• Title/Summary/Keyword: collagenase

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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
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    • v.50 no.4
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    • pp.238-250
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    • 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.

Transcription factor EGR-1 transactivates the MMP1 gene promoter in response to TNFα in HaCaT keratinocytes

  • Yeo, Hyunjin;Lee, Jeong Yeon;Kim, JuHwan;Ahn, Sung Shin;Jeong, Jeong You;Choi, Ji Hye;Lee, Young Han;Shin, Soon Young
    • BMB Reports
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    • v.53 no.6
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    • pp.323-328
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    • 2020
  • Matrix metalloproteinase 1 (MMP-1), a calcium-dependent zinccontaining collagenase, is involved in the initial degradation of native fibrillar collagen. Tissue necrosis factor-alpha (TNFα) is a pro-inflammatory cytokine that is rapidly produced by dermal fibroblasts, monocytes/macrophages, and keratinocytes and regulates inflammation and damaged-tissue remodeling. MMP-1 is induced by TNFα and plays a critical role in tissue remodeling and skin aging processes. However, the regulation of the MMP1 gene by TNFα is not fully understood. We aimed to find additional cis-acting elements involved in the regulation of TNFα-induced MMP1 gene transcription in addition to the nuclear factor-kappa B (NF-κB) and activator protein 1 (AP1) sites. Assessments of the 5'-regulatory region of the MMP1 gene, using a series of deletion constructs, revealed the requirement of the early growth response protein 1 (EGR-1)-binding sequence (EBS) in the proximal region for proper transcription by TNFα. Ectopic expression of EGR-1, a zinc-finger transcription factor that binds to G-C rich sequences, stimulated MMP1 promoter activity. The silencing of EGR-1 by RNA interference reduced TNFα-induced MMP-1 expression. EGR-1 directly binds to the proximal region and transactivates the MMP1 gene promoter. Mutation of the EBS within the MMP1 promoter abolished EGR-1-mediated MMP-1 promoter activation. These data suggest that EGR-1 is required for TNFα-induced MMP1 transcriptional activation. In addition, we found that all three MAPKs, ERK1/2, JNK, and p38 kinase, mediate TNFα-induced MMP-1 expression via EGR-1 upregulation. These results suggest that EGR-1 may represent a good target for the development of pharmaceutical agents to reduce inflammation-induced MMP-1 expression.

THE STUDY ON PERIODONTAL REGENERATION OF REPLANTED TEETH FOLLOWING THE APPLICATION OF RH-BMP-4 (rh-BMP-4가 재식치아 치주조직의 재생에 관한 연구)

  • Lee, Se-Joon;Choi, Kyung-Ku;Park, Sang-Jin;Choi, Ho-Yong;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.25 no.2
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    • pp.170-179
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    • 2000
  • The rh-BMP-4 is a subgroup of TGF-${\beta}$ superfamily. The application of rh-BMP in alveolar bony defect was reported to new alveolar bone and new cementum formation. For minimized complications following tooth replantation, a operator must replant a tooth fast at the pertinent position. This study was to evaluate the effect of rh-BMP-4 on periodontal regeneration and root resorption following tooth replantation in rats. The 50 Sprague-Dawley rats weighting about 130gm were used in this study. The animals were divided into three groups. Group 1 ; immediate replantation after extraction : Group 2 ; replantation stored teeth extraction of first molar, the removal of periodontal ligament with collagenase, and etching with citric acid : Group 3 ; replantation stored teeth with treated rh-BMP-4 in mesial root. Experimental animals were sacrificed 3, 7, 14 days after replantation by heart infusion. The maxillae were removed, fixed, demineralized, dehydrated, infiltrated and embedded with JB-4 mixture. For light microscopic observation, 5 micron sections were cut and stained with toluidine blue. The results of this study were as follows : 1. After experimental 3 days, all groups were observed dead space between periodontum and root. 2. After experimental 7 days, group 1 and group 3 were observed filling periodontal fibers between alveolar bone and root but group 2 were not. 3. After experimental 7 days, group 3 were observed appearance of attached cementoblast like cell on root surface. Group 1 were observed regular arrangement of fibroblasts and collagen fibers at ${\times}400$ observation. 4. After experimental 14 days, all group were observed filling periodontal fibers between alveolar bone and root. Group 1 were observed normal arrangement of periodontal fibers. Group 3 were observed less abnormal arrangement of periodontal fibers. Group 2 were not observed functional normal arrangement of periodontal fibers. 5. After experimental 14 days, group 2 and 3 were observed several root resorption and irregular root surface but group 1 were not. These results suggest that the rh-BMP-4 can stimulate cementogenesis and enhance to attach collagen fibers.

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Antioxidation and Anti-photoaging Effects of White Taraxacum Coreanum Extract by Lactobacillus plantarum (Lactobacillus plantarum에 의한 흰 민들레 발효물의 항산화 및 광노화 억제 효과)

  • Hong, Ji Woo;Park, Ha Young;Kim, Jun Hee;Yeom, Suh Hee;Kim, Jin Woo
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.4
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    • pp.554-562
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    • 2021
  • Lactobacillus plantarum SM4, a strain producing β-glucosidase, was isolated from kimchi and fermented with white Taraxacum coreanum to enhance the production of bioactive compounds. The total polyphenol content(TPC), total flavonoid content(TFC), radical scavenging activity(RSA), tyrosinase inhibitory activities(TIA), and collagenase inhibitory activities(CIA) were measured to evaluate the skin whitening and anti-wrinkle effects of the fermented product. The TPC of fermented white T. coreanum was 41.8±0.26 mg GAE/g DW, which was approximately two times higher than the hot-water extraction of 21.4±0.67 mg GAE/g DW. RSA, an indicator of antioxidant activity, was 65.6±4.7% in fermentation, which is four times higher than that of the hot-water extract. TAI and CAI, which are indicators of the whitening and anti-wrinkle effects, were 87.9±4.73% and 66.7±3.48%, respectively, which were 2.4 and 1.5 times higher than those of hot-water extraction. When comparing the UVA(320 nm) protection effects of fermented and hot-water extraction, the fermented white T. coreanum showed higher protection with an absorption rate of 64.7% and 15.2%, respectively. The white T. coreanum fermented product showed higher bioactive properties and improved skin whitening, anti-wrinkle, and UV protection effects through the production of β-glucosidase from L. plantarum SM4.

Functional Characterization of the Extracts from Nipa Palm, Molokhia, and Finger Root for Cosmetic Ingredients (니파팜, 몰로키아, 핑거루트 추출물의 화장품 소재로서의 기능적 특성 분석)

  • Jun, Yue Jin;Lee, Sohyun;Heo, Sojeong;Jin, Byung Suk
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.3
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    • pp.821-829
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    • 2019
  • This study was conducted to evaluate the applicability of the extracts from nipa palm, molokhia, and finger root in functional cosmetics as a natural active ingredient. The extracts were obtained through the processes of heating under reflux with ethanol, filtration, concentration, and freeze-drying. UV absorption and blocking effects of the extracts were examined by using the UV-vis spectrophotometer equipped with an integrating sphere. Antioxidant activity and its stability between the extracts were compared using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. Also, total polyphenol content in the extracts was determined quantitatively using the Folin-Ciocalteu reagent, with gallic acid as the standard. Antibacterial activity of the extracts was investigated by the disc diffusion test against Staphylococcus aureus (gram-positive) and Escherichia coli (gram-negative). Finally, collagenase inhibitor assay was performed to examine the anti-wrinkle effect of the extracts. From the results of this study, the extract of nipa palm showed the potential for use in cosmetics as an antioxidant and anti-wrinkle agent, and the extract of finger root as a sunscreen and antibacterial agent.

Analysis on the Efficacy of Cosmetic Application of Lijang Snow Tea (Nekemias grossedentata) (리장 설차 (Nekemias grossedentata )의 화장품적 적용 효능 분석)

  • Wen, Ying;Lee, Seol-Hoon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.4
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    • pp.281-287
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    • 2021
  • In this study, we analyzed the cosmetic applicability of extract from snow tea, native to Lijiang, Yunnan-province, China. After confirming the species as N. grossedentata through DNA analysis of Lijiang snow tea, experiments were conducted using representative tea, green tea, and a representative control group for each efficacy analysis. Both teas were extracted using 70% (v/v) ethanol aqueous solution. The polyphenol content in the Lijiang snow tea extract (gallic acid equivalent, 23.9 ± 3.2 mg/mL) was higher than that in green tea extract (16.4 ± 2.3 mg/mL). In contrast, the antioxidant (Radical scavenging, IC50 104 ㎍/mL), tyrosinase enzyme inhibitory (whitening agent, IC50 40.7 ㎍/mL), and Escherichia coli growth inhibitory (preservative) activities (IC50 2.85 mg/mL) were analyzed based on the solid content in the extract, and it was confirmed that the activities of Lijiang snow tea extract were superior to those of green tea extract (radical scavenging, IC50 234 ㎍/mL. It also showed similar efficacy to previously used active substances such as antioxidants (vitamin C, IC50 108 ㎍/mL), whitening agents (vitamin C, IC50 80㎍/mL), and preservatives (methylparaben, IC50 4.35 mg/mL). However, green tea was found to be better in collagenase inhibition activity (anti-wrinkle). Through this study, the cosmetic application potential of Lijiang snow tea is high.

Molecular Characterization of Hanwoo Glucose Transporter 4 Gene (한우 Glucose Transporter 4 유전자의 분자생물학적 해석)

  • Lee, S.M.;Jeong, Y.H.;Kim, H.M.;Park, H.Y.;Yoon, D.H.;Moon, S.J.;Chung, E.R.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1087-1094
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    • 2005
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose transport protein. In the glucose transport protein family, GLUT4 plays a key role in cellular glucose uptake stimulated by insulin in skeletal muscles and adipose tissue in rodents and human. In this studies, we reported the identification, characterization, and expression of Hanwoo GLUT4 gene. The Hanwoo GLUT4 cDNA includes a 1527 bp open reading frame encoding a protein of 509 amino acids. The GLUT4 amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The highest mRNA expression of GLUT4 was detected in heart and lower expression was detected in rib meat, sirloin, and colon. We confirmed the expression of GLUT4 in the subcutaneous and small intestinal adipose tissue using RT-PCR. To investigate the expression of GLUT4 in the bovine intramuscular adipose differentiation, fibroblast-like cells were isolated from the sirloin of Hanwoo bull aged 12 months by collagenase digestion of minced tissue and cultured with activators of PPAR gamma. We identified that GLUT4 mRNA expression decreased during differentiation of preadipocytes into adipocyte in Korean cattle. These results indicated that function of GLUT4 in bovine adipose tissue was different from that of mouse and human.

Skin Protection Effect of Grape Pruning Stem Extract on UVB-induced Connective Tissue Injury (포도전정가지 추출물이 UVB로 유도된 결합 조직 손상에 미치는 피부 보호 효과)

  • Kim, Joung-Hee;Kim, Keuk-Jun
    • Journal of Life Science
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    • v.28 no.2
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    • pp.141-147
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    • 2018
  • This study aimed to analyze the contents of rutin, procyanidin B3, quercetin, and kaempferol, known to have antioxidant, anti-inflammatory, and anti-carcinogenic effects, among the polyphenol types contained in grape pruning stem extracts (GPSE). It utilized grape stems discarded after harvest to measure the effects of GPSE on skin moisture, inhibition of skin cell proliferation, and anti-inflammatory activity on the damaged skin of HR-1 mice induced with ultraviolet B (UVB), and to verify the applicability of GPSE as a material for functional food and functional cosmetics. The polyphenol was extracted from grape pruning stems with 80% EtOH, and then the extract was used while storing at $-20^{\circ}C$, after filtering, concentrating, and freeze-drying it. The content of an active ingredient of GPSE was analyzed using high performance liquid chromatography (HPLC). From 53 kg of the grape pruning stem specimen, 2.34 kg of the EtOH fraction extracts were extracted to achieve a 4.42% yield ratio. Analysis of the active ingredients showed 0.28 mg/g of procyanidin B3, 12.81 mg/g of rutin, 0.51 mg/g of quercetin, and 8.24 mg/g of kaempferol. After UVB irradiation on the dermis, to confirm the degree of inhibition of collagen synthesis, we examined the protein expression of MMP-9 using immunohistochemical staining. The results of this study confirm the existence of active polyphenol types, such as rutin, kaempferol, quercetin, and procyanidin B3, in GPSE. Moreover, the study found that GPSE has anti-collagenase effects and it decreases the effects of UV damage on skin barrier function. GPSE is a functional ingredient with a potential for skin protection effects, and it has high utilization potential as an ingredient for functional cosmetics.

THE STUDY ON EFFECTS OF THE PLATELET-DERIVED GROWTH FACTOR-AA, BB ON THE CELLULAR ACTIVITY OF THE HUMAN PERIODONTAL LIGAMENT CELLS (Platelet-derived growth factor-AA, BB가 치주인대세포의 세포활성에 미치는 영향에 대한 연구)

  • Oh, Sang-Deok;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.303-320
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    • 1994
  • Current acceptable methods For promotin gperiodontal regeneration are base on removal of diseased soft tissue, root treatment, guided tissue regeneration, inteoduction of new graft materials and biological mediators. Platelet-derived growth factor(PDGF) is one of polypeptide growth factor. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of PDGF-AA, BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Author measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis and alkaline phosphatase activity according to the concentration of PDGF-AA and BB(0, 0.1, 1, 10, 100ng/ml). The results were as follows : The DNA synthetic activity was increased dose dependently by PDGF-AA and BB. The maximum mitogenic effect was at the 100ng/ml of PDGF-AA and 10ng/ml of PDGF-BB. The total protein, collagen and noncollagen systhesis was increased dose dependently by PDGF-AA and BB. The % of collagen was slightly decresed according to the concentration of PDGF-AA and BB. The effect of PDGF-AA and BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. The effect of PDGF-AA and BB on alkaline phosphatase activity did not show any significant, meanwhile the alkaline phosphatase activity of 14 days group showed significnat increase. In conclusion, PDGF-AA and BB may have important roles in stimulation of DNA synthesis in human periodontal ligament cells, which means an increase in collagen-synthesizing cells, and may be useful for clinical application in periodontal regenerative procedures.

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Effects of Estradiol and Pituitary Hormones on in vitro Vitellogenin Synthesis in the Eel, Anguilla japonica (뱀장어의 in vitro Vitellogenin 합성에 대한 Estradiol과 뇌하수체 호르몬의 영향)

  • KWON Hyuk-Chu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.2
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    • pp.282-290
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    • 1997
  • Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.

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