• Title/Summary/Keyword: collagen assay

Search Result 310, Processing Time 0.026 seconds

Effect of High glucose on JNK/ERK signaling pathway in UMR106 cells

  • Jung, In-Ok;Jin, Mei-Hua;Kim, Sung-Jin
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 2003.11a
    • /
    • pp.79-79
    • /
    • 2003
  • Recently diabetes has been found to be associated with metabolic bone diseases such as osteoporosis. In the present study, attempts have been made-to explore the effect of high glucose in bone formation. Osteoblast-like UMR 106 cells were treated with high glucose (22mM, 33mM, 44mM) for 1 or 2 days. High glucose significantly inhibited proliferation of UMR106 cells in a time- and dose- dependent manner as evidenced by MTT assay. For the evaluation of collagen synthesis, UMR 106 cells were cultured in high glucose media (44mM) for 24 h and the ratio of collagen content to total protein was measured. In addition, gene expression pattern of type I collagen was assessed by RT-PCR. The high concentration of glucose inhibited a collagen synthesis, a marker of bone formation activity. JNK, c- Jun N-terminal Kinase, is known to play an important role in stress-associated cell death. In this regard, we tested to determine whether high glucose has any effect on JNK activity. It has been found that treatment of high glucose induced phosphorylation of JNK. On the other hand, ERK phosphorylation was inhibited by high glucose in a dose-dependent manner. Taken together, Therefore these results indicate that inhibition of proliferation in UMR 106 cells following high glucose is related to JNK/ERK containing signal pathways. This study showed high glucose concentration could alter the bone metabolism leading to defective bone formation, suggesting that high glucose due to diabetes may playa significant role in the development of metabolic bone disease.

  • PDF

DEVELOPMENT OF POLYETHOXYLATED RETINAMIDE AS AN ANTI-AGING AGENT

  • Song, Young-Sook;Chung, Bong-Yul;Chang, Min-Youl;Park, Mun-Eok;Lee, Sung-Jun;Cho, Wan-Goo;Kang, Seh-Hoon
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.25 no.4 s.34
    • /
    • pp.145-154
    • /
    • 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.

  • PDF

UV Protection and Whitening Effects of Collagen Isolated from Outer Layer of the Squid Todarodes pacificus (동해 오징어 (Todarodes pacificus) 껍질 유래 콜라겐의 UV 보호능 및 미백효과)

  • Kwon, Min-Chul;Qadir, Syed Abdul;Kim, Hyo-Sung;Ahn, Ju-Hee;Cho, Nam-Hyun;Lee, Hyeon-Yong
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.41 no.1
    • /
    • pp.7-12
    • /
    • 2008
  • To develop a whitening agent, cytotoxicity of the soluble collagen isolated from Todarodes pacificus (CIT) was evaluated. CIT tested for cytotoxicity on human dermal fibroblast (CCD-986sk) was exhibited very low cytotoxicity. Because tyrosinase is the key enzyme for melanin biosynthesis, the use of various tyrosinase inhibitors is a common practice for whitening purpose in cosmetics. Tyrosinase inhibitory activity and melanin production assay were measured to confirm the whitening effect. The inhibitory effect of MMP-1 in UV-irradiated human dermal fibroblast was also performed. CIT showed strong inhibition potency on tyrosinase by 51.5% at 0.2 mg/mL which increased the inhibition by increasing the concentration of CIT, and showed 69.1% inhibition at 1.0 mg/mL. CIT showed strong inhibition effect on melanin production with 82% at 1.0 mg/mL. The CIT also reduced about 76% expression of MMP-1 in UV-irradiated CCD-986sk cell at 1.0 mg/mL. From the preliminary observations, we suggest that the collagen isolated from CIT could be a potential source of natural skin-whitening and anti-aging agents for the photo-damaged skin.

Inhibitory Effect of Fucoidan on TGF-β1-Induced Activation of Human Pulmonary Fibroblasts (후코이단에 의한 인간 폐 섬유모세포의 활성 억제 효과)

  • Yim, Mi-Jin;Lee, Dae-Sung;Choi, Grace;Lee, Jeong Min;Choi, Il-Whan
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.49 no.6
    • /
    • pp.807-814
    • /
    • 2016
  • Fucoidan, one of the dominant sulfated polysaccharides extracted from brown seaweed, possesses a wide range of biological activities. Transforming growth $factor-{\beta}$ ($TGF-{\beta}$) plays a pivotal role in the pathogenesis of pulmonary fibrosis, by stimulating the synthesis of profibrotic factors. In this study, we investigated the in vitro effects of fucoidan on collagen synthesis, ${\alpha}-smooth$ muscle actin (${\alpha}-SMA$) expression, and interleukin (IL)-6 production in $TGF-{\beta}$-stimulated human pulmonary fibroblasts. The expression of type I collagen and ${\alpha}-SMA$ was detected by Western blot, and the production of IL-6 by enzyme-linked immunosorbent assay. $TGF-{\beta}1$ treatment of pulmonary fibroblasts enhanced the expression of ${\alpha}-SMA$, type I collagen, and IL-6 whereas these effects were inhibited in cells pretreated with fucoidan. The activation of Smad2/3, p38 mitogen-activated protein kinases (MAPKs), and Akt was also inhibited in fucoidan-pretreated, $TGF-{\beta}1-stimulated$ human pulmonary fibroblasts. These data demonstrate the anti-fibrotic potential of fucoidan in $TGF-{\beta}-induced$ human pulmonary fibroblasts, via the inhibition of Smad2/3, p38 MAPKs, and Akt phosphorylation. Our results suggest the therapeutic potential of fucoidan in the prevention or treatment of pulmonary fibrosis.

Effect of Defatted Torreya nucifera Seed Extract on the Cross-linking of Advanced Glycation End Products to Collagen (비자 유박 추출물이 최종당화산물과 콜라겐 교차결합에 미치는 효과)

  • Son, Dahee;Kim, Minkyung;Park, Deokhoon;Jung, Eunsun
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.45 no.1
    • /
    • pp.19-25
    • /
    • 2019
  • Advanced glycation end products (AGEs) are final products formed by glycation reaction between reducing sugars and proteins, lipids or nucleic acids. These AGEs are related to progress of skin aging. In this study, we evaluate anti-aging activity of Defatted Torreya nucifera seed extract (DTSE) through antioxidant, anti-glycation, anti-elastase and inhibitory and breaking activity on the cross-linking of AGEs to collagen assay. Results showed that DTSE contained polyphenols and flavonoids. The $IC_{50}$ values of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity were $16.4{\mu}g$ (Dried materials, DM)/mL and $16.7{\mu}g\;DM/mL$, respectively. DTSE also inhibited the formation of AGEs, elastase activity and cross-linking of AGEs to collagen as well as broke existing cross-linking of AGEs to collagen in a dose-dependent manner. Consequently, our findings suggest that DTSE could be useful as a cosmetic material with anti-aging activity.

Effects of solvents on the anti-aging activity of Salvia miltiorrhiza extract (추출용매에 따른 단삼 추출물의 항노화 활성)

  • Guo, nan;Lee, Ji-An
    • Journal of Convergence for Information Technology
    • /
    • v.12 no.5
    • /
    • pp.290-297
    • /
    • 2022
  • This study was conducted to determine the cell cytotoxicity, astringency, nitrite oxide scavenging, iNOS protein expression level, pro-inflammatory cytokine, elastase inhibition, and type I pro-collagen synthesis as a functional cosmetics material of Salvia miltiorrhiza root. We prepared the 80% ethanol(SE) and hot-distilled water(SW), respectively. Both SE and SW showed no toxicity from 0.05 to 0.5 mg/mL concentration as a result of MTT assay in NHDF or RAW264.7 cells. In the measurement of astringent effect, SE reveled 74.6% of astringent activity in 10 mg/mL. SE showed that LPS-induced nitric oxide production, iNOS protein expression, and cytokines were inhibited in a dose-dependent manner. Furthermore, two extracts significantly inhibited elastase activity and increased the type I pro-collagen production. Therefore, it is expected that Salvia miltiorrhiza extract is used as a natural material for functional cosmetics that can effectively prevent skin-related inflammation and wrinkles, and aging.

The Effect of Tissue Plasminogen Activator on TGF-${\beta}1$ Pre-Treated Human Mesothelial Cell Line (TGF-${\beta}1$으로 자극한 사람중피세포주에서 조직플라스미노겐 활성제가 미치는 영향)

  • Lee, Jung-Lim;Jeon, Soo-Jin;Yoo, Young-Choon;Kim, Ji-Hye;Lee, Yu-Mi;Kwon, Sun-Jung;Son, Ji-Woong;Choi, Eu-Gene;Na, Moon-Jun
    • Tuberculosis and Respiratory Diseases
    • /
    • v.70 no.5
    • /
    • pp.405-415
    • /
    • 2011
  • Background: In an effort to find alternative therapeutic agents to prevent excessive fibrosis as a sequela to complicated parapneumonic effusion or empyema, we examined the effect of tissue plasminogen activator (tPA) as a fibrinolytic agent combined with talc or transforming growth factor (TGF)-${\beta}1$ in a human pleural mesothelial cell line, MeT-5A. Methods: MeT-5A cells were stimulated with various doses of talc, doxycycline or TGF-${\beta}1$ for 24 h and then were treated with tPA for an additional 24 h. Cell viability was measured by MTT assay. The production of interleukin (IL)-8 and vascular endothelial growth factor (VEGF) in the culture supernatants was measured by ELISA. Real-time PCR was carried out for measurement of type I collagen mRNA. Results: MeT-5A cells treated with talc showed a dose-dependent increase in production of IL-8. Talc also increased production of type I collagen mRNA at low doses, but talc did not influence the induction of VEGF. Addition of tPA to talc-stimulated cells showed further increases in the production of IL-8, but tPA did not influence the production of VEGF or type I collagen mRNA. TGF-${\beta}1$ increased the production of both VEGF and collagen type I mRNA, both of which were effectively inhibited by additional tPA treatment in MeT-5A cells. Conclusion: TGF-${\beta}1$ is a potent inducer of collagen synthesis without induction of IL-8 in MeT-5A cells. Addition of tPA after TGF-${\beta}1$ stimulation inhibited further fibrosis by direct inhibition of collagen mRNA synthesis as well as by inhibition of VEGF production.

Evaluation of the mechanical properties and clinical efficacy of biphasic calcium phosphate-added collagen membrane in ridge preservation

  • Lee, Jung-Tae;Lee, Yoonsub;Lee, Dajung;Choi, Yusang;Park, Jinyoung;Kim, Sungtae
    • Journal of Periodontal and Implant Science
    • /
    • v.50 no.4
    • /
    • pp.238-250
    • /
    • 2020
  • Purpose: This study aimed to evaluate the biocompatibility and the mechanical properties of ultraviolet (UV) cross-linked and biphasic calcium phosphate (BCP)-added collagen membranes and to compare the clinical results of ridge preservation to those obtained using chemically cross-linked collagen membranes. Methods: The study comprised an in vitro test and a clinical trial for membrane evaluation. BCP-added collagen membranes with UV cross-linking were prepared. In the in vitro test, scanning electron microscopy, a collagenase assay, and a tensile strength test were performed. The clinical trial involved 14 patients undergoing a ridge preservation procedure. All participants were randomly divided into the test group, which received UV cross-linked membranes (n=7), and the control group, which received chemically cross-linked membranes (n=7). BCP bone substitutes were used for both the test group and the control group. Cone-beam computed tomography (CBCT) scans were performed and alginate impressions were taken 1 week and 3 months after surgery. The casts were scanned via an optical scanner to measure the volumetric changes. The results were analyzed using the nonparametric Mann-Whitney U test. Results: The fastest degradation rate was found in the collagen membranes without the addition of BCP. The highest enzyme resistance and the highest tensile strength were found when the collagen-to-BCP ratio was 1:1. There was no significant difference in dimensional changes in the 3-dimensional modeling or CBCT scans between the test and control groups in the clinical trial (P>0.05). Conclusions: The addition of BCP and UV cross-linking improved the biocompatibility and the mechanical strength of the membranes. Within the limits of the clinical trial, the sites grafted using BCP in combination with UV cross-linked and BCP-added collagen membranes (test group) did not show any statistically significant difference in terms of dimensional change compared with the control group.

Increased Expression of Type I Collagen in AF-343 Treated Human Skin Fibroblasts (섬유아세포에서 민들레 추출물 함유제(AF-343)에 의한 제 1형 교원질 발현 증가)

  • Cho, Soo-Muk;Kim, Jae-Hyun;Kim, Jong-Keun;Park, Ki-Moon;Cho, Ho-Chan;Kim, In-Su;Kim, Beom-Joon;Chun, Young-Jin;Cho, Jae-We
    • YAKHAK HOEJI
    • /
    • v.56 no.1
    • /
    • pp.9-13
    • /
    • 2012
  • We previously reported that the extract of Taraxacum platycarpum (AF-343) had several biological properties such as skin hydration and anti-inflammatory effects, thereby AF-343 be a promising anti-atopic dermatitis agent. However, few studies have been conducted to evaluate its effect on modulation of extracellular matrix proteins in human skin fibroblasts. The purpose of this study was to investigate the expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 proteins in AF-343-treated human skin fibroblasts. Human skin fibroblasts were treated by various concentrations of AF-343 (0~2 mg/ml). The expressions of type I collagen, matrix metalloproteinase-1 (MMP-1), Smad2/3, and TIMP-1 proteins were analyzed by Western blot analysis. In addition, level of type I collagen mRNA was analyzed by CAT assay. Expression of type I collagen protein was increased in AF-343-treated human skin fibroblasts by dose and time-dependent manners. Consistent with this result, the expressions of phospho-Smad2/3 in skin fibroblasts were increased and MMP-1 expression was decreased by AF-343 treatment. TIMP-1 expression was not significantly changed in AF-343 treated skin fibroblasts. Extract of Taraxacum platycarpum (AF-343)-induced up-regulation of type I collagen expression was through increased expression of phospho-Smad2/3. These results were occurred combined with down-regulation of MMP-1 in skin fibroblasts. Taken together, this study indicated that AF-343 has property of the modulation of ECM in tissue as well as skin hydration and anti-inflammation.

Anti-oxidation and Anti-aging Effect of Mixed Extract from Korean Medicinal Herbs (한약재 복합 추출물이 항산화 및 항노화에 미치는 영향)

  • Hwang, Jae-Gyu;Yun, Jong-Kuk;Han, Kil-Hwan;Do, Eun-Ju;Lee, Jin-Sang;Lee, Eun-Ju;Kim, Jong-Boo;Kim, Mi-Ryeo
    • The Korea Journal of Herbology
    • /
    • v.26 no.1
    • /
    • pp.111-117
    • /
    • 2011
  • Objectives : The present study was designed to investigate effects of mixed extract from korean medicinal herbs (MIX) on oxidation/reduction reaction-related and aging-related enzyme in vitro. Methods : We performed MTT assay, collagenase inhibition assay, elastase inhibition assay, tyrosinase inhibition assay, DPPH free radical scavenging assay, SOD-like activity and xanthine oxidase inhibition assay. Results : Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. The MIX showed 97% inhibition of collagenase activity, and 64% inhibition of elastase activity at 1 mg/ml concentration of MIX, next only to positive control, which indicate good efficacy for anti-wrinkle ingredient. Also it's treatment showed 34% inhibition of tyrosinase activity, to relate whitening effect, at the same dose of MIX. Antioxidant activities were determined by DPPH radical scavenging, xanthine oxidase (XO) inhibiting activity and SOD-like activity. Also these scavenging, XO-inhibiting and SOD-like activities were measured in 91%, 80%, and 63% inhibition, respectively, at a treated dose of 1 mg/ml, compare to control. Conclusions : These results suggest that possibility of mixed korean medicinal herbs as a functional ingredient for anti-wrinkle and whitening, anti-oxidation and anti-aging cosmetic formula.