In vitro and in vivo evaluation of a biomimetic scaffold embedding silver nanoparticles for improved treatment of oral lesions

https://doi.org/10.1016/j.msec.2021.112015Get rights and content

Highlights

  • The distribution of silver nanoparticles was visualised by autometallography.

  • The hybrid biomaterial was in vitro cytocompatible and non-genotoxic.

  • The hybrid biomaterial had angiogenic and regenerative properties in CAM assay.

  • Anti-inflammatory activity was enhanced by embedded silver nanoparticles.

  • The hybrid biomaterial inhibited the growth of bacteria involved in periodontitis.

Abstract

Background

New materials are currently designed for efficient treatment of oral tissue lesions by guided tissue regeneration. The aim of this study was to develop a multifunctional 3D hybrid biomaterial consisting of extracellular matrix components, collagen, chondroitin 4-sulfate and fibronectin, functionalised with silver nanoparticles, intended to improve periodontitis treatment protocols.

Methods

Structural observations were performed by autometallography, scanning and transmission electron microscopy. In vitro tests of 3D constructs of embedded gingival fibroblasts within hybrid biomaterial were performed by MTS and Live/Dead assays. Genotoxicity was assessed by comet assay. In vivo experiments using chick embryo chorioallantoic membrane (CAM) assay analysed the degradation and nanoparticles release, but also angiogenesis, new tissue formation in 3D constructs and the regenerative potential of the hybrid material. Biological activity was investigated in experimental models of inflamed THP-1 macrophages and oral specific bacterial cultures.

Results

Light micrographs showed distribution of silver nanoparticles on collagen fibrils. Scanning electron micrographs revealed a microstructure with interconnected pores, which favoured cell adhesion and infiltration. Cell viability and proliferation were significantly higher within the 3D hybrid biomaterial than in 2D culture conditions, while absence of the hybrid material's genotoxic effect was found. In vivo experiments showed that the hybrid material was colonised by cells and blood vessels, initiating synthesis of new extracellular matrix. Besides the known effect of chondroitin sulfate, incorporated silver nanoparticles increased the anti-inflammatory activity of the hybrid biomaterial. The silver nanoparticles maintained their antibacterial activity even after embedding in the polymeric scaffold and inhibited the growth of F. nucleatum and P. gingivalis.

Conclusion

The novel biomimetic scaffold functionalised with silver nanoparticles presented regenerative, anti-inflammatory and antimicrobial potential for oral cavity lesions repair.

Keywords

Collagen
Fibronectin
Nanosilver
Chorioallantoic membrane
Cytokine
Oral microbiome

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