• Toxicological Research
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• v.14 no.3
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• pp.315-320
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• 1998

In vitro skin iritation of anti-inflammatory drugs was investigated in terms of the cytotoxicity method to human skin fibroblast cells. Five anti-inflammatory drugs (Diclofenac, Naproxen, Meclofenamic acid, Ibuprofen and Fnoprofen) which are commercially available as oral preparations or injections were tested. The cytotoxicity of 5 chemicals was evaluated by using MTT[tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. NRU (neutral red uptake) assay and Alamar Blue assay after fibroblast cells had been exposed to the chemicals for 24 hours or 489 hours. The $IC_{50}$ values of the chemicals showed the comparative strength of cytotoxicity as following order of Meclofenamic acid>Diclofenac>Fenoprofen>Ibuprofen>Naproxen. The values of $IC_{50}$ determined by Alamar Blue assay were lower than those of MTT and NRU assay. These data suggest Alamar Blue assay can be useful method for assessing in vitro skin irritation potential of anti-inflammatory drugs.

• Toxicological Research
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• v.28 no.2
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• pp.113-116
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• 2012

Various kinds of positive effects of green tea extracts had been studied for long time which included anti-inflammation, anti-aging, and cardiometabolic effects. Although topical steroid and non-steroidal calcineurin inhibitors may control clinical symptoms of allergic contact dermatitis, some of patients also present allergic reaction to these topical agents. Therefore, we have tried green tea extracts for managing this skin disorder with expectation of anti-inflammatory effect without potential side effects including skin irritation and toxic responses. The toxicity test of green tea extract also did not show any sign of irritation in the skin throughout the test period. Moderate severity of allergic contact dermatitis presented satisfactory clinical outcome at second week follow-up which was final visit of outpatient. This result mean that green tea extract has a positive effect for managing allergic contact dermatitis but its potency and efficacy seem to be so not strong enough to control moderate severity allergy skin lesion. In this pilot study, we were able to conclude that green tea cell extracts might be applied for potential anti-inflammatory soaking without skin toxicity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.5
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• pp.1125-1131
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• 2009

Lophatheri Herba, Patriniae Radix, Alli tuberosi Herba, and Menthae Herba have long been used externally for treatment of injuries or inflammations in Korea. This study was carried out to examine the anti-dandruff effect of the mixture made with equal parts of 70%-ethanol extracts of Lophatheri Herba, Patriniae Radix, Allii tuberosi Herba, and Menthae Herba (LPAM) and of each one above, compared with 1%-zinc pyrithione (Standard) used commercially as anti-dandruff. The results showed the mixture was more effective than each crude drug. The 10%-LPAM produced no toxicity in skin reaction test and eye irritation test, and showed significant antimicrobial activity against Pityrosporum ovale, the major cause of dandruff, and also showed significant anti-dandruff and itch-improvement effects on head skin.

• Natural Product Sciences
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• v.27 no.4
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• pp.300-306
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• 2021

Chronic ultraviolet (UV) radiation causes photoaging, which represents skin damage, disrupts skin barrier function, and promotes wrinkle formation. We investigated that the polysaccharide extract of an edible basidiomycetous white jelly mushroom, Tremella fuciformis, (TF-Glucan^®) exhibited statistically photoprotective activity by inhibiting matrix metalloproteases (MMPs) and increasing collagen synthesis, and an anti-inflammatory activity by inhibiting nitric oxide and pro-inflammatory cytokines at the concentrations of less than 1000 ㎍/ml, which is not cytotoxic (p < 0.05). Additionally, TF-Glucan^® increased the expression of involucrin and filaggrin to prevent the disruption of UVB-induced barrier function (p < 0.05). TF-Glucan^® was assessed as a safe material by the human primary skin irritation (1, 3, 5%), human repeated insult patch test (no sensitization at 5%), 3T3 NRU phototoxicity assay (no phototoxicity, PIF < 2, MPE < 0.1), eye irritation test test by BCOP (no category, IVIS ≤ 3) and local lymph node assay (negative at 10, 25, 50%) for identifying potential skin sensitizing. These results suggest that TF-Glucan^® may be useful as an anti-photoaging ingredient for developing cosmeceuticals.

• Proceedings of the SCSK Conference
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• 1999.10a
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• pp.145-154
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• 1999

A novel retinol derivative, polyethoxylated retinamide (Medimin A) was synthesized, as an anti-aging agent. Collagen synthesis, skin permeation, stability, and toxicity of Medimin A were evaluated and compared with those of retinol and retinyl palmitate. In vitro collagen synthesis was evaluated by quantitative assay of [$^3H$]-proline incorporation into collagenase sensitive protein in fibroblast cultures. For in vitro skin permeation experiments, Franz diffusion cells (effective diffusion area: $1, 766{\;}\textrm{cm}^2$) and the excised skin of female hairless mouse aged 8 weeks were used The stabilities of retlnoids were evaluated at two different temperature ($25{\;}^{\circ}C$ and $40{\;}^{\circ}C$) and under UV in solubilized state and in OW emulsion. To estimate the safety, acute oral toxicity, acute dermal toxicity, primary skin irritation, acute eye irritation and human patch test were performed The effect of Medimin A on collagen synthesis was similar to that of retinol. The skin permeability of Medimin A was higher than those of retinol and retinyl palmitate. The Medimin A was more stable than retinol and retinyl palmitate. Medimin A was nontoxic in various toxicological tests. These results suggest that Medimin A would be a good anti-aging agent for enhancing bioavailability and stability.

• Toxicological Research
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• v.18 no.3
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• pp.267-273
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• 2002

Fructose 1,6-diphosphate(FDP), a glycolytic metabolite, is reported to ameliorate inflammation and inhibit the nitric oxide production in murine macrophages stimulated with endotoxin. It is also reported that FDP has cytoprotective effects against hypoxia or ischemia/reperfusion injury in brain and heart, and may play a protective role in ultraviolet B (UVB, 280~320 nm)-injured keratinocyte by attenuating prostaglandin (PG)-E$_2$production and cyclooxygenase (COX)-2 expression, which are possibly through blocking the intracellular reactive oxygen species (ROS) accumulation. Therefore FDP is considered to act as a potent antioxidant especially in the skin. We conducted the several safety tests (single-dose toxicity, primary skin irritation test, eye irritation test, skin sensitization test, phototoxicity test, photosenitization test and human patch test) to see if FDP is safe in case used for the skin application. Our data obtained hitherto suggest that FDP is very safe if applied to the skin.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.731-741
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• 1996

ELISA kit for cortisol was developed and then evaluated. Polyclonal antihydrocortisone-3-(o-carboxymethyl)oxime BSA rabbit serum was used to coat the 96-well microplates. The minimum detection limit of the kit was 250pg of cortisol per milliliter. The within-run variation and the day to day variation of the ELISA system were 2.0 and 5.9 at maximum, respectively. The kit was used to determine whether salivary cortisol concentration could replace blood cortisol concentration in dexamathasone suppression test of dogs. Changes of cortisol concentration were measured in serum or saliva after intravenous administration of 0.01mg of dexamethasone per kilogram of body weight. Blood alone, saliva alone or both were collected at 0, 30, 60, 120, 240, and 360 minutes after injection of dexamethasone. The change in blood cortisol concentration was found to be suitable in dexamathasone suppression test of dogs, but the change in salivary cortisol concentration was not. The kit was also used to determine whether salivary cortisol concentration could be a stress index as well as blood cortisol concentration in dogs. Two types of trial were performed to estimate the stress either by blood or salivary cortisol concentration. The first trial was stress experiment by intravenous injection of 0.2IU of PZI-insulin per kilogram body weight. Either blood alone or saliva alone was collected at 0, 30, 60, and 90 minutes after insulin administration. Both blood and salivary cortisol concentration were found to be suitable index in estimating stress from hypoglycemia by injection of insulin. The second trial was stress experiment by electrical irritation. The dogs were irritated with anti-bark device for 10 seconds. Blood was collected before and at 2 and 5 minutes after electrical irritation. Saliva was collected before and at 3 and 6 minutes after electrical irritation. The blood cortisol concentration, but not the salivary cortisol concentration was found to be suitable index in estimating stress from electrical irritation. Cushing syndrome in a dog was also successfully diagnosed with this kit.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.2
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• pp.227-233
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• 2004

Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.25 no.4 s.34
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• pp.145-154
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• 1999

A novel retinol derivative, polyethoxylated retinamide(Medimin A) was synthesized, as an anti-aging agent. Collagen synthesis, skin permeation, stability, and toxicity of Medimin A were evaluated and compared with those of retinol and retinyl palmitate. In vitro collagen synthesis was evaluated by quantitative assay of $[^3H]-proline$ incorporation into collagenase sensitive protein in fibroblast cultures. For in vitro skin permeation experiments, Franz diffusion cells(effective diffusion area: 1,766 $cm^2$) and the excised skin of female hairless mouse aged 8 weeks were used, The stabilities of retinoids were evaluated at two different temperature($25^{\circ}C\;and\;40^{\circ}C$) and under UV in solubilized state and in O/W emulsion. To estimate the safety, acute oral toxicity, acute dermal toxicity, primary skin irritation, acute eye irritation and human patch test were performed. The effect of Medimin A on collagen synthesis was similar to that of retinol. The skin permeability of Medimin A was higher than those of retinol and retinyl palmitate. The Medimin A was more stable than retinol and retinyl palmitate. Medimin A was nontoxic in various toxicological tests. These results suggest that Medimin A would be a good anti-aging agent for enhancing bioavailability and stability.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.12 no.1
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• pp.18-35
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• 1999

Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散) have been as eye washes of inflammatory eye disease in the oriental medicine. Especially Jinpisan(秦皮散) has been used for the disease which is similar to Peudomonas aeruginisa keratitis. This research was attempted to investigate the effect of Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散), on Peudoronas aeruginisa keratitis. Pseudomonas aeruginosa keratitis causes a deep rapid intense ulceration which often leads to perforation of the cornea within 48 hours. In this research, we induced keratits in the rabbits by inoculating Pesudomonas aeruginosa(9027) and observed the effect on the keratitis and the irritation against the external eye. Also we mesured the minimum inhibitory consentration(MIC) of Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散) by agar diliution method and the anti-bacterial activites by disk method. The tested bacteria were as follows : a) Pseudomonas aeruginosa (9027), b) Streptococcus pneumoniae(6303), c) Staphylococcus epidermidis(12228), d) Staphylococcus aureus(6538P). The results were as follows ; 1. The groups which were applied eye washes of Fraxini Cortex, Jinpisan reavealed a significant effect, but the group applied eye wash of Coptidis Rhizoma reveaded no effect on Pseudomonas aeruginosa keratitis. 2. Applying eye washes of Coptidis Rhizoma, Fraxini Cortex, Jinisan revealed an irritation against external eyes. 3. Coptidis Rhizoma showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylucoccus aureus by agar diliution method 4. Coptidis Rhizoma showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by disk method. 5. Fraxini Cortex showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by agar diliution method 6. Fraxini Cortex showed an anti-bacterial activity on Pseudomonas aeruginosa, Sireptococcus pneumoniae, Staphylococcus epidermidis, Staphy1ococcus aureus by disk method. 7. Jinpisan showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by agar diliution method. 8. Jinpisan showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by disk method. According to the above results, Fraxini Cortex, Jinpisan(秦皮散) are recognized to have an effective treatment on the Pesudomonas aeruginosa keratitis, so this experiment is thought to be a basic ingredient in proving the effect of Fraxini Cortex, Jinpisan which is applied many in documents and clinical medicine. In the comparison of anti-bacterial activity and results of treatment on the Pesudomonas aeruginosa keratitis, Jinpisan(秦皮散) was more effective than Coptidis Rhizoma, Fraxini Cortex.