Size- and shape-dependent clinical and mycological efficacy of silver nanoparticles on dandruff

Figures & data

Table 1 Standards for skin irritation study

Skin responses	Score
Erythema and eschar formation
No erythema	0
Very slight erythema (barely perceptible)	1
Well-defined erythema	2
Moderate to severe erythema	3
Severe erythema (beet redness) to slight eschar formation (injuries in depth)	0
Edema formation
No edema	0
Very slight edema (barely perceptible)	1
Slight edema (edges of area well-defined by definite raising)	2
Moderate edema (raised ~1.0 mm)	3
Severe edema (raised >1.0 mm and extending beyond exposure area)	4
Total possible score for irritation	8

Figure 1 TEM micrograph.

Notes: (A) 20 nm spherical-shaped Ag NPs; (B) 50 nm with spherical-shaped Ag NPs; (C) TEM image showing 50 nm of rod-shaped Ag NPs. The insets show the diffraction pattern recorded by aligning the electron beam perpendicular to one of the square faces of an individual nanoparticle. (D) XRD pattern of Ag NR.

Abbreviations: TEM, transmission electron microscopy; NPs, nanoparticles; XRD, X-ray diffraction; NR, nanorod.

Figure 2 Comparison of different antifungal agents and correlation of their concentration with biocidal efficiency.

Notes: (A) Represents the concentration dependent antifungal activity of Ag NPs on M. furfur. (B) Represents the comparative antifungal activity of Ag NPs with standard drugs.

Abbreviations: NPs, nanoparticles; M. furfur, Malassezia furfur.

Table 2 Rat skin irritation test with all nanomaterials and drugs

	Time (h)	Score
		Rat 1 (control)	Rat 2 (formalin)	Rat 3 (20 nm Ag NPs)	Rat 4 (50 nm Ag NPs)	Rat 5 (50 nm Ag NR)
Erythema and scar	6	0	1	0	0	0
	12	0	2	0	0	0
	48	0	3	2	1	0
Edema	6	0	0	0	0	0
	12	0	1	0	0	0
	48	0	2	0	0	0
Primary irritation index (PII)		PII =0/6=0.0	PII =9/6=1.5	PII =2/6=0.33	PII =1/6=0.16	PII =0/6=0.0

Abbreviations: h, hour; NPs, nanoparticles; NR, nanorod.

Table 3 Clinical and mycological evaluation of treatment efficacy of silver nanoparticles in rat models

S no	Group factor	Control	Infected	Treated (itracanzole)	Treated (ketocanzole)	Treated with Ag NPs
						20 nm	50 nm	50 nm
1	Redness	0	3	2	1	1	1	1
2	Ulceration	0	3	2	2	0	0	1
3	Scaling	0	3	2	2	0	1	1
4	Total score	0	9	6	5	1	2	3
5	Efficacy (%)	NA	NA	33.34	44.45	88.89	77.78	66.67

Abbreviations: NPs, nanoparticles; NA, not applicable.

Figure 3 Skin photographs of the dorsal surface of rats.

Notes: (A) Control group I showing no redness, no inflammation; (B) M. furfur diseased group, scales on the skin; (C) and (D) Groups III and IV treated with itracanozole and ketoconazole, respectively, showing skin with decreased scales as compared to Group II with pinkish color; (E) Group V (20 nm spherical Ag NPs) showing complete treatment of M. furfur as compared to Group II; (F) and (G) Groups VI and VII (50 nm spherical- and rod-shaped Ag NPs) also showed normalization of the skin but less as compared to group V.

Abbreviations: M. furfur, Malassezia furfur; NPs, nanoparticles.

Table 4 Results of biochemical activity assays in different groups of studied rats

Groups of animals	CAT (KU/g protein)	SOD (KU/g protein)	GPx (UI/g protein)	MDA (mmol/g tissue)	THP (mmol/g protein)	Total protein (mg/dL)
Group I (control)	74.27±4.64	70.92±11.34	41.22±2.11	18.03±1.97	8.40±1.5	7.91±0.49
Group II (diseased model)	34.98±3.53	17.80±2.60	9.00±1.37	47.47±0.64	2.63±0.20	4.69±1.2
Group III (itracanozole)	49.43±4.65	32.53±3.21	17.02±2.63	31.21±2.81	4.24±1.41	5.53±0.88
Group IV (ketoconazole)	48.28±5.73	34.21±6.11	19.87±3.72	34.72±3.32	4.92±2.1	5.21±1.5
Group V (20 nm spherical Ag NPs)	72.32±3.21	68.87±3.87	38.67±4.11	19.93±4.72	7.87±0.89	7.74±1.56
Group VI (50 nm spherical Ag NPs)	69.54±2.73	64.67±3.63	35.83±3.39	23.21±3.21	7.11±1.1	6.77±3.2
Group VII (50 nm rod Ag NPs)	62.73±3.72	59.32±2.32	32.64±2.79	26.54±5.11	6.93±0.68	6.13±2.22

Abbreviations: CAT, catalase; SOD, superoxide dismutase; GPx, gluthatione peroxidase; MDA, malondidialdehyde; THP, total hydroperoxides; NPs, nanoparticles.

Figure 4 Histopathological microphotographs of different groups of studied rats.

Notes: (A) Control I; (B) M. furfur diseased model group II; antifungal agent-treated groups; (C) III (itracanozole); (D) IV (ketoconazole); (E) V (20 nm spherical-shaped Ag NPs); (F) VI (50 nm spherical), and (G) VII (50 nm rod-shaped Ag NPs).

Abbreviations: M. furfur, Malassezia furfur; NPs, nanoparticles.

Figure S1 Schematic representation of synthesis of Ag nanoparticles and nanorods.

Abbreviation: AOT, aerosol-OT.

Figure S2 UV–vis spectra showing absorption at 406 nm and 425 nm for 20 nm and 50 nm spherical-shaped Ag NPs, respectively, and silver nanorods showed two plasma bands at 531 nm and 432 nm.

Abbreviation: NPs, nanoparticles.

Figure S3 Comparison of ZOI for different antifungal agents in groups of studied rats (A–E).

Notes: (A) Control; (B) aqueous solution of formalin (7%) (irritant); (C) 50 nm rod shaped Ag NPs; (D) 50 nm spherical shaped Ag NPs; (E) 20 nm spherical shaped Ag NPs.

Abbreviation: ZOI, zones of inhibition.

Figure S4 Skin photographs showing irritation studies.

Notes: (A) Comparative ZOI study of Ag Nps with itraconazole; (B) ZOI study of 20 nm spherical Ag Nps; (C) ZOI study of 50 nm spherical Ag Nps; (D) ZOI study of 50 nm rod Ag Nps.

Abbreviations: NPs, nanoparticles; ZOI, zones of inhibition.

Figure S5 The graph showing scratching behavior assessment of rat for scratching tendency and comparison with control untreated rats.

Abbreviations: NP, nanoparticle; NR, nanorod.