IADR Abstract Archives
Dual-cured GelMA-Fibrin hybrid hydrogel for organotypic bone culture model
Objectives: Gelatin methacryloyl (GelMA) a hydrogel made by crosslinking gelatin with methacrylic anhydride, is widely used as a scaffold for tissue engineering and bone regenerative strategies. However, the reduced temporal stability of the GelMA scaffold makes it non-conducive to support osteogenic differentiation. Fibrin is a proangiogenic bioactive polymer, frequently used as a matrix to support physiologically relevant organotypic cultures. Fibrin is the product of crosslinking between fibrinogen and thrombin in the presence of calcium ions. In the present study, we propose the development of dual-cure GelMA-Fibrin hydrogel and investigate its potential to support osteodifferentiation.
Methods: GelMA in different concentrations (G:6%,8%,10%) were mixed with 6% fibrinogen (F) in 1:1 (v/v) ratio to develop a hybrid hydrogel (3H, 4H & 5H). These prepolymer gels were tested for their viscoelastic properties. The gels were later photocured (1625mW/cm2) followed by crosslinking with thrombin and characterized for enzymatic degradation. Cellular viability was evaluated by encapsulating human periodontal ligament-derived stem cells (PDLSC) within the gels followed by live/dead staining and confocal laser microscopy. For osteodifferentiation, the PDLSC encapsulated in hydrogels were cultured in osteogenic media for 14 days and the levels of osteogenic markers were evaluated.
Results: GelMA-Fibrin hydrogels demonstrated shear thinning behavior and reduced viscoelastic properties compared to GelMA hydrogel controls. The incorporation of fibrinogen into GelMA significantly reduced the mass loss percentage and increased its elastic modulus. A significant reduction in the number of dead cells was observed in hybrid hydrogels compared to respective GelMA controls (p<0.05). Dual-cured hybrid hydrogels demonstrated enhanced osteodifferentiation potential as evidenced by the presence of phosphate, carboxyl ions peaks in spectroscopy, and increased levels of alkaline phosphatase levels under osteogenic conditions.
Conclusions: The dual-cured GelMA-fibrin hydrogel demonstrated increased mechanical properties, lower degradation rates, and superior bio-functionality. This offers promising applications in the field of tissue engineering and bone regenerative strategies.
Methods: GelMA in different concentrations (G:6%,8%,10%) were mixed with 6% fibrinogen (F) in 1:1 (v/v) ratio to develop a hybrid hydrogel (3H, 4H & 5H). These prepolymer gels were tested for their viscoelastic properties. The gels were later photocured (1625mW/cm2) followed by crosslinking with thrombin and characterized for enzymatic degradation. Cellular viability was evaluated by encapsulating human periodontal ligament-derived stem cells (PDLSC) within the gels followed by live/dead staining and confocal laser microscopy. For osteodifferentiation, the PDLSC encapsulated in hydrogels were cultured in osteogenic media for 14 days and the levels of osteogenic markers were evaluated.
Results: GelMA-Fibrin hydrogels demonstrated shear thinning behavior and reduced viscoelastic properties compared to GelMA hydrogel controls. The incorporation of fibrinogen into GelMA significantly reduced the mass loss percentage and increased its elastic modulus. A significant reduction in the number of dead cells was observed in hybrid hydrogels compared to respective GelMA controls (p<0.05). Dual-cured hybrid hydrogels demonstrated enhanced osteodifferentiation potential as evidenced by the presence of phosphate, carboxyl ions peaks in spectroscopy, and increased levels of alkaline phosphatase levels under osteogenic conditions.
Conclusions: The dual-cured GelMA-fibrin hydrogel demonstrated increased mechanical properties, lower degradation rates, and superior bio-functionality. This offers promising applications in the field of tissue engineering and bone regenerative strategies.