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

Author(s)

Soares, Igor Paulino Mendes ; Anselmi, Caroline ; Kitagawa, Fernanda Ali ; Ribeiro, Rafael Antonio de Oliveira ; Leite, Maria Luisa ; Costa, Carlos Alberto de Souza ; Hebling, Josimeri

• Document type: Journal article
• Source: Dental Materials; v. 38, n. 6, p. 18-pg., 2022-06-01.

Abstract

Objectives: 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.Methods: 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 (alpha = 5%).Results: 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 x higher for PCL+ 2%nHA. Significance: 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.(c) 2022 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved. (AU)

• FAPESP's process: 19/07400-0 - Polycaprolactone/nano-hydroxyapatite nanofibrous scaffolds for dentin-pulp complex regeneration: synthesis, characterization, and evaluation on human dental pulp cells
• Grantee: Igor Paulino Mendes Soares
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 19/16473-1 - Polycaprolactone-nanohydroxyapatite nanofiber scaffolds for regeneration of the pulp-dentin complex: synthesis, characterization and biological behavior in human teeth culture
• Grantee: Josimeri Hebling Costa
• Support Opportunities: Regular Research Grants