Hyperpigmentation: 0=even skin color, no observable hyperpigmentation, 9=significant (severe) hyperpigmented appearance, involving most of the area, with very strong intensity
Skin tone evenness: 0=even, healthy skin color, 9=uneven, extensive discolored appearance (brown and red colors)
Radiance/Luminosity/Brightness: 0=radiant, luminous, or glowing appearance, 9=full/matte and/or sallow skin appearance
In addition, at weeks 4, 8, and 12, an expert grader assessed the global improvement in overall skin texture (pronounced, extensive visible skin roughness, lines/wrinkles, laxity and crepey/crinkled appearance) on the neck and overall photodamage on the décolleté using a grading scale, where 1=worse, 2=no improvement, 3=mildly improved, 4=moderately improved, and 5=markedly improved.
Standardized Photography
Digital images were taken of each subject’s face, neck, and décolletage (center view, left side, and right side) using Stephens & Associates’ photo station with a Canon Mark II 7D digital SLR camera (Canon Inc, Tokyo, Japan) with a Canon EF-S 60 mm f/2.8 macro lens under visible light. Unfiltered full-spectrum (white) light was provided using Profoto D1 (500W) studio strobes affixed to the photo station. Canon EOS utility software was used for image overlay to ensure post-baseline images match baseline images. Prior to photography, subjects were instructed to remove any jewelry from the area to be photographed, cleansed their face, and equilibrated in the clinic for at least 15 minutes.
Cutometer Measurements
A single measurement was taken of each subject’s left side of the neck, in the center (vertically), approximately left of the laryngeal prominence (“Adam’s appleâ€) using Cutometer MPA 580 (Courage + Khazaka Electronic GmbH, Köln, Germany) to measure the viscoelastic properties of the skin. For each subject, the measured location was marked on a body diagram to ensure the same site was measured at week 12.
Subject Self-Assessment Questionnaire
Subjects completed a self-assessment questionnaire regarding various skin parameters and product texture and attributes at weeks 4, 8, and 12.
Statistical Analysis
Clinical grading scores at weeks 4, 8, and 12 were compared to baseline scores using Wilcoxon signed-rank test and comparisons between treatment groups using Wilcoxon rank sum test. Average percent change from baseline/placebo was calculated for all parameters at each follow-up visit. Comparisons between treatment groups were calculated for all parameters at each follow-up visit. All differences are considered statistically significant at the P≤0.05 level.
Skin tone evenness: 0=even, healthy skin color, 9=uneven, extensive discolored appearance (brown and red colors)
Radiance/Luminosity/Brightness: 0=radiant, luminous, or glowing appearance, 9=full/matte and/or sallow skin appearance
In addition, at weeks 4, 8, and 12, an expert grader assessed the global improvement in overall skin texture (pronounced, extensive visible skin roughness, lines/wrinkles, laxity and crepey/crinkled appearance) on the neck and overall photodamage on the décolleté using a grading scale, where 1=worse, 2=no improvement, 3=mildly improved, 4=moderately improved, and 5=markedly improved.
Standardized Photography
Digital images were taken of each subject’s face, neck, and décolletage (center view, left side, and right side) using Stephens & Associates’ photo station with a Canon Mark II 7D digital SLR camera (Canon Inc, Tokyo, Japan) with a Canon EF-S 60 mm f/2.8 macro lens under visible light. Unfiltered full-spectrum (white) light was provided using Profoto D1 (500W) studio strobes affixed to the photo station. Canon EOS utility software was used for image overlay to ensure post-baseline images match baseline images. Prior to photography, subjects were instructed to remove any jewelry from the area to be photographed, cleansed their face, and equilibrated in the clinic for at least 15 minutes.
Cutometer Measurements
A single measurement was taken of each subject’s left side of the neck, in the center (vertically), approximately left of the laryngeal prominence (“Adam’s appleâ€) using Cutometer MPA 580 (Courage + Khazaka Electronic GmbH, Köln, Germany) to measure the viscoelastic properties of the skin. For each subject, the measured location was marked on a body diagram to ensure the same site was measured at week 12.
Subject Self-Assessment Questionnaire
Subjects completed a self-assessment questionnaire regarding various skin parameters and product texture and attributes at weeks 4, 8, and 12.
Statistical Analysis
Clinical grading scores at weeks 4, 8, and 12 were compared to baseline scores using Wilcoxon signed-rank test and comparisons between treatment groups using Wilcoxon rank sum test. Average percent change from baseline/placebo was calculated for all parameters at each follow-up visit. Comparisons between treatment groups were calculated for all parameters at each follow-up visit. All differences are considered statistically significant at the P≤0.05 level.
RESULTS
In vitro analysis
The efficacy of NCC in supporting skin rejuvenation was assessed at a molecular biology level using quantitative real-time PCR to provide an early indication of potential activity of the cosmetic product. Human 3D skin models were irradiated with ultraviolet (UV) light to mimic extrinsic skin aging prior to the application of NCC. Gene expression results for UV-irradiated, non-treated tissues and UV-irradiated, NCC-treated tissues were calculated as percentage increase compared to non-UV-irradiated, non-treated control tissues (set as 0%). Expression levels of genes encoding collagens (COL1A1, COL3A1, COL6A1, COL7A1), elastic fiber proteins (ELN, FBN1, FBLN5, MFAP1, LOXL1),TGFβ1 and decorin (DCN) were assessed (Figure 1A and 1B). Application of NCC resulted in significant upregulation of all genes tested compared to UV-irradiated, non-treated samples indicating that NCC stimulates ECM components for dermal structure improvement that could translate into firmer skin, as well as improvement of the dermal-epidermal junction (DEJ), which is important for skin elasticity. Additionally, elastase activity assays showed that NCC significantly reduced the level of elastase (an enzyme that breaks down elastin) following UV irradiation suggesting protection against elastic fiber degradation (Figure 1C).
Aging alters essential processes involved in maintaining cellular proteostasis (protein homeostasis), including the proteasome and autophagy systems, leading to an accumulation in cellular debris, protein aggregation, and cellular damage.9 Gene expression levels of key recycling genes involved in proteasome activity (POMP, PSMB5, PSMB6)10 and autophagy (ATG5, ATG7, ATG12, BECN1) along with a heat shock protein that facilitates and stabilizes proper folding of proteins (HSPA8) were increased with NCC treatment (Figure 2), indicating stimulation of the various cellular recycling processes.
Altogether the in vitro pre-clinical data demonstrate the efficacy of NCC in providing overall skin rejuvenation benefits by boosting various ECM and DEJ components as well as supporting systems that help maintain proteostasis.
Clinical Efficacy of NCC
Sixty-nine subjects with Fitzpatrick skin types I–V, completed the 12-week, double-blind, randomized, regimen-controlled study (NCC-treatment group: n=42, control group: n=27). Overall results indicate that twice daily topical application of NCC was effective in improving neck and décolletage skin condition and firmness.
The efficacy of NCC in supporting skin rejuvenation was assessed at a molecular biology level using quantitative real-time PCR to provide an early indication of potential activity of the cosmetic product. Human 3D skin models were irradiated with ultraviolet (UV) light to mimic extrinsic skin aging prior to the application of NCC. Gene expression results for UV-irradiated, non-treated tissues and UV-irradiated, NCC-treated tissues were calculated as percentage increase compared to non-UV-irradiated, non-treated control tissues (set as 0%). Expression levels of genes encoding collagens (COL1A1, COL3A1, COL6A1, COL7A1), elastic fiber proteins (ELN, FBN1, FBLN5, MFAP1, LOXL1),TGFβ1 and decorin (DCN) were assessed (Figure 1A and 1B). Application of NCC resulted in significant upregulation of all genes tested compared to UV-irradiated, non-treated samples indicating that NCC stimulates ECM components for dermal structure improvement that could translate into firmer skin, as well as improvement of the dermal-epidermal junction (DEJ), which is important for skin elasticity. Additionally, elastase activity assays showed that NCC significantly reduced the level of elastase (an enzyme that breaks down elastin) following UV irradiation suggesting protection against elastic fiber degradation (Figure 1C).
Aging alters essential processes involved in maintaining cellular proteostasis (protein homeostasis), including the proteasome and autophagy systems, leading to an accumulation in cellular debris, protein aggregation, and cellular damage.9 Gene expression levels of key recycling genes involved in proteasome activity (POMP, PSMB5, PSMB6)10 and autophagy (ATG5, ATG7, ATG12, BECN1) along with a heat shock protein that facilitates and stabilizes proper folding of proteins (HSPA8) were increased with NCC treatment (Figure 2), indicating stimulation of the various cellular recycling processes.
Altogether the in vitro pre-clinical data demonstrate the efficacy of NCC in providing overall skin rejuvenation benefits by boosting various ECM and DEJ components as well as supporting systems that help maintain proteostasis.
Clinical Efficacy of NCC
Sixty-nine subjects with Fitzpatrick skin types I–V, completed the 12-week, double-blind, randomized, regimen-controlled study (NCC-treatment group: n=42, control group: n=27). Overall results indicate that twice daily topical application of NCC was effective in improving neck and décolletage skin condition and firmness.
