Anti-Candidal Activity And Toxicity Studies Of Alginate-Enclosed Chitosan–Calcium Phosphate-Loaded Fe-Bovine Lactoferrin Nanocapsules

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Date
2018-07
Authors
Khoo, Miew Leng
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Publisher
Universiti Sains Malaysia
Abstract
Lactoferrin has been known to have antimicrobial properties. This research was conducted to investigate the anti-candidal activity and toxicity of Alginate/EUDRAGIT® S 100-enclosed chitosan–calcium phosphate-loaded Fe-bLf nanocapsules (NCs) by in vitro and in vivo study. The screening of NCs for antimicrobial activity showed that NCs has good anti-candidal activity. The minimum inhibitory concentration (MIC) value of NCs using broth microdilution method was found to be 500 μg/mL. The growth profile study demonstrated that the NCs at MIC, two times MIC and four times MIC inhibited the growth of Candida albicans USM-K1 (C. albicans). The electron microscopy studies showed that NCs caused the destruction of yeast cells by disruption of cell wall and cell membrane with increase of incubation time until 36 hours. NCs also inhibited germ tube formation of C. albicans. Evaluation of vital staining of epithelial cell and C. albicans coculture demonstrated that NCs enhanced the killing of C. albicans by epithelial cells. In vivo studies showed that NCs caused the reduction of colony forming unit (CFU) of kidney samples of mice induced with yeast infection from (21.3 ± 1.3) × 104 to (4120.0 ± 6.4) × 10 (p < 0.05). This shows that NCs have in vivo anticandidal effect. Histological examination of kidney samples of mice treated with NCs showed better cell structure and coordination compared to untreated mice. Brine shrimp lethality assay showed that the LC50 value of NCs was more than 1 mg/mL which indicated that NCs was not toxic. MTT cytotoxicity assay also revealed that NCs was not toxic against non-cancerous Vero cell line. Genotoxicity studies by comet assay on Vero cells and Allium cepa root assay revealed that the NCs was destitute of significant genotoxic effect under experimental conditions. In conclusion, NCs has effective anticandidal properties and has the potential as a therapeutic agent against candidiasis.
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Keywords
Lactoferrin has been known to have antimicrobial properties , anti-candidal activity and toxicity of Alginate
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