• Title/Summary/Keyword: skin fibroblast cell

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Comparison of Sensitivity Between Balb/c 3T3 Cell and HaCaT Cell by NRU Assay to Predict Skin Phototoxicity Potential

  • Lee, Jong-Kwon;Lee, Eun-Hee;Lee, Sun-Hee
    • Toxicological Research
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    • v.18 no.3
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    • pp.227-232
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    • 2002
  • In order to find out the appropriate in vitro method for high correlation with in vivo, we com-pared the sensitivities of phototoxicity (PT) in vitro method between in human keratinocytes, HaCaT cells and in 3T3 fibroblast cells derived from Balb/c mice. Both cells were exposed to six known phototoxic chemicals : promethazine, neutral red, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen, or non-phototoxic chemical, ALS (ammonium laureth sulfate) and then irradiated with 5 J/$cm^2$ of UVA. Cell viability ($IC_{50}$ ) was measured by neutral red uptake (NRU) assay. The ratio of $IC_{50}$ value of chemicals in the presence and absence of UVA was determined by the cut-off value. The phototoxic potential of test chemicals in NRU assay was determined by measuring the photoirriation factor (PIF) with a cut-off value of 5. In both 3T3 and HaCaT cells, all known phototoxic chemicals were positive (over 5 of PIF value), except that bithionol was found to be non-phototoxic to HaCaT cells, and ALS, non-phototoxic chemical was negative. These results suggest that Balb/c 3T3 cell was more sensitive than HaCaT cell to predict phototoxicity potential.

Effect of Fermented Soybean on the Proliferation and Growth in HaCaT and Fibroblast Cell (대두 발효물이 인간 유래 피부세포의 증식 및 성장에 미치는 영향)

  • Kim, Eun-Joo;Han, Myung-Ryun;Lee, So-Young;Kim, Ae-Jung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.2
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    • pp.326-335
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    • 2021
  • This study was undertaken to determine the effect of fermented soybean extract and its fractions on skin cell proliferation and growth. The extract was procured by the pepsin and Lactobacillus rhamnosus fermentation of soybean. LC-MS analysis was performed subsequent to soybean fermentation, and cell viability was measured by the WST-1 assay. Cell proliferation was observed to increase after exposing cells to the fermented soybean extract and its fractions at all concentrations tested (0~2,000 ㎍/mL). In particular, compared to the normal control group and 120 % proliferation of the EGF (epidermal growth factor) positive control group, 160~180 % cell proliferation was achieved at 800 ㎍/ml, indicating the excellent potential as an application material for skin aging inhibition and skin cell regeneration. In addition, we also examined the effects of fermented soybean extract and its fractions on wound healing ability, in HaCaT cells and fibroblasts. Our results indicate excellent cell migration abilities after treatment with fermented soybean extract and its fractions, as compared to the control treatment. Similar cell migration abilities were observed in the positive control group (EGF). Taken together, our results indicate that fermented soybean extract and its fractions (F4 and F5) exert amelioratory effects as a natural material for skin.

Effect of Topically Applied Silver Sulfadiazine on Fibroblast Cell Proliferation and Biomechanical Properties of the Wound

  • Lee, Ae-Ri-Cho;Moon, Hee-Kyung
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.855-860
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    • 2003
  • The effect of silver sulfadiazine (SSD) on the proliferation of human dermal fibroblast (HDF) was studied to determine the impact of the drug on the wound healing process and dermal mechanical strength. Human dermal fibroblasts were cultured to 80% confluency using DMEM with 10% FBS and viability of the cell was estimated using neutral red assay. In addition, the $2^{nd}$ degree burn wound was prepared on the anterior part of rabbit ear skin and dressings containing SSD were applied for 96 h. Presence of inflammatory cells and degree of re-epithelialization were investigated in the wound. After 15 day of the induction of burn wounds, the treated area was excised and dermal mechanical strength was quantitatively measured with a constant speed tensiometer. SSD was found to be highly cyto-toxic in cultured HDF cells. The topical application of SSD (2%) could control the infection as evidenced by the lack of accumulation of inflammatory cells in histological evaluation. Therefore, these observations suggested that the impairment of dermal regeneration and decreased mechanical strength of dermal tissue was resulted from the cyto-toxic effect of SSD on dermal cells. Since the decreased mechanical strength may lead to reduction in resilience, toughness and maximum extension of the tissue, the identification of optimum dose for SSD that limits infection while minimizes the cyto-toxic effect may be clinically relevant.

Effect of Torilis Fructus on Procollagen Biosynthesis and Activity of Matrix Metalloproteinase-I(MMP-1) in Human Dermal Fibroblast (사상자(Torilis Fructus)가 섬유아세포의 Procollagen 생합성과 Matrix metalloproteinase-I(MMP-1)의 활성에 미치는 영향)

  • Koo, Bon-Suk;Hwang, Eui-Il;So, Seung-Ho;Lee, Seong-Kye;Han, Gyeong-Ho;Kim, Na-Mi
    • Korean Journal of Pharmacognosy
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    • v.38 no.4
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    • pp.349-353
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    • 2007
  • Skin wrinkle formations are associated with collagen synthesis and matrix metalloproteinase-I(MMP-1) activity. This study was carried out to find out skin wrinkle reducing components in Torilis Fructus. Torilis Fructus were extracted with 70% ethanol and the ethanol extracts were systematically fractionated with n-hexane, ethylacetate, n-butanol and distilled water. Among them, antiwrinkle component from n-hexane fraction was purified by several column chromatographies and HPLC, which identified as torilin by $^1H-NMR,\;^{13}C-NMR$ and ESI-MS. To determine cell viability, collagen biosynthesis and MMP-1 activity, human dermal fibroblast was treated with 1-5 ppm concentrations of Torilis Fructus extract fraction and torilin. Cell viability was showed 84-102% at all group treated with 1-5 ppm. Collagen synthesis was increased in all group, especially torilin-treated group was highest amount. Active forms of MMP-1 were decreased in all group. From these results, we consider that Torilis Fructus have several antiwrinkle components and torilin may be one of the effective components.

Anti-oxidant and Anti-skin-aging Effects of Abalone Viscera Extracts in Human Dermal Fibroblasts (전복내장추출물의 항산화 및 human dermal fibroblasts에 대한 항피부노화 효과)

  • Li, Jinglei;Tong, Tao;Ko, Du-Ock;Chung, Dong-Ok;Jeong, Won-Chul;Kim, Ji-Eun;Kang, Seong-Gook
    • Food Science and Preservation
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    • v.19 no.4
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    • pp.463-469
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    • 2012
  • In this study, the anti-oxidant and anti-elastase activities of four abalone viscera extracts were investigated to screen the most promising extract. This extract was further studied in terms of its anti-skin-aging properties. In the DPPH-scavenging assay, the Tris-HCl extract showed a $58.60{\pm}0.88%$ radical-scavenging activity, which was followed closely by the ethanol extract that had a $55.40{\pm}0.62%$ scavenging activity. In the anti-elastase assay, however, the ethanol extract showed the significantly highest elastase inhibition activity. Furthermore, none of the extracts had a harmful effect on the human dermal fibroblast, as revealed in the MTT assay. In the cell study, the effect of the ethanol extract at various concentrations on the human dermal fibroblast was investigated. At the 10 ${\mu}g/mL$ concentration, the ethanol extract boosted the pro-collagen type I synthesis to $705.30{\pm}3.06$ ng/mL and reduced the MMP-1 to $54.30{\pm}0.80$ ng/mL, which was considered the optimum concentration. This is the first study that focused on the anti-oxidant and anti-skin-aging effects of abalone viscera extract. Its results may provide fundamental data for further study.

In vitro Alternatives to Skin Irritation Test

  • Shin, Dae-Sup;Kim, Dai-Byung;Ryu, Seung-Rel;Lee, Sun-Hee;Koh, Jae-Sook;Park, Won-Sae;Kim, Pu-Young
    • Biomolecules & Therapeutics
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    • v.3 no.3
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    • pp.242-244
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    • 1995
  • In vitro cell culture system has been proposed as a promising alternative model to in vivo skin irritation test. These studies were performed to screen the cytotoxicity effects of surfactants using normal human skin fibroblasts. Cell membrane integrity assessed by the leakage of lactate dehydrogenase (LDH) and mitochondrial integrity by MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromides reduction test were affected in a dose dependent manner. The irritation potential of surfactants to human skin patch test, and the changes of capillary permeability by rabbit intradermal safety test were assessed as in vivo methods. Our results suggest that LDH leakage assay and MTT reduction test using cultured human fibroblasts could be predictive for the irritancy of various surfactants in human, and LDH assay is superior correlated with in vivo test (r=0.886) to MTT test with in vivotest (r=0.757).

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Effect of Campsis grandiflora on Antioxidative Activity in UVB-irradiated Human Dermal Fibroblasts (사람 섬유아세포에서 UVB 조사에 대한 능소화 추출물의 항산화 효과)

  • Kim, Jin-Hwa;Lee, Bum-Chun;Zhang, Yong-He;Pyo, Hyeong-Bae
    • YAKHAK HOEJI
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    • v.49 no.2
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    • pp.174-179
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    • 2005
  • The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmen tal facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Campsis grandiflora extract on the UVB-irradiated human dermal fibroblasts (HDFs). We tested free radical and superoxide scavenging effect in vitro. C. grandiflora extracts had potent radical scavenging effect by 82% at $100{\mu}g/ml$, respectively. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB 20 $MJ/cm^2$ after treatment of C.grandiflora extracts. The results showed that oxidation of CM-DCFDA was inhibited by C.grandiflora extracts effectively and C.grandiflora extracts has a potent free radical scavenging activity in UVB- irradiated HDFs. In ROS imaging using confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our results suggest that C.grandiflora can be effectively used for the prevention of UV-induced adverse skin reactions such as radical production, and skin cell damage.

Antioxidant potential of silk protein sericin against hydrogen peroxide-induced oxidative stress in skin fibroblasts

  • Dash, Rupesh;Acharya, Chitrangada;Bindu, P.C.;Kundu, S.C.
    • BMB Reports
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    • v.41 no.3
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    • pp.236-241
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    • 2008
  • The antioxidant potential of silk protein sericin from the non-mulberry tropical tasar silkworm Antheraea mylitta cocoon has been assessed and compared with that of the mulberry silkworm, Bombyx mori. Skin fibroblast cell line (AH927) challenged with hydrogen peroxide served as the positive control for the experiment. Our results showed that the sericin obtained from tasar cocoons offers protection against oxidative stress and cell viability is restored to that of control on pre-incubation with the sericin. Fibroblasts pre-incubated with non-mulberry sericin had significantly lower levels of catalase; lactate dehydrogenase and malondialdehyde activity when compared to untreated ones. This report indicates that the silk protein sericin from the non-mulberry tropical tasar silkworm, A. mylitta can serve as a valuable antioxidant.

Effects of Lavender, Lemon and Eucalyptus Essential Oil on Th2 related Factors of DNCB-induced Atopy dermatitis in NC/Nga Mice Model (라벤더, 레몬, 유칼립투스 혼합 에센셜오일이 아토피 피부염 동물 모델의 Th2 관련인자에 미치는 영향)

  • Kim, Hyeon-Ah;Yun, Mi-Young;Song, Hyang-Hee;Cheong, Kwang-Jo;Yoo, Hwa-Seung
    • Journal of Pharmacopuncture
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    • v.13 no.1
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    • pp.53-61
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    • 2010
  • Purpose : To investigate the effects of the lavender, lemon and eucalyptus oil mixture on the atopy dermatitis skin lesions induced on NC/Nga Mice by dinitrochlorobenzene (DNCB). Material and Method : For this purpose, we fabricated the oil mixture blending three essential oils (lavender, lemon, eucalyptus : ELL) with one carrier oil (jojoba) and apply it on the atopic dermatitis skin lesions of NC/Nga Mice. Atopic dermatitis in NC/Nga Mice was induced by DNCB treatment on the dorsal skin of mice for 8 weeks. The mixture of ratio of each essential oil drop was 1 (eucalyptus) : 2 (lemon) : 2 (lavender) and this mixture was blended with jojoba oil 50ml (0.025%). The ELL-ointment was supplied for 8 weeks. We evaluated the effects of ELL on cell viability of mouse lung fibroblast, clinical skin features and severity, the level of serum Immunoglobulin (Ig) E & Ig G1, Interleukin (IL)-4, IL-13 and Interferon (IFN)-$\gamma$. Results : ELL showed safety on the cell viability of mouse lung fibroblast compared with control group. The cell viability was measured by SRB method. The effects of ELL on clinical skin features and severity in DNCB-induced dermatitis model of NC/Nga mice was significant compared with control group. EEL also showed significant effects on clinical symptom score compared with control group. Serum IgE & IgG1 level and development of atopy dermatitis skin lesions were evaluated. Serum IgE & IgG1 production was significantly down-regulated in EEL group compared with control group. ELL also down-regulated the levels of IL-4 and IL-13, and up-regulated the level of IFN-$\gamma$ compared with control group significantly. Conclusion : ELL was effective on atopy dermatitis by modulating Th2 related factors.

Inhibitory activity of Lactobiocin on the skin inflammation and acnes (Lactobiocin의 피부 염증 및 여드름 저해효과에 관한 연구)

  • 김광수;오세종;김기환;홍진천;이승화
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.28 no.1
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    • pp.150-165
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    • 2002
  • The purpose of this study was to evaluate bacteriocin activity against human flora. Lactobiocin, a bacteriocin produced by Lactococcus sp. HY 449, inhibited the growth of Starphylococcus epidermidis, Starphylococcus aureus, Streptoccoccus pyogenes and Propionibacterium acnes. When crude bacteriocin was added to indicator cells during logarithmic growth, the optical density(O.D 650nm) of cells without bacteriocin increase after 5h of incubation. Whereas in the presence of bacteriocin, the O.D of cell suspensions decreased. The similar patterns were observed for absorbance readings at 280 nm and 260 nm. The release of cellular components when cell were treated with Lactobiocin suggests some degree of membrane damage or cell lysis. Scanning electron microscopy of cells following treatments with Lactobiocin in PBS buffer revealed disruptures of cell morphology. These results indicate that bacteriocin appears to cause cell lysis of tested strains. In cytotoxicity on human fibroblast, LD$\_$50/ of Lactobiocin was ca. 50 mg/ml and no change was observed cell proliferation at the same concentration. Any irritation and allergic reaction did not observed when evaluated by human patch test for Lactobiocin.