In order to achieve successful in vitro production of embryo, it is necessary to establish intrauterine environment during in vitro culture. Thus, this study was investigated to establish embryo culture system using co-incubated collagen matrix gel (CM) with endometrial epithelial cells (EC). Endometrial epithelial cells were isolated from porcine endometrium at follicular phase, the cells seeded in insert dish for co-incubation with CM-coated culture dish. Then, culture media treated with/without 2.0 IU/ml hCG or 10 ng/ml IL-1β. After incubation for 24 h, the co-incubated insert dishes were removed from CM-coated culture dish before embryo culture. Embryos at 48 h after in vitro fertilization (IVF) were cultured on the dish for 120 h with porcine zygote medium. We determined PTGS-2 expression in the ECs, VEGF protein in co-incubated CM with EC and observed cleavage rate and blastocyst development of embryos at 168 h after IVF. In result, expression of PTGS-2 was higher at co-incubated EC with hCG and IL-1β groups than EC without hCG and IL-1β. The VEGF protein was detected at co-incubated CM with EC, EC treated with hCG and IL-1β groups higher than CM group. Also, cleavage rate was no significantly difference among all group, however, blastocyst development was significantly higher in co-incubated CM with EC treated with hCG group than un-treated groups (p<0.05). Therefore, we suggest that novel embryo culture system using co-incubated collagen matrix gel with endometrial epithelial cells treated with IL-1β is beneficial and useful for enhancing the production of porcine blastocysts in vitro.

This experiment was carried out to find the quality characteristics of yanggaeng added antioxidant rich collagen powder in the color, sugar contents, texture, radical scavenging activity and electronic nose detected flavor. L value and a value kept decreased as the more collagen powder was added in the yanggaeng while b value was increased significantly at the same time. Sugar contents was increased significantly from 43 °Brix in the control to 48% in the 10% treatment (p<0.05). Hardness in the 10% treatment of collagen powder increased biggest among the treatments but springiness decreased when hardness increased. And adhesiveness, cohesiveness, gumminess and chewiness were also increased with more the collagen powder in the treatments, therefore 10% treatment showed biggest result significantly. Radical scavenging activity kept increased with higher addition of collagen powder and it resulted 60% of radical scavenging activity in the 10% treatment. In the flavor test under electronic nose experiment, 10% of collagen powder treatment was believed it flavored most for it was found in the furthest from the AIR location. Control was found also in the nearest from the AIR, which meant it has least flavor than any other treatment. Also 2.5% and 5% of collagen powder added treatments showed little differences of flavor from control, which meant 5% of treatment was believed idea condition in the treatments. With the above experimental results, 5% of collagen powder treatment in the yanggaeng manufacturing was chosed as the best mixture ratio in the test.

This study was performed to improve the efficiency of guided bone regeneration by adapting the tissue-engineered bone regeneration approach. The PHBV-collagen nonofibrous membranes cultured with rat calvarial periosteum derived cells in osteogenic culture medium for 7 days were applied to 5 x 2mm size artificial rat tibial defects for 3weeks and evaluated its efficiency as an alternative cell engineered membrane for effective guided bone regeneration by routine histological observation. The defects covered with cell attached PHBV-collagen nanofibrous membrane complexes showed favorable bone repair compared to both membrane non-covered control and membrane only covered group without specific side effects. These findings suggest that the favorable guided bone generation can be achieved by adapting the tissue engineered bone regeneration approach using PHBV-Collagen nanofibrous membrane scaffold.

In this research we extracted water-soluble collagen peptide from flatfish skin and compared it with commercially available collagen peptide extracted from Tilapia scale currently placed on the market in the aspect of physiochemical property. The physical property and nutritional components of FSCP appeared almost similarly to those of TSCP, and also in calorie, FSCP marking 3.82 Kcal showed no differences from TSCP marking 3.84 Kcal. As for forming amino acids, in aspartic acid, serine, histidine, tyrosine, methionine, FSCP had higher content than TSCP, but in OH-proline, proline and alanine FSCP had lower content than TSCP. Especially the content of essential amino acids of FSCP marked 22.74% with a higher content compared with 13.64% of TSCP. In the distribution of molecular weight FSCP with 1,000 Da showed omparatively low compared with TSCP, and in emulsion property and stability FSCP and TSCP showed similar excellent trend.

콜라겐 함유량이 높은 한식메뉴에 대한 미생물학적 위해 평가를 위해 S. aureus와 Salmonella를 삼계탕, 어글탕, 오돌뼈 볶음, 전약, 족발에 접종하여 다양한 저장온도에서 세균수를 관찰하였다. 4oC의 저장온도에서는 5가지 콜라겐 함유식품에서 세균의 성장이 관찰되지 않았고, 10oC의 경우 삼계탕과 족발이 다른 식품에 비해 세균이 생장하는 패턴을 나타냈다. 저장온도 20oC에서는 전약을 제외한 나 머지 식품에서 세균이 생장하였고, 30oC에서도 전약만 세균수가 감소하는 것으로 관찰되었다. 5가지 콜라겐 함유 식품 중 전약만 고온 저장온도에서 위해세균의 활성이 관찰되지 않고 감소하는 경향을 나타냈기 때문에 전약에 첨 가된 정향, 대추, 계피, 생강, 후추에 대한 항균활성을 실험하였다. 그 결과 정향, 계피, 후추, 생강이 S. aureus에 대해 항균성을 보였고, Salmonella에 대해서는 정향과 계피만 항균력이 나타났으며, 대추는 항균능력이 없는 것으로 나타났다.

콜라겐 성분을 최대로 보호하면서 안정하게 체내로 흡수 될 수 있도록 고순도 수첨 포스 파티딜콜린과 용매사출방법을 이용하여 콜라겐 리포좀을 제조하였다. 리포좀 막의 안정성을 높이기 위해 포스파티딜콜린에 콜레스테롤을 혼합하여, 에탄올과 프로필렌글리콜 혼합용매에 용해하였으며, 이온의 안정화를 위하여 PBS Buffer를 사용하였다. 다양한 변수에 의해 제조된 콜라겐 리포좀의 특성은 동적광산란광도계(DLS), 주사현미경(SEM), 편광현미경(POM)로 분석하였다.

Calcineurin-binding protein 1 (Cabin1) regulates calcineurin phosphatase activity as well as the activation, apoptosis, and inflammatory responses of fibroblast-like synoviocytes (FLSs), which actively participate in the chronic inflammatory responses in rheumatoid arthritis (RA). However, the mechanism of action of Cabin1 in FLS apoptosis is not clear. The aim of this study was to define the regulatory role of Cabin1 in FLSs of mice with collagen-induced arthritis (CIA). Transgenic mice overexpressing human Cabin1 in joint tissues, under the control of a type II collagen promoter, were generated. hCabin1 expression in joints and FLSs was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of cytokines, matrix metalloproteinases (MMPs), and apoptosis-related genes in FLSs was determined by enzyme- linked immunosorbent assay, gelatin zymography, and RT-PCR, respectively. Joints were histologically examined after H&E and TRAP staining. hCabin1-transgenic CIA mice had less severe arthritis than wild-type CIA mice, based on hind paw thickness and histology. This was accompanied by significantly enhanced apoptosis in transgenic mice, evidenced by significantly more TUNEL-positive cells in synovial tissues. The expression of inflammatory cytokines and MMPs was reduced, and the transgenic CIA mice exhibited decreased Akt activation and increased expression of p53, caspase-3, caspase-9, and Bax. hCabin1 plays a critical role in promoting apoptosis of FLSs and in attenuating inflammation and the destruction of cartilage and bone in RA. These findings help elucidate the pathogenic mechanisms of RA and suggest that Cabin1 is a potential target for RA treatment.

The anti-inflammatory effect of PHBV/Collagen (PHCP) was examined in a mouse model of lipopolysaccharide (LPS)-induced skin inflammation. Vascular permeability on the back skin was measured by the local accumulation of Evan’s blue dye after subcutaneous injection of LPS (30 µg site^-1 ). Dye leakage in the skin showed a significant increase at 2 h after injection of LPS. This LPS-induced dye leakage was also completely inhibited by HO-1 inhibitor, ZnPP, and antioxidants, including methyl gallate, trolox, and mannitol. To study the possible mechanisms underlying the in vivo anti-inflammatory effect of PHCP against LPS-induced inflammation, we also examined the effects of PHCP on malondialdehyde (MDA) and glutathione levels in skin tissues and found that pretreatment with PHCP resulted in inhibited MDA elevation and a remarkable reduction of glutathione level. In addition, similar results were obtained after pretreatment with antioxidants, including trolox and mannitol, and HO-1 inhibitor, ZnPP. Histopathologically, an influx of neutrophils into the skin dermis was detected between 24 h and 72 h after LPS injection (30, 100 µg site^-1), compared to control animals after injection of saline. This increase was greater in mice treated with 100 µg of LPS than in those treated with 30 µg of LPS and was significantly suppressed by pretreatment with PHCP, antioxidants, and HO-1 inhibitor. These results collectively suggest that PHCP has an anti-inflammatory effect against LPS-induced inflammation model in vivo and may be a good candidate for the skin tissue engineering biomedical application primarily through manipulation of the redox state.

In this study, a nanofibrous scaffold was obtained by co-electrospinning poly (3-hydroxybutyrate- co-3-hydroxyvalerate) (PHBV) and collagen in 2,2,2-trifluoroethanol at a ratio of 3/7. The fiber diameters were in the range of 250-600 nm. It was found that PHBV/Collagen (PHCP) nanofibrous scaffold showed greater proliferation than the PHBV nanofibrous scaffold induced by oxidant in NIH3T3 cells. Otherwise, in the early-stage wound-healing mouse model, wound closure was evaluated according to wound size reduction and histology of regenerated skin on the backs of mice. Each of the tissues removed on day 0, 3, 6, 9, 12, 15, and 18 was used for analysis of biochemical and pathological changes. None of the nanofiber-attached mice showed significant difference on the third day, however, from the third day until the ninth day, significantly faster healing was observed in PHCP-attached mice, compared to control wounds in epithelialization, wound contraction, and histopathological examinations. These results strongly support the beneficial effects of biomedical application of PHCP nanofiber in acceleration of the initial phase of wound healing through α-SM actin contraction.

EGCG has inhibitory effect on a variety of cancers by inducing apoptosis and cell cycle arrest or inhibiting angiogenesis and metastasis. EGCG has been found to induce apoptosis in salivary gland carcinoma cells. But the potential anti-invasive effect of EGCG in salivary gland cancer has not been studied yet. The aim of this study is to evaluate the effect of EGCG on salivary gland adenocarcinoma SGT cell adhesion and migration to Type I collagen treatment. Western blot, adhesion and migration assay were performed to evaluate the impacts of EGCG on the expression of MMP-2/-9 and its upstream signaling molecules after treatment of type I collagen. SGT cell adhesion to type I collagen is significantly suppressed by EGCG. EGCG decreased expression of β1 integrin, phosphorylation of FAK, MMP-2/-9 compared with type I collagen treatment. In addition, EGCG inhibited the migration of SGT cells treated with type I collagen. These results suggest that EGCG could effectively inhibit the invasion and migration of human SGT cells by downregulating the expression of β1 integrin and MMP-2/-9 and phosphorylation of FAK, Akt, and Erk. Adhesion and migration to type I collagen of SGT cells can be influenced through EGCG.

We have previously shown that 5’-nitro-indirubinoxime (5’-NIO) has potent anti-tumor effect in various human cancer cells. But the potential anti-invasive effect of 5’-NIO in salivary gland cancer has not been studied yet. The goal of this study is to evaluate the effect of 5'-NIO on salivary gland adenocarcinoma SGT cell adhesion and migration to Type I collagen treatment. Western blot, adhesion and migration assay were performed to evaluate the impacts of 5’-NIO on the expression of MMP-2/-9 and its upstream signaling molecules after treatment of type І collagen. SGT cell adhesion to type I collagen is significantly suppressed by 5’-NIO. 5’-NIO decreased expression of β1 integrin, phosphorylation of FAK, MMP-2/ -9 compared with type I collagen treatment. In addition, 5’-NIO inhibited the migration of SGT cells treated with type I collagen. These results suggest that 5’-NIO could effectively inhibit the invasion and migration of human SGT cells by downregulating the expression of β1 integrin and MMP-2/-9 and phosphorylation of FAK, Akt, and Erk. Adhesion and migration to type I collagen of SGT cells can be influenced through 5’-NIO..

This study is manufacturing method and analysis of feasibility about collagen peptide from fish scale. This is processed by enzyme hydrolysis, isolating and refining etc. The results of analysis of nutritional composition showed protein content of collagen peptide. In the analysis of constitutive amino acids, the ratio of contents of hydroxyproline and glycine, the characteristics of collagen peptides appeared similar and the contents of glutamic acid and aspartic acid which are involved in protein metabolism. As a result of measurement of total polyphenol content and total flavonoid, it showed that collagen peptide had more contents generally, and the effect of bioactivity of pig-skin collagen peptide appeared higher although different kinds of scale collagen peptide showed a little DPPH radical scavenging ability, total antioxidant capacity by ABTS, ACE inhibitory.

Gingival overgrowth can cause dental occlusion and seriously interfere with mastication, speech, and dental hygiene. It is observed in 25 to 81% of renal transplant patients treated with cyclosporine A (CsA). CsA-induced gingival overgrowth (CIGO) is caused by quantitative alteration of the extracellular matrix components, particularly collagen. However, the molecular mechanisms involved in the pathogenesis of CIGO remain poorly understood, despite intense clinical and laboratory investigations. The aim of the present work is to identify differentially expressed genes closely associated with CIGO. Human gingival fibroblasts were isolated by primary explant culture of gingival tissues from five healthy subjects (HGFs) and two patients with the CIGO (CIGO-HGFs). The proliferative activity of CsA-treated HGFs and CIGO-HGFs was examined using the MTT assay. The identification of differentially expressed genes in CsA-treated CIGO-HGF was performed by differential display reverse transcriptase-polymerase chain reaction (RT-PCR) followed by DNA sequencing. CsA significantly increased the proliferation of two HGFs and two CIGO-HGFs, whereas three HGFs were not affected. Seven genes, including the beta subunit of prolyl 4-hydroxylase (P4HB) and testican 1, were upregulated by CsA in a highly proliferative CIGO-HGF. The increased P4HB and testican-1 mRNA levels were confirmed in CsA-treated CIGO-HGFs by semiquantitative RT-PCR. Furthermore, CsA increased type I collagen mRNA levels and suppressed MMP-2 mRNA levels, which are regulated by P4HB and testican-1, respectively. These results suggest that CsA may induce gingival overgrowth through the upregulation of P4HB and testican-1, resulting in the accumulation of extracellular matrix components.

Human gingival fibroblasts are necessary for oral homeostaslS These cells are fundamental in tissue healing and tissuc remodeling processes under a response to physiological actions such as mastication, Collagen and elastin, that are extracell ular glycoprotein of gingival fibroblast, are found in all animals, '1γpe 1 collagen is most dominant protein found in human gingival fibroblasts , Matrix metalloproteinase-1(MMP-1) has a role play in destruction of metabolism of extracellular matrix(ECM) and MMP-1 can destroy many ECMs as well as non-ECM molecules MMP-1’s local activation is conytolled by tissue inhibitor of metalloproteinase-1(TIMP-1) , Therefore, it is important to have a balance between in both s ituations MMPs and TIMPs of increased 0 1' decreased extracelluar matrix molecules, The purpose of trus study is to find out the effect of physical stimulus to human gingival fibroblast on mRNA, proteins of collagen 1, elastin, MMP-1, and TIMP-1 Healthy human gingival fibroblasts were separated and cultur때 in DMEM(Dulbeco’s Modified Eagle’s Medium) , When the sample reached to confluence state, it was separated with 0,25% t rypsin and 0‘ 53mM ethylendiaminetetraacetic aCld Separated cells were centrifuged in a cell culturing flask at 1000rpm for 30, 60, 120 mmutes Then it was forced by 35g/cm' continuously, The obtained results that expression of mRNA using histological study and Reverse Transcription Polymerase Chain Reaction (RT-PCR) , expression of protein using Enzyme-Linked Immunosorbent Assay(ELISA) for this study, At 30minutes after cen trifuging, there were s pindl e shaped gingival fibroblasts with long processes parallel to other cells in the control group , However the cell density was simil ar to compared group, At 60minutes after centrifuging, spindle shaped human gingival fibroblast with relatively long process, less densely packed, At 120minutes after centrifuging, cell processes were lengthened 2-3 times‘ and cell density was lower, At 30-60 minutes after centrifuging, it was increased by 1,3-1,7 times in expressoin of collagen 1 mRNA as compared with comparison group, However, there was no change in elastin, TIMP-l, and MMP-1, At 120 minutes after centrifuging, The revealed collagen 1 mRNA was increased 3 times as compared with comparison group, It was increased 2 times in elastin , 12 times in TIMP-1 as compared with comparison group, However, there was no change in MMP-l. At 30-60 minutes after centrifuging, it was increased by 1.1 times in revealing of protein revealing in collagen 1, TIMP-1 But there was no cha nge in elastin, MMP-1 At 120 minutes after centrifuging, it was increased by 1,2 times in revealing collagen 1 protein, 11 times in elastin, 12 times in TIMP-l, but there was no change in MMP-l. ln conclu s ion, it increased in revelation of collagen 1 ,elastin and TIMP-1 by continous stimulus in human gi ngival fibroblast, But there was no change in revelation of MMP-l Therefore, th is type of pressu re is one of the components for healing of gingiva l fibroblast