Antiaging effects of a novel facial serum containing L-Ascorbic acid, proteoglycans, and proteoglycan-stimulating tripeptide: ex vivo skin explant studies and in vivo clinical studies in women

Figures & data

Figure 1 Resazurin levels, indicating cell viability, in ex vivo photoaging and photo-oxidation study.

Notes: Expressed as a percentage compared to NIC levels (control considered 100%), mean ± standard deviation in NIC, IC, and IST.
Abbreviations: IC, irradiated control; IST, irradiated and serum treated; NIC, nonirradiated control; RES, resazurin.

Table 1 Ex vivo study, antioxidant, and antiphotoaging parameters

Parameter	IST	IC	NIC
Antioxidant parameters
ROS, % of positive cells	16.44 (3.30)	32.58 (4.34)	14.68 (4.04)
Glutathione, nmol/L	97.94 (2.56)	79.67 (2.39)	113.61 (5.32)
CPD, ng/mL	1.23 (0.32)	3.30 (0.75)	0.81 (0.51)
ECM parameters
Collagen, µg/mg	1.69 (0.34)	1.20 (0.53)	2.01 (0.54)
Collagen, % level with respect to healthy group	83.97 (16.87)	59.61 (26.19)	100 (2.01)
Elastin, µg/mg	8.45 (0.90)	6.79 (0.84)	8.56 (0.83)
Elastin, % level with respect to healthy group	98.84 (10.54)	79.40 (9.79)	100 (7.51)

Note: Mean ± standard deviation (SD).

Abbreviations: CPD, cyclobutane pyrimidine dimer; ECM, extracellular matrix; IC, irradiated control; IST, irradiated and serum treated; NIC, nonirradiated control; ROS, reactive oxygen species.

Figure 2 Antiphotoaging study, mean collagen, and elastin levels at the end of the study (day 10).

Note: *P<0.05 compared with IC.
Abbreviations: IC, irradiated control; IST, irradiated and serum treated; NIC, nonirradiated control.

Figure 3 Masson–Goldner staining of dermal tissue in the ex vivo photoaging study, with connective fibers stained blue-green, fibroblast nuclei stained dark brown-black, and fibroblast cytoplasm stained light red-pink (magnification 200×).

Notes: (A) Untreated control tissue; (B) irradiated control tissue; and (C) irradiated and serum-treated tissue.

Table 2 P-values from statistical analysis in photoaging and photo-oxidation study: unpaired t-test comparisons between experimental groups

Groups compared	P-values
	Collagen	Elastin
IC vs NIC	0.027*	0.002*
IC vs IST	0.092	0.006*
NIC vs IST	0.253	0.554

Note:

* Statistically significant.

Abbreviations: IC, irradiated control; IST, irradiated and serum treated; NIC, non-irradiated control.

Figure 4 Markers of oxidation in the antioxidant and antiphotoaging study.

Notes: (A) CPDs; (B) percentage of ROS-positive cells; and (C) total glutathione levels in NIC, IC, and IST. *P<0.05 compared with IC.
Abbreviations: CPDs, cyclobutane pyrimidine dimers; IC, irradiated control; IST, irradiated and serum treated; NIC, nonirradiated control; ROS, reactive oxygen species.

Table 3 P-values from statistical analysis in photoaging and photo-oxidation study: unpaired t-test comparisons between experimental groups

Groups compared	P-values
	CPDs	ROS	Glutathione
IC vs NIC	0.002*	<0.001*	<0.001*
IC vs IST	0.001*	<0.001*	<0.001*
NIC vs IST	0.130	0.417	<0.001*

Note:

* Statistically significant.

Abbreviations: CPDs, cyclobutane pyrimidine dimers; IC, irradiated control; IST, irradiated and serum treated; NIC, nonirradiated control; ROS, reactive oxygen species.

Figure 5 Resazurin levels, indicating cell viability, throughout the hydrocortisone aging study.

Note: Expressed as a percentage compared to nonaged control levels (control considered 100%), mean ± standard deviation.
Abbreviations: HC, hydrocortisone control; HST, hydrocortisone and serum treated; NHC, non-HC; RES, resazurin.

Figure 6 Masson–Goldner staining of dermal tissue in the ex vivo hydrocortisone study, with connective fibers stained blue-green, fibroblast nuclei stained dark brown-black, and fibroblast cytoplasm stained light red-pink (magnification 200×).

Notes: (A) Untreated control tissue; (B) hydrocortisone control tissue; and (C) hydrocortisone- and serum-treated tissue.

Table 4 Outcome variables for antiaging efficacy in the hydrocortisone ex vivo study

Antiaging parameters	HST	HC	NHC
Collagen, µg/mg	3.34 (0.58)	2.56 (0.41)	3.25 (0.48)
Collagen, % level with respect to healthy group	102.63 (17.94)	78.61 (12.63)	100.00 (14.86)
Elastin, µg/mg	10.71 (0.66)	8.41 (0.39)	11.04 (1.46)
Elastin, % level with respect to healthy group	93.72 (5.19)	72.76 (3.39)	100.00 (11.11)
Proteoglycan	0.76 (0.06)	0.64 (0.09)	0.82 (0.07)
Proteoglycan, % with respect to healthy group	92.56 (7.70)	78.76 (11.61)	100.00 (8.79)
Hyaluronic acid, µg/mg skin	0.04 (0.03)**	0.18 (0.04)	0.27 (0.06)
Hyaluronic acid, % with respect to healthy group	14.72 (9.96)**	64.83 (13.90)	100 (23.82)

Note:

** In the hyaluronic assay, the study product interfered with the colorant, so results were inconclusive in the serum-treatment group. Data are presented as mean (SD).

Abbreviations: HC, hydrocortisone control; HST, hydrocortisone and serum treated; NHC, nonhydrocortisone control.

Figure 7 Alcian blue-stained dermal tissue from ex vivo hydrocortisone study.

Notes: Proteoglycans are stained in deep blue (magnification 200×). (A) Untreated control tissue; (B) hydrocortisone control tissue; and (C) hydrocortisone- and serum-treated tissue.

Table 5 Clinical studies subject characteristics

Study	Number of subjects	Women, n (%)	Age, years (min–max)
Clinical study 1: hydration kinetics	20	20 (100)	49.2 (21–67)
Clinical study 2: efficacy and tolerability under dermatological control	30	30 (100)	37 (21–45)
Clinical study 3: consumer study	105	105 (100)	35.7 (18–45)

Figure 8 Hydration kinetics study of skin moisture levels over time as measured with corneometer.

Note: Serum treated vs control.

Figure 9 Hydration kinetics study of skin moisture levels over time as measured with Skicon hygrometer.

Note: Serum treated vs control.

Table 6 Results from subjective efficacy questionnaire under dermatological control and consumer test questionnaire

Questionnaire item	% of subjects in agreement
	Efficacy and tolerability under dermatological control	Consumer test
The product provided immediate hydration	83	94
The product had an immediate antifatigue effect (flash effect)	70	87
Skin feels more hydrated day by day	87	91
Wrinkles, if present, decreased or softened	77	78
Lines, if present, decreased or softened	70	78
The skin of the face looks brighter	83	95
Skin feels more rested	87	90
Skin feels healthier	87	91
Liked the product overall	80	95
Would like to continue using the product	55	89
Would buy the product independently of the price	60	82
Overall you liked this product	80	95