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The role of the skin barrier in modulating the effects of common skin microbial species on the inflammation, differentiation and proliferation status of epidermal keratinocytes.

Duckney P, Wong HK, Serrano J, Yaradou D, Oddos T, Stamatas GN - BMC Res Notes (2013)

Bottom Line: S. aureus induced complete keratinocyte cell death.On the contrary, topically applied S. epidermidis and P. acnes caused no inflammatory response even when tested at high concentrations, while topical S. aureus induced a weak reaction.None of the tested species were able to alter the expression of keratinocyte differentiation or expression markers, when applied topically.

View Article: PubMed Central - HTML - PubMed

Affiliation: Johnson & Johnson Santé Beauté France, 1 rue Camille Desmoulins, Issy-les-Moulineaux 92787, France. GStamata@its.jnj.com.

ABSTRACT

Background: Skin resident microbial species are often thought of either as pathogenic or commensal. However, little is known about the role of the skin barrier in modulating their potential for causing disease. To investigate this question we measured the effects of three microbial species commonly found on the skin (Staphylococcus epidermidis, Staphylococcus aureus, and Propionibacterium acnes) on a reconstructed human epidermal model by either applying the bacteria on the model surface (intact barrier) or adding them to the culture medium (simulating barrier breach).

Results: When added to the medium, all of the tested species induced inflammatory responses and keratinocyte cell death with species-specific potency. P. acnes and S. epidermidis induced specific alterations in the expression of keratinocyte differentiation and proliferation markers, suggesting a barrier reparation response. S. aureus induced complete keratinocyte cell death. On the contrary, topically applied S. epidermidis and P. acnes caused no inflammatory response even when tested at high concentrations, while topical S. aureus induced a weak reaction. None of the tested species were able to alter the expression of keratinocyte differentiation or expression markers, when applied topically.

Conclusions: We show that the skin barrier prevents the effects of common skin bacteria on epidermal keratinocyte inflammation, differentiation and proliferation and highlight the importance of skin barrier in defending against the pathogenic effects of common skin bacteria.

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(A) S. epidermidis and (B) P. acnes added to the medium of reconstructed human epidermal equivalents induce dose-dependent changes in the expression of genes involved in differentiation (FLG, TG1, CLD, OCCL) and proliferation (PCNA). After 24 hrs incubation with bacteria at 1 × 107, 1 × 108 and 1 × 109 CFU, RNA from the RHEs was extracted and reverse transcribed. The mRNA expression levels of a range of genes were determined by quantitative reverse transcription polymerase chain reaction. The amount of target transcripts was normalized using an 18S RNA normalization gene. Data are expressed as percentage of untreated control (UnC = 100%) and are presented as mean ± one standard deviation. Asterisks indicate statistically significant differences (P < 0.05) vs the untreated control.
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Figure 3: (A) S. epidermidis and (B) P. acnes added to the medium of reconstructed human epidermal equivalents induce dose-dependent changes in the expression of genes involved in differentiation (FLG, TG1, CLD, OCCL) and proliferation (PCNA). After 24 hrs incubation with bacteria at 1 × 107, 1 × 108 and 1 × 109 CFU, RNA from the RHEs was extracted and reverse transcribed. The mRNA expression levels of a range of genes were determined by quantitative reverse transcription polymerase chain reaction. The amount of target transcripts was normalized using an 18S RNA normalization gene. Data are expressed as percentage of untreated control (UnC = 100%) and are presented as mean ± one standard deviation. Asterisks indicate statistically significant differences (P < 0.05) vs the untreated control.

Mentions: We studied the effects of bacteria added to the RHE medium on the keratinocyte expression of the genetic differentiation and proliferation markers: FLG, TG1, OCCL, CLD and PCNA. Addition of S. aureus in the medium induced keratinocyte cell death and degradation of marker mRNA, making it impossible to collect useful data. However, addition of P. acnes and S. epidermidis induced common patterns of changes in gene expression indicating a conserved response of the keratinocytes (Figure 3). In both cases, adding the bacteria to the medium induced decreased FLG and PCNA expression and increased OCCL and TG1 expression. The expression of CLD was not significantly affected by the presence of S. epidermidis or P. acnes in the RHE growth medium. As observed with the release of proinflammatory cytokines, S. epidermidis induced a stronger response than P. acnes in the induction of transcriptional changes of the RHE keratinocytes. S. epidermidis was able to induce significant changes in the expression of FLG, TG1, OCCL and PCNA when tested at only 1 × 107 CFU, whereas P. acnes was only able to induce significant increases in the affected differentiation markers at 1 × 109 CFU and PCNA at 1 × 108 CFU.


The role of the skin barrier in modulating the effects of common skin microbial species on the inflammation, differentiation and proliferation status of epidermal keratinocytes.

Duckney P, Wong HK, Serrano J, Yaradou D, Oddos T, Stamatas GN - BMC Res Notes (2013)

(A) S. epidermidis and (B) P. acnes added to the medium of reconstructed human epidermal equivalents induce dose-dependent changes in the expression of genes involved in differentiation (FLG, TG1, CLD, OCCL) and proliferation (PCNA). After 24 hrs incubation with bacteria at 1 × 107, 1 × 108 and 1 × 109 CFU, RNA from the RHEs was extracted and reverse transcribed. The mRNA expression levels of a range of genes were determined by quantitative reverse transcription polymerase chain reaction. The amount of target transcripts was normalized using an 18S RNA normalization gene. Data are expressed as percentage of untreated control (UnC = 100%) and are presented as mean ± one standard deviation. Asterisks indicate statistically significant differences (P < 0.05) vs the untreated control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4225502&req=5

Figure 3: (A) S. epidermidis and (B) P. acnes added to the medium of reconstructed human epidermal equivalents induce dose-dependent changes in the expression of genes involved in differentiation (FLG, TG1, CLD, OCCL) and proliferation (PCNA). After 24 hrs incubation with bacteria at 1 × 107, 1 × 108 and 1 × 109 CFU, RNA from the RHEs was extracted and reverse transcribed. The mRNA expression levels of a range of genes were determined by quantitative reverse transcription polymerase chain reaction. The amount of target transcripts was normalized using an 18S RNA normalization gene. Data are expressed as percentage of untreated control (UnC = 100%) and are presented as mean ± one standard deviation. Asterisks indicate statistically significant differences (P < 0.05) vs the untreated control.
Mentions: We studied the effects of bacteria added to the RHE medium on the keratinocyte expression of the genetic differentiation and proliferation markers: FLG, TG1, OCCL, CLD and PCNA. Addition of S. aureus in the medium induced keratinocyte cell death and degradation of marker mRNA, making it impossible to collect useful data. However, addition of P. acnes and S. epidermidis induced common patterns of changes in gene expression indicating a conserved response of the keratinocytes (Figure 3). In both cases, adding the bacteria to the medium induced decreased FLG and PCNA expression and increased OCCL and TG1 expression. The expression of CLD was not significantly affected by the presence of S. epidermidis or P. acnes in the RHE growth medium. As observed with the release of proinflammatory cytokines, S. epidermidis induced a stronger response than P. acnes in the induction of transcriptional changes of the RHE keratinocytes. S. epidermidis was able to induce significant changes in the expression of FLG, TG1, OCCL and PCNA when tested at only 1 × 107 CFU, whereas P. acnes was only able to induce significant increases in the affected differentiation markers at 1 × 109 CFU and PCNA at 1 × 108 CFU.

Bottom Line: S. aureus induced complete keratinocyte cell death.On the contrary, topically applied S. epidermidis and P. acnes caused no inflammatory response even when tested at high concentrations, while topical S. aureus induced a weak reaction.None of the tested species were able to alter the expression of keratinocyte differentiation or expression markers, when applied topically.

View Article: PubMed Central - HTML - PubMed

Affiliation: Johnson & Johnson Santé Beauté France, 1 rue Camille Desmoulins, Issy-les-Moulineaux 92787, France. GStamata@its.jnj.com.

ABSTRACT

Background: Skin resident microbial species are often thought of either as pathogenic or commensal. However, little is known about the role of the skin barrier in modulating their potential for causing disease. To investigate this question we measured the effects of three microbial species commonly found on the skin (Staphylococcus epidermidis, Staphylococcus aureus, and Propionibacterium acnes) on a reconstructed human epidermal model by either applying the bacteria on the model surface (intact barrier) or adding them to the culture medium (simulating barrier breach).

Results: When added to the medium, all of the tested species induced inflammatory responses and keratinocyte cell death with species-specific potency. P. acnes and S. epidermidis induced specific alterations in the expression of keratinocyte differentiation and proliferation markers, suggesting a barrier reparation response. S. aureus induced complete keratinocyte cell death. On the contrary, topically applied S. epidermidis and P. acnes caused no inflammatory response even when tested at high concentrations, while topical S. aureus induced a weak reaction. None of the tested species were able to alter the expression of keratinocyte differentiation or expression markers, when applied topically.

Conclusions: We show that the skin barrier prevents the effects of common skin bacteria on epidermal keratinocyte inflammation, differentiation and proliferation and highlight the importance of skin barrier in defending against the pathogenic effects of common skin bacteria.

Show MeSH
Related in: MedlinePlus