Anticandidal Activity of Capsaicin and Its Effect on Ergosterol Biosynthesis and Membrane Integrity of Candida albicans

Jawad M Behbehani 1, Mohammad Irshad 2 3, Sheikh Shreaz 2, Maribasappa Karched 2

Affiliations

06 May 2023

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doi: 10.3390/ijms24021046

Abstract

Oral candidiasis is an infection of the oral cavity commonly caused by Candida albicans. Endodontic treatment failure has also been found to be persistent from C. albicans in the root canal system. Despite the availability of antifungal drugs, the management of Candida oral infection is difficult as it exhibits resistance to a different class of antifungal drugs. Therefore, it is necessary to discover new antifungal compounds to cure fungal infections. This study aimed to examine the antifungal susceptibility of Capsaicin, an active compound of chili pepper. The susceptibility of Capsaicin and Fluconazole was tested against the Candida species by the CLSI (M27-A3) method. The effect of Capsaicin on the fungal cell wall was examined by the ergosterol inhibitory assay and observed by the scanning electron micrograph. The MIC range of Capsaicin against Candida isolates from oral (n = 30), endodontic (n = 8), and ATCC strains (n = 2) was 12.5−50 µg/mL. The MIC range of Fluconazole (128- 4 µg/mL) significantly decreased (2- to 4-fold) after the combination with Capsaicin (MIC/4) (p < 0.05). Capsaicin (at MIC) significantly reduced the mature biofilm of C. albicans by 70 to 89% (p < 0.01). The ergosterol content of the cell wall decreased significantly with the increase in the Capsaicin dose (p < 0.01). Capsaicin showed high sensitivity against the hyphae formation and demonstrated a more than 71% reduction in mature biofilm. A fluorescence microscopy revealed the membrane disruption of Capsaicin-treated C. albicans cells, whereas a micrograph of electron microscopy showed the distorted cells’ shape, ruptured cell walls, and shrinkage of cells after the release of intracellular content. The results conclude that Capsaicin had a potential antifungal activity that inhibits the ergosterol biosynthesis in the cell wall, and therefore, the cells’ structure and integrity were disrupted. More importantly, Capsaicin synergistically enhanced the Fluconazole antifungal activity, and the synergistic effect might be helpful in the prevention of Fluconazole resistance development and reduced drug-dosing.

Keywords: Capsaicin; anticandidal; chili pepper; ergosterol.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1 Effect of Capsaicin on the mature biofilm of C. albicans. The standard deviations (SD) of each sample are shown in the graph. The mean differences between the control and test molecules at the MIC level were statistically significant (p-level, 0.01). MIC/2—half of the MIC concentration; Endo—endodontic isolates.

Figure 1 Effect of Capsaicin on the mature biofilm of C. albicans. The standard deviations (SD) of each sample are shown in the graph. The mean differences between the control and test molecules at the MIC level were statistically significant (p-level, 0.01). MIC/2—half of the MIC concentration; Endo—endodontic isolates.

Figure 2 Effect of Capsaicin (MIC) on the hyphae growth of C. albicans (Endo-902). The live cells photograph was recorded by the cell observer microscope (20×). The photograph of untreated cells (1a–1d) and treated cells (2a–2d) were recorded at three-hour time points, respectively.

Figure 2 Effect of Capsaicin (MIC) on the hyphae growth of C. albicans (Endo-902). The live cells photograph was recorded by the cell observer microscope (20×). The photograph of untreated cells (1a–1d) and treated cells (2a–2d) were recorded at three-hour time points, respectively.

Figure 3 Spectrophotometric sterol profiles of C. albicans (Endo-904, Endo-906, and Endo-908). Sterols were extracted from the cells and spectral profiles between 230 and 300 nm were determined.

Figure 3 Spectrophotometric sterol profiles of C. albicans (Endo-904, Endo-906, and Endo-908). Sterols were extracted from the cells and spectral profiles between 230 and 300 nm were determined.

Figure 4 Confocal scanning laser microscopy (CSLM) images of endodontic C. albicans cells (Endo) treated with Capsaicin (MIC) (right panel) and untreated control cells (left panel). To observe membrane damage, cells were stained with propidium iodide (red signals).

Figure 4 Confocal scanning laser microscopy (CSLM) images of endodontic C. albicans cells (Endo) treated with Capsaicin (MIC) (right panel) and untreated control cells (left panel). To observe membrane damage, cells were stained with propidium iodide (red signals).

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