METHODS: NIH 3T3 mouse fibroblasts were cultured in Dulbecco's Modified Eagle's Medium and incubated for 3 days. The cells (3×104) were seeded on the pulpal side of dentine discs and the occlusal side of the discs were treated with different cavity disinfectants: Group 1: de-ionized water (control); Group 2: 2% chlorhexidine (CHX); Group 3: 2% QAS; Group 4: 5% QAS, and Group 5: 10% QAS. Cell morphology of NIH 3T3 cells was examined using scanning electron microscopy (SEM) and cell viability was assessed using Trypan blue assay. The eluates were collected and applied on cells seeded in 24-well plates. The total protein production, alkaline phosphatase activity and deposition of mineralized nodules were evaluated after 7 and 14 days. Immunofluorescence staining was performed on the samples with primary antibodies of CD68+, CD80+, and CD163+ assessing the macrophage M1/M2 phenotypes. The macrophages were imaged using a confocal scanning light microscope with an excitation wavelength of 488nm.
RESULTS: No significant difference in cell viability (p<0.0001), total protein production (p<0.01) and mineralized nodule production (p<0.05) was found between 2% QAS and the control, which was significantly higher than 2% CHX, 5% and 10% QAS after 14 days. Alkaline phosphatase production of 2% QAS was significantly lower than the control (p<0.001), but higher than 2% CHX at 14 days. The M1/M2 macrophage ratio was also significantly lower in the 2% and 10% QAS groups (p<0.05) compared to the control and 2% CHX groups.
SIGNIFICANCE: The 2% QAS cavity disinfectant does not have cytotoxic effects on 3T3 NIH mouse fibroblast cells and the predominance of the anti-inflammatory phenotype after its application may stimulate healing and tissue repair.
RESULTS: Minimal inhibitory concentration was determined at 0.625% of the concentration of ACV against S. mutans and E. faecalis and 1.25% of the concentration of ACV against L. casei with two-fold serial dilutions. A concentration of 5 × 10-1% with 10-fold serial dilutions was found to be the MIC value for all three bacteria. No significant differences were found when compared with the positive control (NaOCl) (p = 0.182, p = 0.171, and p = 0.234), respectively, for two-fold serial dilutions and (p = 1.000, p = 0.658, and p = 0.110), respectively for 10-fold serial dilutions. MBC was observed to be 5% ACV for both E. faecalis and S. mutans. However, positive microbial growth was observed on the agar plate when cultured with L. casei. An independent sample t-test showed no significant differences (p > 0.05) in the antimicrobial activities between 5% ACV and 5% pure AA. TEM revealed cell wall and cytoplasmic membrane disruptions on all three bacteria at MIC value.
CONCLUSION: Apple cider vinegar has antimicrobial activities against Enterococcus faecalis, Streptococcus mutans, and Lactobacillus casei at their respective MIC values.
CLINICAL SIGNIFICANCE: Apple cider vinegar can be an alternative antimicrobial dental pulp disinfectant to sodium hypochlorite. Apple cider vinegar can be used safely, especially in children's dental pulp therapy and deep caries management, when adequate tooth isolation is not readily achievable. Thus, adverse reactions commonly associated with other frequently used chemical disinfectants can be avoided.