• Nutrition Research and Practice
• /
• 제10권3호
• /
• pp.265-273
• /
• 2016

BACKGROUND/OBJECTIVES: The inhibitory effect of Hijikia fusiforme (HF) extracts on degenerative osteoarthritis was examined in primary cultured rat cartilage cells and a monosodium iodoacetate (MIA)-induced osteoarthritis rat model. MATERIALS/METHODS: In vitro, cell survival and the expression of matrix metalloproteinases (MMPs), collagen type I, collagen type II, aggrecan, and tissue inhibitor of metalloproteinases (TIMPs) was measured after $H_2O_2$ ($800{\mu}M$, 2 hr) treatment in primary chondrocytes. In vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats, and then RH500, HFE250 and HFE500 were administered orally once a day for 28 days. To determine the anti-inflammatory effects of HFE, nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) expression were measured. In addition, real-time PCR was performed to measure the genetic expression of MMPs, collagen type I, collagen type II, aggrecan, and TIMPs. RESULTS: In the in vitro assay, cell survival after $H_2O_2$ treatment was increased by HFE extract (20% EtOH). In addition, anabolic factors (genetic expression of collagen type I, II, and aggrecan) were increased by HFE extract (20% EtOH). However, the genetic expression of MMP-3 and 7, known as catabolic factors were significantly inhibited by treatment with HFE extract (20% EtOH). In the in vivo assay, anabolic factors (genetic expression of collagen type I, II, aggrecan, and TIMPs) were increased by oral administration of HFE extract. However, the genetic expression of MMP-3 and 7, known as catabolic factors, and production of NO and $PGE_2$ were significantly inhibited by treatment with oral administration of HFE extract. CONCLUSION: HFE extract inhibited articular cartilage degeneration through preventing extracellular matrix degradation and chondrocyte injury.

• 대한의용생체공학회:의공학회지
• /
• 제17권4호
• /
• pp.479-490
• /
• 1996

To utilize collagen as an implantable biomateriall the mcct widely used bovine skin origin Type I collagen was investigated Pepsin treated, Type I atelocollagen was extracted and crosslinked by the ultraviolet(W) ray with wavelength of 254nm or by various concentrations of glutaraldehyde to produce collagen membranes. The crosslink rates of the specimens were observed by a polarized light microscope, a scanning electron microscope, and a Fourier transform infrared (FT-lR) spectrometer. The followings are concluded 1. The collagen membranes produced by both 2.5% glutaraldehyde solution and 254nm UV ray irra- diation demonstrated similar morphologies on polarized light microscopic and scanning electron microscopic views. 2. The chemical structures of the crosslinked membranes by glutaraldehyde over 2.5% in concentrations revealed similar intensities to that of the UV ray irradiated one in FT-lR investigation. 3. To obtain optimal croulink in bovine stalin origin Type I atelocollagen, 2.5% glutaraldehyde solution or UV ray irradiation with 254nm wavelength is acceptable.

• Archives of Plastic Surgery
• /
• 제32권2호
• /
• pp.143-148
• /
• 2005

Chemotactic migration of bone forming cell, osteoblast, is an important event during bone formation, bone remodeling, and fracture healing. Migration of cells is mediated by adhesion receptors, such as integrins, that link the cell to extracellular matrix ligands, type I collagen, fibronectin, laminin and depend on interaction between integrin and extracellular ligand. Our study was designed to investigate the effect of extracellular matrix like fibronectin, laminin, type I collagen on migration of osteoblast. Migration distance and speed of MC3T3-E1 cell on extracellular matrix-coated glass were measured for 24 hours using 0.01% type I collagen, 0.01% fibronectin, 100 microliter/ml laminin. The migration distance and speed of MC3T3-E1 cell was compared using a video-microscopy system. To determine migration speed, cells were viewed with a 4 phase- contrast lens and video recorded. Images were captured using a color CCD camera and saved in 8-bit full-color mode. The migration distance on 0.01% type I collagen or 0.01% fibronectin was longer than that on $100{\mu}l/ml$ laminin-coated glass. The migration speed on fibronectin-coated glass was 68 micrometer/hour which was fastest. The migration speed on type I collagen-coated glass was similar with that on fibronectin-coated glass. The latter two migration speeds were faster than that on no-coated glass. On the other hand, the average migration speed on laminin-coated glass was 37micrometer/hour and not different from that of control group. In conclusion, the extracelluar matrix ligands such as type I collagen and fibronectin seem to play an important role in cell migration. The type I collagen or fibronectin coated scaffold is more effective for migration of osteoblast in tissue engineering process.

• Restorative Dentistry and Endodontics
• /
• 제29권4호
• /
• pp.370-377
• /
• 2004

The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 ($1{\;}{\times}{\;}10^5{\;}cells/ml/well$) were cultured at 12-well plate at $37^{\circ}C$ for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin. According to this study, the results were as follows: 1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05), 2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days. 3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively). 4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05). 5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels. This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제34권2호
• /
• pp.119-130
• /
• 2008

Absorbable atelo-collagen sponge $TERUPLUG^{(R)}$, Termo Co. Tokyo, Japan) is inserted in the extraction wound where alveolar bone is exposed. It protects wounds and promotes the formation of granulation. This is made of atelo-collagen, to minimize antigenicity, which is cross-linked by heat treatment for biocompatibility. $TERUPLUG^{(R)}$ consists of between 85 and 95 % of collagen type I and between 5 to 15 % of collagen type III. The raw material for the collagen is derived from bovine skin. It features a sponge block design and is shaped for easy insertion in the extraction wound. This study was designed to find out the bone healing capacity of $TERUPLUG^{(R)}$. We implanted $TERUPLUG^{(R)}$ (experimental group I) and $TERUPLUG^{(R)}$ with rhBMP-2 (experimental group II) in the rabbit cranium defect and then histologically analysed the specimen. The results were as follows. 1. In the 4 weeks, a lot of the newly formed collagen fibers around material of the experimental group I implanted $TERUPLUG^{(R)}$ were observed. But, in the experimental group II implanted $TERUPLUG^{(R)}$ with rhBMP-2, a little of newly formed collagen fibers around material were observed. The cell proliferating activity and apoptosis of the experimental group I, II was positive in and around the implanted material. 2. In the 8 weeks, the amount of newly formed and matured bone in the experimental group II was more observed than the experimental group I and control group. The results of this study indicate that absorbable atelo-collagen sponge ($TERUPLUG^{(R)}$) is relatively favorable bone void filler with biocompatibility and has the better bone healing capacity in case of application with rhBMP-2.

• BMB Reports
• /
• 제40권5호
• /
• pp.825-831
• /
• 2007

Tumor cell invasion and metastasis are often associated with matrix metalloproteinases (MMPs), among which MMP-2 and MMP-9 are of central importance. We previously showed that H-Ras, but not N-Ras, induced invasion of MCF10A human breast epithelial cells in which the enhanced expression of MMP-2 was involved. MMP-2 is produced as a latent pro-MMP-2 (72 kDa) to be activated resulting the 62 kDa active MMP-2. The present study investigated if H-Ras and/or N-Ras induces pro-MMP-2 activation of MCF10A cells when cultured in two-dimensional gel of type I collagen. Type I collagen induced activation of pro-MMP-2 only in H-Ras MCF10A cells but not in N-Ras MCF10A cells. Induction of active MMP-2 by type I collagen was suppressed by blocking integrin ${\alpha}2$, indicating the involvement of integrin signaling in pro-MMP-2 activation. Membrane-type (MT)1-MMP and tissue inhibitor of metalloproteinase (TIMP)-2 were up-regulated by H-Ras but not by N-Ras in the type I collagen-coated gel, suggesting that H-Ras-specific up-regulation of MT1-MMP and TIMP-2 may lead to the activation of pro-MMP-2. Since acquisition of pro-MMP-2 activation can be associated with increased malignant progression, these results may help understanding the mechanisms for the cell surface matrix-degrading potential which will be crucial to the prognosis and therapy of breast cancer metastasis.

• 약학회지
• /
• 제56권1호
• /
• pp.9-13
• /
• 2012

We previously reported that the extract of Taraxacum platycarpum (AF-343) had several biological properties such as skin hydration and anti-inflammatory effects, thereby AF-343 be a promising anti-atopic dermatitis agent. However, few studies have been conducted to evaluate its effect on modulation of extracellular matrix proteins in human skin fibroblasts. The purpose of this study was to investigate the expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 proteins in AF-343-treated human skin fibroblasts. Human skin fibroblasts were treated by various concentrations of AF-343 (0~2 mg/ml). The expressions of type I collagen, matrix metalloproteinase-1 (MMP-1), Smad2/3, and TIMP-1 proteins were analyzed by Western blot analysis. In addition, level of type I collagen mRNA was analyzed by CAT assay. Expression of type I collagen protein was increased in AF-343-treated human skin fibroblasts by dose and time-dependent manners. Consistent with this result, the expressions of phospho-Smad2/3 in skin fibroblasts were increased and MMP-1 expression was decreased by AF-343 treatment. TIMP-1 expression was not significantly changed in AF-343 treated skin fibroblasts. Extract of Taraxacum platycarpum (AF-343)-induced up-regulation of type I collagen expression was through increased expression of phospho-Smad2/3. These results were occurred combined with down-regulation of MMP-1 in skin fibroblasts. Taken together, this study indicated that AF-343 has property of the modulation of ECM in tissue as well as skin hydration and anti-inflammation.

• BMB Reports
• /
• 제49권6호
• /
• pp.331-336
• /
• 2016

In murine collagen-induced arthritis (CIA), self-reactive T cells can recognize peptide antigens derived from type-II collagen (CII). Activation of T cells is an important mediator of autoimmune diseases. Thus, T cells have become a focal point of study to treat autoimmune diseases. In this study, we evaluated the efficacy of recombinant MHC class II molecules in the regulation of antigen-specific T cells by using a self peptide derived from CII (CII260-274; IAGFKGEQGPKGEPG) linked to mouseI-A^q in a murine CIA model. We found that recombinant I-A^q/CII260-274 molecules could be recognized by CII-specific T cells and inhibit the same T cells in vitro. Furthermore, the development of CIA in mice was successfully prevented by in vivo injection of recombinant I-A^q/CII260-274 molecules. Thus, treatment with recombinant soluble MHC class II molecules in complex with an immunodominant self-peptide might offer a potential therapeutic for chronic inflammation in autoimmune disease such as rheumatoid arthritis.

• 대한치과교정학회지
• /
• 제26권4호
• /
• pp.455-467
• /
• 1996

견인력에 의한 치아 이동시 전반적인 치주 조직의 변화를 관찰하고 특히 제1형 교원질의 발현 정도 및 분포의 변화를 알아보고자, Sprague-Dawley계 백서 21마리를 대상으로 대조군(3마리)과 실험군(18마리)으로 나누었으며, 실험군은 양 중절치 사이에 견인력(75g)을 가한 후 12시간, 1일, 4일, 7일, 14일, 28일째에 각각 3마리씩 희생시켜, 시간에 따른 제1형 교원질의 발현과 조직학적 변화를 면역조직화학적 및 조직병리학적으로 관찰한 바 다음과 같은 결과를 얻었다. 견인력을 가한 후 28일째까지, 견인측의 치주인대 섬유는 신장되어 있었고, 압박측의 치주인대 섬유는 압축되어 있었으며 치주인대 섬유의 배열은 완전히 회복되지 않았다. 대조군에서의 제1형 교원질 발현은 구강 상피, 전상아질, 치수와 치주인대에서 경미하였으나, 치조골에 연한 조골세포, 치근단의 무세포성 백악질, 조백악세포와 악간봉합 부위에서 약양성의 발현을 보였다. 실험군에서의 제1형 교원질의 발현은, 무세포성 백악질에서 치아 이동 1일째에 중등도, 7일째부터 강양성의 발현을 보였으며, 정중 구개 봉합 부위에서는 치아 이동 1일째에 중등도, 14일째부터 강양성 발현을 보였다. 치주인대에서는 치아 이동 4일째에서 견인측이 압박측보다 제1형 교원질의 발현이 많아져 7일째에서 최고조에 달하다가 14일째부터는 측간의 차이가 없었다. 골조직에서는 조골세포가 붙어 있는 골 주변부에서 강하게 반응을 보여 대조군과 구별되었으며 특히 7일째 이후에 많은 것으로 관찰되어 골개조 현상과 관련하여 주목되었다. 이러한 관찰로 교정적 치아 이동 후 골재형성 과정은 조직 형성 세포의 분화와 이들 세포에서 형성되는 제1형 교원질과 밀접한 관계가 있음을 확인할 수 있었다.

• Archives of Plastic Surgery
• /
• 제33권4호
• /
• pp.427-432
• /
• 2006

Purpose: Noninflammatory synovial fibrosis has been noted for main causal factor of carpal tunnel syndrome (CTS). Recently, there are some reports that heparin have not only anti-coagulative effect but also anti-inflammatory and anti-fibrotic potential and have an effect on interstitial pulmonary fiborosis. Authors examined whether heparin affects pathogenesis of CTS. Methods: First, heparin was administered to fibroblast that was cultured from patient's transverse carpal ligament. Secondly, we evaluated the expression from genes of type I, III collagen, TGF ${\beta}$ isoforms and MMP. Fibroblasts were isolated and cultured from transverse carpal ligaments of 5 patients with CTS. Heparin (0, 1, 10,$100{\mu}g/ml$) was administered to cultured fibroblast and reverse transcription PCR for mRNA expression of type I, III collagen, TGF-${\beta}$ isoforms and MMP was done. Results: Heparin suppressed gene expression of type I, III collagen and TGF-${\beta}1$, ${\beta}3$ but promoted gene expression of TGF-${\beta}2$ and MMP-2. Conclusion: Heparin directly suppress gene expression of type I, III collagen. But, It is undetermined that heparin can present it's effect mediated by TGF ${\beta}$ isoforms or MMP.