Pre-menopausal dilemma: how to reduce Acne and Aging signs and still maintain a balanced microbiome?

Although acne is usually associated with adolescence, the truth is that this condition can be present throughout life. In women it is specially noted in pre-menopausal and menopausal phases. In this period, female reproductive hormones (estrogen and progesterone) decline while androgen levels remain elevated, decreasing gradually in latter menopause. This hormonal change with relative androgen excess favours acne appearance. In parallel, this is a period in women's life when the appearance of wrinkles, loss hydration and firmness are intensified.1Mature skin with acne is a combination not easy to address. This is because traditional anti-acne treatments usually promote dryness making aging signs more noticeable, whereas anti-aging products tend to be oily, promoting pore-clogging, which worsens acne conditions. All of this makes complicated deciding the most adequate skincare routine, a situation known as the pre-menopausal dilemma.However, if we consider the mechanism behind each condition and focus on reestablishing the functions of the skin that triggers the main cycles, offering the skin the nourishment, cleaning, and support for microbiome equilibrium, it is possible to reach the right balance.Acne is a condition of the pilosebaceous unit that starts with an excessive sebum production. This leads to the proliferation of pathogenic microorganisms causing a microbiome dysbiosis. The microorganisms break down sebum, producing inflammatory metabolites that impair the barrier function. This, alongside the hyperkeratinisation of the pilosebaceous unit results in acne breakouts and inflammation, leading to skin imperfections that also intensify the aging signs (Figure 1).2Figure 1Currently, the role of the microbiome on the skin's health is in the forefront of several research groups. The traditional belief that acne was caused due to a proliferation of Cutibacterium Acnes was challenged in more recent studies that report the commensal behaviour of C. Acnes in sebum-regulated skin. In 2019 Su et. al. reported that the microbiome of healthy skin is composed of around 50-60% of C. Acnes.3 Brillet et. al. reported in 2021 the role of C. Acnes in maintaining the homeostasis and skin health by producing short chain fatty acids, predominantly propionic acid, through triglycerides hydrolisis.4 Short chain fatty acids are key in the hydrolipidic mantle, regulating the skin's pH. This is very important in aging and acne skin as increased pH has been observed, among other conditions. 5On the other hand, when metabolizing sebum C. Acnes produces porphyrins that are pro-inflammatory metabolites. In 2019 Maia et. al. showed a direct correlation between the amount of sebum on the skin and porphyrin concentration6, which seems to indicate that C. Acnes behaves differently according to the environment, and that the sebum content is key to its commensal or pathogenic behaviour.Another species present in the skin's microbiome is Staphylococcus Epidermis, which has been proven to have an important role on the induction of inflammation-related markers.7 This is in line reports show that the concentration of Staphylococcus increases with acne severity.8All of this suggests that in order to restore the microbiome equilibrium of mature oily skin, we should pay close attention to the levels of sebum and the presence of these bacteria in the skin.Classic approaches to treat acne include the combination of aggressive molecules such benzoyl peroxide and topical antibiotics. However, the use of these substances commonly worsen skin inflammation and dryness and may cause bacterial resistance.9 A more modern and safe approach consist in feeding the skin commensal microbes with postbiotics, which would naturally promote homeostasis. 10Ageing oily skin also needs to be carefully hydrated and structured to recoup its barrier function, and a grand finale can be obtained with a gentle cell renewal, so dead cells are removed, acne lesions softened and skin per se rejuvenates.In terms of formulation, a fine equilibrium has to be strategically addressed in order to promote oil control and still improve ageing signs and balance the microbiome.Bicosome® is a double encapsulation technology that mimics skin lamellar bodies in composition, structure, and function. It is a dual system composed of two types of lipid structures: the internal smart nanodisks (about 20 nm, white arrows in Figure 2) and the external vesicle (black arrow in Figure 2) that encapsulates the disks, in structures ranging from 250 to 500 nm in total size.11 With his technology it is possible to encapsulate molecules in different compartments and target deliver them in adequate skin sites, allowing for the development of multi-active delivery systems.It appear 2In this paper, we describe the development of Bicobiome®. This Bicosome® system was specially designed for mature skin prone to acne as an anti-ageing system with oil control. The Bicobiome® strategy focuses on re-stablishing the skin environment so that the natural homeostasis can be restored by the skin's own natural processes. In addition, Bicobiome® nourishes the skin with compounds that hydrate, restore the skin barrier and promote cell renewal, giving youth back to the skin.Experimental designProduct designBicobiome focuses on four pillars: sebum regulation, microbiome re-balance, restoring the barrier function and cell renewal. This multi-active skin delivery system includes a combination of sebostatic, anti-inflammatory, post-biotic and keratolytic agents that are targeted into different skin layers, thanks to the Bicosome® Technology.The sebostatic, anti-inflammatory and moisturising agent Niacinamide is target delivered in the Epidermis and follicle promoting sebum regulation, reducing inflammation and reinforcing skin hydration. The postbiotic, Lactobacillus ferment, is delivered into the first layers of the stratum corneum to favour the growth of commensal microorganisms and re-balance the microbiome. This active is also known for its soothing and healing benefits. The skin lipid structure is reinforced by the Bicosome® disks per se that fill up the lipid matrix spaces with organised bilayers, strengthening the skin barrier. Finally, the Salicylic acid is delivered in the skin surface to promote a mild desquamation, cell renewal, preventing follicle hyperkeratinisation and softening acne lesions, wrinkles and other skin imperfections.The key for the success of this multi-active technology as an anti-ageing system with oil and acne control is the capacity of Bicosomes® to incorporate the molecules in adequate compartments within its structure, preserving their physicochemical characteristic, their stability and to deliver each active in the right levels of the skin, where they need to work, boosting their efficacy and their synergistic effect.The efficacy of the Bicobiome® system in mature skin prone to acne is presented in our clinical study.MethodsTo evaluate the efficacy of Bicobiome® an in vivo study has been carried out. The study involved 20 female volunteers over 35 years old with oily skin, mild-medium grade acne and signs of aging. The volunteers applied the Bicobiome® 1% serum for 56 days twice daily on clean and dry skin. The measurements were carried out at initial time 0 (control), 14, 28 and 56 days. The improvement of the skin condition was evaluated by monitoring sebum amount, porphyrin presence, microbiome, acne lesions, cell renewal, wrinkles and firmness through different techniques described as follows.Sebum assessmentThe quantity of sebum on the chin area was evaluated using sebum collector foils Sebufix® F16 (CKelectronic, Cologne, Germany). The foils were applied to the chin surface, and the absorbed sebum became visible as transparent spots of various sizes. The number and size of the spots indicated the activity of the sebaceous glands. Then the foils were evaluated with Visioscan® and its software. The sebum area and the number of oily spots were analysed.Porphyrin reductionPorphyrins were assessed using a Visiopor® PP34 featuring a high-resolution sensor and a ring-shaped UV light source for uniform skin illumination of areas with acne lesions. The red-orange fluorescence indicates the presence of porphyrins.Analysis of the skin microbiome:Samples of the skin microbiota of the volunteers were taken before the treatment (Day 0) and at the end of the treatment (Day 56) by using a sterile, dry swab. The DNA from the samples was extracted and used as a template to amplify 7 hypervariable regions of the bacterial rRNA gene.12 These amplicons were used to construct the genomic libraries, which were subsequently templated in an Ion 530™ chip and sequenced by the Ion S5 plus™. The resulting reads were subjected to quality control trimming and classified within the lowest taxonomic range possible using the reference library Curated MicroSEQ ® 16S v2013.1 and the database Curated Greengenes v13.5. The species present on the skin were analysed.Acne assessmentA board-certified dermatologist graded acne of volunteers according to a reported comprehensive acne severity scale (CASS) developed by Tan et al13 which classifies lesions from clear (0) to very severe (5).Cell renewalCell renewal was assessed by the skin desquamation index on the cheek. The number and size of corneocytes were measured on the initial day, 2 hours after application, after 14 and 56 days of product use. The tape Corneofix® F20 plus (CKelectronic, Cologne, Germany)) with collected corneocytes was placed on the Visioscan® VC 20 plus (CKelectronic, Cologne, Germany) probe which takes an image and processes it with a determined colour grading, depending on the desquamation level. The colour scale ranges from cool to warm colours. The software also analyses the number, size, and thickness of the attached corneocytes to the Corneofix® tape and gives the desquamation index (DI).Wrinkles reductionThe skin topography was measured by image analysis, with Visioscan VC 20 plus (Courage & Khazaka electronic). The principle for measuring wrinkles is the grey scale photographic analysis of skin topography. The basis for the calculation is the grey distribution histogram. Wrinkles were measured in the forehead, between eyebrows and in the nasolabial folds.Firmness improvementR0 values were obtained using a Cutomer® dual MPA 580. The elasticity of the skin surface is measured with suction in the cheekbone (negative pressure 450 mbar). In the measurement 10 cycles, with two seconds of suction and two seconds of relaxation per cycle were performed to tire the skin. The resistance of the skin to suction after 10 cycle is defined by firmness, assessing its ability to return elastically to original position.Results and discussionOil and acne control Sebum assessmentThe acne cycle begins with an increase of sebum production, for this reason when developing a product for acne prone skin, it is necessary to closely follow this parameter, since its decrease will start a cascade of events relevant to the reestablishment of skin equilibrium. Figure 3 shows Sebufix® images of a volunteer in three time points. At the beginning of the study (T0), numerous coloured areas of sebum can be observed. After 56 days the number and sizes of the coloured areas are clearly decreased, indicating a decrease in sebum production of about 22% (Figure 4).Figura 3Niacinamide is a well-known sebum-regulator active with excellent anti-inflammatory and moisturising properties.14 The use of the Bicosome® technology to target deliver it to the follicle, where the sebaceous gland is, maximizes its effect regulating sebum at its source.Figura 4Porphyrin reductionFollowing the cascade, a decrease in sebum will also decrease the production of pro-inflammatory metabolites by the C. Acnes, as mentioned before.6 As Niacinamide is also an anti-inflammatory molecule, this effect is boosted. Indeed, Figure 5 shows images of a volunteer skin before and after treatment with Bicobiome 1% serum. At the beginning of the study the fluorescence in the pores indicate a high porphyrin presence. After 56 days of treatment the fluorescence is clearly reduced indicating the reduction of inflammation.Figure 5The graph of Figure 6 shows the improvement of pro-inflammatory porphyrins by 21% in 56 days compatible with the 22% sebum-regulation observed in Figure 4.Figura 6Skin MicrobiomeThe decrease of sebum and inflammation is clearly reflected in the microbiome population. At day 0 we found the average amount of C Acnes to be below healthy levels, around 37% and high presence of Staphylococcus, around 21%. These values are compatible with inflamed skin, with high oil content.After 56 days of treatment with the Bicobiome 1% serum these species re-balanced (Figure 7). This is explained by the previous results. The decrease of sebum stopped the trigger for C. Acnes to produce porphyrins, returning to its commensal behaviour. The decrease of the pro-inflammatory metabolites downregulated the presence of Staphylococcus spp to around 7%. In addition, the delivery of the postbiotic Lactobacillus ferment to the superficial Stratum Corneum layers promoted the growth of commensal C. Acnes to a healthy 60%.Figure 7Acne assessmentRestoring a healthy skin environment with an adequate sebum regulation, decrease of pro-inflammatory molecules and microbiome rebalance provided the skin with the nourishment needed. Finally, the fourth step was the delivery of Salicylic acid to the skin surface. The action of this molecule was boosted, and its potential irritating effected was reduced by its encapsulation in the Bicosome® structures, so the volunteers could benefit from an effective, mild, and safe desquamation that released dead cells and reduced hyperkeratinisation of the pilosebaceous unit. This had a visible impact on the skin of the volunteers by eliminating acne lesions (Figure 8).It appear 8 After 56 days, the acne severity decreased by 30% (Figure 9).It appear 9Anti-aging benefits Cell renewalThe cell renewal effect of Bicobiome® 1% serum is effective in just 2 hours right after the first application as a soft peeling-like effect of 28% of cell renewal, with no associated irritation. This effect is still intense in the first two weeks, showing 25% of desquamation. This is because at the beginning of the study the Stratum Corneum layer was thickened, with reduced cell renewal and keratinisation of the pilosebaceous unit. As the treatment progressed the cell renewal was optimised resulting in a smoother skin texture. (Figure 10).Figure 10Visioscan® images depicted at Figure 11, evidence the changes in the skin surface. The colour scale evolves from warm (thickened SC) to cool colours (smooth skin texture). The dark blue colours seen in day 56 image denotes a smoother skin surface, more hydrated and uniform.Figure 11Reduction of wrinklesAll Bicosome® actives are anti-ageing agents including the Bicosome® lipid structures. Therefore, it is not surprising that the use of this multi-active complex also impacts the appearance of wrinkles (Figure 12). It is well known that Niacinamide boosts skin hydration and barrier function14, and that Lactobacillus ferment has protective and soothing effects15. The Bicosome® nano disks have a filling-up effect in the stratum corneum layer, placing new bilayers in the inter-corneocytes spaces; and Salicylic acid promotes cell renewal. But what makes real magic about this product is its capacity to place every component right where they need to work. This not only adds up the effects of the different actives but make them synergic.Figure 12The use of Bicobiome® 1% serum decreased 38% of the wrinkles in between eyebrows, 19% in the nasolabial area 17% in the forehead after 56 days of use (Figure 12). An image of one volunteer with improvement in the forefront is shown on Figure 13It appear 13FirmnessThe recoup of skin equilibrium is ultimately seen with the improvement of firmness. After restoring skin balance, stopping triggers for inflammation, filling the lipid layers, and cleaning up the skin restoring cell renewal the skin defensive functions can finally "relax", and the biological processes of the deeper layers return optimised. The result is a firmer skin, moisturised and with a more homogeneous surface as seen in Figure 14.It appear 14The firmness improvement was quantified to be 16% in 28 days and 18% in 56 days (Figure 15).It appear 15ConclusionBicobiome® multi-active delivery system proved to be effective to promote oil and acne control, improve ageing signs and balance the microbiome, therefore constituting a promising tool to the pre-menopausal skincare dilemma. The key aspects for the success of this new strategy are the focus on re-stabilising a healthy sebum-regulated environment while providing nourishing molecules, so the skin could auto-regulate and restore its biological processes naturally in a positive cascade back to wellness.

Pre-menopausal dilemma: how to reduce Acne and Aging signs and still maintain a balanced microbiome?

In this article we describe the development of Bicobiome® which was specially designed for mature, acne-prone skin.

Lucyanna Barbosa

CEO And Co-Founder of BICOSOME