• Title/Summary/Keyword: collagen assay

Search Result 310, Processing Time 0.025 seconds

Water Extract of Taraxaci Radix Improves Rheumatoid Arthritis Induced by Type-II Collagen in Animal Models (민들레 뿌리 물 추출물의 류마티스 관절염 동물 모델에 대한 개선 효과)

  • Nho, Jong Hyun;Lee, Hyun Joo;Jang, Ji Hun;Yang, Beo Dul;Kim, A Hyeon;Woo, Kyeong Wan;Hwang, Tae Yeon;Seo, Jae Wan;Cho, Hyun Woo;Jung, Ho Kyung
    • Korean Journal of Medicinal Crop Science
    • /
    • v.27 no.1
    • /
    • pp.38-44
    • /
    • 2019
  • Background: Taraxacum platycarpum has been used in traditional medicine in Korea to treat intoxication and edema and as a diuretic. According to previous reports, it has anti-cancer, anti-gastritis, and anti-inflammation effects. However, the improvement effect of T. platycarpum on rheumatoid arthritis has not been investigated. The anti-oxidative and anti-inflammation effects of the aerial parts of T. platycarpum are different from those of its subterranean parts. Thus, we evaluated the effect of the water extracts of Taraxaci radix (WTR) on type II collagen-induced rheumatoid arthritis (CIA) in animal models. Methods and Results: Rheumatoid arthritis was induced by type II collagen. WTR (100 mg/kg and 500 mg/kg) was administered to the animal models. Methotrexate was used as the positive control. The levels of interleukin-6, TNF-alpha, and type II collagen IgG in the animals were measured by using enzyme-linked immunosorbent assay. Treatment with 500 mg/kg WTR decreased the serum levels of interleukin-6, TNF-alpha, and collagen IgG in the CIA models. Moreover, treatment with WTR diminished the arthritisinduced swelling of the hind legs and monocyte infiltration in the bloodvessels of the animal models. Conclusions: These results indicate that WTR has the potential to improve rheumatoid arthritis by reducing the levels of inflammatory cytokines such as interleukin-6 and TNF-alpha. However, further experiments are required to elucidate the influence of WTR on signal transduction in vitro and in vivo.

Effects of Polygoni Multiflori Radix on the Elastase, and Collagenase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts

  • Kim, Myung-Gyou;Leem, Kang-Hyun
    • The Korea Journal of Herbology
    • /
    • v.29 no.1
    • /
    • pp.7-12
    • /
    • 2014
  • Objectives : Polygoni Multiflori Radix (PMR), the roots of Polygonum multiflorum Thunberg, is used to nourish the blood and yin and used for preventing premature greying of the hair. There are some articles on its preventing effects on the melanogenesis. However, there is no report about its effects on the collagen and elastin. The present study was designed to investigate its effects on collagen metabolism and elastase activity. Methods : The effects of PMR on type I procollagen production and collagenase activity in human normal fibroblasts Hs68 after UVB (312 nm) irradiation were measured by ELISA method. Cells were pretreated with the PMR for 24 hours prior to UVB irradiation. After UVB irradiation, cells were retreated with the sample and incubated for additional 24 hours. The amount of collagen type I was measured with a procollagen type I C-peptide assay kit. The activity of collagenase was measured with a MMP-1 human biotrak ELISA system. The elastase activities after treatment of PMR were measured as well. Results : In the present study, the collagen production was not increased. However, the increased collagenase activity after UVB damage was significantly recovered to $50.2{\pm}14.5%$, $8.2{\pm}3.1%$, and $10.0{\pm}3.3%$ (10, 30, and $100{\mu}g/ml$). The elastase activities (10, 100, and $1000{\mu}g/ml$) significantly reduced to $75.2{\pm}5.2%$, $40.3{\pm}1.2%$, and $27.0{\pm}1.9%$, respectively. Conclusion : PMR showed the inhibitory effects on collagenase and elastase activity. These results suggest that PMR may have potential as an anti-aging ingredient in cosmetic herbal treatment.

Effects of Draconis Resina on the Collagenase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts, and Tyrosinase Activity

  • Kim, Tae Yeon;Leem, Kang-Hyun
    • The Korea Journal of Herbology
    • /
    • v.30 no.6
    • /
    • pp.1-6
    • /
    • 2015
  • Objectives : Draconis Resina (DR), the resin of Daemonorops draco Bl., is used to circulate the blood and to stop bleeding. It also has been used to generate flesh including ulceration. The present study investigated the effects of DR extract on collagen metabolism in human fibroblasts and tyrosinase activity in mushroom tyrosinase.Methods : The effect of DR extract on type I procollagen production (collagen type I synthesis) and collagenase (matrix metalloproteinase-1, henceforth referred as MMP-1) activity in human normal fibroblasts cell line. Hs68 cells after ultraviolet B (UVB, 312 nm) irradiation was measured using the enzyme - linked immunosorbent assay (ELISA). The tyrosinase activity was also measured to find out the whitening effects in mushroom tyrosinase by ELISA method.Results : There was no cytotoxicity at DR extract at concentrations of 10 μg/ml, 30 μg/ml, and 100 μg/ml. DR extract significantly inhibited the increase of collagenase activity, whereas it did not show on the reduction of type I procollagen in UVB damaged Hs68 cells. DR extract did not reduce the L - DOPA oxidation. However, it significantly reduced the tyrosinase activity by DR extract at concentraions of 0.1 mg/ml, 1 mg/ml and 10 mg/ml.Conclusions : In conclusion, DR showed the anti-wrinkle and whitening effects via the inhibition of collagenase production and the tyrosinase activity. These results suggest that DR may have potential as an anti-aging ingredient in cosmetic herb markets.

Fabrication of gelatin-amorphous CaP nano fibrous mat forusing as fast bone healing material

  • Sarkar, Swapan Kumar;Song, Ho-Yeon;Lee, Byong-Taek
    • Proceedings of the Materials Research Society of Korea Conference
    • /
    • 2009.11a
    • /
    • pp.40.2-40.2
    • /
    • 2009
  • Using the favorable resorption behavior of amorphous Calcium phosphate (CaP) we fabricated a gelatin basednano fibrous mat by electrospinning for using as a fast healing patch for minorbone defects. Bone is predominantly formed by an inorganic phase of nano-crystalline HAp materials and nano fibrous protein material of collagen. The osteoblast cells, which are the bone formation cells and are key to the new bone formation, receive these materials to form new bone. Taking these considerations we make a new nano fibrous mat of amorphous CaP and gelatin, which is derived from collagen itself. A polymer carrier of poly caprolactone(PCL) was used in the system to stabilize the materials in biological condition. The electrospinning conditions were optimized for smooth mat without any droplet formation. The fabricated mat was characterized for its morphologyby SEM. Mechanical properties like tensile strength was evaluated. To investigate the bio-compatibility we performed the MTT assay and investigated its resorption behavior and apatite formation behavior by SBF immersion.

  • PDF

Effects of Silkworm Hemolymph and Cartilage-specific Extracellular Matrices on Chondrocytes and Periosteum-derived Progenitor Cells

  • Shin, Hyun-Chong;Choi, Yong-Soo;Lim, Sang-Min;Lee, Chang-Woo;Kim, Dong-Il
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.11 no.4
    • /
    • pp.364-367
    • /
    • 2006
  • In order to enhance the repair of defects in articular cartilage via cell therapy with autologous chondrocytes, as well as with periosteum-derived progenitor cells (PDPCs), silkworm hemolymph (SH) and a variety of cartilage-specific extracellular matrices (ECMs) including type II collagen, proline, chondroitin 4-sulfate, and chondroitin 6-sulfate were assessed with regard to their efficacy as media supplements. SH, a known anti-apoptotic agent, was found to enhance cell growth, as was shown by the results of a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to the results of reverse transcriptase polymerase chain reaction (RT-PCR) analyses, the cartilage-specific ECMs were found to stimulate the expression of hyaline cartilage-specific genes, most notably type II collagen and Sox9, in monolayer cultures of PDPCs.

Antithrombotic Effect of Galla Rhois (오배자의 항혈전 효과)

  • Song, Gyu-Yong;Park, Byung-Jun;Kim, Sung-Hoon
    • Korean Journal of Pharmacognosy
    • /
    • v.33 no.2 s.129
    • /
    • pp.120-123
    • /
    • 2002
  • The possibility of Galla Rhois(GR) water extract as an antithrombotic agent was investigated. The effect of GR on platelet aggregation in human platelet-rich plasma(PRP) induced by collagen and ADP in vitro and coagulation parameters in a pathological model induced by endotoxin and hydrocortisone acetate(HA) in vivo were examined. In platelet aggregation assay, GR extract significantly inhibited platelet aggregation induced by collagen and ADP in a dose-dependent manner. GR extract significantly increased the number of platelet and shortened prothrombin time(PT) and activated thromboplastin time(APTT) as compared with the control in pathological model induced by endotoxin and HA. Also, GR extract significantly increased fibrinogen level as compared with the control in a pathological model induced HA. These results suggest that GR may be a promising antithrombotic agent.

An improved method to determine hydroxyproline in an immortalized rat liver stellate cell line (HSC-T6)

  • Lee, Eun-Su;Kwak, Wie-Jong;Lee, Eung-Seok;Soh, Yun-Jo
    • Proceedings of the PSK Conference
    • /
    • 2003.04a
    • /
    • pp.154.1-154.1
    • /
    • 2003
  • Hydroxyproline (HYP) is a post-translational product of proline hydroxylation catalyzed by an enzyme prolyl 4-hydroxylase which plays a crucial role in the synthesis of all collagens, because the 4-hydroxyproline residues are essential for the folding of the newly synthesized collagen polypeptide chains into triple-heical molecules. Considering the role of collagen and its significance in many clinically important diseases such as liver cirrhosis, a great deal of attention has been directed toward the development of an assay at cell-based system. (omitted)

  • PDF

TWO COLORIMETRIC ASSAYS VERIFY THAT CALCIUM SULFATE PROMOTES PROLIFERATING ACTIVITY OF HUMAN GINGIVAL FIBROBLASTS

  • Chae, Min;Kim, Su-Yeon;Kim, Soo-Yeon;Lee, Suk-Won
    • The Journal of Korean Academy of Prosthodontics
    • /
    • v.45 no.3
    • /
    • pp.382-388
    • /
    • 2007
  • Statement of problem. The role of calcium sulfate in stimulating the growth of gingival soft tissue has been reported in few studies. Such a unique property of calcium sulfate could serve as a trouble-solving broker in compensating for the lack of soft tissues in various oral surgeries. Purpose. The purpose of this study was to compare the proliferating activities of human gingival fibroblasts seeded on various bone graft barrier materials of calcium sulfate, collagen, and polytetrafluorethylene (PTFE). Material and methods. Two calcium sulfates ($CAPSET^{(R)}$. and $CalForma^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA), a resorbable natural collagen ($Bio-Gide^{(R)}$, Geistlich Pharma Ag., Wolhusen, Switzerland), and a non-resorbable PTFE ($TefGen-FD^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA) served as the human gingival fibroblasts' substrates and comprised the four experimental groups, whereas the untreated floors of culture plastics were used in the control group, in this study. Cells were trypsinized, seeded, and incubated for 48 h. The proliferating activities of fibroblasts were determined by XTT and SRB assay and absorbance (optical density, OD) was measured. One-way ANOVA was used to analyze the differences in the mean OD values between the groups of CAPSET, CalForma, Bio-Gide, TefGen, and the control (p<0.05). Results. From the XTT assay, the mean OD value of the control group, the highest, was significantly greater than that of any of the four experimental groups followed by CalForma, CAPSET, TefGen, and Bio-Gide. Further, the mean OD value of CalForma, was significantly greater compared to that of Bio-Gide. From the SRB assay, Calforma showed the highest mean OD value, which was significantly greater than that of any other groups, followed by the control, CAPSET, Bio-Gide, and TefGen. The mean OD values of both the control and CAPSET were significantly greater compared to that of TefGen (p<0.05). Conclusion. Assessment of the viability and proliferation of cultured fibroblasts seeded and incubated for 48 h on various barrier-material substrates using XTT and SRB assay showed that calcium sulfate $CalForma^{(R)}$ promotes the proliferating activity of human gingival fibroblasts.

Antioxidant and Antiwrinkle Effects of Persimmon Leaves extract (시엽(Persimmon Leaves) 에탄올 추출물의 항산화와 항주름 효과)

  • Sung-Hee Kim;Dong-Hee Kim;Wi-Hye Yeon;Jin-Tae Lee;Young-Ah Jang
    • Journal of the Korean Applied Science and Technology
    • /
    • v.40 no.3
    • /
    • pp.534-546
    • /
    • 2023
  • In this study, we investigated the antioxidant and anti-winkle activity in human fibroblast cell (CCD-986sk) of Persimmon Leaves (PL) as a cosmetic ingredient. As a result of investigating antioxidant activity through electron-donating ability and ABTS+ radical scavenging assay, the PL showed concentration-dependent antioxidant activity similar to ascorbic acid, a control group, at a concentration of 1,000 ㎍/ml. As a result of investigating the anti-wrinkle effect through elastase inhibition and collagenase inhibition assay, the PL showed concentration-dependent antioxidant activity similar to epigallocatechin gallate, a control group, at a concentration of 1,000 ㎍/ml. As a result of measuring the synthesis rate of pro-collagen type I and the inhibition rate of MMP-1 in UVB-induced CCD-986sk cells, the control group EGCG showed a 90.2% pro-collagen synthesis rate at 20 ㎍/ml and PL showed an 88.5% synthesis rate at 30 ㎍/ml. In addition, the inhibition rate of MMP-1 of 33.0% and 40.8% were confirmed in EGCG 20 ㎍/ml and PL 30 ㎍/ml, respectively. As a result of measuring the protein expression of pro-collagen type I and MMP-1 in the PL through western blot, it was confirmed that the protein expression of pro-collagen type I increased, and MMP-1 decreased when the PL was treated together compared to the UVB alone group. According to the above experimental results, it is expected to be used as a natural product material for cosmetics by confirming that the PL prevent photoaging caused by UVB stimulation and have antioxidant and anti-wrinkle effects.

In vitro Angiogenic Activity of Aloe vera Gel on Calf Pulmonary Artery Endothelial (CPAE) Cells

  • Lee, Myoung-Jin;Lee, Ok-Hee;Yoon, Soo-Hong;Lee, Seung-Ki;Chung, Myung-Hee;Park, Young-In;Sung, Chung-Ki;Choi, Jae-Sue;Kim, Kyu-Won
    • Archives of Pharmacal Research
    • /
    • v.21 no.3
    • /
    • pp.260-265
    • /
    • 1998
  • Angiogenic activity of Aloe vera gel was investigated by in vitro assay. We obtained the most active fraction from dichloromethane extract of Aloe vera gel by partitioning between hexane and 90% aqueous methanol. The most active fraction (F3) increased the proliferation of calf pulmonary artery endothelial (CPAE) cells. In addition, F3 fraction induced CPAE cells to invade type I collagen gel and form capillary-like tube through in vitro angiogenesis assay, and increased the invasion of CPAE cells into matrigel through in vitro invasion assay. Furthermore, the effect on the MRNA expression of proteolytic enzymes which are key participants in the regulation of extracellular matrix degradation was investigated by northern blot analysis. F3 fraction enhanced mRNA expression of urokinase-type plasminogen activator (u-PA), matrix metalloproteinase-2 (MMP-2), and membrane-type MMP (MT-MMP) in CPAE cells whereas the expression of plasminogen activator inhibitory (PAl-1) mRNA was not changed.

  • PDF