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

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Abstract

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.

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Funding

The work was supported by financial support from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (grant 001), the Fundação de Amparo à Pesquisa do Estado de São Paulo (grants 2019/11192–4 and 2019/16473–1) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (grant 303391/2019–7).

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Authors and Affiliations

  1. Department of Morphology and Pediatric Dentistry, School of Dentistry, São Paulo State University (UNESP), Araraquara, Brazil

    Caroline Anselmi & Josimeri Hebling

  2. Department of Dental Materials and Prosthodontics, School of Dentistry, São Paulo State University (UNESP), Araraquara, Brazil

    Igor Paulino Mendes Soares, Maria Luísa Leite & Fernanda Ali Kitagawa

  3. Department of Physiology and Pathology, School of Dentistry, São Paulo State University (UNESP), Araraquara, Brazil

    Carlos Alberto de Souza Costa

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  1. Caroline Anselmi
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  2. Igor Paulino Mendes Soares
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  3. Maria Luísa Leite
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  4. Fernanda Ali Kitagawa
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  5. Carlos Alberto de Souza Costa
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  6. Josimeri Hebling
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Caroline Ansemi, Igor Paulino Mendes Soares, Maria Luísa Leite, and Fernanda Ali Kitagawa. Founding acquisition, resources, and supervision were performed by Carlos Alberto de Souza Costa and Josimeri Hebling. The first draft of the manuscript was written by Caroline Anselmi and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Josimeri Hebling.

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The study approval was obtained from the ethics committee of School of Dentistry of Araraquara (CAAE: 18119319.9.0000.5416).

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Anselmi, C., Mendes Soares, I.P., Leite, M.L. et al. Cytocompatibility and bioactivity of calcium hydroxide-containing nanofiber scaffolds loaded with fibronectin for dentin tissue engineering. Clin Oral Invest 26, 4031–4047 (2022). https://doi.org/10.1007/s00784-022-04372-6

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