Bioactive and biomimetic 3D scaffolds for bone tissue engineering using graphitic carbon nitride as a sustainable visible light photoinitiator.

Graphitic carbon nitride (g-C3N4) is explored as a novel sustainable visible light photoinitiator for the preparation of biomimetic 3D hydrogel scaffolds comprising gelatin methacrylamide (GelMA) and dopamine methacrylamide for use in tissue engineering. The initiator efficiency was assessed by comparing the swelling behavior and the stability of photopolymerized hydrogels prepared with GelMA of different degrees of functionalization and different comonomer compositions. Bioactive composite hydrogels with a 50 wt% nanohydroxyapatite (nHAp) content, to closely mimic the actual bone composition, were successfully obtained by the introduction of nHAp in the prepolymer solutions followed by photopolymerization. The composite hydrogels demonstrated enhanced mechanical properties and excellent stability in PBS verifying the preparation of robust 3D scaffolds for use in cancellous or pre-calcified bone tissue engineering applications. The in vitro cell response of the composite scaffolds exhibited high cell viability and enhanced differentiation of pre-osteoblasts to mature osteoblasts, demonstrating their osteogenic potential. This work establishes, for the first time, the excellent properties of g-C3N4 as a sustainable, visible light initiator, fully satisfying the principles of green chemistry, for the preparation of robust and biologically relevant hydrogels, and proposes a new approach to overcome the main challenges of conventional photoinitiators in cell scaffold fabrication, such as photobleaching, high cost and non-scalable synthesis employing toxic organic precursors and solvents.

Kappa-carrageenan/chitosan/gelatin scaffolds enriched with potassium chloride for bone tissue engineering.

Kappa-carrageenan is a biocompatible natural polysaccharide able to form hydrogels for tissue regeneration. In bone tissue engineering, achieving a bioactive microenvironment with appropriate mechanical properties in polysaccharide-based scaffolds remains a challenge. This study aims to fabricate 3D scaffolds comprising kappa-carrageenan, chitosan and gelatin, crosslinked with KCl, and evaluate their mechanical and biological properties for bone tissue engineering. The produced scaffolds include kappa-carrageenan/chitosan (KC), kappa-carrageenan/chitosan/gelatin (KCG), kappa-carrageenan/chitosan/gelatin enriched with KCl (KCG-KCl), and chitosan/gelatin (CG). All scaffolds present degradation rates ranging from 30% weight loss on day 21, pore size distribution in the range of 100-160 µm and porosity above 80%. The Young modulus values range from 9 to 256 kPa, with the KCl-containing KCG scaffolds demonstrating the highest values, validating the role of KCl in the coil to helix transition of kappa-carrageenan leading to firmer structures. In vitro biological evaluation indicates that pre-osteoblasts proliferate significantly from day 3 up to day 14 on all scaffold compositions. The alkaline phosphatase activity shows a significant increase up to day 14. The calcium production displays a constant increase from day 14 up to day 28, proving that all scaffold compositions support the osteogenic differentiation potential.

Fibro/chondrogenic differentiation of dental stem cells into chitosan/alginate scaffolds towards temporomandibular joint disc regeneration.

Tissue engineering (TE) may provide effective alternative treatment for challenging temporomandibular joint (TMJ) pathologies associated with disc malpositioning or degeneration and leading to severe masticatory dysfunction. Aim of this study was to evaluate the potential of chitosan/alginate (Ch/Alg) scaffolds to promote fibro/chondrogenic differentiation of dental pulp stem cells (DPSCs) and production of fibrocartilage tissue, serving as a replacement of the natural TMJ disc. Ch/Alg scaffolds were fabricated by crosslinking with CaCl2 combined or not with glutaraldehyde, resulting in two scaffold types that were physicochemically characterized, seeded with DPSCs or human nucleus pulposus cells (hNPCs) used as control and evaluated for cell attachment, viability, and proliferation. The DPSCs/scaffold constructs were incubated for up to 8 weeks and assessed for extracellular matrix production by means of histology, immunofluorescence, and thermomechanical analysis. Both Ch/Alg scaffold types with a mass ratio of 1:1 presented a gel-like structure with interconnected pores. Scaffolds supported cell adhesion and long-term viability/proliferation of DPSCs and hNPCs. DPSCs cultured into Ch/Alg scaffolds demonstrated a significant increase of gene expression of fibrocartilaginous markers (COLI, COL X, SOX9, COM, ACAN) after up to 3 weeks in culture. Dynamic thermomechanical analysis revealed that scaffolds loaded with DPSCs significantly increased storage modulus and elastic response compared to cell-free scaffolds, obtaining values similar to those of native TMJ disc. Histological data and immunochemical staining for aggrecan after 4 to 8 weeks indicated that the scaffolds support abundant fibrocartilaginous tissue formation, thus providing a promising strategy for TMJ disc TE-based replacement.