Nano-hydroxyapatite-incorporated polycaprolactone nanofibrous scaffold as a dentin tissue engineering-based strategy for vital pulp therapy

Targeting a tissue engineering-based vital pulp therapy (VPT), this study investigated the incorporation of nano-hydroxyapatite (nHA) into polycaprolactone (PCL) nanofibers, and the metabolism of human dental pulp cells (HDPCs) seeded on the scaffolds. PCL-based solutions (10% w/v) containing nHA (0 – control; 0.5; 1.0; or 2.0% w/v) were electrospun into nanofibrous scaffolds. The scaffolds were characterized for morphology and composition (MEV/EDS), solubility, the release of calcium/phosphate (C/P), and modulation of medium pH. Then, HDPCs were seeded on the scaffolds and evaluated for cell viability (alamarBlue and live/dead), adhesion and spreading (F-actin), total protein (TP; Lowry), alkaline phosphatase activity (ALP; thymolphthalein assay), expression of odontogenic genes (RT-qPCR), and formation of a mineralized matrix (Alizarin Red). Data were analyzed with ANOVA and post-hocs (α = 5%). Higher nHA concentrations roughened fiber surfaces, whereas PCL+ 2%nHA increased the interfibrillar spaces. PCL+ 1%nHA or PCL+ 2%nHA significantly released more C/P but the medium pH was maintained below 8.0. HDPCs viability was not affected by nHA, while cell adhesion/spreading was favored, especially for PCL+ 2%nHA. Higher protein content and ALP activity were seen for scaffolds incorporated with nHA, after 21 days. PCL+ 1%nHA and PCL+ 2%nHA upregulated the expression of DSPP and DMP1 in 14 days, and COL1A1, ALPL, and DMP1 in 21 days. The formation of a mineralized matrix was nHA concentration-dependent, and it was about 9 × higher for PCL+ 2%nHA. nHA-incorporated PCL nanofibrous scaffolds are cytocompatible and can stimulate the adhesion and odontogenic potential of HDPCs. PCL+ 2%nHA formulation is a bioactive tissue engineering-based cell-homing strategy for VPT. •PCL/nHA formulations produced interconnected fibers that released bioactive ions keeping the neutral pH of the medium.•The presence of nHA enhanced human dental pulp cells spreading and odontogenic differentiation.•PCL scaffolds functionalized with 2% nHA have great potential to mediate dentinogenesis by human dental pulp cells.