• Journal of Life Science
• /
• v.13 no.3
• /
• pp.308-313
• /
• 2003

The collagenase gene from Vibrio parahaemolyticus 04 was subcloned into an expression vector pET-29b. The recombinant collagenase was expressed in Escherichia coli BL2l(DE3) and partially purified by Hi-Trap affinity and Sephacryl S-100 size exclusion chromatographies. The recombinant enzyme was purified by 43.7-fold and the yield was 73%. SDS-PAGE revealed that the molecular weight of the enzyme was approximately 35 kDa. Substrate specificity study of the enzyme displayed that the enzyme showed the highest activity with the type I collagen and the synthetic peptide, Z-GPGGPA, indicating that the enzyme was indeed a collagenase. The enzyme showed broad pH optimum around pH 6-12 and was stable between pH 5.5 and 11.5. The optimum temperature for the type I collagen degradation was $35^{\circ}C$. The thermostability measurement of the enzyme indicated that the enzyme was stable up to $55^{\circ}C$, but the activity was diminished quickly above $60^{\circ}C.$

• Development and Reproduction
• /
• v.11 no.1
• /
• pp.1-11
• /
• 2007

Specific endometrial preparation should occur during periimplantation period. That is a progress of serial differentiation and is absolute in implantation of embryo and successful pregnancy. Remodeling of tissues shown during embryogenesis is regulated by various factors including extracellular matrix (ECM). Marked changes during pregnancy are including embryo migration, decidual response, and differentiation of placenta in placental animals including human. These changes to successful implantation in embryo and uterus have to prepare the competence for attachment of embryo and uterus, and invasion defense of uterus. During these changes, ECM dramatically changes for maintaining the uterine and embryonic functions. The major component of most connective tissue is collagens. It is very complex and hard to explore the mechanisms for ECM modulation. Recently using high throughput methodology, PCR-select cDNA subtraction method, microarray, many candidate genes have been identified. Steroid hormones have fundamental role in implantation and maintenance of pregnancy. Dermatopontin, a regulator of collagen accumulation, is regulated spatio-temporally in the uterus by primarily progesterone through progesterone receptors at the time of implantation. Modulation of extracellular matrix is critically regulated by cascade of gene net-works which are regulated by cascade of sex steroid hormones. Pathological regulation of uterine extracellular matrix reported in diabetic patients. To know the extracellular modulation is essential to understanding implantation, feto-placental development and overcome the paths involved in female reproduction. Though ECM composed with very various components and it is complex, the present review focused on the fate of collagens during periimplantation period.

• Journal of the Korean Applied Science and Technology
• /
• v.40 no.5
• /
• pp.1010-1021
• /
• 2023

In this study, we would like to confirm the collagen recovery effect through cell experiments of the flavonoid apigenin and baicalein and propose results that can support the collagen recovery effect through comparative transcriptome analysis. As a result of the study, it was confirmed that apigenin and baicalein were effective in recovering type I collagen damaged by UV in the HS68 cell line, and that both substances resulted in changes in the expression of ECM-related genes. Not only did it show a common mechanism for regulating the ECM, but it also showed the results of changing different categories of genes, making it possible to predict that it would have various effects on cells.

• Journal of Life Science
• /
• v.32 no.2
• /
• pp.175-188
• /
• 2022

Relaxin has been demonstrated to have regulatory functions on both the smooth muscle and extracellular matrix (ECM) of blood vessels and fibrotic organs. The diverse mechanisms by which relaxin acts on small resistance arteries and fibrotic organs, including the bladder, are reviewed here. Relaxin induces vasodilation by inhibiting the contractility of vascular smooth muscles and by increasing the passive compliance of vessel walls through the reduction of ECM components, such as collagen. The primary cellular mechanism whereby relaxin induces arterial vasodilation is mediated by the endothelium-dependent production of nitric oxide (NO) through the activation of RXFP1/PI3K, Akt phosphorylation, and eNOS. In addition, relaxin triggers different alternative pathways to enhance the vasodilation of renal and mesenteric arteries. In small renal arteries, relaxin stimulates the activation of the endothelial MMPs and EtB receptors and the production of VEGF and PlGF to inhibit myogenic contractility and collagen deposition, thereby bringing about vasodilation. Conversely, in small mesenteric arteries, relaxin augments bradykinin (BK)-evoked relaxation in a time-dependent manner. Whereas the rapid enhancement of the BK-mediated relaxation is dependent on IK_Ca channels and subsequent EDH induction, the sustained relaxation due to BK depends on COX activation and PGI₂. The anti-fibrotic effects of relaxin are mediated by inhibiting the invasion of inflammatory immune cells, the endothelial-to-mesenchymal transition (EndMT), and the differentiation and activation of myofibroblasts. Relaxin also activates the NOS/NO/cGMP/PKG-1 pathways in myofibroblasts to suppress the TGF-β1-induced activation of ERK1/2 and Smad2/3 signaling and deposition of ECM collagen.

• Korean Chemical Engineering Research
• /
• v.60 no.3
• /
• pp.386-391
• /
• 2022

This study is about a technique for obtaining collagen by extracting fat by treating collagen-containing liposuction effluent in the presence of supercritical fluid. Using a supercritical solvent, a collagen extract could be obtained from animal-derived fat in a short time (about 6 hours), and about 2-3% of collagen by mass compared to the raw material could be obtained. The presence of collagen in the extract obtained by supercritical extraction was confirmed by SDS-PAGE, and it was confirmed that it was type 1 collagen having a relatively large molecular weight. In addition, the growth factors of IGF-1, bFGF, VEGF and NGF were analyzed to find out which growth factors were present in the collagen obtained by supercritical extraction, and it was found that these growth factors were contained in the extract. There was no significant difference in DNA content per mg of sample before and after supercritical treatment. Further in-depth studies are likely to be needed on decellularization technology using the supercritical process. In conclusion, the extracellular matrix obtained through the solvent extraction process using a supercritical fluid contains growth factors above a certain amount even after decellularization and removal of fat, so that it was found that not only biocompatibility is greatly increased, but also tissue regeneration can be rapidly induced.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.47 no.2
• /
• pp.147-153
• /
• 2021

Collagen hydrolysate (CH) is known to prevent skin aging by stimulating skin dermal fibroblasts to promote synthesis of extracellular matrix such as collagen and elastin. Recently, among the various factors that cause skin aging, advanced glycation end products (AGEs) have received particular attention. However, the effect of CH on AGE accumulation has not been studied. Since CH could affect AGE accumulation by promoting production of skin structural proteins, clinical trial was performed using low molecule collagen peptide (LMCP), which were CH containing 25% tripeptide and 4% Gly-Pro-Hyp. Skin autofluorescence (SAF) values were measured using an AGE reader to evaluate accumulation of AGE in skin. As a result of applying 0.5% and 1.0% LMCP solutions to the subject's forearm for 8 weeks, the SAF value at the test site significantly decreased compared to the control site. Additionally, in vitro test was performed using CCD-986sk to evaluate the promotion of collagen synthesis in skin fibroblasts by LMCP. As a result, 800 ㎍/mL of LMCP significantly increased synthesis of human pro-collagen Iα1 (COL1A1) in CCD-986sk. Through this study, we have confirmed that tropical LMCP applications can promote collagen synthesis to help anti-glycation effects, suggesting that LMCP has potential as an anti-aging cosmetic material.

• The Journal of Korean Academy of Prosthodontics
• /
• v.49 no.3
• /
• pp.254-262
• /
• 2011

Purpose: This study aims to evaluate and prospect for current research trend and developmental perspectives via analyzing recent biomaterial coated-implants study. Materials and methods: To investigate each subject respectively, several biomaterials that are using for implant surface coating were set as 'keywords'. By these keywords, major research groups in each subject were chosen, and research trend of them was analyzed. Trend of In vivo studies that examined selected biomaterials were analyzed to evaluate commercial potential. Results: The collagen research accounted for 40% of total implant study, which was the highest, and fibronectin, BMPs (bone morphogenetic proteins) and RGD (Arg-Gly-Asp) peptides followed, which were ranked in descending order. Furthermore, figures of all four research subjects were also increased with time, especially a sharp increase in RGD research. According to the results of major research group, collagen that was combined with other organic and inorganic biomaterials was mostly examined, rather than using collagen only. Major research groups investigating BMPs mostly focused on rhBMP-2. In animal studies, collagen was used as resorbable membrane in guided bone regeneration (GBR) or drug carrier, while BMPs were used with bone graft materials or coating material for titanium implant surface. Conclusion: There is not consistency of results even in identical subjects research field. Many studies are ongoing to optimize combination between mechanical surface treatment and biomaterials such as extracellular matrix component and growth factors.

• Journal of Life Science
• /
• v.6 no.4
• /
• pp.241-249
• /
• 1996

A collagenase was isolated from the culture filtrate of Vibrio mimicus (ATCC 33658). The enzyme was purified to homogeneity by ammonium sulfate precipitation and DEAE-Sephadex A-50 chromatography, which an activity recovery of 22%. The molecular weight of the purified enzyme was estimated to be 42 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indication a monomer structure. The optimum pH and temperature od the enzyme for insoluble collagen (Type I) were around 7.75 and 28$\circ$C, respectively. Some chelating agents and serine protease inhibitor inactivated the enzyme, but L-cysteine and histidine did not affect the activity. The amino acid composition indicated that the collagenase contained high amounts of amino acid residues of glycine and alanine. The K$_{m}$ and R$_{cat}$/K$_{m}$ values for the collagenase, using insoluble collagen (type I) as substrate, were 2.86 mg/ml and 972.28 U/mg-protein, respectively.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.40 no.4
• /
• pp.365-371
• /
• 2014

To identify new active anti-wrinkle ingredients, this study investigated the anti-wrinkle effects of Schizandra chinensis Baillon fermented with Lactobacillus plantarum (SCF) by assessment of cytotoxicity of human dermal fibroblast, collagen biosynthesis, matrix metalloproteinase-I (MMP-1) inhibition and elastase inhibition. S. chinensis was fermented with L. rhamnosus for 1 day at $37^{\circ}C$. The cytotoxicity of SCF was evaluated by a cytopathic effect reduction method. Effects on collagen biosynthesis and matrix metalloproteinase-I (MMP-1) of SCF were evaluated by previous reported method using procollagen type-IC peptide EIA kit and Matrix Metalloproteinase-1 Biotrack activity Assay Kit, respectively. Elastase inhibition assay was conducted by reaction of enzyme and substrate using N-Suc-$(Ala)_3$-nitroanilide as the substrate. As the results, SCF didn't show cytotoxicity against human dermal fibroblast at concentration of $100{\mu}g/mL$. Also, SCF was increased collagen synthesis and showed inhibitory effect of MMP-1 (p < 0.05). In the elastase inhibition assay, the $IC_{50}$ of SCF was $36.4{\mu}g/mL$. Therefore, our results indicated that SCF possesses anti-wrinkle effects and can be used practically for anti-wrinkle care of skin.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.41 no.4
• /
• pp.351-359
• /
• 2015

Acute and chronic ultraviolet (UV) irradiation triggers severe skin photoaging processes, which directly disrupt the normal three-dimensional integrity of skin. UV light stimulates the expression of matrix metalloproteinases (MMPs) which degrade constituents of extracellular matrix (ECM) proteins. These MMPs reduce collagen synthesis and decrease skin elasticity and integrity, resulting in wrinkle formation. In this study, we identified Rosa multiflora hip extract (RME) as an effective anti-photoaging ingredient. First, cell proliferation activity of RME was verified using Hs68 human dermal fibroblast cell line. RME downregulated MMPs expression through the inhibition of activator protein (AP)-1. In addition, type I and IV collagen expressions were increased with RME treatment and UVB-induced inflammatory responses were also reduced after RME treatment. In conclusion, R. multiflora hip extract may effectively improve UVB-induced skin aging and wrinkle formation which may provide as an anti-aging, anti-wrinkle, and anti-inflammation ingredient in cosmetic industry.