Received: 20 July 2020 | Accepted: 28 July 2020

DOI: 10.1111/jocd.13676

ORIGINAL CONTRIBUTION

A randomized, triple-blind, placebo-controlled, parallel study to

evaluate the efficacy of a freshwater marine collagen on skin
wrinkles and elasticity

Malkanthi Evans DVM, PhD | Erin D. Lewis PhD | Nisrine Zakaria PhD |
Tetyana Pelipyagina MD | Najla Guthrie BSc

KGK Science Inc., London, Ontario, Canada

Abstract
Correspondence Background: Collagen is the primary component in human skin. With age, there is
Malkanthi Evans, KGK Science Inc., 255
Queens Ave, London, Ontario N6A 5R8 loss of skin elasticity and collagen, resulting in wrinkle formation and reduction in
Canada. skin appearance.
Email: [email protected]
Aims: The objective of this randomized, triple-blind, placebo-controlled study was to
Funding information evaluate the safety and efficacy of a hydrolyzed marine collagen (Vinh Wellness Collagen,
Vinh Hoan Corporation
VWC) on aspects of skin health and quality in women between 45 and 60 years of age.
Patients/Methods: Assessments of skin wrinkles, elasticity, and self-reported appear-
ance were conducted using the VISIA skin analysis system, Cutometer®, and Skin Quality
Visual Analogue Scale. Outcomes were assessed at weeks 0 (baseline), 6, and 12.
Results: After 12 weeks, participants supplemented with VWC had a significant 35%
reduction in wrinkle score (P = .035) from baseline. Participants in the VWC group
showed a 24% greater reduction in wrinkles on the right side of the face than those on
placebo. A planned subgroup analysis based on age showed women 45-54 years had a
significant 20% and 10% improvement in cheek skin elasticity from baseline to week 6
(P = .016) and 12 (P = .022), respectively. At week 12, participants in the VWC group
reported greater percentage improvements in overall skin score (9%) and wrinkle (15%),
elasticity (23%), hydration (14%), radiance (22%), and firmness (25%) scores vs placebo.
Conclusion: Supplementation with VWC was found to be safe and well-tolerated.
The results of this study support the use of fish-derived hydrolyzed collagen for the
improvement of skin health in an aging population.

KEYWORDS

hydration, hydrolyzed collagen, skin elasticity, wrinkles

1 | I NTRO D U C TI O N This process slows with age, compromising the quality of collagen
fibres.1 Collagen, which constitutes 95% of human skin, 2 and elastin
Skin is composed of an outer (epidermis) and inner (dermis) layer. are the two primary components of the dermis. These two fibres
The nonvascular epidermis relies on the dermis to receive nutrients. work synergistically to form the structure of the skin. Aging damages

This is an open access article under the terms of the Creative Commons Attribution-­NonCommercial-­NoDerivs License, which permits use and distribution in
any medium, provided the original work is properly cited, the use is non-­commercial and no modifications or adaptations are made.
© 2020 Vinh Hoan Corp. Journal of Cosmetic Dermatology published by Wiley Periodicals LLC.

J Cosmet Dermatol. 2021;20:825–834. wileyonlinelibrary.com/journal/jocd | 825

https://doi.org/10.1111/jocd.13676
826 | EVANS et al.

the elastic capacity of the skin, and thus, aging skin is marked by lack Harmonization of Technical Requirements for Pharmaceuticals for
3
of elasticity, fragmentation, and collagen bundle fragility. Human Use (ICH) guidelines and Good Clinical Practice Current
Skin appearance is influenced by nutrition as well as endogenous Step 4 Version, dated June 10, 1996. The trial was registered at
and environmental factors, including the exposure to chemicals, Clinicaltrials.gov (NCT04449159).
smoking, or ultraviolet radiation. Skin aging is due to changes in the The study design was a randomized, triple-blind, placebo-con-
deeper layer of the skin. Oral supplements, in contrast to topical ap- trolled, parallel study conducted at the KGK Science Inc clinic site
plications, represent a practical approach to the prevention of skin (London, Ontario, Canada) from November 25, 2016, to July 18,
aging because they can be delivered to the dermis through the circu- 2017. Written informed consent was obtained from all participants
lation.4 The ability of nutritional supplements to enhance skin char- prior to any study procedures being initiated.
acteristics has received increasing attention as the North American Participants were included if they were females between the
population continues to age.5 ages of 45-60 with a BMI 20.0-29.9 kg/m2 and displayed visible
Collagen is the most abundant protein in mammals and is cur- signs of natural and photoaging on their face, as assessed by the
rently being promoted by nutrition, biomedicine, and cosmetics in- Fitzpatrick questionnaire at screening. All participants agreed to
6
dustries. Gelatin, a protein which is used extensively in the food avoid prolonged exposure to ultraviolet (UV) radiation for the dura-
sector, is a hydrolyzed analog of collagen.6 Subsequent enzymatic tion of the study.
degradation of gelatin results in the generation of hydrolyzed colla- Individuals were excluded if they suffered from an acute or chronic
gen, which contains peptides ranging in molecular weights between skin disease or dermatological disorder; used natural health supple-
<500 Da and 6 KDa, depending on the processing conditions.7 ments for improving the skin; were on a low protein diet; had planned
Orally administered hydrolyzed collagen is absorbed in the small in- or unavoidable exposure to UV radiation; had tattoos on or near the
testine and into the blood stream as peptides and free amino acids test area; used systemic corticosteroids or applied topical alpha hy-
and distributed into the dermis for up to 14 days.8 In the dermis, hy- droxyl acids near the test site within 4 weeks of enrolment; used top-
drolyzed collagen provides amino acids for the formation of collagen ical medications near the test area within 6 weeks of enrolment; had
and elastin fibres, in addition to stimulating endogenous production Botulinum toxin A (Botox) treatment or filler injection (collagen, hyal-
of new collagen, elastin, and hyaluronic acid.9 uronic acid, etc) near the test sites within 2 years of enrolment; were
Hydrolyzed collagen is commonly derived from cow, pig, and cognitively impaired and/or unable to give informed consent; or had
chicken sources. In recent years, collagen derived from fish has any other condition which in the medical investigator's opinion may
emerged as an alternative source due to lower environmental im- adversely affect the individual's ability to complete the study or its
pact and risk of disease transmission. Further, marine fish collagen measures or which may pose significant risk to the individual.
and collagen peptides have a high degree of homology to human
structure and bioavailability through the gastrointestinal barrier.10
Although a number of preclinical studies provide evidence for the 2.2 | Investigational product
beneficial effect of hydrolyzed collagen on skin health, less infor-
mation is known about the clinical benefits.11,12 Previous stud- The investigational product was Vinh Wellness Collagen (VWC), a
ies have demonstrated improvements in skin wrinkles, elasticity, hydrolyzed collagen powder derived from Pangasius hypophthalmus,
and hydration following supplementation with hydrolyzed colla- a tropical and sustainable freshwater fish (Vinh Hoan Corporation).
gen.12,13 Fish collagen peptides have been shown to significantly The placebo was matched to the investigational product using malto-
reduce crow's feet 14 and periorbital wrinkles in women.15 Further, dextrin powder (Qinhuangdao Lihua Starch Co., Ltd). Participants
hydrolyzed collagen supplementation was reported to improve were instructed to consume 10 g of hydrolyzed collagen or placebo
skin elasticity and moisture while reducing evaporation.16 The aim powder daily, in the morning, on an empty stomach for 12 weeks. It
of this study was to evaluate the clinical benefits of a 12-week sup- was required that the product was dissolved it in at least 100 mL of
plementation of a fish-derived collagen peptide on skin wrinkles water. In the event a dose was missed participants were to consume
and elasticity, and self-reported skin appearance. it as soon as they remembered and were instructed not to exceed
10 g of the investigational product or placebo per day.

2 | M E TH O DS
2.3 | Randomization and blinding
2.1 | Participants
Eligible participants were assigned a randomization number by a
This study was approved by the institutional review board (IRB blinded investigator per the order of the randomization list (www.
Services, Aurora) on July 12, 2016 (Pro00018179), and by Health rando​mizat​ion.com). Each randomization code represented an allo-
Canada on June 13, 2016 (219 935). The clinical trial was performed cation to a dosing arm of the study. Following randomization, par-
according to the ethical guidelines detailed in the Declaration of ticipants were identified by their initials and date of birth and were
Helsinki (2008) and complied with the International Council for assigned a participant number at their screening visit.
EVANS et al. | 827

The investigational product and placebo were sealed in sachets participants by using standardized photographs of nasolabial wrin-
that were identical in appearance and labelled per the ICH-GCP re- kles alone and assessment of wrinkle depth.
quirements and applicable local regulatory guidelines. Unblinded
personnel who were not involved in any study assessments labelled
the investigational product. The investigators, statistician, other site 2.7 | Skin elasticity analysis
personnel, and participants were blinded to the products.
Skin elasticity measurements were performed using the Cutometer ®
dual MPA 580 (Courage Khazaka electronic GmbH). A cheek free
2.4 | Compliance from skin damage or the cheek with the least damage (eg, scars,
burns or cuts) was chosen at baseline and used for subsequent visits
Product compliance was assessed by counting the returned unused (week 6 and week 12), and a single Cutometer ® measurement was
product at week 6 and week 12 and determined by the number of performed. Participants did not wash their face within 2 hours prior
dosage units taken divided by the number expected, multiplied by to testing, and creams were not to be used prior to measurements.
100. In the event of a discrepancy between the information in the A higher score from the Cutometer® measurement indicated greater
study diary, compliance was based on the product returned unless skin elasticity.
an explanation for the loss was provided.

2.8 | Skin quality VAS questionnaire

2.5 | Outcome evaluation
Skin quality was self-assessed by using a Visual Analogue Scale (VAS)
The study investigated the changes in skin wrinkles and elasticity, questionnaire adapted from Gold et al 2013.17 Using a scale from
and self-reported skin appearance after 12 weeks of collagen or 0 (no improvement) to 100 (great improvement), participants were
placebo supplementation. Clinical and qualitative assessments were asked to report their skin health based on skin elasticity, hydration,
conducted at screening, week 0 (baseline), week 6, and week 12 (end- radiance, firmness, wrinkles, and overall feel.
of-study). At all visits, participants’ diaries were reviewed for concom-
itant therapies, adverse events, and study product use. Assessments
including evaluation of nasolabial wrinkles and cheek elasticity and 2.9 | Laboratory analyses
skin quality questionnaire are described below. Vital signs (blood
pressure and heart rate) and anthropometric measurements (weight Safety parameters were analyzed by LifeLabs using standardized
and BMI) were taken at each visit, while laboratory parameters for all procedures. Analysis of hemoglobin, hematocrit, platelet count,
safety endpoints were assessed at screening and week 12. red blood cell count (RBC), red blood cell indices, red cell distribu-
tion width (RDW), white blood cell count (WBC) and differentials
(neutrophils, lymphocytes, monocytes, eosinophils, basophils), liver
2.6 | Skin wrinkle analysis function tests (AST, ALT, bilirubin), and kidney function tests (Na, K,
Cl, creatinine, eGFR) were performed. Urine pregnancy tests were
The skin wrinkle analysis was performed using the 6th Generation conducted at the KGK Clinic for participants of childbearing poten-
VISIA skin analysis system (Canfield Imaging Systems). The VISIA tial at screening and baseline visits.
skin analysis system assesses the number of visible wrinkles and
provides a score ranging from 0 to 100, with a lower score indicative
of fewer wrinkles. The left and right sides of the face were analyzed 2.10 | Adverse events
separately.
Prior to testing, participants were provided a facial wipe and Participants recorded any adverse events (AEs) in their daily diary.
asked to remove all makeup, after which the face was dried with a AEs were documented in the study record and were classified based
lint-free towel or allowed to air dry. Hair was clipped away from the on the description, duration, intensity, frequency, and outcome. The
face when applicable. The participant placed their face on the chin Qualified Investigator assessed all AEs and determined causality and
(resting) platform of the machine to perform the analysis. Manual intensity. The Medical Dictionary for Regulatory Activities terminol-
masks were created for the nasolabial area of the face of each partic- ogy (MEDRA) version 17.0 was used for coding.
ipant at baseline and were used at each subsequent visit.
The Modified Fitzpatrick Wrinkle Scale (MFWS) is a validated
questionnaire evaluating skin wrinkle severity. The MFWS is com- 2.11 | Statistical analyses
posed of 3 major classes, in which definitions are based on a set of
reference photographs and descriptions, as well as 3 interclasses The sample size calculation was based on the primary efficacy objec-
based only on descriptions. Trained clinic staff applied this scale to tive of the comparison of 12-week skin elasticity changes between
828 | EVANS et al.

the collagen and placebo groups. With an estimated 80% power, 5% (ANOVA) with group as a fixed effect. A repeated-measures analy-
significance and 20% attrition, 50 participants were required (n = 25 sis of covariance (ANCOVA) modeling was employed to assess the
per group). This calculation assumed between group variability of changes from baseline between groups. The model included base-
0.080 R2 parameter units based on an earlier clinical trial.12 As line value as a covariate with fixed effects for group, study day,
standard deviations of intervention outcomes tend to increase with and group by study day interaction. Linear contrast statements
time and the proposed study was longer than Proksch et al, 2014, from this model were constructed to provide between group
the sample size was conservatively increased by 25%. p-values at each time point.
The safety population consists of all participants who received A planned subgroup analysis was carried out based on age. This
any amount of either study product and on whom any postrandom- subgroup was created from the PP population, and all analyses were
ization safety information was available. The per-protocol (PP) pop- conducted in the same manner as the whole set.
ulation consists of all participants who consumed at least 80% of Probabilities ≤.05 were considered statistically significant. All
either study product doses, did not have any major protocol viola- statistical analyses were completed using SAS version 9.3.1 (Cary,
tions, and completed all study visits and procedures connected with NC) for Microsoft Windows.
measurement of the primary variable. Only observed values were
used for the analysis of the PP population. No imputation was per-
formed for missing values of variables. 3 | R E S U LT S
For continuous endpoints measured on multiple study visits
(week 0, week 6, and week 12), descriptive statistics were pre- A total of 85 volunteers were screened and 50 eligible partici-
sented for each study day and for the changes from baseline to pants were enrolled, with 25 participants randomized into each
each study day. Within-group changes from baseline were as- study group (Figure 1). Forty-five participants completed the study.
sessed using the Wilcoxon signed rank test. Possible differences Four participants in the VWC group and 1 participant in the pla-
between groups at baseline were assessed by analysis of variance cebo group withdrew consent. The PP population consisted of all

FIGURE 1 Disposition of participants

in the trial
EVANS et al. | 829

participants who consumed at least 80% of their assigned product TA B L E 2 Average skin quality scores for the per protocol
dose, did not have any major protocol violations, and who completed population as assessed by the skin quality VAS questionnaire

all study visits and procedures related to measurements of cheek Baseline

skin elasticity. Participant demographic and lifestyle characteristics between-
are presented in Table 1. Participant demographics were similar be- VWC Placebo Group
Mean ± SD (n) Mean ± SD (n) P-valuea
tween both groups.
Overall Score
Baseline 0.59 ± 1.97 (17) 2.63 ± 11.47 (19) 0.538 (r)
3.1 | Efficacy of VWC supplementation on Week 6 46.41 ± 29.40 (17) 38.42 ± 30.63 (19)
skin quality Week 12 54.94 ± 31.82 (17) 52.42 ± 34.28 (19)
Elasticity Score
There was no significant difference in skin quality VAS scores be- Baseline 0.41 ± 1.28 (17) 2.63 ± 11.47 (19) 0.538 (r)
tween participants supplemented with VWC or placebo. Notably, at Week 6 38.71 ± 30.76 (17) 31.26 ± 28.22 (19)
baseline, participants in the VWC group reported lower VAS scores
Week 12 50.35 ± 30.38 (17) 43.26 ± 37.10 (19)
compared to those in placebo (78% lower for VAS overall, 85% for
Hydration Score
elasticity, 82% for hydration, 69% for radiance, 64% for firmness, and
Baseline 0.47 ± 1.50 (17) 2.63 ± 11.47 (19) 0.538 (r)
80% for wrinkle). However, at week 12, participants supplemented
Week 6 41.35 ± 30.12 (17) 37.32 ± 31.31 (19)
with VWC reported higher scores in all VAS parameters vs partici-
Week 12 50.24 ± 30.65 (17) 46.37 ± 34.75 (19)
pants on the placebo (Table 2). Although not significant, participants
Radiance Score
Baseline 0.41 ± 1.28 (17) 1.32 ± 5.74 (19) 0.538 (r)
TA B L E 1 Demographics for participants in the per-protocol
population Week 6 42.06 ± 30.15 (17) 35.47 ± 31.31 (19)
Week 12 49.35 ± 29.43 (17) 41.47 ± 31.19 (19)
VWC Placebo Between-Group
(n = 17) (n = 19) P-Valueb Firmness Score

a Baseline 0.47 ± 1.50 (17) 1.32 ± 5.74 (19) 0.538 (r)

Age (years) 53.2 ± 3.6 55.5 ± 3.2 0.054
[Mean ± SD] Week 6 41.76 ± 31.91 (17) 31.58 ± 32.65 (19)

Gender [n (%)] Week 12 48.18 ± 31.01 (17) 39.37 ± 32.87 (19)

Male 0 (0%) 0 (0%) 1.000 Wrinkle Score

Female 17 (100%) 19 (100%) Baseline 0.53 ± 1.74 (17) 2.63 ± 11.47 (19) 0.538 (r)

Smoking Status [n (%)] Week 6 39.24 ± 26.04 (17) 28.21 ± 27.46 (19)

Ex-Smoker 3 (18%) 4 (21%) 1.000 Week 12 46.41 ± 32.27 (17) 42.53 ± 36.57 (19)

No 14 (82%) 15 (79%) Note: Probability values P ≤ .05 are statistically significant

Engagement in Regular Exercise [n (%)] Abbreviations: n, number; SD, standard deviation.
a
No 3 (18%) 3 (16%) 1.000 For Baseline (Day 0), between group p-value generated by ANOVA
with Group as a fixed effect (r) indicates values were ranked prior to
Yes 14 (82%) 16 (84%)
generating ANOVA or ANCOVA
Ethnicity [n (%)]
Hispanic or Latino 0 (0%) 1 (5%) 1.000
Not Hispanic or 17 (100%) 18 (95%)
Latino
Race [n (%)]
East Asian 1 (6%) 0 (0%) .457
Eastern European 1 (6%) 0 (0%)
White
Middle Eastern 0 (0%) 1 (5%)
Western European 15 (88%) 18 (95%)
White

Note: Probability values P ≤ .05 are statistically significant.

Abbreviations: %, percentage; n, number; SD, standard deviation.
a
Between-group comparisons were made using the independent F I G U R E 2 Percentage improvement from baseline (week 0)
Student's t test. to week 6 and week 12 in skin quality VAS scores for participants
b
Between-group comparisons were made using the Fisher's exact test. taking VWC over participants taking placebo
830 | EVANS et al.

taking VWC showed improvements from baseline to week 12 over face for participants supplemented with VWC compared to placebo
the placebo in the VAS overall score (9%), particularly in the elastic- (P = .035) (Figure 3C).
ity (23%), hydration (14%), radiance (22%), firmness (25%), and wrin-
kle (15%) scores (Figure 2).
3.3 | Efficacy of VWC supplementation on
skin elasticity
3.2 | Efficacy of VWC supplementation on wrinkle
count and appearance There was no significant difference in cheek skin elasticity be-
tween VWC and placebo groups after 12 weeks. Participants sup-
A typical image taken by the VISIA to determine nasolabial wrinkle plemented with VWC experienced an 11% improvement in cheek
count and score is shown in Figure 3A. From baseline to week 12, elasticity from baseline to week 6 (P = .032). However, this was
there were significant reductions in wrinkle score on both the left not maintained as there was a 6% reduction in elasticity from
(17%, P = .009) and right (35%, P = .005) sides of the face for partici- weeks 6 to 12. Participants on the placebo showed a 5% improve-
pants supplemented with VWC (Figure 3B and C). After 12 weeks, ment from baseline to week 6 and a 3% improvement from week
there was a 24% reduction in wrinkle score on the right side of the 6 to week 12.

F I G U R E 3 Nasolabial wrinkle analysis of participants supplemented with VWC or placebo. (A) Representative VISIA images at baseline
and after 12 weeks of supplementation. (B) Change in the average absolute wrinkle score for the left side of the face. (C) Change in average
absolute wrinkle score for the right side of the face. n = 11 (placebo); n = 12 (VWC). + Within group p-values generated by the Wilcoxon
signed rank test. Probability values P ≤ .05 are statistically significant
EVANS et al. | 831

Notably, in a planned subgroup analysis based on age, partici- supplemented with VWC showed a significant 24% improvement in
pants between 45 and 54 years of age in the VWC group showed a the absolute wrinkle score on the right side of the face compared to
significant 20% and 10% improvement in cheek skin elasticity from placebo. On both sides of the face, there was a significant decrease
baseline to week 6 (P = .032) and week 12 (P = .027), respectively in the wrinkle score from baseline to week 12 for participants sup-
(Figure 4). plemented with VWC.
The improvements in skin health observed in the current study
are consistent with previous studies examining collagen supple-
3.4 | Safety evaluation of VWC supplementation mentation. Supplementation with 3 g of collagen peptide was as-
sociated with a higher report of participant satisfaction compared
A total of 32 AEs were recorded in this study, with 19 unique partici- to placebo.13 Similar improvements in facial wrinkles in this study
pants experiencing these events. Of these, 18 AEs were reported by were reported following 30 days of hydrolyzed collagen supplemen-
participants in the VWC group, and 14 in the placebo. Of the 18 AEs tation in females.18 Bonnet al. (2017) found that following 12-week
in the VWC group, 17 were reported as being not related or unlikely supplementation with 10 g of fish collagen peptides, women had a
to be related. One report of mild nausea was the only AE reported significant 10% reduction in periorbital wrinkles.19 As the improve-
as being possibly related to the investigational product. All 14 of the ment found by Bonnet et al (2017) was for periorbital wrinkles, a
AEs in the placebo were reported as being not related or unlikely to direct comparison to the current study cannot be drawn. However,
be related. There was no difference in the number of AEs reported one possible reason for the greater improvement in wrinkle score
between the groups and all were resolved by the end of the study. observed in the current study may be due to higher concentration
There was no between-group differences in hematological of anti-oxidant amino acids (glycine, proline, hydroxyproline) found
and clinical chemistry parameters at screening or after 12 weeks in VWC.
(Table 3), and all participants were deemed healthy. For both Although not significant, the improvement in cheek skin elas-
groups, all hematology, clinical chemistry, electrolytes, and liver and ticity over the 12-week VWC supplementation period is consistent
kidney function markers remained within healthy clinical reference with previously published research that utilized similar doses and
ranges. Any changes in these safety parameters were deemed not study populations. Korean females and males experienced a signifi-
clinically significant. There were no significant differences in vital cant improvement in elasticity following 12-week supplementation
13
signs or anthropometric measurements between VWC and placebo with 3 g of collagen peptide and vitamin C or 3 g hydrolyzed
20
(data not shown). fish collagen combined with astaxanthin. Supplementation with
10 g of collagen peptides combined with vitamins A, C, and E and
zinc significantly improved gross cheek elasticity in females aged
4 | D I S CU S S I O N 40-60 years after 90 days. 21 This suggests collagen peptides may
act synergistically with other nutrients to improve skin elasticity.
This randomized, triple-blind, placebo-controlled study evaluated Future studies should consider investigating the potential syner-
the efficacy and safety of Vinh Wellness Collagen on face wrin- gistic effects of VWC with other skin enhancing nutrients.
kles, elasticity, and self-reported improvements in skin health. This is the first study to report a differential response between
Participants in the VWC group reported greater improvements in right and left facial areas with collagen supplementation. As the abso-
overall in skin elasticity, hydration, radiance, firmness, and wrinkle lute wrinkle score reflects several factors, both photoaging and mel-
score, suggesting that VWC supplementation had a beneficial ef- anoma are generally more prevalent on the left side of the face due
fect on self-perceived skin appearance. After 12 weeks, participants to sun exposure while driving.22 Sleeping habits causing micro-pres-
sure over an extended time have also been suggested to contribute
to aging lines and faster aging on the left side of the face. 23 It may be
that the left cheek was more photoaged and more affected by sleep
lines than the right, making the determination of product efficacy
challenging. This is one possible explanation for the significant results
between VWC and placebo for the right cheek, but only significant
changes from baseline to week 12 for the VWC group for the left.
This suggests that longer usage of the product may result in signif-
icant differences between VWC and placebo on the left side of the
face; however, this warrants further investigation in future studies.
A planned subgroup analysis based on age found that females
F I G U R E 4 Gross cheek skin elasticity after supplementation
between 45 and 54 years supplemented with VWC had significant
with VWC for participants 45-54 years of age (n = 12), assessed
improvements in skin elasticity at week 12. Compared to baseline,
by the Cutometer®. Higher values indicate more elastic skin. +
Within group p-values generated by the Wilcoxon signed rank test. these women had 20% and 10% improvements in the cheek skin gross
Probability values P ≤ .05 are statistically significant elasticity at weeks 6 and 12, respectively. There were no significant
832 | EVANS et al.

TA B L E 3 Hematology and clinical chemistry analysis at screening and week 12 for participants in the safety population

VWC Placebo Between Group

Mean ± SD (n) Mean ± SD (n) P-value

Hemoglobin Concentration (g/L)

Screening 131.5 ± 7.4 (25) 132.3 ± 6.9 (25) .696a
Week 12 129.9 ± 8.7 (23) 131.1 ± 8.2 (24) .636a
Hematocrit (L/L)
Screening 0.39 ± 0.02 (25) 0.39 ± 0.02 (25) .660a
Week 12 0.39 ± 0.02 (23) 0.39 ± 0.02 (24) .486a
White Blood Cell Count (×109/L)
Screening 5.91 ± 1.50 (25) 6.10 ± 1.91 (25) .688a
Week 12 5.66 ± 1.56 (23) 6.08 ± 1.62 (24) .373a
Red Blood Cell Count (×E12/L)
Screening 4.36 ± 0.22 (25)) 4.45 ± 0.37 (25) .289a
Week 12 4.33 ± 0.29 (23) 4.44 ± 0.37 (24) .261a
Mean Corpuscular Volume (fL)
Screening 90.1 ± 3.3 (25) 89.1 ± 5.8 (25) .459a
Week 12 90.0 ± 3.8 (23) 89.2 ± 5.3 (24) .560a
Mean Corpuscular Hemoglobin (pg)
Screening 30.16 ± 1.15 (25) 29.83 ± 1.97 (25) .466a
Week 12 30.04 ± 1.48 (23) 29.63 ± 2.07 (24) .445a
Mean Corpuscular Hemoglobin Concentration (g/L)
Screening 335.1 ± 5.1 (25) 335.1 ± 6.5 (25) 1.000a
Week 12 334.1 ± 5.9 (23) 332.2 ± 7.6 (24) .341a
Red Cell Distribution Width (%)
Screening 13.39 ± 0.54 (25) 13.36 ± 0.64 (25) .867a
Week 12 13.37 ± 0.50 (23) 13.48 ± 0.67 (24) .545a
Platelet Count (×109/L)
Screening 264 ± 54 (25) 257 ± 46 (25) .618a
Week 12 257 ± 58 (23) 256 ± 45 (24) .956a
9
Neutrophil Count (×10 /L)
Screening 3.41 ± 1.09 (25) 3.49 ± 1.39 (25) .813a
Week 12 3.18 ± 1.03 (23) 3.33 ± 1.09 (24) .638a
Lymphocyte Count (×109/L)
Screening 1.81 ± 0.57 (25) 1.92 ± 0.78 (25) .636a,c
Week 12 1.75 ± 0.63 (23) 2.08 ± 0.80 (24) .120a,c
9
Monocyte Count (×10 /L)
Screening 0.504 ± 0.149 (25) 0.472 ± 0.159 (25) .466a
Week 12 0.522 ± 0.124 (23) 0.483 ± 0.143 (24) .333a
Eosinophil Count (×109/L)
Screening 0.148 ± 0.145 (25) 0.172 ± 0.209 (25) .558b
Week 12 0.174 ± 0.163 (23) 0.167 ± 0.087 (24) .443b
9
Basophil Count (×10 /L)
Screening 0.020 ± 0.041 (25) 0.012 ± 0.033 (25) .454b
Week 12 0.022 ± 0.042 (23) 0.017 ± 0.038 (24) .673b
Creatinine Concentration (μmol/L)
Screening 72.4 ± 12.6 (25) 71.5 ± 10.0 (25) .766a

(Continues)
EVANS et al. | 833

TA B L E 3 (Continued)

VWC Placebo Between Group

Mean ± SD (n) Mean ± SD (n) P-value

Week 12 75.5 ± 12.0 (23) 73.2 ± 10.7 (24) .505a

Sodium Concentration (mmol/L)
Screening 141.60 ± 2.06 (25) 142.28 ± 2.07 (25) .250a
Week 12 141.39 ± 2.48 (23) 141.88 ± 2.46 (24) .505a
Potassium Concentration (mmol/L)
Screening 4.60 ± 0.46 (25) 4.65 ± 0.49 (25) .701a
Week 12 4.66 ± 0.35 (23) 4.76 ± 0.53 (24) .443a
Chloride Concentration (mmol/L)
Screening 101.92 ± 2.08 (25) 101.88 ± 2.30 (25) .949a
Week 12 102.22 ± 2.24 (23) 101.92 ± 2.00 (24) .629a
Bilirubin Concentration (μmol/L)
Screening 6.3 ± 3.1 (25) 7.3 ± 4.1 (25) .583a,c
Week 12 7.04 ± 2.82 (23) 6.62 ± 2.50 (24) .679a,c
2
Estimated Glomerular Filtration Rate(mL/min/1.73m )
Screening 83.0 ± 14.5 (25) 82.8 ± 12.7 (25) .975a
Week 12 79.0 ± 14.2 (23) 81.2 ± 14.0 (24) .593a
Aspartate Transaminase (U/L)
Screening 19.2 ± 4.7 (25) 20.6 ± 4.7 (25) .315a,c
Week 12 19.5 ± 6.0 (23) 19.7 ± 5.0 (24) .733a,c
Alanine Transaminase (U/L)
Screening 18.6 ± 6.4 (25) 19.8 ± 7.1 (25) .556a,c
Week 12 18.9 ± 10.0 (23) 18.6 ± 8.5 (24) .890a,c

Note: Probability values P ≤ .05 are statistically significant.

Abbreviaions: n, number; SD, standard deviation.
a
Between-group comparisons were made using the independent Student's t test.
b
Between-group comparisons were made using the Mann-Whitney U test.
c
Logarithmic transformation was required to achieve normality.

differences observed in women between the ages of 55 and 60 years. the confounding factor of hysteresis. The test-re-rest reliability of
These results are in contrast to a previous study demonstrating im- skin elasticity measurements while avoiding cofounding has been
provements in skin elasticity were more pronounced in women over described in a previous study. 24 Given this information, a poten-
the age of 50 compared to women under 50 years old.12 However, tial solution would be to perform baseline measurements of cheek
these conflicting results may be due to differences in the location of skin elasticity on both cheeks, such that any measurement out of
measurement, dosage used, and the population studied. One possi- range can be performed on the other cheek without hindering the
ble explanation to glean from this observation is that oral collagen statistical significance of the results. Recognizing that it may not
supplementation may affect distinct areas of the body differently be possible to use the values from one cheek to replace another,
and could have greater efficacy on certain body regions. A larger previous studies have included a rest period of 45 minutes between
sample size powered based on the findings of this study, particularly repeated measurements in order to avoid consequences of hyster-
within the 45-54 age group, should be considered in future studies. esis. 24 Further, repeated measurements, up to three times, 25 would
A limitation of measurements and specifically re-measurements provide a mean score and potentially accommodate out of range or
of any viscoelastic property is the phenomenon of hysteresis. As missing values. Future work should incorporate these methods to
such, repeated measurements of viscoelastic material are discour- reduce missing data.
aged as they cannot account for the inability of the property (ie, skin)
to return to its prestrained state prior to the next measurement cycle
and the impression is then carried forward into subsequent mea- 5 | CO N C LU S I O N
surements. In the current study, 9 Cutometer ® measurements for
gross elasticity were either out of range or missing. Unfortunately, In conclusion, participants supplemented for 12 weeks with Vinh
these measurements could not be repeated without introducing Wellness Collagen showed improvements in wrinkle scores on
834 | EVANS et al.

both sides of the face, cheek skin hydration and self-reported elas- 12. Proksch E, Schunck M, Zague V, Segger D, Degwert J, Oesser S.
Oral intake of specific bioactive collagen peptides reduces skin
ticity, hydration, radiance, firmness, and wrinkle scores. Overall,
wrinkles and increases dermal matrix synthesis. Skin Pharmacol
the current study demonstrates that VWC was safe and well tol- Physiol. 2014;27(3):113-119.
erated in a healthy female population as a similar number of ad- 13. Choi SY, Ko EJ, Lee YH, et al. Effects of collagen tripeptide sup-
verse events were reported between groups; none were classified plement on skin properties: A prospective, randomized, controlled
study. J Cosmet Laser Ther. 2014;16(3):132-137.
as probably related or related to VWC and all blood safety pa-
14. Luc Duteil CQR, Bruno-Bonnet C, Lacour JP. Effect of low dose
rameters for the complete blood count, electrolytes; and liver and type I fish collagen peptides combined or not with silicon on skin
kidney markers were within normal clinical ranges. Future stud- aging signs in mature women. JOJ Case Stud. 2018;6(4):001–005.
ies are needed to examine potential synergistic effects of VWC 15. Koizumi S, Inoue N, Shimizu M, Kwon C-J, Kim H-Y, Park KS. Effects
with other skin enhancing nutrients and the mechanism of action of Dietary Supplementation with Fish Scales-Derived Collagen
Peptides on Skin Parameters and Condition: A Randomized,
associated with collagen supplementation and improvements of
Placebo-Controlled, Double-Blind Study. Int J Peptide Res Ther.
skin health. The results of this study support the use of fish-de- 2018;24(3):397-402.
rived hydrolyzed collagen for the improvement of skin health in 16. Sumida E, Hiorata A, Kuwaba K. The Effect of Oral Ingestion of
an aging population. Collagen Peptide on Skin Hydration and Biochemical Data of Blood.
J Nutr Food. 2004;7:45-52.
17. Gold MH, Biron JA. Safety and Cosmetic Effects of Photodynamic
DATA AVA I L A B I L I T Y S TAT E M E N T Therapy using Hexyl Aminolevulinate and Intense Pulsed Light: A
Data available on request from the authors. Pilot Study Conducted in Subjects with Mild-to-moderate Facial
Photodamage. J Clin Aesthet Dermatol. 2013;6(10):27-31.
18. Zhou S-L, Wang H-Y, Yue D-X. Clinical Effects and Safety of Oral
ORCID
Treatment with Low-Molecular Fish Collagen Hydrolysate on
Malkanthi Evans https://orcid.org/0000-0003-0832-677X Female Facial Skin Properties. J Pract Dermatol. 2011;4(3).
Erin D. Lewis https://orcid.org/0000-0002-1760-3073 19. Bonnet D.Introduction to Naticol® marine collagen peptides for
anti-aging and overview of clinical studies. https://pdfs.seman​ticsc​
holar.org/fed1/fa27d​e3676​b 4626​3 43d3​5 ea1d​1567c ​a 3f216.pdf
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