Bilayer gelatin-methacryloyl scaffold for pulp inflammation suppression and dentin-like tissue regeneration

Balancing pulp tissue inflammation and dentin regeneration is essential for maintaining tooth responsiveness and reparative ability in vital pulp therapy (VPT). Here, we describe a photocrosslinkable gelatin methacryloyl (GelMA) biomaterial electrospun into a bilayer fibrous scaffold with clinically relevant functions. It comprises a 10 % (w/v) GelMA layer loaded with ibuprofen (IBP, 10 % or 20 % w/w) for immunomodulation, and a 20 % GelMA layer containing amorphous magnesium phosphate (AMP, 5 % or 15 % w/w) to promote biomineralization. Fiber morphology, chemical composition, mechanical properties, swelling, enzymatic degradation, and release kinetics of IBP and Mg2+/PO43- ions were evaluated. Bioactivity was assessed with human dental pulp stem cells (DPSCs), macrophages, an LPS-challenged artificial pulp chamber (APC) model, and subcutaneous implantation. All fibers were bead-free and porous, and photocrosslinking yielded pulp-like stiffness. IBP-loaded fibers showed an initial burst followed by sustained release over 14 days; AMP fibers released ions continuously for 7 days. IBP layer reduced IL-1α, TNF-α, and IL-6 secretion, inhibiting NF-κB activation without cytotoxicity. AMP layer increased ALP activity, mineral deposition, and expression of COL1A1, RUNX2, and ALPL. In the APC model, the bilayer scaffold downregulated IL1A, IL1B, and TNF within 3 h; after 14 days, it upregulated ALPL, DSPP, and OCN compared to controls under LPS. Subcutaneous implantation confirmed biocompatibility: IBP decreased M1 polarization, while AMP induced transient RUNX-2 followed by ALP and osteocalcin, indicating ongoing mineralization. The dual functionality of this bilayer scaffold suggests it may serve as a promising next-generation VPT biomaterial capable of controlling inflammation and guiding dentin-like tissue regeneration.

Resumo (idioma não especificado)

Balancing pulp tissue inflammation and dentin regeneration is essential for maintaining tooth responsiveness and reparative ability in vital pulp therapy (VPT). Here, we describe a photocrosslinkable gelatin methacryloyl (GelMA) biomaterial electrospun into a bilayer fibrous scaffold with clinically relevant functions. It comprises a 10 % (w/v) GelMA layer loaded with ibuprofen (IBP, 10 % or 20 % w/w) for immunomodulation, and a 20 % GelMA layer containing amorphous magnesium phosphate (AMP, 5 % or 15 % w/w) to promote biomineralization. Fiber morphology, chemical composition, mechanical properties, swelling, enzymatic degradation, and release kinetics of IBP and Mg2+/PO43- ions were evaluated. Bioactivity was assessed with human dental pulp stem cells (DPSCs), macrophages, an LPS-challenged artificial pulp chamber (APC) model, and subcutaneous implantation. All fibers were bead-free and porous, and photocrosslinking yielded pulp-like stiffness. IBP-loaded fibers showed an initial burst followed by sustained release over 14 days; AMP fibers released ions continuously for 7 days. IBP layer reduced IL-1α, TNF-α, and IL-6 secretion, inhibiting NF-κB activation without cytotoxicity. AMP layer increased ALP activity, mineral deposition, and expression of COL1A1, RUNX2, and ALPL. In the APC model, the bilayer scaffold downregulated IL1A, IL1B, and TNF within 3 h; after 14 days, it upregulated ALPL, DSPP, and OCN compared to controls under LPS. Subcutaneous implantation confirmed biocompatibility: IBP decreased M1 polarization, while AMP induced transient RUNX-2 followed by ALP and osteocalcin, indicating ongoing mineralization. The dual functionality of this bilayer scaffold suggests it may serve as a promising next-generation VPT biomaterial capable of controlling inflammation and guiding dentin-like tissue regeneration.