Abstract
The aims of this study were to synthesize highly positively charged chitosan nanoparticles (Ch-Np) using the electrospraying technique, and to test their antimicrobial activity against endodontic pathogens, and cytotoxicity against fibroblast cells. Ch-Np were synthesized from low molecular weight chitosan (LMW-Ch) using the electrospraying technique, and characterized. The antimicrobial activity was evaluated against Streptococcus mutans, Enterococcus faecalis, and Candida albicans in their planktonic state using a Time-Kill Test performed by using broth micro-dilution technique, and against biofilm biomass using a microtiter plate biofilm assay. The cytotoxicity was evaluated using Balb/c 3T3 fibroblast cells with the standard MTT assay. Electrospraying of LMW-Ch produced Ch-Np with an average size of 200 nm, and a surface charge of 51.7 mV. Ch-Np completely eradicated S. mutans and E. faecalis in the planktonic state and showed fungistatic activity against C. albicans. Furthermore, it significantly reduced the biofilm biomass for all the tested microbial species [S. mutans (p = 0.006), E. faecalis (p < 0.0001), and C. albicans (p = 0.004)]. When tested for cytotoxicity using 3T3 cells, Ch-Np showed no cytotoxicity. In conclusion, the highly positively charged, colloidal dispersion of Ch-Np are effective as a biocompatible endodontic antimicrobial agent.
Highlights
The aims of this study were to synthesize highly positively charged chitosan nanoparticles (Ch-Np) using the electrospraying technique, and to test their antimicrobial activity against endodontic pathogens, and cytotoxicity against fibroblast cells
Electrospraying of low molecular weight chitosan (LMW-Ch) dispersed in Trifluoroacetic acid (TFA) at room temperature with a flow rate of 0.4 mL/h and 25 kV potential resulted in the deposition of a layer of nanoparticles upon the collectors that was placed 12 cm away from the tip of the needle and perpendicular to it, which were collected and further characterized using the following characterization methods
Following dispersion of LMW-Ch in TFA and electrospraying, forming Ch-Np, the particle size was decreased by an order of magnitude to an average size of 419 nm when the dried particles were suspended in water for the particle size measurement (Fig. 1a, b)
Summary
The aims of this study were to synthesize highly positively charged chitosan nanoparticles (Ch-Np) using the electrospraying technique, and to test their antimicrobial activity against endodontic pathogens, and cytotoxicity against fibroblast cells. Ch-Np completely eradicated S. mutans and E. faecalis in the planktonic state and showed fungistatic activity against C. albicans It significantly reduced the biofilm biomass for all the tested microbial species [S. mutans (p = 0.006), E. faecalis (p < 0.0001), and C. albicans (p = 0.004)]. The main cause of endodontic and periapical diseases is microbial contamination of the root canal system. One of the main shortcomings of the current root canal treatment modalities is the inability to completely eradicate persistent pathogens. Some microbial species such as Enterococcus faecalis were resistant to commonly used antimicrobial root canal irrigants and medicaments[12]. The limitation of using the physical cross-linking method to produce Ch-Np is the tendency of the Ch-Np to aggregate upon the addition of another polymer to produce the nanoparticles[22]
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