Biological properties of modified bioactive glass on dental pulp cells

doi:10.1016/j.jdent.2019.01.017

Journal of Dentistry

Volume 83, April 2019, Pages 18-26

https://doi.org/10.1016/j.jdent.2019.01.017 Get rights and content

Highlights

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Fabricating a new material combining chitosan and silver-contained bioactive glass.
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CS/Ag-BG possesses biocompatibility and induces differentiation of dental pulp cells.
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CS/Ag-BG suppresses inflammation through inactivation of NF-κB pathway.
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Potentially applied in pulp-capping of irreversible pulpitis.

Abstract

Dental caries is a bacteria-caused condition classified among the most common chronic diseases worldwide. Treatment of dental caries implies the use of materials having regenerative and anti-bacterial properties, and controlling inflammation is critical for successful endodontic regeneration.

Objectives

The aim of this study was to fabricate and characterize a novel composite incorporating sol-gel derived silver-doped bioactive glass (BG) in a chitosan (CS) hydrogel at a 1:1 wt ratio(Ag-BG/CS).

Methods

The effect of Ag-BG/CS on dental pulp cells (DPCs) proliferation was analyzed by CCK-8 assay, whereas the adhesion of DPCs was evaluated by confocal microscopy. The physical morphology of Ag-BG/CS was analyzed by scanning electron microscope. The anti-inflammatory effect of Ag-BG/CS was investigated by quantitative polymerase chain reaction (qPCR). Moreover, the effect of Ag-BG/CS on odontogenic differentiation of DPCs was studied by immunochemical staining, tissue-nonspecific alkaline phosphatase staining, qPCR, and western blot analyses. The antibacterial activity against dental caries key pathogenic bacteria was also evaluated.

Results

The results of this study showed that Ag-BG/CS did not affect the proliferation of DPCs, it down-regulated the inflammatory-associated markers (IL-1β, IL-6, IL-8, TNF-α) of DPCs treated with Escherichia coli lipopolysaccharide (LPS) by inhibiting NF-κB pathway, and enhanced the in vitro odontogenic differentiation potential of DPCs. Furthermore, Ag-BG/CS strongly inhibited Streptococcus mutans and Lactobacillus casei growth.

Conclusions

This novel biomaterial possessed antibacterial and anti-inflammatory activity, also enhanced the odontogenic differentiation potential of LPS-induced inflammatory-reacted dental pulp cells. The material introduced in this study may thus represent a suitable dental pulp-capping material for future clinical applications.

Introduction

Dental pulp vitality is essential for root development in immature permanent teeth and reparative dentin formation to respond to external cues. Vital pulp therapy (VPT) is an effective approach to protect the injured pulp, in order to reduce inflammation and promote formation of reparative hard tissue that shields the pulp tissue from injurious agents. An ideal pulp-capping material should stimulate tertiary reparative dentinogenesis and possess biocompatibility, bactericidal, and anti-inflammatory capabilities, among the others [1]. Besides calcium hydroxide and mineral trioxide aggregate (MTA), which are the most common pulp-capping materials, a variety of materials, including bioactive ones, have been investigated in recent studies (Appendix. Table 1). However, no ideal dental pulp capping material for the repair of inflamed pulp has been identified to date.

Following the synthesis of 45S5 Bioglass® [2] by Hench and coworkers, many in vitro and in vivo studies carried out in the recent decades confirmed bioglass as a bioactive material capable of bonding to mineralized and soft tissue [[2], [3], [4], [5]]. Silver is a common element used in medical materials together with bioglass, due to its established antibacterial activity [6,7]. Ag-doped bioactive glass (Ag-BG) was fabricated and confirmed strong bonding to dentin [8], and demonstrated the odontogenic properties in vivo [9]. Chitosan (CS)-based thermogelling solution, which undergoes sol-gel transition upon temperature increase, possesses multiple intrinsic properties such as biocompatibility and bacteriostatic effects [10]. It has been extensively investigated in the past decade [11] and offered more predictable hydrogel properties. Here, CS was added to Ag-BG materials to enhance their operability as dental pulp capping agents. The hypothesis of the study was that this novel biomaterial possessed antibacterial and anti-inflammatory activity, also enhanced the odontogenic differentiation potential of inflamed dental pulp cells. The purpose of this study is to evaluate the biological properties of a hybrid system produced by incorporating Ag-BG into injectable chitosan hydrogel(Ag-BG/CS).

Section snippets

Synthesis of Ag-BG/CS gel

Ag-doped bioactive glass micro-size particles were provided by Professor Papagerakis from University of Saskatchewan, they were fabricated according to a previously reported procedure [12]. Briefly, the fabrication protocol was on the basis of the sol-gel process by mixing the solution stage of the 58S sol-gel bioactive glass (in the system SiO₂ 58-CaO 33-P₂O₅ 9 wt.%), with the respective solution stage of the sol-gel glass in the system SiO₂ 60-CaO 6-P₂O₅ 3-Al₂O₃ 14-Na₂O 5-K₂O 5- Ag₂O 7 wt.%.

Statistical analysis

The statistical significance of differences within the CCK-8 assay and quantitative PCR results were analyzed by one-way ANOVA (Dunnett test) and Student t-test using the PRISM 6 software, at significance levels of p < 0.05*, p < 0.01**, and p < 0.001***.

Effects of Ag-BG/CS on the proliferation of DPCs

The CCK-8 assay was used to investigate the influence of Ag-BG/CS on DPCs proliferation. After treatment with Ag-BG/CS extract liquid, DPCs showed stable growth (Fig. 1B), the proliferation rate was slightly lower compared to control group (Fig. 1B). At day3, 5, 7, there was significantly difference in the numbers of viable cells between two groups (p < 0.01) (Fig. 1A).

Effects of Ag-BG/CS on the adhesion of DPCs

The result of scanning electron microscope(SEM) analysis showed that Ag-BG/CS possessed porous characteristics, with pore

Discussion

Dental pulp capping is a common clinical approach aiming to preserve dental pulp tissue and to prevent invasive endodontic procedures [13] that may result in tooth necrosis and tooth loss. Innovative bioactive/biomimetic ion-releasing materials are required for preserving the vitality of dental pulp tissue in immature permanent teeth and primary teeth, by blocking the progress of inflammation and inducing the formation of reparative tissue. DPCs is a mixed population of mesenchymal cells

Conclusion

In the present work, an anti-inflammatory, bactericidal, and remineralizing material, consisted of Ag-doped bioactive glass particles incorporated in a chitosan gel, has been proven to be able to suppress the expression of inflammatory cytokines, promote the proliferation and differentiation of iDPCs, and inhibit S. mutans and L. Casei bacteria growth. The injectable Ag-BG/CS exhibits various bioactive properties and thus represents a promising vital pulp conservation material to improve

Acknowledgements

Dr. Zhu would like to give her acknowledgment to Dr. YongXiang Xu, for his help of fabricating the Ag-BG/CS material. This study has been funded by a grant from the National Natural Science Youth Foundation of China (No. 81500837). The authors deny any conflicts of interest.

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Biological properties of modified bioactive glass on dental pulp cells

Highlights

Abstract

Objectives

Methods

Results

Conclusions

Introduction

Section snippets

Synthesis of Ag-BG/CS gel

Statistical analysis

Effects of Ag-BG/CS on the proliferation of DPCs

Effects of Ag-BG/CS on the adhesion of DPCs

Discussion

Conclusion

Acknowledgements

Acta Biomater.

Mater. Sci. Eng. C

Eur. J. Pharm. Biopharm.

J. Eur. Ceram. Soc.

Int. J. Biol. Macromol.

Drug Discov. Today

Acta Biomater.

Acta Biomater.

Acta Biomater.

Recent advances in pulp capping materials: an overview

J. Clin. Diagn. Res.

Bioglass: a short history and bibliography

J. Aust. Ceram. Soc.

Mineralization of dentin induced by treatment with bioactive glass S53P4 in vitro

Acta Odontol. Scand.

Ectopic bone formation in and soft-tissue response to P(CL/DLLA)/bioactive glass composite scaffolds

Clin. Oral Implants Res.

Control of Ag nanoparticle distribution influencing bioactive and antibacterial properties of Ag-doped mesoporous bioactive glass particles prepared by spray pyrolysis

J. Biomed. Mater. Res. B Appl. Biomater.