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P6581Quantification and visualization of cellular NADP(H) as a skin cell energybiomarker with in vivo 2-photon tomography
Kukizo Miyamoto, PhD, P&G Innovation GK, Kobe, Japan; Hiroyuki Kudoh, P&GInnovation GK, Kobe, Japan
Background: In vivo 2-photon tomography is a novel noninvasive, 3-dimensionaloptical in vivo skin imaging technology with subcellular resolution that enables thesensitive detection of endogenous fluorophores. One of these fluorophores,NADP(H) (a coenzyme that plays an important role in the release of free energyduring glycolysis and influences filaggrin and lipid synthesis) can be selectivelydetected in keratinocyte (granular cells) with 2-photon tomography. The quantifi-cation of NADP(H) levels in subsurface skin may be important for skin condition andaging characterization.
Methods: Total of 30 healthy Asian females were enrolled in this study, having 2subgroups of 1:17 subjects of age 20s (average age, 26.2 years; SD 6 3.7) and 2:13subjects of age 60s (average age, 63.4; SD6 2.7). Skin measurement was performedon the facial skin by near infrared femtosecond laser pulses at low pico Joule pulseenergies using the in vivo 2-photon tomography equipment(DermaInspect/MPTflex). The laser beam scans a skin field of interest in slices of300 3 300 �m2 (512 3 512 pixels) with a typical beam well time of \40microseconds per pixel. The time between 2 laser pulses was 12 nanoseconds. Thisoptical skin biopsy image scanning with 2-photon tomography carried out from skinsurface (on top of stratum corneum) to 40-�m depth in the epidermis layer, by 2-�mdepth pitch. The near infrared laser pulses excite endogenous fluorophores (NADP[H]). Image processing was performed to obtain high resolution autofluorescenceimages (optical biopsies) and to quantify the fluorescent gray scale of NADP(H)levels and compared average values in 2 groups of young (age 20s) and elder (age60s) subjects.
Results: Mean of fluorescent gray scale of NADP(H) at age 20s was 38.8(SD 6 12.39), and that of age 60s was 32.7 (SD 6 12.47). NADP(H) level at age20s was significantly larger than age 60s (P ¼ .0078).
Discussion: NADP(H) was produced in younger granular cells, and declinedsignificantly with age. It is assumed NADP(H) production would be less activatedbecause of chronologic aging and environmental damage (eg, photodamage). Thisstudy shows that NADP(H) levels in granular cells in the epidermal layer can be usedas a biomarker of skin aging. It is also suggests that increasing NADP(H) productionmay be a novel approach to maintaining a good skin condition and caring for agingskin.
AB32
nsored by Procter and Gamble Company.
100% spoP6986Skin bioenergy and antiaging responses to a complex of cosmetic ingre-dients with dill seed extract
Robert Carver, PhD, BASF, Stony Brook, NY, United States; Lisa Mullins, MS, TheProcter and Gamble Company, Cincinnati, OH, United States; Mary Johnson, MS,The Procter and Gamble Company, Cincinnati, OH, United States; RosemarieOsborne, PhD, The Procter and Gamble Company, Cincinnati, OH, United States
Background: One approach to enable the renewal of aging skin is restoration ofcellular energy capacity. Niacinamide (NA) increases NADH levels and provides skinbenefits, such as improved barrier, texture, and uneven pigmentation. In the currentwork, we evaluated a complex containing NA, a wrinkle reducing peptide Pal-KTTKS, and an olive derivative that boosts the antioxidant capacity of skin. Theeffects of the complex on ATP, NADH, and procollagen 1 (PC-1) were determined inhuman dermal fibroblasts (Fbs) derived from aging adult skin as compared to youngskin. The complex was evaluated also with dill seed extract, which has been shownto boost LOXL levels and skin elasticity.
Methods: Human dermal Fbs were treated with the complex of NA, Pal-KTTKS andan olive derivative (olive oilederived fatty acids modified with PEG-7) for 24 to 72hours, without or with addition of dill seed extract. Dill seed extract alone wasevaluated further in dermal Fbs and skin equivalent cultures (MatTek EFT 400). ATPwas measured by luminescence (Promega), total NAD+/NADH by colorimetry(Cayman), PC-1 by ELISA (Takara), and mRNA by RT-PCR and Affymetrix arrays.
Results: The complex of cosmetic ingredients, alone and in combination with dillseed extract, produced a significant increase in ATP and NAD+/NADH in aging adultFbs to levels comparable to those in younger Fbs. Dill seed extract itself promotedincreased expression of genes related to energy metabolism and antioxidantprotection in dermal Fbs. The addition of dill seed extract to the complex boostedPC-1 production to levels comparable to neonatal Fbs, and significantly greater thancomplex alone. A stimulatory effect of dill seed extract on PC-1 was detected also inaging dermal Fbs and tissue engineered skin equivalent cultures.
Conclusion: A complex of cosmetic ingredients including dill seed extract stimu-lated adult dermal Fbs to produce levels of ATP and NAD+/NADH comparable toyounger cells. This effect on cellular energy sources was paralleled by an increase inproduction of natural collagen, confirming the vital role of bioenergy in productionof skin structure components. The effects of dill seed extract, previously shown toincrease skin elasticity, on collagen suggests broader effects of this extract on overalldermal matrix than previously known. The complex of ingredients plus dill seedextract addresses multiple approaches to renew aging skin cells and provides apromising new direction for cosmetic products.
k was sponsored 100% by BASF and P&G Beauty.
This worJ AM ACAD DERMATOL
P7019Skin biomarkers confirm skin antioxidant activity of an olive derivative
Rosemarie Osborne, PhD, Procter and Gamble Beauty, Cincinnati, OH, UnitedStates; Akira Date, PhD, Procter and Gamble Beauty, Cincinnati, OH, United States;Bradley Jarrold, MS, Procter and Gamble Beauty, Cincinnati, OH, UnitedStates; Deborah Finlay, PhD, Procter and Gamble Beauty, Cincinnati, OH, UnitedStates; Heather Matheny, MS, Procter and Gamble Beauty, Cincinnati, OH,United States; Lisa Mullins, MS, Procter and Gamble Beauty, Cincinnati,OH, United States; Makio Tamura, PhD, Procter and Gamble Beauty, Cincinnati,OH, United States
Background: Reactive oxygen species (ROS) play important roles in UV-induced skindamage, photoaging, andmelanogenesis. The antioxidant response element (ARE) is atranscriptional control element that meditates a family of phase II enzymes andantioxidant proteins, under control of the transcription factor NRF2. NRF2 up-regulates the family of antioxidant and repair proteins in response to ROS and freeelectrophiles in the skin. Therefore, the induction of ARE-dependent genes plays animportant role in protection of cells against oxidative damage. We have reportedpreviously thatNRF2 expression is down-regulated in intrinsically and photoaged skin.In parallel, protective enzymes under the control of the ARE are decreased inexpression, while markers of oxidative stress are increased. The objective of thecurrentworkwas todetermineantioxidant effectsofa topically appliedolivederivativeon skin antioxidant biomarkers related to ARE and associated enzymes and proteins.
Methods: Human skin keratinocytes (KC), fibroblasts (Fb), skin equivalents, andexplants were treated with the olive derivative (olive oilederived fatty acidsmodified with PEG-7). ARE activity was assayed using a reporter cell line (ARE-32,CXR-Biosciences) and confirmed in skin models by enzyme activity (HO-1) andmRNA (RT-PCR and Affymetrix arrays).
Results: The olive derivative stimulated dose-dependent ARE activity in a reportercell line. This response was confirmed by HO-1 activity in skin KC, Fb, and explants.Bioinformatics analysis of significantly overregulated gene ontology terms from genearray analysis of skin KC and Fb treated with the olive-derived material indicated areversal of the down regulation of antioxidant genes occuring in intrinsically andphotoaged skin. This finding was confirmed by RT-PCR analysis of specific ARE-regulated genes and proteins (eg, SOD). In parallel, there was increased expressionof genes related to epidermal and dermal structure and mitochondrial metabolism.
Conclusion: The olive-derived material affected antioxidant biomarkers in humanskin models, and therefore enhance the ability of skin to protect against damagefrom ROS. This was confirmed by the positive effects of the material on skinstructure and mitochondrial metabolism markers. Therefore, the antioxidantproperties of the olive derivative can provide powerful protection and repair foraged skin against the continual assault of UV, environmental, and aging-induced ROS.
d by Procter and Gamble Beauty.
SupporteARTS, HISTORY, AND HUMANITIESOF DERMATOLOGY
P6152An exploration of how culture plays a role in one’s perception of acne
Jennifer A. Ledon, University of Miami Miller School of Medicine Department ofDermatology and Cutaneous Surgery, Miami, FL, United States; Anna Chacon,MD, University of Miami Miller School of Medicine Department of Dermatologyand Cutaneous Surgery, Miami, FL, United States; Katlein Franca, MD, MS,University of Miami Miller School of Medicine Department of Dermatology andCutaneous Surgery, Miami, FL, United States; Keyvan Nouri, MD, University ofMiami Miller School of Medicine Department of Dermatology and CutaneousSurgery, Miami, FL, United States
‘‘Acne’’ can be traced back to the Ancient Greek word ‘‘akme,’’ meaning ‘‘point.’’While the Greeksmay have coined theword for this infamous blemish, many authorshave exploredwhether culture or ethnicity influences one’s perception of their acneand if those same views power treatment. Chinese traditional medicine, for example,purports that acne is thought to be a result of disharmony leading to excess heat ordampness. In Ayurvedic medicine, it is thought that reducing one’s ‘‘vata,’’ which isthe ‘‘dosha’’ representing movement, minimizes the scarring associated with acne.Furthermore, current Native Americanmedicine incorporates the use of Kanwa Clayto treat blemishes, and one nonprofit Russian organization even advocates the use ofvodka. For these reasons, various investigators have explored how these influencesplay a role in perceptions of acne in today’s diverse societies. In a study looking at 2Nigerian universities, only 5.2% of students had sought medical care for their acne atone point in time, yet an American study by Cheng et al found that black participantswho reported moderate severity acne were more likely to seek professional help fortheir acne as compared to white participants. In the same study, psychologicalperception of acne also differed between ethnicities; a greater number of Hispanicscompared to non-Hispanics felt that acne is the patient’s fault. Furthermore,questionnaires linking acne to quality of life have been validated in other languages,such as French, suggesting that despite the differences between cultures andethnicities, certain themes break cultural boundaries. Acne has been shown to havea broad negative psychological and emotional impact regardless of race (white,black, Hispanic, or Asian), and a surprisingly high percentage of those sufferers havenever sought professional help for their disease. This presentation will discuss someof the cultural differences in the perception of acne and how current literaturesupports the notion that acne is a source of insecurity and distress throughout allcultures and races. By understanding how culture influences the patient’s percep-tion of acne, clinicians may be able to better comprehend, encourage, and improvetreatment for those suffering from the disease.
cial support: None identified.
CommerAPRIL 2013
