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ORIGINAL ARTICLE |
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| Year : 2022 | Volume
: 67
| Issue : 6 | Page : 667-673 |
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| Development of novel topical anti-acne cream containing postbiotics for mild-to-moderate acne: An observational study to evaluate its efficacy |
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Hongchang Cui, Congrui Feng, Chaoqun Guo, Zhi Duan
From the Qingdao Vland Biotech Group Co., Ltd. 596-1 Jiushui East Road, Laoshan District, Qingdao, Shandong Province, China
| Date of Web Publication | 23-Feb-2023 |
Correspondence Address:
Zhi Duan
596-1 Jiushui East Road, Laoshan District, Qingdao, Shandong Province
China
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/ijd.ijd_655_22
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 Abstract | | |
Background: Acne is one of the most common skin diseases in adolescents and results in high healthcare costs and psychological burdens severely affecting individuals. Treatments other than contraceptives, antibiotics, and photodynamic therapies are needed to prevent and/or improve acne's onset and evolution. Aims: The purpose of this study was to investigate the efficacy of a fermentation lysate of Lactiplantibacillus plantarum VHProbi® V22 in ameliorating acne. Methods: An anti-acne skincare cream containing fermentation culture lysate was applied to subjects with mild-to-moderate acne vulgaris for 4 weeks. The assessments were evaluated based on instrumental measurements using Visia®-CR and CK-MPA® systems. Results and Conclusions: The anti-acne skincare cream was found to be safe and not cause any irritation. Significant improvements in the acne lesion proportion (P < 0.01), transepidermal water loss (P < 0.001), and sebum secretion (P < 0.05) were observed in comparison to the baseline in the subjects. The analysis of the statistical data after 4 weeks of treatment showed a positive decrease in skin tone, stratum corneum hydration, and superficial pH without statistical significance, compared with the baseline. The results of this study suggest that the topical application of the anti-acne skincare cream was effective and safe in subjects with mild-to-moderate acne and could represent an optional complement for acne treatments.
Keywords: Acne, Cutibacterium acnes, Lactiplantibacillus plantarum, probiotics, postbiotic
How to cite this article:
Cui H, Feng C, Guo C, Duan Z. Development of novel topical anti-acne cream containing postbiotics for mild-to-moderate acne: An observational study to evaluate its efficacy. Indian J Dermatol 2022;67:667-73 |
How to cite this URL:
Cui H, Feng C, Guo C, Duan Z. Development of novel topical anti-acne cream containing postbiotics for mild-to-moderate acne: An observational study to evaluate its efficacy. Indian J Dermatol [serial online] 2022 [cited 2023 Mar 23];67:667-73. Available from: https://www.e-ijd.org/text.asp?2022/67/6/667/370335 |
| Introduction | |  |
Acne disorders is a kind of chronic inflammatory disease of the sebaceous gland caused by an androgen-induced increase in sebum production, altered keratinization, and the bacterial colonization of hair follicles on the neck, chest, face, and back by Cutibacterium acnes (C. acnes, formerly Propionibacterium acnes) or Staphylococcus epidermidis.[1] According to Goodarzi, acne is one of the 10 most prevalent disorders in the world, affecting 47–90% of adolescents 14 years of age and older.[2] The psychological side effects of acne are associated with an increase in the risk of anxiety, insomnia, depression, hyperactivity disorders, and attention deficits.[3] Topical agents such as antibiotics, benzoyl peroxide, and retinoids are usually used individually or in combination to control mild-to-moderate acne vulgaris. However, antimicrobial resistance is a challenge to topical macrolides reported in over 50% of the C. acnes strains, which has decreased their effectiveness.[4–6] Benzoyl peroxide is an effective over-the-counter preparation but may cause initial local irritation.[7] Moreover, given the need for long-term therapeutics, the side effects and difficulties in adhering to treatments, promising and safe therapeutics with fewer or no complications are needed to improve facial skin health and well-being in these individuals.
As defined by a joint Food and Agriculture Organization of the United Nations/World Health Organization (FAO/WHO) expert consultation on the health and nutritional properties of probiotics in food, probiotics are live microorganisms, which, when consumed in adequate amounts, can confer a health benefit on the host.[8] The wide application range for probiotics, involving gastrointestinal functions, has been extensively elucidated during the last few decades. Investigating the beneficial effects of probiotics on skin disease in atopic dermatitis, eczema, skin aging, and bacterial and fungal infections has yielded many promising results.[9–12] The administration of probiotics has proven beneficial when addressing antibiotic resistance and inhibiting inflammation in serious cases.[13] However, the application of live bacteria on the skin poses several challenges,[14] and skincare products containing probiotics are generally illegal in China. The term “postbiotics” was defined as the “preparation of inanimate microorganisms and/or their components that confers a health benefit on the host” by the International Scientific Association of Probiotics and Prebiotics in 2019.[15] Effective postbiotics contain inactive microbial cells or cell components that contribute to observed health benefits.
In our previous study, we isolated a bacterial strain named Lactiplantibacillus plantarum VHProbi® V22 from fecal samples of infants, who have not consumed probiotic dietary supplements within 3 months. Here, we conducted a trial to determine whether a skincare cream containing a bacterial lysate of L. plantarum VHProbi® V22 could reduce the severity in subjects with mild-to-moderate acne.
| Materials and Methods | | |
In vitro studies
Bacterial strains
C. acnes ATCC 11827 and ATCC 6919 were purchased from American Type Culture Collection, (Manassas, VA, USA). C. acnes ATCC 11827 and ATCC 6919 were maintained on a Reinforced Clostridial Medium (RCM, cat. no.: HB0316, Hope Bio-Technology Co., Ltd) and incubated in an anaerobic jar (Sigma-Aldrich®) with an oxygen absorber CO2 generator (AnaeroPack C-1, MGC®, Japan) at 37°C for 48 h.
Antimicrobial activity of L. plantarum VHProbi® V22
The inhibitory activity of L. plantarum VHProbi® V22 against C.acnes ATCC 11827 and ATCC 6919 was evaluated using a double-layer agar plate assay and agglutination test, as described previously with some modifications.[16] Briefly, bacterial spots were inoculated with 100 μL of test bacterium culture (109 CFU/mL) in triplicate plus a blank on the RCM agar surface in a Petri dish and incubated at 37°C for 48 h. Then, the plate was overlaid with 8 mL of RCM soft agar (0.75%, 1% Tween 80) which had been inoculated with 106 CFU/mL of mixed cultures of C. acnes ATCC 11827 and ATCC 6919 and incubated at 37°C for 72 h. The plate was examined for the presence or absence of an inhibitory halo around the spots. In the agglutination test, 300 μL of the test bacterium culture (109C FU/mL) was co-incubated with 300 μL of mixed cultures of C. acnes ATCC 11827 and ATCC 6919 (106 CFU/mL) in 24-well plates at 37°C in a thermostatic oscillation incubator (HUXI®, HW-400TG). At the same time, the test bacterium culture was co-incubated with Tris-HCI buffer (Sigma) and used as negative control simultaneously. After 24 h, the plates were then checked for the presence or absence of tiny white floccules.
Bacterial lysate
The starter cultures of L. plantarum VHProbi® V22 were inoculated in a de-Man-Rogosa-Sharpe (MRS) broth (Cat CM187, BEIJING LANDBRIDGE) and incubated under anaerobic conditions (AnaeroPack C-1, MGC®, Japan) at 37°C for 72 h. Afterward, the seed cultures were inoculated into a fermentation medium with 1% (V/V) and incubated under anaerobic conditions at 37°C for 24 h. The fermentation medium contained 3% brown sugar, 3% collagen, 0.3% yeast extract, 0.25% (NH4) 2HPO4, and 1 L of distilled water. The fermentation cultures were processed using a homogenizer device (SPXFLOW, APV-1000) under 1000 bar 3 times; then, the lysate was harvested and used as a functional ingredient. This ingredient has the following International Nomenclature Cosmetic Ingredient code: V22 ferment lysate.
Anti-acne skincare cream
The weight percentage of contents in the anti-acne skincare cream were as follows: 8% V22 ferment lysate, 0.05% EDTA-2Na, 1% polydimethylsiloxane, 3% caprylic, 2.5% isohexadecane, 4% glycerin, 0.8% butyrospermum parkii, 0.6% sorbitan olivate, 0.4% cetearyl alcohol, 0.4% 1,2-hexanediol, 1.5% cetearyl olivate, 0.3% hyroxyacetophenone, 0.35% polyacrylate crosspolymer-6, and 0.2% phenoxyethanol.
Safety assessments
The safety assessments were performed based on the Safety and Technical Standards for Cosmetics (2015 version) issued by the National Medicine Product Administration.[17] The human skin patch tests and acute dermal toxicity tests were commissioned by the Pony Test International Group. The microbial tests, physical and chemical inspections, and toxicological tests were commissioned by the Health Analysis and Test Center of Nanjing Medical University.
Study population
A total of 20 volunteers of both genders aged 18–25 years were enrolled in this study between April 2021 and July 2021. The demographic information of all the subjects was provided in [Table S1]. Only subjects with mild-to-moderate acne vulgaris, with a score of 2 or 3 according to the global acne assessment scale [Table 1] were included.[18] All subjects gave their informed consent for inclusion before they participated in the study. The authors confirm that the ethical policies of the journal, as noted on the journal's author guidelines page, have been adhered to. No ethical approval was required as the cosmetic products involved in this study have passed the toxicological tests. The main exclusion criteria included pregnant or breastfeeding women, the current use of cosmetic products or having had any facial cosmetic procedures for acne, or any known diseases that could have interfered with the clinical assessments.[19]
The global acne assessment scale was used to categorize the subjects as having mild, moderate, and severe acne vulgaris in the current study. To qualify for this study, the subjects were required to have mild-to-moderate acne vulgaris as determined by the investigative staff (defined as a score of 2 or 3).
Study design and outcome measurements
All the subjects were treated with the anti-acne skincare cream twice daily (morning and night) on their forehead, cheek, nose, and jaw after cleansing the face for 4 weeks. Clinical evaluation was performed and digital photographs were taken at baseline and during follow-up visits at 1, 2, 3, and 4 weeks from the initiation of treatment using the Visia®-CR system (Canfield Scientific Inc). The clinical assessments included the determination of the acne lesion proportion and skin tone. The stratum corneum hydration (SCH), transepidermal water loss (TEWL), sebum production, and skin surface pH were measured using the CK-MPA® system (CK-MPA10, Courage + Khazaka electronic GmbH) at baseline and during follow-up visits at 2 and 4 weeks. The subjects were asked to rest for 20 min before the tests in an environment with a temperature of 21°C ± 1°C and a relative humidity of 50% ± 10%.
Data analysis
The Shapiro–Wilk test was used to assess the normality of data distribution; Single-factor repeated measures analysis of variance was used for efficacy assessment from baseline to different visit times. All the statistical analyses were performed using Statistical Package for Social Sciences ver. 20 (SPSS Inc., Chicago, IL, USA). P < 0.05 was considered to indicate statistical significance. The analysis also included comparing the improvement rates between the baseline and each follow-up visit by using the formula below
% IRs = (Wi−W0)/W0×100%
Where Wi is the assessment value at week i (i = 1, 2, 3, and 4) and W0 is the assessment value at baseline.
| Results | | |
In vitro microbial study
The double-layer plate assay showed a clear halo around the spots in the plate with a diameter of 26.33 ± 0.58 mm [Figure 1]a. This indicated that the bacterial strain L. plantarum VHProbi® V22 can inhibit the growth of C. acnes. In the agglutination assay, the L. plantarum VHProbi® V22 did not show self-agglutination after 72 h of incubation [[Figure 1]b, left]. The first time of observation of a tiny white floccule was at 18 h of incubation [[Figure 1]b, right]. | Figure 1: Microbial activity of investigated strain. (a) Double-layer plate assay: control: blank; experimental group: 1, 2, and 3. (b) Agglutination test: left, blank; right, experimental group
Click here to view |
Safety assessments
The microbial test showed that the total numbers of microbial colonies, mycetes and yeasts, thermotolerant coliforms, Staphylococcus aureus, and Pseudomonas aeruginosa in the anti-acne skincare cream were within the ranges of normal values. The contents of heavy metals including Hg, Pb, As, and Cr were below the detection limits. A toxicological test was conducted on the New Zealand rabbits according to the skin irritation test protocol. No irritating effect occurred in the skin of the rabbits' back. An acute dermal toxicity test result showed that the median lethal dose (LD50) in rats exceeded 2180 mg/kg body weight. The anti-acne skincare cream can be classified as being of a low poison grade. None of the 30 volunteers showed reactions with a value of “1” or greater in the human skin patch test. Overall, the anti-acne skincare cream is mild, causes no irritation, and shows favorable safety.
Acne lesion proportion
The statistical results for the determination of the lesion proportions from baseline to each follow-up visit are presented in [Table S2]. In week 3 and week 4, there was a statistically significant decrease in the proportion of acne lesions from baseline (P < 0.05 and P < 0.05, respectively). [Figure 2]a shows the changes in the lesion proportion over the 4-week study period. At week 4, 70.0% of the subjects showed decreases in the proportion of superficial lesions; 78.6% of them showed a 50% reduction in the lesion proportion. Representative photographic evaluations of the subjects' improvements in the lesion proportion at baseline and each follow-up visit are shown in [Figure 3].
 | Figure 2: Clinical assessments of subjects at different times. (a) Acne lesion proportion, (b) Skin tone, (c) Stratum corneum hydration (SCH), (d) Transepidermal water loss (TEWL), (e) Sebum content, and (f) pH. W0: week 0, W1: week1; W2: week 2; W3: week 3; W4: week 4
Click here to view |
 | Figure 3: Representative photographic evaluation of the improvement in the subjects in acne lesion. W0: week 0; W1: week 1; W2: week 2; W3: week 3; W4: week 4
Click here to view |
Skin tone
The statistical results of the skin tone from baseline to each follow-up visit are presented in [Table S3]. At each follow-up visit, there was no statistically meaningful difference from baseline to each follow-up visit [see [Figure 2]b].
Stratum corneum hydration
The statistical results of the SCH from baseline to each follow-up visit are presented in [Table S4]. Although 35% of the subjects demonstrated an increase in the SCH at week 4, they did not show a statistically significant increase from baseline to each follow-up visit [see [Figure 2]c].
Transepidermal water loss
The statistical results of the TEWL from baseline to each follow-up visit are presented in [Table S5]. At each follow-up visit, there was a statistically significant decrease in the TEWL from baseline (P < 0.05 and P < 0.001, respectively) [see [Figure 2]d]. At week 4, 85% of the subjects showed a decrease in the TEWL, 35.3% of these subjects showed a 40% reduction.
Sebum production
The statistical results of sebum production from baseline to each follow-up visit are presented in [Table S6]. At each follow-up visit, subjects showed a significant decrease in the sebum production compared with baseline (P < 0.05, P < 0.01, respectively) [See [Figure 2]e]. At week 4, 80% of the subjects achieved a decrease from baseline in the sebum production.
Skin surface pH
The statistical results for the changes in skin surface pH from baseline to each follow-up visit were presented in [Table S7]. At each follow-up visit, no statistically significant difference in the skin surface pH compared with baseline was observed [see [Figure 2]f]. However, 55% of the subjects showed a decrease in the skin surface pH.
| Discussion | | |
Acne is a multifactorial inflammatory disease, generally known as a disorder affecting teenagers during puberty, which has both physical and psychological side effects impacting their health.[20] According to Kanlayavattanakul et al., the availability, cost and side effects limit the use of conventional therapeutics for the management of acne. Thus, alternative therapeutics with minimal risks and the holistic management of the problem are in high demand.[21],[22]
In recent years, the characterization of specific probiotic strains used for preventing and treating skin disorders has progressed significantly, especially as there is growing evidence in support of probiotics as a treatment for acne.[23] Such beneficial health effects are believed to stem from their antimicrobial properties and their ability to maintain hydration and barrier functions.[24–29] For example, acne lesion was associated with an increased proportion of C. acnes as a skin commensal bacterium; probiotics can inhibit C. acnes due to their antibacterial proteins and bacteriocin-like inhibitory substances.[30],[31]
Kang et al. conducted a clinical study of a lotion that has demonstrated that bacteriocin produced by Enterococcus faecalis SL-5 could significantly ameliorate acne lesions or pustules in patients with mild-to-moderate acne lesions caused by C. acnes compared with placebo.[32] Majeed M et al. conducted a clinical study of cream containing extracellular metabolites secreted by Bacillus coagulans MTCC 5856 as a postbiotic antimicrobial treatment for acne, which demonstrated a significant reduction in noninflammatory lesions in both open and closed comedones and an improvement in inflammatory signs such as redness and skin elevation.[33] In the present study, we used a bacterial lysate homogenized by L. plantarum VHProbi® V22 fermentation cultures as a postbiotic functional ingredient for acne treatment. Firstly, the strain L. plantarum VHProbi® V22 exhibited a strongly inhabitation against two strains of C. acnes in vitro study. This may play a fundamental role for its effecting in ameliorating the severity of acne. Toxicological test and acute dermal toxicity test confirmed the investigational products can be uesd for human trial. The anti-acne skincare cream met the requirements of the Safety and Technical Standards of Cosmetics and was, deemed, overall safe. In the intervention scenario, the anti-acne skincare cream significantly reduced the acne lesions proportions and sebum secretion throughout the treatment period and improved skin barrier integrity such as TEWL. Although some indicators including the SCH and superficial pH did not show a significant difference, a positive trend was observed after treatment.
This study has some limitations, such as the relatively small population, the use of a single site, the short duration of the intervention, and low evidence level because of the before-after study in the same subjects. Large randomized, placebo-control trials are needed to further evaluate this anti-acne cream. Postbiotics are soluble, bioactive, nonliving microbial cell components that have many practical advantages over probiotics, as they show bioactivity in a nonviable state and are easier to formulate due to their favorable physicochemical properties.[34] In China, cosmetic products containing live probiotics are prohibited by legislation due to unpredictable risks. Hence, postbiotics may have promising applications as a novel acne-control ingredient in skincare products.
| Conclusion | | |
The results from this study suggest that the topical treatment with an anti-acne skincare cream containing V22 ferment lysate in subjects with mild-to-moderate acne is safe, well-tolerated and effective after 4 weeks of treatment. Thus, postbiotics produced by L. plantarum VHProbi® V22 are highly suitable for treating subjects with mild-to-moderate acne vulgaris. Further large-scale population studies are needed to substantiate our findings.
Statements
This study strictly complied with the declaration of Helsinki. All of the subjects recruited in this study have provided informed written consent and got appropriate medical protection measures. The protocol was in accordance with the safety and technical standards of cosmetics issued by the national medical products administration.
Acknowledgements
The authors would like to thank the trial investigators at Jiangnan University.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3]
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