Poly(Ɛ-caprolactone) (PCL) is a biocompatible polymer with a high potential to be used in tissue engineering especially in tight tissues. In the current study, cold atmospheric plasma (CAP) is used as a promising method for immobilization of gelatin as a functional biomacromolecule on PCL nanofibrous substrates. The CAP surface modification leads to oxidation of chemical groups existing on the PCL surface without doing any damage to the bulk properties of biomaterials for gelatin biomacromolecule grafting. The water contact angle (WCA) of the CAP-treated surface and gelatin-grafted PCL using CAP indicates an effective increment in the hydrophilicity of the PCL surface. Also to achieve the highest levels of gelatin grafting on the PCL surface, two different grafting methods and gelatin concentration diversity are utilized in the grafting process. The immobilization of gelatin biomacromolecules onto the CAP surface-modified PCL nanofibers is investigated using scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR). The gelatin-modified PCL substrates revealed uniform nanofibrous morphology with increased average fiber diameter. The results of FTIR spectra, including hydroxyl groups, NH groups, and amide II of gelatin-grafting peaks, confirm the gelatin immobilization on the surface of nanofibers. The metabolic activity of cultured mesenchymal stem cells (MSCs) on the surface-modified scaffolds is evaluated using MTT analysis (
P ≤
0.05). The results of metabolic activity and also SEM and DAPI staining observations indicate proper attachment on the surface and viability for MSCs on the surface-immobilized nanofibrous scaffolds. Therefore, CAP treatment would be an effective method for biomacromolecule immobilization on nanofibers towards the enhancement of cell behavior.
Vulvovaginal candidiasis (VVC) is a common mucosal infection, mainly caused by Candida albicans. The use of common antifungal drugs in treatment of VVC is limited due to emergence of resistant fungal strains and severe side effects. Cold atmospheric plasma (CAP) as a novel therapeutic approach is proven to display strong antifungal activity against C. albicans. In the present study, the effects of CAP treatment on virulence and pathogenicity of C. albicans in a murine model was investigated. Candida albicans was treated with CAP at different time exposures. Fungal cell morphology and the expression profile of CaMCA1 gene in CAP-treated fungus was evaluated using electron microscopy and quantitative RT-PCR. Moreover, the mice model of VVC was developed using CAP-treated and non-treated C. albicans and characterized in terms of vaginal fungal burden, the rate of hyphae formation in the vaginal tissue and fluid and the inflammation degree of mice vaginal tissue. Significant reduction in CaMCA1 expression and remarkable mitochondrial degradation were observed in CAP-treated C. albicans cells. The lowest fungal burden, reduced hyphae formation, poor adherence of yeast cells to vaginal epithelium, and the low degree of inflammation were observed in mice infected with CAP treated C. albicans. Suppression of CaMCA1 gene and mitochondrial degradation in CAP-treated C. albicans yeast cells may diminish yeast to hyphae transition and reduce fungal pathogenicity in murine model of VVC. CAP treatment can be considered as a novel and efficient therapeutic strategy against C. albicans and related Candida infections in practice.
Lay Abstract
CAP was successfully used to inhibit fungal growth and CaMCA1 gene expression in C. albicans. It caused morphological alterations in membranous structures of the yeast cells and finally led to the cell death. CAP meaningfully reduced C. albicans virulence and pathogenicity in a murine model of VVC.
Trichophyton rubrum, a major human pathogenic dermatophyte, is responsible for the most recurrent dermatophytoses as globally important superficial fungal infections. Typical chemotherapy is used to handle such infections; however, emerging drug resistance and side effects necessitate the new remedial method development. Cold atmospheric plasma (CAP) is an emerging technology, consisted of neutral and charged particles and photons newly developed as a potent and safe antimicrobial technique to combat drug-resistant microbial pathogens. In the present study, the vast effects of CAP irradiation containing oxygen (2%) and helium (98%) on T. rubrum growth and pathogenicity were explored. After exposure of T. rubrum to CAP jet for 90, 120, 150, 180, and 210 s in 96-well microtiter plates, cell morphology and viability, ergosterol content of fungal hyphae, HSP90 gene expression, and the pattern of drug susceptibility were studied by using electron microscopy, RT-qPCR, spectrophotometry, disk diffusion and CLSI microbroth dilution methods. CAP irradiation significantly inhibited the fungal growth by 25.83 to 89.10%, reduced fungal cell viability by 11.68 to 87.71%, disrupted cellular membranous organelles and structures of the fungal hyphae, and suppressed efficiently the expression of HSP90 gene by 2 folds in 210 s exposure. Taken together, our results demonstrated that CAP is an efficient tool with potential in-vivo therapeutic applications against chronic dermatophytosis caused by T. rubrum due to its effectiveness, harmless, and ease of access.
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