Synergistic Inhibition of Tumor Necrosis Factor-Alpha-Stimulated Pro-Inflammatory Cytokine Expression in HaCaT Cells by a Combination of Rapamycin and Mycophenolic Acid

BACKGROUND: Keratinocytes release various pro-inflammatory cytokines, chemokines, and adhesion molecules such as intercellular adhesion molecule 1 (ICAM-1) in response to cytokines such as tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma. Rapamycin and mycophenolic acid (MPA) have potent immunosuppressive activity because they inhibit lymphocyte proliferation. OBJECTIVE: We investigated the effects of rapamycin and MPA on the expression of inflammation-related factors such as ICAM-1 and inducible nitric oxide synthase (iNOS), pro-inflammatory cytokines and chemokines, and related signaling pathways in TNF-alpha-stimulated HaCaT cells. METHODS: The viability of HaCaT cells treated with rapamycin and MPA was confirmed using MTT assay. The expression of various cytokines such as interleukin (IL)-1beta, IL-6, and IL-8; inflammation-related factors such as ICAM-1 and iNOS; and the activation of mitogen activated protein kinase (MAPK) signaling pathways mediated by extracellular signal-related kinases (ERK), p38, and c-Jun N-terminal kinases (JNK) in TNF-alpha-stimulated HaCaT cells were confirmed using reverse transcription-polymerase chain reaction and western blotting. RESULTS: Combined treatment of TNF-alpha-induced HaCaT cells with rapamycin and MPA decreased ICAM-1 and iNOS expression and ERK and p38 activation more than treatment with either drug alone. The most significant decrease was observed with a combination of rapamycin (80 nM) and MPA (20 nM). These results show that co-treatment with these agents has a synergistic anti-inflammatory effect by blocking the activation of the ERK/p38 MAPK signaling pathway and thus suppressing the TNF-alpha-induced expression of ICAM-1 and iNOS. CONCLUSION: The combination of rapamycin and MPA could potentially be used as a therapeutic approach in inflammatory skin diseases.

Safety Evaluation of Stamp Type Digital Microneedle Devices in Hairless Mice

BACKGROUND: Microneedles provide a minimally invasive means to transport molecules into the skin. A number of specific strategies have been employed to use microneedles for transdermal delivery. OBJECTIVE: The purpose of this study was to investigate the safety of two new digital microneedle devices (Digital Hand(R) and Digital Pro(R); Bomtech Electronics Co., Ltd., Seoul, Korea) for the perforation of skin in skin-hairless-1 mice. This device replaces conventional needles and is designed specifically for intradermal delivery. METHODS: We used two newly developed digital microneedle devices to perforate the skin of skin-hairless-1 mice. We conducted a comparative study of the two digital microneedle devices and DTS(R) (Disk type-microneedle Therapy System; DTS lab., Seoul, Korea). To evaluate skin stability, we performed visual and dermatoscopic inspections, measurements of transepidermal water loss, and biopsies. RESULTS: The two novel digital microneedle devices did not induce significant abnormalities of the skin on visual or dermatoscopic inspection, regardless of needle size (0.25~2.0 mm). No significant histopathological changes, such as inflammatory cell infiltration, desquamation of the stratum corneum, or disruption of the basal layer, were observed. The digital microneedle devices and microneedle therapy system produced similar results on measures of skin stability. CONCLUSION: These two novel digital microneedle devices are safe transdermal drug delivery systems.

The Safety Evaluation of a Potent Angiogenic Activator, Synthetic Peptide (SFKLRY-NH2) for the Skin Application

A novel synthetic hexapeptide (SFKLRY-NH2) that displays angiogenic activity has been identified by positional scanning of a synthetic peptide combinatorial library (PS-SPCL). This study was carried out to investigate the irritation of the SFKLRY-NH2 on the skin. The tests were performed on the basis of Korea Food and Drug Administration (KFDA) guidelines. In results, cell toxicity is not appeared for SFKLRY-NH2 in HaCaT cells and B16F10 cells. SFKLRY-NH2 induced no skin irritation at low concentration (10 microM), mild irritation at high concentration (10mM). We consider that this result is helpful for saying about the safety of SFKLRY-NH2 in clinical use.

The Evaluation of Skin Safety and Skin Cell Toxicity for Scutellaria baicalensis Georgi Extract according to Extraction Conditions / 대한피부과학회지

BACKGROUND: Scutellaria baicalensis Georgi extract is used as a traditional herbal medicine. The efficacy of Scutellaria baicalensis Georgi extract is known for antioxidative activity, antiinflammation effect, antibacterial effect, inhibitory effect of melanin synthesis, sun protection effect, antiallergy effect, and etc. OBJECTIVE: We confirmed the cell viability or inhibitory effect of melanin synthesis in HaCaT (human keratinocyte cell line) and B16F10 (murine melanoma cell line) cells and the skin safety test through a clinical test (dermal irritation study) for Scutellaria baicalensis Georgi extract, according to the extraction methods. METHODS: We checked the cell viability, using MTT assay and inhibitory effect of melanin synthesis in B16F10 cells or HaCaT cells for thirty one Scutellaria baicalensis Georgi extract, according to the extraction methods. Then, we evaluated the skin safety for selected eight Scutellaria baicalensis Georgi extract through a primary dermal irritation test. RESULTS: Among the thirty one Scutellaria baicalensis Georgi extracts, according to the extraction methods, we selected eight Scutellaria baicalensis Georgi extracts that were not detected with cell toxicity in HaCaT cells and B16F10 cells, and could have inhibited the melanin synthesis in B16F10 cells. The selected eight Scutellaria baicalensis Georgi extracts identified the skin safety through a primary dermal irritation test. CONCLUSION: We expect clinical trials for whitening efficacy based on inhibitory effect of melanin synthesis and human skin safety for Scutellaria baicalensis Georgi extracts.

Successful Treatment of Alopecia Areata with Topical Calcipotriol

Alopecia areata (AA) is an inflammatory hair loss of unknown etiology. AA is chronic and relapsing, and no effective cure or preventive treatment has been established. Vitamin D was recently reported to be important in cutaneous immune modulation as well as calcium regulation and bone metabolism. It is well known that areata is common clinical finding in patients with vitamin D deficiency, vitamin D-resistant rickets, or vitamin D receptor (VDR) mutation. The biological actions of vitamin D3 derivatives include regulation of epidermal cell proliferation and differentiation and modulation of cytokine production. These effects might explain the efficacy of vitamin D3 derivatives for treating AA. In this study, we report a 7-year-old boy with reduced VDR expression in AA, recovery of whom was observed by topical application of calcipotriol, a strong vitamin D analog.

Inhibitory Effect of Vitamin U (S-Methylmethionine Sulfonium Chloride) on Differentiation in 3T3-L1 Pre-adipocyte Cell Lines

BACKGROUND: S-methylmethionine sulfonium chloride was originally called vitamin U because of its inhibition of ulceration in the digestive system. Vitamin U is ubiquitously expressed in the tissues of flowering plants, and while there have been reports on its hypolipidemic effect, its precise function remains unknown. OBJECTIVE: This study was designed to evaluate the anti-obesity effect of vitamin U in 3T3-L1 pre-adipocyte cell lines. METHODS: We cultured the pre-adipocyte cell line 3T3L1 to overconfluency and then added fat differentiation-inducing media (dexamethasone, IBMX [isobutylmethylxanthine], insulin, indomethacin) and different concentrations (10, 50, 70, 90, 100 mM) of vitamin U. Then, we evaluated changes in the levels of triglycerides (TGs), glycerol-3-phosphate dehydrogenase (G3PDH), AMP-activated protein kinase (AMPK), adipocyte-specific markers (peroxisome proliferator-activated receptor gamma [PPAR-gamma], CCAAT/enhancer-binding protein alpha [C/EBP-alpha], adipocyte differentiation and determination factor 1 [ADD-1], adipsin, fatty acid synthase, lipoprotein lipase) and apoptosis-related signals (Bcl-2, Bax). RESULTS: There was a gradual decrease in the level of TGs, C/EBP-alpha, PPAR-gamma, adipsin, ADD-1 and GPDH activity with increasing concentrations of vitamin U. In contrast, we observed a significant increase in AMPK activity with increasing levels of vitamin U. The decrease in bcl-2 and increase in Bax observed with increasing concentrations of vitamin U in the media were not statistically significant. CONCLUSION: This study suggests that vitamin U inhibits adipocyte differentiation via down-regulation of adipogenic factors and up-regulation of AMPK activity.

A Study on Tests of Skin Safety and Inhibition of Atopic Dermatitis Using a StoneTouch(R) Infrared Scanner in a Mouse Model / 대한피부과학회지

BACKGROUND: Atopic dermatitis is a chronic inflammatory skin disease. It is caused by immunological abnormalities, abnormalities of the skin barrier, environmental factors and genetic factors. Atopic dermatitis destroys the skin barrier and passes through the skin, triggering an immune response. To treat atopic dermatitis, we anticipate use of hypoallergenic cures to hydrate skin that has been dried by destruction of the skin barrier. OBJECTIVE: We did a preclinical trial to identify inhibitory effects of the StoneTouch(R) infrared scanner on atopic dermatitis. We conducted skin safety tests, comparing the use of infrared energy to drug treatment. We then confirmed the effects of the StoneTouch(R) infrared scanner through animal tests using Nc/Nga mice as a model of atopic dermatitis in order to identify any inhibition of the immune response in atopic dermatitis. METHODS: We irradiated Nc/Nga mice using a StoneTouch(R) infrared scanner under a variety of conditions. During skin safety tests of the StoneTouch(R) infrared scanner on hairless mice, we assessed immune response and burn risk in irradiated mouse skin. We identified any inhibitory effects on atopic dermatitis using Dermoscope assessments, measurements of transepidermal water loss (TEWL) and IgE levels, measurements of pro-inflammatory cytokines, H&E staining and immunofluorescence staining (IF) of substance P and CGRP as neurotransmitters on the backs and ears of Nc/Nga mice irradiated by the StoneTouch(R) infrared scanner. RESULTS: We did not observe any skin abnormalities after using the StoneTouch(R) infrared scanner on Nc/Nga mice. We confirmed the inhibitory effect of the StoneTouch(R) infrared scanner irradiation on atopic dermatitis. We found that irradiated epidermis was thinner than that of the epidermis in Nc/Nga mice in which atopic dermatitis was induced. We observed no significant between groups differences in expression level of substance P. The expression of CGRP in mice with atopic dermatitis was decreased, but, the increased irradiation led to greater expression of CGRP in irradiated skin. CONCLUSION: The StoneTouch(R) infrared scanner does not as a function of irradiation dosage. It inhibits the development of atopic dermatitis.

A Study of Potential Application of Rubus coreanus Miquel Extract for Seborrheic Dermatitis Treatment / 대한의진균학회지

BACKGROUND: Seborrheic dermatitis is chronic relapsing inflammatory skin disorder. Bokbunja (Rubus coreanus Miquel) is a wild berry to Rosaceae genus and also known to have an anti-inflammation effect. OBJECTIVE: We were to determine the effect of Rubus coreanus Miquel extract for seborrheic dermatitis in vivo and in vitro. METHODS: Seven patients with mild seborrheic dermatitis were enrolled in this study. PCR and culture were performed to identify subtypes of six Malassezia species (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa). Topical application of Rubus coreanus Miquel Extract was applied twice daily for 2 weeks. Clinical improvement and safety assessment were performed initially and 2 weeks later. Minimum inhibitory concentration (MIC) was evaluated on Malassezia globosa comparing with ketoconazole and itraconazole. Sebum production was also checked prior the experiment and 2 weeks later. RESULTS: Five of seven patients showed improvement. No significant adverse effects were found during the clinical trial. Mild dryness was reported in 2 patients but they resolved spontaneously without any treatment. Rubus coreanus Miquel Extract didn't show antimicrobial effect to Malassezia globosa. However, Rubus coreanus Miquel Extract showed anti-inflammatory effect. CONCLUSION: In this study, we were verified that Rubus coreanus Miquel Extract can be applied for seborrheic dermatitis treatment. And this action mechanism is not related with antimicrobial effect.

Distribution of Malassezia Species on the Scalp in Korean Seborrheic Dermatitis Patients

BACKGROUND: Malassezia species play an important role in the pathogenesis of seborrheic dermatitis. In particular, M. restricta and M. globosa are considered to be the predominant organisms in seborrheic dermatitis of Western countries. However, species distribution of Malassezia in seborrheic dermatitis has not been clearly determined yet in Asia. OBJECTIVE: To identify the distribution of Malassezia species on the scalp of seborrheic dermatitis patients in Korea using 26S rDNA PCR-RFLP analysis. METHODS: A total of 40 seborrheic dermatitis patients and 100 normal healthy volunteers were included in this study. For the identification of Malassezia species, the scalp scales of the subjects were analyzed by 26S rDNA PCR-RFLP analysis. RESULTS: The most commonly identified Malassezia species were M. restricta in the seborrheic dermatitis patients, and M. globosa in the normal controls. In the seborrheic dermatitis group, M. restricta was identified in 47.5%, M. globosa in 27.5%, M. furfur in 7.5%, and M. sympodialis in 2.5% of patients. In the healthy control group, M. globosa was identified in 32.0%, M. restricta in 25.0%, M. furfur in 8.0%, M. obtusa in 6.0%, M. slooffiae in 6.0%, and M. sympodialis in 4.0% of subjects. CONCLUSION: M. restricta is considered to be the most important Malassezia species in Korean seborrheic dermatitis patients.

The Development and Evaluation of Multiplex PCR Technique for Identification of Malassezia Yeast / 대한의진균학회지

BACKGROUND: Malassezia yeasts as major pathogenic fungi causes the common skin diseases including dandruff, psoriasis, seborrheic dermatitis and atopic dermatitis etc. various molecular techniques were developed to identify and classify the Malassezia species until now. But, these methods were discovered the problems. So, the development of the better molecular methods required to identify and classify of Malasseiza species. OBJECTIVE: We sought to develop of molecular techniques to identify and classify of six Malassezia species (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa). METHODS: We designed primers about ITS1 (Internal transcribed space 1) region that were well-known region useful to identify of Malassezia species. Because, ITS1 region that is located between 18S and 5.8S rDNA of ribosomal DNA was comparatively mutated quickly. The mono PCR using ITS1 primers was performed to confirm the specificity of ITS1 primers with six Malassezia standard strains. Then, Malassezia Multiplex detection kit was developed on the basis of technique using ITS1 regions. Malassezia Multiplex detection kit was used to perform multiplex PCR with six Malassezia standard strains and clinical isolates. RESULTS: The results of mono PCR using ITS1 primers about six Malassezia standard strains was detected each Malassezia standard strains. Also, the multiplex PCR using developed Malassezia Multiplex detection kit was confirmed to classify about six Malassezia standard strains and clinical isolates. CONCLUSION: In this study, we verified that six Malassezia yeasts was classified using Malassezia Multiplex detection kit from Malasszia standard strains and clinical isolates. And we anticipate that Malassezia Multiplex detection kit is able to do accurate diagnosis about six Malassezia yeasts (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa).

Effect of Anti-inflammation and Skin Hydration of AF-343 on Macrophage Raw 264.7 Cells and NC/Nga Mice with Atopic Dermatitis / 대한의진균학회지

BACKGROUND: Inflammatory response on LPS and IFN-gamma induced Macrophage Raw 264.7 cells was secreted NO (nitric oxide) and PGE2 (prostaglandin E2) though expression of iNOS and COX-2. And many pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6 etc.) was secreted on LPS and IFN-gamma induced Macrophage Raw 264.7 cells, too. Atopy dermatitis was inflammatory skin disease with pruritus, xeroderma and specific eczema. OBJECTIVE: We sought to effect of anti-inflammation and skin hydration of AF-343 on Macrophage Raw 264.7 cells and NC/Nga mice with Atopic Dermatitis. METHODS: The immune response of Raw 264.7 cells were induced by LPS and IFN-gamma. Then LPS and IFN-gamma induced Raw 264.7 cells was measured NO, PGE2 production after treatment of different concentrations for AF-343. The related genes (iNOS, COX-2) for NO, PGE2 production were detected using Western blot in LPS and IFN-gamma induced Raw 264.7 cells after treatment of different concentrations for AF-343. Pro-inflammatory cytokines were detected, too. NC/Nga mice as Atopy dermatitis model was induced atopy dermatitis. Then NC/Nga mice with atopy dermatitis were performed oral administration of AF-343 for 1weeks. After oral administration of AF-343, TEWL was measured on skin tissues of NC/Nga mice with atopy dermatitis according to whether were performed oral administration of AF-343 or not. And pro-inflammatory cytokines and IgE was measured in serum, protein of skin tissues of NC/Nga mice. Skin tissues of NC/Nga mice were performed H&E staining, immunohistochemical staining for PCNA, Involucrin and filaggrin. RESULTS: LPS and IFN-gamma induced Raw 264.7 cells was decreased NO, PGE2 production in dose-dependent after treatment of different concentrations for AF-343. The expression level of iNOS, COX-2 protein was decreased in dose-dependent, too. The related pro-inflammatory cytokines in media with LPS and IFN-gamma induced Raw 264.7 cells were decreased after treatment of different concentrations for AF-343. TEWL level of NC/Nga mice skin (back, ear) with atopy dermatitis according to whether were performed oral administration of AF-343 or not was decreased in NC/Nga mice with atopy dermatitis group was performed oral administration by AF-343. When NC/Nga mice group with atopy dermatitis was performed oral administration by AF-343, induced pro-inflammatory cytokines and IgE expression in serum, protein of back, ear skin tissues of each NC/Nga mice group was decreased. H&E stained Skin tissues of NC/Nga mice was confirmed that thickness of epidermis, dermis were decreased in NC/Nga mice group with atopy dermatitis was performed oral administration by AF-343 than NC/Nga mice group with atopy dermatitis. The expression of PCNA, involucrin and filaggrin were decreased in NC/Nga mice group with atopy dermatitis was performed oral administration by AF-343 than NC/Nga mice group with atopy dermatitis as results of immunihistochemical staining using specific antibodies such as PCNA as cell proliferation marker, involucrin and filaggrin as keratinocytes differentiation markers for skin tissues (back, ear) of NC/Nga mice. CONCLUSION: We confirmed effect of anti-inflammation and skin hydration of AF-343 on Macrophage Raw 264.7 cells and NC/Nga mice with Atopic Dermatitis. In conclusion, AF-343 is expecting as therapeutics for atopic dermatitis.

A Study on the Antimicrobial Effect of Bamboo (Phyllosrachys bambusoides) Essential Oil on Malassezia / 대한의진균학회지

BACKGROUND: Malassezia is considered as major factor in dandruff of human scalp. OBJECTIVE: In order to develop an antimicrobial agent, bamboo oil was extracted by high temperture suction from dried bamboo truk abd then antimicrobial activities against Malassezia are investigated. METHODS: Minimum inhibitory concentration and antimicrobial activity were measured in Malassezia species. RESULTS: 1. Minimum inhibitory concentration of the Bamboo (Phyllosrachys bambusoides) Essential Oil Malassezia furfur standard, Malassezia furfur patient, Malassezia sympodialis standard, Malassezia sympodialis patient, Malassezia dermatis standard, Malassezia dermatis patient were 10 microliter/ml, 5 microliter/ml, 5 microliter/ml, 10 microliter/ml, 5 microliter/ml and 10 microliter/ml respectively. 2. Minimum inhibitory concentration of the Itraconazole Malassezia furfur standard, Malassezia furfur patient, Malassezia sympodialis standard, Malassezia sympodialis patient, Malassezia dermatis standardntia, Malassezia dermatis patient were 10 microgram/ml, 10 microgram/ml, 10 microgram/ml, 0.1 microgram/ml, 1 microgram/ml, and 0.01 microgram/ml, respectively. 3. Minimum inhibitory concentration of the ketoconazole Malassezia furfur standard, Malassezia furfur patient, Malassezia sympodialis standard, Malassezia sympodialis patient, Malassezia dermatis standard, Malassezia dermatis patient were 0.01 microgram/ml, 10 microgram/ml, 10 microgram/ml, 0.1 microgram/ml, 0.01 microgram/ml, and 0.01 microgram/ml, respectively. 4. Malassezia furfur standard, Malassezia furfur patient, Malassezia sympodialis patient and Malassezia dermatis patient showed the strongest antimicrobial effect on bamboo oil > ketoconazole > itraconazole. 5. Malassezia sympodialis standard, Malassezia sympodialis patient and Malassezia dermatis standard strongest antimicrobial effect on ketoconazole > bamboo oil > itraconazole. CONCLUSION: Bamboo oil might be applied as antidandruff treatment modality by its anti-malassezial effect.

Anti-inflammatory and Anti-oxidant Effects of Sophora flavescens Root Extraction in Lipopolysaccharide-activated Raw 264.7 Cells / 대한의진균학회지

BACKGROUND: The macrophages activated by lipopolisaccharide produce numerous molecules and proteins, such as tumor necrotic factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-1beta, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and free radicals, associated with inflammation. The response was occurred by intracellular cascaded, NF-kappaB pathway. So, the regulation of this pathway is very important in control of inflammation. OBJECTIVE: In this study, the anti-inflammatory and anti-oxidant effects of Sophora flavescens that is used empirically in oriental medicine and folk remedy were evaluated and the mechanism of the effects was studied. METHODS: By using the root extracts of Sophora flavescens, we performed experiment in LPS and interferon-gamma (IFN-gamma)-activated Raw 264.7 cells. We measured the production of NO, PGE2 and expression of iNOS and COX-2 in activated Raw 264.7 cells with Sophora flavescens root extract. Also, we tested anti-oxidant effect of Sophora flavescens root extracts by ELISA kit in activated Raw 264.7 cells, and the free radical scavenging effect of material itself by DPPH assay. RESULTS: The Sophora flavescens root extracts decreased the production of NO (p<0.001) and PGE2 (p<0.01) in Raw 264.7 cells activated by LPS and IFN-gamma. The expression of proteins, iNOS and COX-2, suppressed along with the elevated concentration of Sophora flavescens root extracts. The result of DPPH assay was that the test material itself had scavenging effect for free radical (p<0.001). And the antioxidant activity in activated Raw 264.7 cells was increased with the level of the Sophora flavescens root extracts (p<0.05). CONCLUSION: The Sophora flavescens root extracts suppressed the production of NO and PGE2 through the decreased expression of iNOS and COX-2. And the Sophora flavescens root extracts had the scavenging effect about free radicals itself and increased the antioxidant activity in activated macrophages.

Optimal Culture Condition for Antifungal Susceptibility Tests of Malassezia globosa / 대한의진균학회지

BACKGROUND: Although numerous culture conditions for Malassezia species were suggested, there were not so many objective evaluation articles in the literature. OBJECTIVES: We examined the various culture conditions for Malassezia globosa. METHODS: Malassezia globosa culture conditions were assessed by Dixon's agar, modified Leeming-Notman medium in diverse oil content and temperature conditions. RESULTS: Maximum growth rate of Malassezia globosa was achieved at 3% olive oil. The optimal temperatures for the maximal growth of M. globosa were observed at 32~34degrees C. CONCLUSION: In this study, we established the optimal culture condition for M. globosa, and confirmed its excellent utility for the antifungal susceptibility tests for M. globosa and M. restricta. Our results can help the investigators plan to do the prospective researches involving Malassezia species, such as the susceptibility test for newly developed antifungal agents.

Development of an Imaging System for Use in Diagnosing Dermatologic Diseases / 대한피부과학회지

BACKGROUND: The imaging system that's currently being used in the field of dermatology is based on such instruments as the dermoscope, phototrichograph and camera. In recent years, the use of an image magnification system based on polarization has become popular. OBJECTIVE: In this study, optical quantification was performed based on the multiwavelength imaging analysis of the structures that form dermatologic diseases, and an attempt was made to enhance the image quality by using polarization technology. METHODS: The lesions of three patients who were clinically diagnosed with cherry angioma, melanocytic nevus and inflammatory lesions in acne and freckles were measured at the outpatient clinic of the Department of Dermatology of the authors' hospital. All the patients were female, and their mean age was 29.3 years. RESULTS: The optical characteristics of the patients' various skin lesions, including cherry angioma, melanocytic nevus and inflammatory lesions in acne and freckles, were distinguishable by their wavelength. CONCLUSION: The use of different kinds of information may be helpful for measuring and diagnosing various skin lesions that have not been differentiated with using the previous modalities. Further, if the various environmental factors that may be generated during the measurement process can be controlled, then these study results can be applied to a standard diagnostic modality in the field of dermatology.