The development of a dental light curable PRFe-loaded hydrogel as a potential scaffold for pulp-dentine complex regeneration: An in vitro study

Abstract

Aim: The study aimed to develop a bicomponent bioactive hydrogel formed in situ and enriched with an extract of platelet-rich fibrin (PRFe) and to assess its potential for use in pulp-dentine complex tissue engineering via cell homing.

Methodology: A bicomponent hydrogel based on photo-activated naturally derived polymers, methacrylated chitosan (ChitMA) and methacrylated collagen (ColMA), plus PRFe was fabricated. The optimized formulation of PRFe-loaded bicomponent hydrogel was determined by analysing the mechanical strength, swelling ratio and cell viability simultaneously. The physical, mechanical, rheological and morphological properties of the optimal hydrogel with and without PRFe were determined. Additionally, MTT, phalloidin/DAPI and live/dead assays were carried out to compare the viability, cytoskeletal morphology and migration ability of stem cells from the apical papilla (SCAP) within the developed hydrogels with and without PRFe, respectively. To further investigate the effect of PRFe on the differentiation of encapsulated SCAP, alizarin red S staining, RT-PCR analysis and immunohistochemical detection were performed. Statistical significance was established at p < .05.

Results: The optimized formulation of PRFe-loaded bicomponent hydrogel can be rapidly photocrosslinked using available dental light curing units. Compared to bicomponent hydrogels without PRFe, the PRFe-loaded hydrogel exhibited greater viscoelasticity and higher cytocompatibility to SCAP. Moreover, it promoted cell proliferation and migration in vitro. It also supported the odontogenic differentiation of SCAP as evidenced by its promotion of biomineralization and upregulating the gene expression for ALP, COL I, DSPP and DMP1 as well as facilitated angiogenesis by enhancing VEGFA gene expression.

Conclusions: The new PRFe-loaded ChitMA/ColMA hydrogel developed within this study fulfils the criteria of injectability, cytocompatibility, chemoattractivity and bioactivity to promote odontogenic differentiation, which are fundamental requirements for scaffolds used in pulp-dentine complex regeneration via cell-homing approaches.

Keywords: injectable hydrogel; methacrylated chitosan; methacrylated collagen; platelet-rich fibrin; visible light crosslinking.