Stratum Corneum Barrier Function Research Articles

A Stratum corneum lipid model as a platform for biophysical profiling of bioactive chemical interactions at the skin level

Stratum Corneum (SC), the outermost layer of mammalian skin, provides the first and most challenging barrier to skin permeation of cosmetic and pharmaceutical compounds. SC barrier function relies primarily on the complex structure and organization of the intercellular lipid matrix. This matrix consists mainly of ceramides (CER), free fatty acids (FFA), and cholesterol (Chol) in an equimolar ratio, forming a multilamellar structure with short- and long-periodicity phases. Along with permeation studies, it becomes paramount the molecular investigation of the interaction between the lipid matrix and the compounds, towards a comprehensive picture of their biophysical impact, including lipid packing impairment, which can result in skin damage, and in consequences for their permeation and mechanism of action. In this study, a lipid-based model (CER[EOS]:CER[AP]:Chol:FFA) was developed to accurately resemble the barrier properties of the intercellular lipid matrix. In a monolayer assembly, this SC lipid model was characterized and applied as an in vitro platform to explore interactions with model compounds – caffeine and testosterone – following OECD recommendations. The SC model successfully mimicked different barrier properties of the SC’s lipid matrix, and its interactions with the model compounds were discussed closely aligned with existing data, including in vitro permeation studies using human and animal membranes. This SC model can be a step forward towards the development of new in vitro platforms for investigating the skin barrier function and the interactions with external molecules. Moreover, the composition of these platforms can be easily modified to mimic impaired lipid matrices to study different skin conditions, providing valuable insights for both cosmetic and pharmaceutical applications.

The multi‐factorial modes of action of urease in the pathogenesis of incontinence associated dermatitis

AbstractBackgroundIncontinence Associated Dermatitis (IAD) is a type of skin inflammation caused by chronic exposure to urine and/or faeces. Current treatment strategies involve creating a barrier between the skin and urine/faeces rather than targeting specific irritants. Urease expressing pathogens catalyse the conversion of urea, present in urine, into ammonia. The accumulation of ammonia causes an elevation in skin pH which is believed to activate faecal enzymes which damage skin, and opportunistic pathogens, which lead to secondary infections.ObjectivesTo develop a better, multi‐factorial model of IAD pathogenesis, including the effect of urease‐expressing bacteria on skin, mechanism of damage of urease and urease‐triggered activity of faecal enzymes and secondary pathogens. To study the effect of urease inhibition on preventing IAD skin damage.MethodsFive separate studies were made using ex vivo porcine skin and in vivo human skin models. Measurements of the change in skin barrier function were made using skin impedance, trans‐epidermal water loss (TEWL), stratum corneum moisture and pH. Skin was exposed to artificial urine, inoculated with various microbes, enzymes and chemicals to examine the influence of: 1) urease‐positive Proteus mirabilis 2) ammonia, 3) combination of P. mirabilis and a faecal enzyme, trypsin, 4) combination of P. mirabilis and opportunistic pathogens, Candida albicans and Staphylococcus aureus, 5) inhibition of urease using acetohydroxamic acid (AHA) on barrier function.ResultsThe urease‐mediated production of ammonia had two principal effects: it elevated skin pH and caused inflammation, leading to significant breakdown in skin (stratum corneum) barrier function. Urease was found to further increase the activity of faecal enzymes and opportunistic pathogens, due to elevated skin pH. The urease inhibitor, AHA, was shown to have significantly reduced damage to skin barrier function, measured as its electrical resistance.ConclusionsTargeted therapeutic strategies should be developed to prevent the manifestation of IAD, rather than creating a generic barrier between skin and urine/faeces. Urease has been identified as a crucial component in the manifestation of IAD, due to its role in the production of ammonia. Urease inhibition provides a promising therapeutic target to halt the progression of IAD.

Effect of Penetration Enhancer on the Structure of Stratum Corneum: On-Site Study by Confocal Polarized Raman Imaging.

Keratin and lipid structures in the stratum corneum (SC) are closely related to the SC barrier function. The application of penetration enhancers (PEs) disrupts the structure of SC, thereby promoting infiltration. To quantify these PE-induced structural changes in SC, we used confocal Raman imaging (CRI) and polarized Raman imaging (PRI) to explore the integrity and continuity of keratin and lipid structures in SC. The results showed that water is the safest PE and that oleic acid (OA), sodium dodecyl sulfate (SDS), and low molecular weight protamine (LMWP) disrupted the ordered structure of keratin, while azone and liposomes had less of an effect on keratin. Azone, OA, and SDS also led to significant changes in lipid structure, while LMWP and liposomes had less of an effect. Establishing this non-invasive and efficient strategy will provide new insights into transdermal drug delivery and skin health management.

Epidermal barrier function in dry, flaky and sensitive skin: A narrative review.

The stratum corneum (SC)-the outermost layer of the epidermis-is the principal permeability and protective barrier of the skin. Different components of the SC, including corneocytes, natural moisturizing factor, a variety of enzymes and their inhibitors, antimicrobial peptides and lipids, work interactively to maintain barrier function. The main barrier properties of the SC are the limitation of water loss and the prevention of infection and contact with potentially harmful exogenous factors. Although the SC functions consistently as a protective barrier throughout the body, variations in functions and morphology occur across body sites with age and skin type. Healthy SC function also depends on the interplay between the chemosensory barrier, the skin's microbiome and the innate immune system. Dysregulation of SC barrier function can lead to the development of skin disorders, such as dry, flaky or sensitive skin, but the complete underlying pathophysiology of these are not fully understood. This review provides insight into the current literature and emerging themes related to epidermal barrier changes that occur in the context of dry, flaky and sensitive skin. Additional studies are needed to further elucidate the underlying aetiology of dry, flaky and sensitive skin and to provide tailored treatment.

Calcitriol, an Active Form of Vitamin D3, Mitigates Skin Barrier Dysfunction in Atopic Dermatitis NC/Nga Mice.

Atopic dermatitis and psoriasis are prevalent chronic inflammatory skin diseases that are characterized by dysfunctional skin barriers and substantially impact patients' quality of life. Vitamin D3 regulates immune responses and keratinocyte differentiation and improves psoriasis symptoms; however, its effects on atopic dermatitis remain unclear. Here, we investigated the effects of calcitriol, an active form of vitamin D3, on an NC/Nga mouse model of atopic dermatitis. We observed that the topical application of calcitriol decreased the dermatitis scores and epidermal thickness of NC/Nga mice with atopic dermatitis compared to untreated mice. In addition, both stratum corneum barrier function as assessed by the measurement of transepidermal water loss and tight junction barrier function as evaluated by biotin tracer permeability assay were improved following calcitriol treatment. Moreover, calcitriol treatment reversed the decrease in the expression of skin barrier-related proteins and decreased the expression of inflammatory cytokines such as interleukin (IL)-13 and IL-33 in mice with atopic dermatitis. These findings suggest that the topical application of calcitriol might improve the symptoms of atopic dermatitis by repairing the dysfunctional epidermal and tight junction barriers. Our results suggest that calcitriol might be a viable therapeutic agent for the treatment of atopic dermatitis in addition to psoriasis.

740 Effect of molecular chirality of ceramide to barrier function of stratum corneum

740 Effect of molecular chirality of ceramide to barrier function of stratum corneum

Morphological evaluation of stratum corneum barrier function by tape stripping keratinocytes in dog models of acute stratum corneum barrier dysfunction

Morphological evaluation of stratum corneum barrier function by tape stripping keratinocytes in dog models of acute stratum corneum barrier dysfunction

257 Clinical skin moisturizing property of the AVENE thermal spring water is associated with water capture and higher lipid organization in the stratum corneum

One of the main functions of the skin is to provide an optimal water level for normal physiological function and maintain its biophysical properties, including elasticity. The aim of this study was to demonstrate clinically the moisturizing effect of AVENE thermal spring water (TSW) and explain the mechanism of action involved, using biophysical analysis of the stratum corneum (SC) approaches. Skin electric capacitance was used to assess the hydration level of the skin outermost layers of 24 volunteers, using corneometry, after 6 daily applications of AVENE TSW for 7 days. The persistence of this effect was analyzed for 20 hours after the last application. In addition, ex vivo raman spectroscopy was performed on isolated SC. Hydration level and barrier function were directly monitored by following the different vibrational contributions of water and lipids in the SC Raman spectra. We clinically observed a significant improvement in hydration of the upper layers of the epidermis which persisted for 20 hours after the last application compared to baseline and to untreated area. Indeed, skin was less dehydrated for 20 hours after the last application. In addition, Raman experiments showed that AVENE TSW application on SC contributes to high global water content and retention. Besides the water content, the water structure and mobility were of high importance, and the high relative amount of partially bound water compared to tightly bound and unbound water can be related to a better capacity of the water to interact with SC components. In parallel, it maintains high conformational order of the lipid matrix, beneficial for skin barrier function and hydration. Altogether, these data demonstrated how the high moisturizing properties of AVENE TSW topically applied on skin subjects, is associated with SC hydration and barrier function improvement.

Modulation of function and structure of stratum corneum in sphingomyelin synthase 2-deficient mice

Sphingomyelin synthase (SMS) synthesizes sphingomyelin (SM) from ceramide (Cer), a precursor of Cer. The effects of SMS deficiency on stratum corneum (SC) barrier function and SC lamellar structure are unknown. In this report, permeation of hydrophilic and lipophilic compounds through full-thickness skin or stripped skin of SMS2-knockout (KO) and wild-type (WT) mice was examined. Furthermore, small-angle and wide-angle X-ray scattering (SAXS and WAXS) measurements of the SC were performed as a function of temperature to analyze the lamellar structure and hydrocarbon chain packing, where a SC sample was changed from 10 °C to 120 °C at 2 °C/min and the X-ray diffraction profile in the small-angle region and the wide-angle region was observed. Skin permeability of the hydrophilic compound increased significantly for SMS2-KO mice when compared with that of WT mice. In contrast, no difference was observed in the penetration of lipophilic compounds in the skin of both SMS2-KO and WT mice. In SC of SMS2-KO mice, two sharp SAXS peaks were observed due to the lamellar structure with a repetition period of 4.8 nm. The WAXS revealed that the intensity ratio R0.42/0.37 of the 0.42 nm peak at 2.4 nm–1 to the 0.37 nm peak at 2.7 nm–1 was smaller in the SMS2-KO mouse than in the WT mouse. Due to the temperature dependence of the WAXS, the peaks of 2.4 and 2.7 nm−1 remained until the higher temperatures in SMS2-KO mouse SC than those in WT mouse SC. The results of X-ray diffraction suggest that deficiency of SMS2 may cause the appearance of highly ordered structures of SC, which in turn may reduce the barrier function of SC.

The Nanostructured lipid carrier gel of Oroxylin A reduced UV-induced skin oxidative stress damage

Oxidative stress damage caused by sun exposure damages the appearance and function of the skin, which is one of the essential inducements of skin aging and even leads to skin cancer. Oroxylin A (OA) is a flavonoid with excellent antioxidant activity and has protective effects against photoaging induced by UV irradiation. However, the strong barrier function of the skin stratum corneum prevents transdermal absorption of the drug, which limits the application of OA in dermal drug delivery. Studies have shown that nanostructured lipid carriers (NLC) can promote not only transdermal absorption of drugs but also increase drug stability and control drug release efficiency, which has broad prospects for clinical applications. In this paper, NLC loaded with OA (OA-NLC) was prepared in order to improve the skin permeability and stability of OA. In vitro studies revealed that OA-NLC had better therapeutic effects than OA solution (OA-Sol) in the cellular model of UVB radiation. OA-Sol and OA-NLC were immobilized in a hydrogel matrix to facilitate application to the dorsal skin of mice. It was found that OA-NLC-gel showed significant antioxidant and anti-apoptotic activity compared to OA-Sol-gel, which was able to protect against skin damage in mice after UV radiation. These results suggest that OA-NLC can improve the deficiencies of OA in skin delivery and show better resistance to UV-induced oxidative damage. The application of OA-NLC to skin delivery systems has good prospects and deserves further development and investigation.

Performance and Acceptability of a New Dexpanthenol-Containing Hand Cream in Subjects with Sensitive and Very Dry Skin: A Randomized Controlled Study

A new dexpanthenol-containing hand cream (ND-HC) was developed for people with dry, sensitive, and/or environmentally stressed hands. To explore the performance and acceptability of ND-HC, we conducted a randomized, intraindividual comparison study in 40 healthy adult subjects with sensitive and very dry skin on the hands. Instrumental measurements determined the effects on stratum corneum (SC) hydration and transepidermal water loss (TEWL) after single and/or 4 weeks’ use of ND-HC. Single and continued at least four times daily applications of ND-HC to very dry skin of the hand for 4 weeks triggered significant increases in SC hydration. On day 29, the mean change in skin capacitance from baseline was significantly greater when ND-HC was applied to the test area compared with the untreated area on the contralateral hand (12.41 vs. 4.46 a.u.; p < 0.001). Upon use of ND-HC over 4 weeks, mean TEWL decreased significantly (bilateral difference: −1.8 vs. 1.0 g/m2/h; p = 0.003), indicating an improvement in SC barrier function. A reduction in dry hand symptoms was observed over the study course. ND-HC was well tolerated and achieved a high level of acceptance and satisfaction. Our findings suggest that ND-HC complies with the required features of a state-of-the-art hand cream.

Cholesterol sulfate fluidizes the sterol fraction of the stratum corneum lipid phase and increases its permeability

Desulfation of cholesterol sulfate (CholS) to cholesterol (Chol) is an important event in epidermal homeostasis and necessary for stratum corneum (SC) barrier function. The CholS/Chol ratio decreases during SC maturation but remains high in pathological conditions, such as X-linked ichthyosis, characterized by dry and scaly skin. The aim of this study was to characterize the influence of the CholS/Chol molar ratio on the structure, dynamics, and permeability of SC lipid model mixtures. We synthesized deuterated CholS and investigated lipid models with specifically deuterated components using 2H solid-state NMR spectroscopy at temperatures from 25°C to 80°C. Although the rigid acyl chains in ceramides and fatty acids remained essentially rigid upon variation of the CholS/Chol ratio, both sterols were increasingly fluidized in lipid models containing higher CholS concentrations. We also show the X-ray repeat distance of the lipid lamellar phase (105 Å) and the orthorhombic chain packing of the ceramide’s acyl chains and long free fatty acids did not change upon the variation of the CholS content. However, the Chol phase separation visible in models with high Chol concentration disappeared at the 50:50 CholS/Chol ratio. This increased fluidity resulted in higher permeabilities to model markers of these SC models. These results reveal that a high CholS/Chol ratio fluidizes the sterol fraction and increases the permeability of the SC lipid phase while maintaining the lamellar lipid arrangement with an asymmetric sterol distribution.

LB736 Wearing N95 masks does not disrupt the facial skin microbiome

The COVID-19 pandemic has elevated concern about mask-related skin problems like so-called ‘maskne’ which is likely rooted in the skin microbiome. We therefore determined if wearing an N95 mask affects the skin microbiome in a 3-day controlled study. On Day 1, subjects (n=10) followed their normal office routine without a mask. On Days 2 and 3, subjects wore an N95 mask (3M Model 8210) from morning to late afternoon (6 hours). The same mask was used both days. Microbiome diversity and composition (16S rRNA amplicon sequencing, V1-V3), stratum corneum (SC) barrier function (TEWL), SC hydration, skin redness and follicular porphyrins were measured on the cheek (masked site) and forehead (control site) each morning and afternoon.

Racial/Ethnic Variations in Skin Barrier: Implications for Skin Care Recommendations in Skin of Color.

Genetic and environmental factors influence stratum corneum (SC) barrier properties and function. Researchers increasingly focus on biophysical studies that may help clinicians provide their patients with an informed choice on tailormade skincare. This literature review on skin barrier properties comparing different ethnic populations aims to offer insights into the information's clinical relevance. A literature review followed by panel discussions and an online review process aimed to answer the questions: Are there racial/ethnic differences in the SC barrier structure and healthy skin barrier function? Is there a need for specific cleansers and moisturizers? Ethnic categories based on race and ethnicity are often not well defined and inconsistent across different studies. Studies comparing ethnic groups' physical and biochemical skin barrier properties have reported differences in transepidermal water loss (TEWL), skin lipid levels, pH, and mast cell granule size. However, these studies frequently had methodological flaws, mainly were small, and demonstrated conflicting results. The literature suggests racial/ethnic variations in ceramide content, SC structure, and filaggrin mutations. Furthermore, studies have shown a greater burden of pruritus and atopic dermatitis among Black populations. Data on barrier properties in Hispanic/LatinX and South Asian populations are lacking. Robust comparative studies are needed to understand these basic concepts to help tailor skincare and skin of color patients' education. J Drugs Dermatol. 2021;20(9):932-938. doi:10.36849/JDD.6312.

Interferon-γ downregulates tight junction function, which is rescued by interleukin-17A.

Although atopic dermatitis (AD) has been reported to be a typical type 2 immune response disease, it is also an inflammatory skin disease that involves cytokines, such as Th1, Th17 and Th22. However, little is known about the mechanism by which the candidate cytokines, alone or in combination, are involved in AD pathology. Differences in cytokine balance, which contribute to the complexity of AD pathology, may influence the stratum corneum barrier function through tight junction (TJ) functional stability and contribute to disease severity. To confirm the regulatory mechanism of TJ protein expression in AD, we investigated the Th1 and Th17 pathways, which are the initiation factors of chronic AD pathology. We examined the effects of these cytokines on TJ protein expression in normal human epidermal keratinocytes in vitro, and also examined their function in a human skin equivalent model. We observed a time‐ and dose‐dependent inhibitory effect of IFN‐γ on claudin‐1 expression via the IFN‐γ receptor/JAK/STAT signalling pathway. IFN‐γ impaired TJ function in a human skin equivalent model. Moreover, we investigated co‐stimulation with IL‐17A, which is highly expressed in AD skin lesions and found that IL‐17A restores IFN‐γ‐induced TJ dysfunction. This restoration of TJ function was mediated by atypical protein kinase C zeta activation without recovery of TJ protein expression. These results are informative for personalized AD treatment via systemic therapies using anti‐cytokine antibodies and/or JAK inhibitors.

Direct assessment of individual skin barrier components by electrical impedance spectroscopy.

Expression of the tight junction proteins Cldn1 and 4 is altered in skin diseases such as atopic dermatitis, and Cldn1 deficiency affects skin barrier formation. Impedance spectroscopy (IS) has been proven to allow detection of alterations in the skin barrier but is currently unable to separate effects on viable epidermis (VE) and stratum corneum (SC). Effects of siRNA-mediated Cldn1 and 4 knockdown in reconstructed human epidermis (RHE) on VE and SC barrier function were investigated with Ussing chamber-based IS. Barrier components were sequentially altered, employing iron oxide nanoparticles and EGTA, to identify their contribution to the impedance spectrum. Resistance changes due to apically applied hyperosmolar electrolyte were used to identify barrier defects non-invasively. IS of RHE yielded two relaxation frequencies, representing the barrier properties of the SC (~1000Hz) and VE (~100Hz). As proof of concept, it was shown that the Cldn1 knockdown-induced resistance drop arises from the impairment of both SC and VE, indicated by a shift of both relaxation frequencies. Hyperosmolar electrolyte penetration allowed non-invasive detection of Cldn1 knockdown via time-dependent frequency shifts. The absence of Cldn4 knockdown-induced changes revealed the weaknesses of transepithelial electrical resistance analysis. In conclusion, the present technique allows to separately measure the barrier properties of SC and VE and further evaluate the Cldn1 and 4 knockdown impact on the skin barrier. As the measurement with agarose-embedded electrolyte allowed non-invasive identification of the Cldn1 knockdown, this opens the way to detailed in vivo skin barrier assessment.

ARTICLE: Compromised Skin Barrier and Sensitive Skin in Diverse Populations.

The most important function of the stratum corneum (SC), the uppermost layer of the human epidermis, is the formation of the epidermal permeability barrier. Lipids, particularly ceramides, cholesterol, and free fatty acids, together form lamellar membranes in the extracellular spaces of the SC that limit the loss of water and electrolytes. In addition to preventing water and electrolyte loss, the SC as a permeability barrier prevents the entry of harmful irritants, allergens, and microorganisms into the skin. Disruption of the epidermal barrier leads to skin that is irritated, more reactive, and more sensitive than normal skin. SC thickness, lipid profile, and barrier function vary with different ethnic groups, which is also reflected the differences in prevalence and manifestation of diverse skin conditions related to the skin barrier function such as atopic dermatitis and sensitive skin. In addition to these compromised skin barrier related conditions, we are just now starting to understand the direct and indirect impact of COVID-19 on the skin and how current preventative measures are contributing to skin barrier disorders. Our understanding of various approaches for restoration of skin barrier, especially the role of topically applied mixtures of cholesterol, ceramides, and essential/nonessential free fatty acids (FFAs) allows for the strengthening of the compromised skin barrier and alleviation of symptoms and discomfort associated with skin barrier disorders. Ceramide containing products on the market are commonly available and offer protection and reparative benefits to the skin barrier. J Drugs Dermatol. 20(4 Suppl):17-22. doi:10.36849/JDD.S589C.

Effects of petrolatum, a petrolatum depositing body wash and a regular body wash on biomarkers and biophysical properties of the stratum corneum.

An important trend in the personal care industry involves the development of body wash products that not only clean the skin without damage but deposit conditioning ingredients to improve skin barrier function. The objective of this study was to develop skin biomarker measures to quantify the treatment effects of body wash products. We employed analysis of structural proteins (keratin 1,10,11 and involucrin), a natural moisturizing factor (pyrrolidone carboxylic acid) and an inflammatory mediator (IL-1ra/IL-1α) from adhesive discs with dry skin grading, TEWL and capacitance measurements to compare the effects of direct application of petrolatum, a high petrolatum depositing body wash, and a regular body wash on dry leg skin in a standard leg-wash treatment protocol. High depositing body wash and petrolatum had positive effects on stratum corneum barrier function as judged by biomarker analysis, biophysical measurements and skin grading compared to the regular body wash product. The results clearly indicate that a combination of biomarker and biophysical property measurements is effective for determining the skin benefits of moisturizing body wash products.

ARTICLE: Compromised Skin Barrier and Sensitive Skin in Diverse Populations

The most important function of the stratum corneum (SC), the uppermost layer of the human epidermis, is the formation of the epidermal permeability barrier. Lipids, particularly ceramides, cholesterol, and free fatty acids, together form lamellar membranes in the extracellular spaces of the SC that limit the loss of water and electrolytes. In addition to preventing water and electrolyte loss, the SC as a permeability barrier prevents the entry of harmful irritants, allergens, and microorganisms into the skin. Disruption of the epidermal barrier leads to skin that is irritated, more reactive, and more sensitive than normal skin. SC thickness, lipid profile, and barrier function vary with different ethnic groups, which is also reflected the differences in prevalence and manifestation of diverse skin conditions related to the skin barrier function such as atopic dermatitis and sensitive skin. In addition to these compromised skin barrier related conditions, we are just now starting to understand the direct and indirect impact of COVID-19 on the skin and how current preventative measures are contributing to skin barrier disorders. Our understanding of various approaches for restoration of skin barrier, especially the role of topically applied mixtures of cholesterol, ceramides, and essential/nonessential free fatty acids (FFAs) allows for the strengthening of the compromised skin barrier and alleviation of symptoms and discomfort associated with skin barrier disorders. Ceramide containing products on the market are commonly available and offer protection and reparative benefits to the skin barrier. J Drugs Dermatol. 20(4 Suppl):17-22. doi:10.36849/JDD.S589C

Temporal Variation of the Facial Skin Microbiome: A 2-Year Longitudinal Study in Healthy Adults.

The human skin microbiome is highly personalized, depending on, for example, body site, age, gender, and lifestyle factors. The temporal stability of an individual's skin microbiome-its resiliency and robustness over months and years-is also a personalized feature of the microbiome. The authors measured the temporal stability of the facial skin microbiome in a large cohort of subjects. In addition to measuring microbiome dynamics, they tracked facial skin condition using noninvasive, objective imaging and biophysical measures to identify significant facial features associated with temporal changes in microbiome diversity and composition. The authors used 16S ribosomal RNA amplicon sequencing to track cheek and forehead skin microbiome diversity and composition annually over a 2-year period (2017-2019) in 115 healthy adult men and women. Skin metadata included facial features, such as wrinkles, hyperpigmentation, porphyrins, and skin color tone, as well as biophysical parameters for stratum corneum barrier function, pH, hydration, and elasticity. Across the subject population, the facial skin microbiome composition and diversity were relatively stable, showing minor variation over the 2-year period. However, for some subjects, composition, diversity, and relative abundance of specific organisms showed substantial changes from one year to the next, and these changes were associated with changes in stratum corneum barrier function and follicular porphyrins. For healthy people, facial skin microbiome diversity and composition are relatively stable from year to year. Tracking the temporal changes in the microbiome along with skin phenotypic changes allows for a deeper understanding of the skin microbiome's role in health and disease. These results should be helpful in the design of longer-term intervention trials with microbiome-based skin care treatments.