Design and synthesis of mucoadhesive nanogel containing farnesol: investigation of the effect on HWP1, SAP6 and Rim101 genes expression of Candida albicans in vitro

Figures & data

Table 1. Primer sequences of genes in quantitative real-time PCR assay.

Gene name	Sequence	Amplicon size (bp)	Accession no.
ACT1	5′-GCGGTAGAGAGACTTGACCAACC-3′ 5′-GACAATTTCTCTTTCAGCACTAGTAGTG-3′	200	XM019475182.1
SAP6	5′-TTACGCAAAAGGTAACTTGTATCAAGA-3′ 5′-CCTTTATGAGCACTAGTTAGACCAAAC-3′	101	XM714012.1
HWP1	5′-ACTGCTCAACTTATTGCTATCGC-3′ 5′-ACCGTCTACCTGTGGGACAG-3′	69	XM704869.2
RIM101	5′-ATTACAACATTCATCCCGATAA-3′ 5′-CCAACATGGTCGTCACAC-3′	81	XM709954.1

Figure 1 Scanning electron microscopic image: (A) chitosan nanogel contain farnesol and (B) alginate nanogel contain farnesol.

Figure 2 Fourier-transform infrared spectra: chitosan nanogel contain farnesol.

Figure 3 Fourier-transform infrared spectra: alginate nanogel contain farnesol.

Figure 4 (A) XRD spectra of chitosan nanogel contain farnesol and (B) XRD spectra of alginate nanogel contain farnesol.

Figure 5 Farnesol release kinetics. Farnesol-loaded chitosan and alginate nanogels at pH 7.

TABLE 2. Minimum inhibitory concentration 90 (MIC90) of farnesol on C. albicans (ATCC 10231) growth rate.

Concentration of farnesol (µM)	Optical density (OD) C. albicans (ATCC 10231)
5	0.053 ± 0.006
10	0.051 ± 0.005
20	0.045 ± 0.001
50	0.037 ± 0.011
75	0.035 ± 0.015
150	0.028 ± 0.011
300	0.003 ± 0.002
Control	0.23 ± 0.00

Minimal inhibitory concentration 90 (MIC90) defined as the lowest concentration of the farnesol that led to 90% reduction absorbance compared to control, without farnesol treatment. MIC values are in (µM). This experiment is performed in triplicate; mean and standard deviation (m±SD) was measured.

TABLE 3. Inhibitory zone of farnesol and nanogels contains farnesol in pure plat assay.

Treatment	Inhibitory zone (mm)^a C. albicans (ATCC 10231)
Farnesol (300 µM)	6.65 ± 0.35
Chitosan + farnesol	8.5 ± 0.5
Alginate + farnesol	ND^b

^aInhibitory zone performed in triplicate, mean and standard deviation (m±SD) was measured. ^bND: not detected.

Figure 6 Cytotoxicity effect of farnesol and farnesol-loaded nanogels on SW480 cells by MTT assay during 12, 24, 48 and 72 h treatment. Results indicated no differences significant cell viability per cent between each of treatment compared to control without treatment. *The difference between groups is statistically significant.

Figure 7 Comparison of gene expression pattern of HWP1, SAP6 and Rim101 of C. albicans compared with ACT1 gene (housekeeping gene) before and after treatment with farnesol and nanogels containing farnesol. Changes in gene expression of HWP1, SAP6 and Rim101 are presented, compared to treatment with farnesol and nanogel. *The difference between groups is statistically significant.