• Title/Summary/Keyword: collagen assay

Search Result 306, Processing Time 0.026 seconds

Study on the Effect of Yipahnsan(立安散) on Angiogenic Inhibition Mechanism (입안산(立安散)이 Angiogenesis 억제기전(抑制棋戰)에 미치는 영향(影響))

  • Lee, Gi-Ryong;Choi, Seung-Hoon;Ahn, Kyoo-Seok
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
    • /
    • v.4 no.1
    • /
    • pp.177-197
    • /
    • 1998
  • This experimental study was carried out to evaluate the effect of Yipahnsan on angiogenic inhibition mechanism. This study investigates the effects of Yipahnsan on angiogenic inhibition mechanism evaluate cell adhesive inhibition effect, DNA fragmantaion analysis, nuclear condensation assay, FACScan analysis, angiogenic lumen formation assay, immunocytochemistry analysis, RT-PCR for mRNA expression, western blot analysis, confocal analysis for $Ca^{2+}influx$. The results were summarized as follows : 1. The cell adhesive inhibition ability was strongly increased from $5{\mu}g/ml$ on ECV304 cell line and ECVPAR cell line. 2. YY water extract caused $G_0/G_1$ arrest peak to existed on the ECV304 cell line. 3. YY water extract caused inhibition of proliferation and inducement of apoptosis on the collagen coated plate in ECV304 cell line. 4. YY water extract inhibited the lumen formation on the matrigel coated plate in ECV304 cell line. 5. YY water extract inhibited the expressions of LFA-1 and ELAM-1 on ECV304 cell line and ECVPAR cell line. 6. YY water extract inhibited the expressions of MMP-9 and uPA on ECV304 cell line and ECVPAR cell line. 7. YY water extract inhibited the expression of integrin ${\alpha}_v{\beta}_3$ on ECV304 cell line and ECVPAR cell line. 8. YY water extract decreased the change of $Ca^{2+}$ in intracellular on ECV304 cell line and ECVPAR cell line. According to the results, Yipahnsan showed to be a key antagonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA were blocked under the angiogenesis model. Thus, we suggested that Yipahnsan blocks angiogenesis by inducing apoptosis in ECV304 and ECVPAR cell lines, and another oriental herbal medicine that treats qi-stagnation and blood-stasis type also has angiogenic inhibition effects.

  • PDF

The Comparison of Sponges and PLGA Scaffolds Impregnated with DBP on Growth Behaviors of Human Intervertebral Disc Cells (DBP 스폰지와 DBP/PLGA 지지체에서의 인간 디스크세포 거동분석 비교)

  • Lee, Seon-Kyoung;Hong, Hee-Kyung;Kim, Su-Jin;Kim, Yong-Ki;Song, Yi-Seul;Ha, Yoon;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
    • /
    • v.34 no.5
    • /
    • pp.398-404
    • /
    • 2010
  • We fabricated sponge and poly(lactide-co-glycolide)(PLGA) scaffolds impregnated demineralized bone particle(DBP)(DBP/PLGA) and investigated proper condition to proliferation and phenotype maintenance of intervertebral disc(IVD) cells by comparison between DBP/PLGA scaffold and DBP sponge. DBP/PLGA scaffolds were prepared by solvent casting/salt leaching. Human IVD cells were seeded in scaffolds of two types. Cell viability and proliferation according to scaffolds were analyzed by WST assay and SEM. RT-PCR was assessed to measure mRNA expression of aggrecan and type II collagen of human IVD cells. In WST assay results, cell viability in scaffolds impregnated DBP/PLGA scaffold were higher than DBP sponge. We could observe that disc cell mRNA expressed better in DBP/PLGA scaffold than DBP sponge. We concluded that the using of DBP/PLGA in terms of scaffold fabrication for bio-disc with human IVD cells is helpful growth of disc cells maintenance of phenotypes.

Study of Cosmeceutical Activities of Hovenia dulcis var. koreana Nakai Extracts (헛개나무 추출물의 화장품 생리활성에 관한 연구)

  • Kim, Sea-Hyun;Jun, Dong-Ha;Jang, Min-Jung;Lee, Jin-Tae;Lee, Chang-Eon;Han, Jin-Gyu;Kim, Jin-Chul;Lee, Do-Hyung
    • Journal of Korean Society of Forest Science
    • /
    • v.99 no.6
    • /
    • pp.836-842
    • /
    • 2010
  • Hovenia dulcis var. koreana Nakai has been reported to liver function improvement effect as functional materials for food and medicine. On these facts, biological activity and safety test were conducted to evaluate biological activities of the fruit petiole and root extracts of H. dulcis as a potential cosmeceutical ingredient. Cosmeceutica activities of different extracts were examined by l.l-diphenyl-2-picrylhydrazyl (DPPH) radical generation, the ABTS+ cation decolorization, tyrosinase activity, collagenase activity and elastase activity compared with the properties of the commercial antioxidant butylated hydroxytoluene (BHT) and L-ascorbic acid (AA). The antioxidant activities HDFW, HDFE, HDRW and HDRE were 83.6%, 39.6%, 85.9% and 74.5% in DPPH assay, 99.5%, 13.7%, 96.4% and 88.6% in ABTS assay. Tyrosinase inhibitiory activities HDFW were 56.0% at 1,000 ppm. Measured the inhibition effect of the H. dulcis about collagenase and elastase where break the peptide bonds in collagen and enzyme from the class of proteases where exists in the dermis. The H. dulcis was inhibition the two kind enzymesm, collagenase activities being on a high scale inhibition, was same concentration. Uses the anti oxidation effect and a anti-wrinkle effect of this resultant H. dulcis and with the functional cosmetics use is thought with the fact that will be possible.

Inhibitory Effect of YBR on Hepatic Fibrogenesis (YBR의 간섬유화(肝纖維化)억제 효과(效果)에 관한 연구(硏究))

  • Seung, Hyun-Seok;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
    • /
    • v.31 no.2
    • /
    • pp.314-330
    • /
    • 2010
  • Objective : This study was performed to investigate the anti-fibrogenic effect and changes of inflammation-related genes by YBR I and YBR II (YBR I: Arteisiae Capillaris Herba, Atractylodis Rhizoma Alba, Hoelen/ YBR II: YBR I +Sanguisorbae Radix, Biotae Cacumen, Cirsii Japonici Herba) on HSC(hepatic stellate cells)-T6 and TAA-induced rat liver tissue. Materials and Methods : HSC-T6 were treated with various concentrations of distilled-water extract YBR I and YBR II extract for 24, 48 and 72 hours. After the treatment, cell viability, proliferation, procollagen levels and IL-6 levels were measured by using MTT Assay, BrdU Assay, Procollagen Type 1 C-peptide EIA kit, and Murine IL-6 ELISA Development kit. Rat liver fibrosis was induced by intraperitoneal TAA injection of 150mg/kg 3 times a week for 6 weeks. After the treatment, body weight, liver & spleen weights, liver function test, complete blood cell count and change of portal pressure were studied. In addition, gene expressions of ASMA, IL-6, MMP-2, TIMP-1 and TIMP-2, all of which are known to be associated with liver fibrosis, were analyzed by using Real-Time PCR. After YBR I and YBR IItreatment, percentages of collagen in TAA-induced rat liver tissue were measured. Results : The viability and proliferation of the HSC-T6 decreased as the concentration increased. The production of procollagen decreased as the concentration increased. The production of IL-6 was little influenced by YBR I and YBR II. There was no difference in rat body weight between the TAA-only group and the YBR groups. Compared with rat liver weight of TAA-only group, that of the YBR groups increased. In the YBR I group, the serum level of AST elevated by TAA injection significantly decreased and in the YBR I and II group, the serum level of ALP and ALT elevated by TAA injection decreased. In the YBR I group, white blood cell count elevated by TAA injection decreased but platelets increased. In the YBR I group, the portal pressure elevated by TAA injection significantly decreased. Decreases in the gene expression of ASMA and MMP-2 were observed in the YBR I group. The gene expression of IL-6 was little influenced by YBR I and YBR II -treated groups. In the histological finding, TAA injections caused severe fibrosis, but YBR I and YBR II treatment significantly reduced the amounts of hepatic collagens. Conclusions : These results suggest that YBR I and II have inhibitory effects on the hepatic fibrogenesis.

Effect of microgrooves and fibronectin conjugation on the osteoblast marker gene expression and differentiation

  • Park, Su-Jung;Leesungbok, Richard;Ahn, Su-Jin;Im, Byung-Jin;Lee, Do Yun;Jee, Yu-Jin;Yoon, Joon-Ho;Cui, Taixing;Lee, Sang Cheon;Lee, Suk Won
    • The Journal of Advanced Prosthodontics
    • /
    • v.7 no.6
    • /
    • pp.496-505
    • /
    • 2015
  • PURPOSE. To determine the effect of fibronectin (FN)-conjugated, microgrooved titanium (Ti) on osteoblast differentiation and gene expression in human bone marrow-derived mesenchymal stem cells (MSCs). MATERIALS AND METHODS. Photolithography was used to fabricate the microgrooved Ti, and amine functionalization (silanization) was used to immobilize fibronectin on the titanium surfaces. Osteoblast differentiation and osteoblast marker gene expression were analyzed by means of alkaline phosphatase activity assay, extracellular calcium deposition assay, and quantitative real-time PCR. RESULTS. The conjugation of fibronectin on Ti significantly increased osteoblast differentiation in MSCs compared with non-conjugated Ti substrates. On the extracellular calcium deposition assays of MSCs at 21 days, an approximately two-fold increase in calcium concentration was observed on the etched 60-${\mu}m$-wide/10-${\mu}m$-deep microgrooved surface with fibronectin (E60/10FN) compared with the same surface without fibronectin (E60/10), and a more than four-fold increase in calcium concentration was observed on E60/10FN compared with the non-etched control (NE0) and etched control (E0) surfaces. Through a series of analyses to determine the expression of osteoblast marker genes, a significant increase in all the marker genes except type I collagen ${\alpha}1$ mRNA was seen with E60/10FN more than with any of the other groups, as compared with NE0. CONCLUSION. The FN-conjugated, microgrooved Ti substrate can provide an effective surface to promote osteoblast differentiation and osteoblast marker gene expression in MSCs.

Skin Anti-aging and Anti-wrinkle Effects of Pinus koreaiensis Seed Oil (해송자 오일의 피부 항노화 및 주름 개선 효과)

  • Kim, Hyung-Mook;Kim, Tae-Jun;Im, Dong-bin;Ha, Sun-Bong;Kim, Ee-Hwa;Cha, Byung-Sun;Heo, Hyo-Jin;Brito, Sofia;Lee, Yong-Moon;Bin, Bum-Ho;Kwak, Byeong-Mun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
    • /
    • v.34 no.4
    • /
    • pp.1-11
    • /
    • 2021
  • Objectives : The aim of this study was to investigated the anti-aging and anti-wrinkles effects of the pinus koreaiensis seed(PKS) oil. Methods : The anti-oxidant effect was performed by beta-carotene bleaching assay and the intracellular proteome was analyzed expression of each 15 proteins by 2-D electrophoresis. And fatty acid was analysed by gas chromatography. Anti-wrinkle effect was analyzing human skin by the PRIMOS system. Results : Fatty acid analysis of PKS oil has shown oleic acid was 49.7% and linoleic acid was 34.1%. And the antioxidant effect was about 125% compared with alpha-tocoperol(0.1%) by beta carotene bleaching assay. In 2D PAGE analysis, fifteen protein changes in five mechanisms which was collagen synthesis pathway, MMPs, ECM-cell interaction, cytokine, antioxidant enzymes were analyzed. In case of anti-wrinkle effect was proved in vivo by analyzing human skin by the PRIMOS system. The analysis results of eye wrinkles for 4 weeks showed an improvement effect of over 6%. Conclusions : In this study, the amount of protein change in the five mechanism through the cell experiment and the skin anti wrinkle efficacy by the human in vivo test were investigated. As a result pinus koreaiensis seed oil by supercritical extraction could be used as a anti-aging and anti-wrinkle substance for the skin.

Evaluation of skin improvement efficacy of herbal medicine extracts on skin keratinocytes stimulated with fine dust PM10 (미세먼지 PM10으로 손상을 유도한 피부각질형성세포에서 한약재 추출물의 피부 개선 효능 평가)

  • Dong-Hee Kim;Yun Hwan Kang;Bo-Ae Kim
    • Journal of the Korean Applied Science and Technology
    • /
    • v.40 no.4
    • /
    • pp.856-867
    • /
    • 2023
  • Due to the increase in fine dust caused by environmental pollution, oxidative damage and aging of the skin are accelerated. In this study, the antioxidant, hyaluronic acid, filaggrin, MMP-1, and ROS level of selected herbal extracts were evaluated to confirm the protective efficacy of keratinocytes treated PM10. As a result, the antioxidant capacity of 1,1-diphenyl-2-picrylhydrazyl(DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid(ABTS), and FRAP assay increased in a concentration-dependent manner. Keratinocytes the group treated with 300 ㎍/ml of PM10, hyaluronic acid and filaggrin decreased by more than 50%, and increased in the group treated with extracts of Alpinia officinarum, Ulmus macrocarpa, and Ulmus macrocarpa but decreased when the extract was treated, which is evaluated as inhibiting the degradation of collagen and elastin. In addition, in the case of ROS measurement using zebrafish embryos, it was confirmed that the extract was reduced when the extract was treated 25 ㎍/ml, the intensity of fluorescence similar to the negative control was shown, confirming that the production of ROS was significantly reduced. Through this study, the selected oriental medicinal materials, Alpinia officinarum, Ulmus macrocarpa, and Ulmus macrocarpa, protect the skin from fine dust. It is thought that it can be used as an anti-aging product for skin improvement as a material that can be improved or improved.

Effects of Imperatae Rhizoma Extract on T helper 2 cell differentiation (백모근 추출물의 T helper 2 세포 분화 조절 효과)

  • Kim, Bok-Kyu;Lim, Jong-Soon;Kil, Ki-Jung
    • The Korea Journal of Herbology
    • /
    • v.29 no.6
    • /
    • pp.27-33
    • /
    • 2014
  • Objectives : The aims of this study were to exploring the therapeutic effect of Imperatae Rhizoma Extract(IRE) on Asthma. Methods : To investigate biological modulation activities of IRE, we conducted the cell-based assay whether IRE could regulate T helper 2 cells activity with EL4 T cells and mouse splenocytes, and followed animal study to conform the efficacy of their therapeutic potential on OVA-induced asthmatic mouse. Results : In cell study, IRE suppress the nuclear translocation of GATA binding protein-3 protein in phorbol 12-myristate 13-acetate/Ionomycin-stimulated EL4 T cells and Interleukine(IL)-4, IL-5 and IL-13 production in splenocytes at concentration dependent manner. In animal study, IRE-treated groups both 100mg/ml and 200mg/ml improve airway hypersensitibility reaction(AHR) response to methacholine about 30% and 40% with positive control group. Peritoneal blood analysis reveal that eosinophil number and ovalbumin-specific IgE is reduced by IRE treatment. Cell number of eosinophil is also reduced in bronchoalveolar lavage of IRE group like to peritoneal cell and real time-polymerase chain reaction data show that expression levels of IL-4, IL-5 and IL-13 were down regulated in lung tissue. Finally, histological analysis indicate that IRE protect the bronchial tissue damages through the accumulation of inflammatory cells and collagen, and these effect may be cause by interfering Th2 cells activity. Conclusions : Our data represent that IRE potentiates therapeutic activities to the allergic diseases such as asthma by regulating Th2 cells differentiation.

Anti-Vascular Endothelial Growth Factor (Bevacizumab) Therapy Reduces Hypertrophic Scar Formation in a Rabbit Ear Wounding Model

  • Kwak, Do Hoon;Bae, Tae Hui;Kim, Woo Seob;Kim, Han Koo
    • Archives of Plastic Surgery
    • /
    • v.43 no.6
    • /
    • pp.491-497
    • /
    • 2016
  • Background Hypertrophic scarring is a pathological condition that occurs after trauma or surgery. Angiogenesis occurs more often with hypertrophic scarring than with normotrophic scarring. The regulation of angiogenesis is one of the key factors in hypertrophic scar management. Vascular endothelial growth factor (VEGF) is an essential factor in the angiogenetic response. This study investigated whether decreasing the level of VEGF is effective for treating hypertrophic scarring. Methods Ten 8-week-old female New Zealand white rabbits were included. Four defects were created on each ear by using a 6-mm punch. Bevacizumab (Avastin, Roche Pharma, Basel, Switzerland) was administered in one ear and normal saline was administered in the other ear. Treatment was administered starting on day 2, every 2 days, until day 14. The levels of VEGF were measured using enzyme-linked immunosorbent assay on day 10 and histologic results were analyzed on day 40. Results Bevacizumab induced-defects showed less hypertrophic scarring when compared with the control group as measured by the scar elevation index (SEI) and loose collagen arrangement. The SEI in the experimental group was $1.89{\pm}0.13$, compared to $1.99{\pm}0.13$ in the control group (n=30, P=0.005). Additionally, the VEGF level was lower ($38.72{\pm}11.03pg$ vs. $82.50{\pm}21.64pg$, n=10, P=0.001) and fewer vessels existed ($8.58{\pm}0.76$ vs. $7.2{\pm}1.20$, n=10, P=0.007). Conclusions Preventing excessive angiogenesis is effective for preventing scar formation, especially with hypertrophic scarring. Although it is not an approach that is sufficient alone for the management of scarring, it may be one of several important strategies for scar treatment.

Enhanced Bone-Regenerative Performance of Porous Hybrid Scaffolds by Surface Immobilization of Nano-Hydroxyapatite

  • Lee, Sang-Cheon
    • Proceedings of the Materials Research Society of Korea Conference
    • /
    • 2009.11a
    • /
    • pp.12.1-12.1
    • /
    • 2009
  • Nano-hydroxyapatite (N-HAp)has shown the pivotal role in producing bone-regenerative materials since it has similarity to natural bone minerals in terms of size, morphology, and the composition. Currently, the combination of biopolymers and N-HAp is recognizedas an attractive approach in generating hybrid scaffolds for bone tissueengineering. Surface engineering is an important issue since it determines whether cells can effectively adhere and proliferate on porous scaffolds. We aim to develop a synthetic approach to porous 3D scaffolds by immobilizing N-HAp on pore surfaces. The discrete nano-level anchoring of N-HAp on the scaffold pore surface is achieved using surface-repellent stable colloidal N-HAp with surface phosphate functionality. This rational surface engineering enables surface-anchored N-HAp to express its overall intrinsic bioactivity,since N-HAp is not phase-mixed with the polymers. The porous polymer scaffolds with surface-immobilized N-HAp provide more favorable environments thanconventional bulk phase-mixed polymer/N-HAp scaffolds in terms of cellular interaction and growth. In vitro biological evaluation using alkalinephosphatase activity assay supports that immobilized N-HAp on pore surfaces of polymer scaffolds contributed to the more enhanced in vitro osteogenicpotential. Besides, the scaffolds with surface-exposed N-HAp provide favorable environments for enhanced in vivo bone tissue growth, estimated by characteristic biomarkers of bone formation such as collagen. The results suggest that newly developed hybrid scaffolds with surface-immobilized N-HApmay serve as a useful 3D substrate with pore surfaces featuring excellent bonetissue-regenerative properties. Acknowledgement. This research was supported by a grant (code #: 2009K000430) from 'Center for Nanostructured Materials Technology' under '21st Century Frontier R&D Programs' of the Ministry of Education, Science and Technology, Korea.

  • PDF