Cytocompatibility and bioactivity of calcium hydroxide-containing nanofiber scaffolds loaded with fibronectin for dentin tissue engineering

• Published in: Clinical oral investigations Vol. 26; no. 5; pp. 4031 - 4047
• Main Authors: Anselmi, Caroline, Mendes Soares, Igor Paulino, Leite, Maria Luísa, Kitagawa, Fernanda Ali, de Souza Costa, Carlos Alberto, Hebling, Josimeri
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-05-2022; Springer Nature B.V
• Subjects: Actin; Adhesives; Alkaline phosphatase; Biocompatibility; Biological activity; Calcium hydroxide; Calcium Hydroxide - pharmacology; Cell Differentiation; Cell viability; Cells, Cultured; Collagen (type I); Dental Pulp; Dentin; Dentistry; Fibronectin; Fibronectins - pharmacology; Gene expression; Humans; Medicine; Mineralization; Nanofibers; Original Article; Polycaprolactone; Tissue Engineering; Tissue Scaffolds; Cell survival; Calcium; Nanofibers; Gene expression; Fibronectin; Dental pulp
• ISSN: 1436-3771; 1432-6981; 1436-3771
• DOI: 10.1007/s00784-022-04372-6

Objectives
The aim of this study was to characterize polycaprolactone-based nanofiber scaffolds (PCL) incorporated with calcium hydroxide (CH) and evaluate their bioactivity on human dental pulp cells (HDPCs) when loaded with fibronectin (FN).
Materials and methods
CH (0.1%; 0.2%; 0.4% w/v; or 0%) was incorporated into PCL (10% w/v) scaffolds prepared by electrospinning. Morphology and composition were characterized using SEM/EDS. HDPCs were seeded on the scaffolds and evaluated for viability (alamarBlue; Live/Dead), and adhesion/spreading (F-actin). Next, scaffolds containing 0.4% CH were loaded with FN (20 µg/mL). HDPCs were evaluated for viability, adhesion/spreading, migration (Trans-well), gene expression (RT-qPCR), alkaline phosphatase activity (ALP), and mineralization nodules (Alizarin Red). Data were submitted to ANOVA and post-hoc tests (α = 5%).
Results
Nanofibers with larger diameter were seen as CH concentration increased, while there was no effect on interfibrillar spaces. An increase in cell viability was seen for 0.4% CH, in all periods. Incorporation of CH and FN into the scaffolds increased cellular migration, spread, and viability, all intensified when CH and FN were combined. ALPL and DSPP expression, and ALP activity were not affected by CH and FN. COL1A1 was downregulated in all groups, while DMP1 was upregulated in the presence of CH, with no differences for the groups loaded with FN. CH increased the formation of mineralized matrix, which was not influenced by FN.
Conclusions
In conclusion, the incorporation of CH enhanced the odontogenic potential of HDPCs, irrespective of the presence of FN. The PCL + 0.4% CH formulation may be a useful strategy for use in dentin tissue engineering.
Clinical relevance
A change in the form of presentation of calcium hydroxide-based materials used for direct pulp capping can increase biocompatibility and prolong the vitality of dental pulp.