Magnesium ascorbyl phosphate vesicular carriers for topical delivery; preparation, in-vitro and ex-vivo evaluation, factorial optimization and clinical assessment in melasma patients

Figures & data

Table 1. Experimental runs, independent and dependent variables of the 3² full factorials experimental design of Magnesium ascorbyl phosphate-loaded ethosomes.

Runs*	Factors (Independent variables)				Responses (Dependent variables)
	Ethanol % (v/v) (mL/100 mL)		PC % (w/v) (g/100 mL)		Y1:E.E (%)	Y2: Particle size (nm)	Y3:PDI	Y4:ZP (mV)	Y5: % permeated (8 h)
FE1	(−1)	30	(−1)	1	51.45 ± 2.34	148.62 ± 15.27	0.496 ± 0.071	−32.24 ± 5.24	38.12 ± 2.61
FE2	(−1)	30	(0)	2	65.56 ± 1.56	158.73 ± 18.53	0.482 ± 0.054	−33.42 ± 3.45	33.24 ± 1.93
FE3	(−1)	30	(1)	3	60.71 ± 2.04	167.42 ± 12.37	0.480 ± 0.123	−31.47 ± 5.34	31.80 ± 2.52
FE4	(0)	40	(−1)	1	63.43 ± 3.17	157.27 ± 17.63	0.415 ± 0.142	−29.37 ± 3.74	51.62 ± 2.14
FE5	(0)	40	(0)	2	70.32 ± 2.86	163.47 ± 10.46	0.375 ± 0.085	−26.72 ± 2.64	42.83 ± 3.51
FE6	(0)	40	(1)	3	66.67 ± 3.46	183.37 ± 20.46	0.262 ± 0.053	−30.28 ± 4.72	40.92 ± 2.13
FE7	(1)	50	(−1)	1	83.43 ± 2.23	160.57 ± 13.74	0.257 ± 0.008	−35.91 ± 4.08	68.47 ± 3.60
FE8	(1)	50	(0)	2	86.14 ± 1.58	180.63 ± 11.42	0.237 ± 0.014	−32.92 ± 3.81	61.91 ± 2.94
FE9	(1)	50	(1)	3	76.47 ± 3.51	193.52 ± 10.58	0.222 ± 0.047	−36.64 ± 3.64	57.52 ± 2.50

*MAP (0.3% w/v (0.333 g/100 mL)) and PG (10% v/v (10 mL/100 mL)) were kept constant in all formulations, volume was completed up to 100 mL with distilled water. EE: Entrapment Efficiency, PDI: Polydispersity index, ZP: Zeta potential.

Table 2. Experimental runs, independent and dependent variables of the 2³ full factorials experimental design of Magnesium ascorbyl phosphate-loaded niosomes.

Runs*	Factors (Independent variables)					Responses (Dependent variables)
	Surfactant type	Surfactant molar ratio		Cholesterol molar ratio		Y1:EE (%)	Y2: Particle size (nm)	Y3:PDI	Y4:ZP (mV)	Y5: % permeated (8 h)
FN1	Span 60	(−1)	1	(−1)	1	69.80 ± 3.56	218.60 ± 22.52	0.253 ± 0.124	−26.83 ± 3.34	36.67 ± 3.64
FN2	Span 60	(−1)	1	(+1)	2	78.63 ± 2.47	258.23 ± 16.85	0.281 ± 0.092	−32.42 ± 2.58	30.14 ± 1.96
FN3	Span 60	(+1)	2	(−1)	1	86.82 ± 4.52	138.43 ± 17.84	0.312 ± 0.112	−27.56 ± 2.67	54.42 ± 2.35
FN4	Span 60	(+1)	2	(+1)	2	92.13 ± 3.09	169.32 ± 12.22	0.292 ± 0.163	−31.42 ± 2.43	46.37 ± 3.24
FN5	Span 80	(−1)	1	(−1)	1	53.85 ± 4.52	362.43 ± 24.63	0.415 ± 0.131	−25.82 ± 4.52	33.34 ± 2.05
FN6	Span 80	(−1)	1	(+1)	2	61.24 ± 2.68	384.62 ± 14.52	0.281 ± 0.082	−32.63 ± 3.84	28.46 ± 2.98
FN7	Span 80	(+1)	2	(−1)	1	68.32 ± 2.97	278.40 ± 10.58	0.354 ± 0.028	26.08 ± 3.06	50.58 ± 2.68
FN8	Span 80	(+1)	2	(+1)	2	87.32 ± 3.12	315.62 ± 11.23	0.382 ± 0.036	−30.52 ± 2.41	40.42 ± 3.21

*MAP (one molar ratio) was kept constant in all formulations. EE: Entrapment Efficiency, PDI: Polydispersity index, ZP: Zeta potential.

Figure 1. Effect of different independent factors; (X1): Ethanol concentration, (X2): PC concentration on: (a) EE%, (b) PS, (c) PDI, (d) ZP, (e) MAP % permeated (8 h) of the prepared MAP Ethosomes.

Figure 2. Effect of different independent factors; (X1): Ethanol concentration, (X2): PC concentration on: (a) EE%, (b) PS, (c) PDI, (d) ZP, (e) MAP % permeated (8 h) of the prepared MAP Niosomes.

Figure 3. Transmission electron microscopy (TEM) photographs of: (a) MAP- loaded ethosomes (FE7), (b) MAP- loaded niosomes (FN3).

Figure 4. MAP skin permeation and skin retention from MAP solution, MAP ethosomes (FE7), MAP ethosomal (FE7) gel, MAP niosomes (FN3) and MAP niosomal (FN3) gel.

Table 3. Effect of storage on selected ethosomes (FE7) and niosomes (FN3) formulations.

Formulation	Month	Stored at 4 °C		Stored at room temperature
		EE%	PS (nm)	EE%	PS (nm)
FE7	0	83.43 ± 2.23	160.57 ± 13.7	83.43 ± 2.23	160.57 ± 13.7
	1	83.12 ± 1.89	163.54 ± 8.67	81.67 ± 2.34	171 ± 11.57
	2	82.63 ± 2.06	167.85 ± 10.69	79.53 ± 1.85	183 ± 9.38
	3	82.17 ± 2.18	170.52 ± 12.85	76.52 ± 1.93	189.32 ± 13.08
FN3	0	86.82 ± 4.52	138.43 ± 17.84	86.82 ± 4.52	138.43 ± 17.84
	1	86.53 ± 3.46	136.62 ± 22.54	85.48 ± 5.39	140.32 ± 19.40
	2	85.72 ± 4.08	141.31 ± 19.42	82.57 ± 3.62	153.72 ± 23.43
	3	85.03 ± 3.94	141.54 ± 24.62	81.44 ± 4.51	159.89 ± 22.24

EE: Entrapment Efficiency, PS: Particle size.

Table 4. Demographic and clinical data.

	N = 40
Gender
Women	23 (57.5%)
Men	17 (42.5)
Age
Mean ± SD	41.90 ± 9.097
Range	25–55
Skin type
II	1 (2.5%)
III	24 (60%)
IV	15 (37.5)
Rt side Average melanin level before Treatment
Mean ± SD	0.799 ± 0.1190
Range	0.5–1
Rt side Average melanin level After 1 month
Mean ± SD	0.776 ± 0.1158
Range	0.5–1
Rt side Average melanin level After 6 months
Mean ± SD	0.748 ± 0.1608
Range	0.1–1
Lt side Average melanin level before treatment
Mean ± SD	0.792 ± 0.1321
Range	0.6–1
Lt side Average melanin level After 1 month
Mean ± SD	0.772 ± 0.1234
Range	0.6 – 1
Lt side Average melanin level After 6 months
Mean ± SD	0.758 ± 0.1619
Range	1

Figure 5. Samples of Antera 3D^® Camera photos of before and after treatment with: (A) Right side (MAP niosoma gel) and (B) Left side (MAP ethosoal gel).

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article.