich secretion profile, with their secretome used in regenerative medicine, including bone tissue engineering. Gelatin methacryloyl (GelMA), among various biomaterials, stands out for its biocompatibility, tunability, and functionality, being often selected as a scaffolding material. Nevertheless, its curing process by high visible energy light potentially deteriorates general cell health. By using GelMA of Good Manufacturing Practice (GMP) grade, this study aimed: 1) to achieve a comprehensive understanding of the biological effects of photo-curing process on DPSCs in GelMA, with a specific focus on oxidative stress; and 2) to develop a strategy to mitigate the adverse effects by employing DPSC-conditioned medium (DPSC-CM).
Methods: DPSC-CM was produced by culturing cells from three donors in serum-free medium for 48 hours, followed by purification through centrifugation. Mass spectrometry-based proteomics was performed to identify the protein content, including antioxidants, in DPSC-CM. The effects of oxidative stress induced by photo-curing on encapsulated DPSCs, as well as the rescue potential of DPSC-CM, were evaluated by bulk RNA sequencing, RT-qPCR, cell kinetics assays (cell viability, apoptosis, senescence, motility, differentiation), and ex ovo chorioallantoic membrane assay.
Results: Following photo-curing, pathways related to oxidative phosphorylation and DNA repair were enriched in the presence of DPSC-CM, which carried 91 plausible antioxidants, while the control samples exhibited enrichment in inflammatory pathways. DPSC-CM significantly reduced the degree of cellular oxidation and stress responses post-curing, resulting in improved cell viability, growth, motility, and osteogenic differentiation, as well as fewer apoptotic and senescent cells. The deteriorated biocompatibility of freshly crosslinked DPSC-laden GelMA hydrogel was confirmed by the disrupted vasculature of chorioallantoic membranes in chicken embryos, which was prevented by the DPSC-CM.
Conclusions: This study demonstrates the robust antioxidative effects of DPSC-CM and its high compatibility and prominent properties for tissue engineering applications.
Methods: DPSC-CM was produced by culturing cells from three donors in serum-free medium for 48 hours, followed by purification through centrifugation. Mass spectrometry-based proteomics was performed to identify the protein content, including antioxidants, in DPSC-CM. The effects of oxidative stress induced by photo-curing on encapsulated DPSCs, as well as the rescue potential of DPSC-CM, were evaluated by bulk RNA sequencing, RT-qPCR, cell kinetics assays (cell viability, apoptosis, senescence, motility, differentiation), and ex ovo chorioallantoic membrane assay.
Results: Following photo-curing, pathways related to oxidative phosphorylation and DNA repair were enriched in the presence of DPSC-CM, which carried 91 plausible antioxidants, while the control samples exhibited enrichment in inflammatory pathways. DPSC-CM significantly reduced the degree of cellular oxidation and stress responses post-curing, resulting in improved cell viability, growth, motility, and osteogenic differentiation, as well as fewer apoptotic and senescent cells. The deteriorated biocompatibility of freshly crosslinked DPSC-laden GelMA hydrogel was confirmed by the disrupted vasculature of chorioallantoic membranes in chicken embryos, which was prevented by the DPSC-CM.
Conclusions: This study demonstrates the robust antioxidative effects of DPSC-CM and its high compatibility and prominent properties for tissue engineering applications.