Evaluating the antifungal effectiveness, leaching characteristics, flexural strength, and impact strength of polymethyl methacrylate added with small-scale silver nanoparticles – An in vitro study
Authors: Palaskar Jayant N. , Hindocha Amit Dhansukhlal , Mishra Aditi , Gandagule Ranjeet , Korde Sayali 
Published: April 2024

Aim:
(1) To assess the release of stable silver nanoparticles (AgNPs) of small scale dimension added to heat polymerized polymethyl methacrylate (PMMA) in 6 months. (2) Assessing the influence of incorporating minimal concentrations of stable AgNPs with nanoscale dimensions into heat polymerized PMMA over a 6 month period on its antifungal efficacy (AF), flexural strength (FS), and impact strength (IS).

Settings and Design:
Incorporating nanoparticles with a very small scale may have minimal impact on mechanical properties due to their diminutive size. However, the influence of these small scaled nanoparticles on antimicrobial efficacy and potential escalation in toxicity to host cells through leaching remains unexplored. AgNPs were prepared using an Ultrasonic Probe sonicator and the addition of ammonia to obtain stabilized AgNPs (< 0.01 nm) of small scale dimension. The characterization of these AgNPs involved ultraviolet visible spectroscopy, X ray diffraction, Zetasizer, and transmission electron microscopy with energy dispersive spectroscopy (TEM).

Materials and Methods:
The prepared AgNPs were then added in various percentages by weight (0%–0.5%) to fabricate 252 modified PMMA samples of sizes 10 mm × 3 mm (AF, n = 108), 65 mm × 10 mm × 3 mm (FS, n = 72), and 65 mm × 10 mm × 2.5 mm (IS, n = 72) as per ADA specification no. 12. These samples underwent testing for leaching out of AgNPs and efficacy against Candida albicans for 6 months. The effect on FS and IS was evaluated using the three point bending test and Charpy’s Impact Tester, respectively.

Statistical Analysis Used:
Intergroup comparison of CFU between various concentrations of AgNP was done using the Kruskal–Wallis ANOVA test succeeded by Mann–Whitney test for pair wise comparisons. Difference in CFU of various concentrations over 6 months was seen using one way ANOVA test. Intergroup comparison of FS and IS was performed using a one way ANOVA test, followed by a post hoc Tukey’s test for pair wise comparisons.

Results:
Repeated tests showed no leaching out of AgNPs from the denture base resin into the storage medium. All concentrations of AgNPs incorporated in resin showed inhibition of Candida growth. Intergroup comparison of FS and IS revealed highly statistically significant differences (F = 15.076, P < 0.01 and F = 28.266, P < 0.01) between the groups showing a reduction in strength.

Conclusion:
The AgNPs of small scale dimension incorporated into the denture base resin imparted a strong antifungal effectiveness against C. albicans, which did not decline during the study period and did not cause any release of nanoparticles. 0.5% showed the best antifungal efficacy. This may prove to be a viable and highly effective treatment for the prevention of Candida associated denture stomatitis. However, the inclusion of these particles resulted in a decrease in both FS and IS, and this reduction was directly proportional to the percentage of added AgNPs, with 0.5% demonstrating the least IS and FS.