• Title/Summary/Keyword: gelatin matrix

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TISSUE CHANGE AFTER EMBEDDING GELATIN MATRIX IMPLANT(FFIBREL®) IN SUBCUTANEOUS TISSUE OF RATS;HISTOLOGIC, IMMUNOHISTOCHEMICAL AND SCANNING ELECTRON MICROSCOPIC STUDY (백서의 피하조직에 Gelatin Matrix Implant (Fibrel®) 매식시 조직변화에 관한 연구)

  • Kim, Hong-Jin;Lee, Chong-Heon;Kim, Kyung-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.20 no.4
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    • pp.341-354
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    • 1998
  • GMI (Fibrel${(R)}$) is one of the dermal filling substances which have been successfully used for the treatment of depressed cutaneous scar and wrinkles. It's major components are; Gelatin powder, which provides a framework for the clot to form and remains stable under the scar, and ${\varepsilon}$-aminocaproic acid, which inhibits the production of fibrinolysin, and Plasma, which provides the necessary ingredients for collagen synthesis. GMI has advantages of low immunogenicity and increased longevity. It has been known to induce fibroblast activity and promote new collagen synthesis. We used 34 Sprague-Dawley rats which were bred under the same condition and duration. 18 of experimental animals were undergone cardiac puncture, and their blood were collected, centrifugated, and stored in freezer. Out of 16 animals, control group were injected with 2ml plasma into the subcutaneous tissue of Lt. scapular, while experimental group were implanted of 2 ml GMI into the Rt. same area. Experimental animals were sacrificed at the 3rd day, 5th day, 1st week and 2nd week respectively after implantation of GMI. To observe the histopathologic change of GMI and surrounding tissue reaction of GMI, we had examined with H&E staining, immunohistochemical staining with vimentin, ${\alpha}$-SMA, S-100 under LM and SEM. The obtained results were as follows ; 1. In LM study, the inflammatory cell infiltrations and granulation tissue formation were observed, and muscle tissues were well attached with adipose tissues in the control group. In the experimental group, inflammatory cell infiltrations had been observed by the 2nd week and irregular adipiose tissues and well differentiated mesenchymal tissues were examined. 2. In immunohistochemical study, the experimental group of ${\alpha}$-SMA study, there were a prominent positive response on endothelial development of granulation tissues and mesenchymal tissues compare with the control group. In vimentin study, positive response on mescenchymal fibroblast continued to 2nd week, but negative in the control group. In S-100 study, both groups were positively responded on irregular adipose tissues. 3. In SEM study, collagen fibers were embedded by the plasma by the 5th day in the control group, and in the 3rd day experiment GMI were resorved but communited with collagen fiber till the 1st week. Collagen fibers were infilt-rated into GMI at the 2nd week and the infilltrated GMI were conglomerated with the mature adipose cells and the collagen fibers. From the above results, GMI implantation in the subcutaneous tissue of Sprague-Dawley rat, the mild infiltration of inflammatory cells were showed till 2nd week and the granulation tissues were observed. GMI were nearly resorbed till 2nd week, but well attached with adipose tissue and collagen fibers. The endothelium and fibroblasts were actively proliferated. Adipose tissues and mesenchymal tissue cells were observed. As already expressed, GMI showed resorptive change in course of time without any early immune reaction, and seemed to induce fibroblast activity and promote new collagen synthesis.

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Effects of Persimmon leaf on the Photoaging Skin Improvement(2) (감잎의 광노화 피부 개선에 미치는 효과(2))

  • Lee, Chang Hyun;Kim, Nam Seok;Choi, Dong Seong;Oh, Mi Jin;Ma, Sang Yong;Kim, Myoung Soon;Ryu, Seung Jeong;Kwon, Jin;Shin, Hyun Jong;Oh, Chan Ho
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.1
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    • pp.35-44
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    • 2014
  • This study was performed to investigate the anti-photoaging effects of Persimmon leaf tea(PLT) in hairless mice(SKH-1) exposed to UVB radiation. The animals were divided into non-treated group (normal, N) and UV-radiated groups. UV-radiated groups were divided into only UV-radiated group(control, C) and UV-radiated and PLT treated experimental groups[first extraction treated group(PLT-I), second extraction treated groupe(PLT-II), and third extraction treated group(PLT-III)]. Three PLT treated experimental groups of mice were treated with both oral administration(300mg/Kg B.W./day) and topical application (100 ul of 2% conc./mouse/day) for 4 weeks. Anti-photoaging effects of Persimmon leaf were evaluated by MTT assay, anti oxidative reaction, MMP immunohistochemistry, gelatin zymography assay and RT-PCR observations. Treatment with Persimmon leaf tea(PLT)-I, and -III groups decreased immunohistochemical density of matrix metalloproteinases(MMP)-3 and -9 related to degradation of extracellular matrix in skin. Especially, immunohistochemical density of MMP-2 decreased in PLT-I, -II and -III groups in skin. On the effects of antioxidant function on the treatment with Persimmon leaf tea(PLT), treatment of HaCaT cells with extracts of PLT-I and PLT-II had also significantly reduced intracellular ROS produced by UVB irradiation in a dose dependent manner(PLT-I, p<0.05, p<0.01, p<0.001; PLT-II, p<0.01, p<0.001). Gelatin zymography assay revealed that PLT-II and PLT-III (200 ug/ml) had inhibitory effect on MMP-9 expression in UVB-radiated HaCaT cells. Western blot analysis revealed that PLT-1, -II and -III groups down-regulates the expression of inflammatory associated genes(IL-$1{\beta}$) and PLT-1 and -II groups down-regulates the expression of TNF-${\alpha}$ in a dose dependent manner. Our study suggests that Persimmon leaf tea(PLT) extracts participates in inhibitory effects on the morphological and molecular experiments related to photoaging skin on UVB irradiated hairless mice.

Effect of Extracelluar Matrix on Cell-Surface Interactions and Growth of Cardiomyocyte(HL-1) (세포외 기질 물질에 따른 심근세포(HL-1)의 성장 연구)

  • Hong, Yoon-Mi;Anwar, Khalid;Kim, Sun-Min
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.34 no.1
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    • pp.83-87
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    • 2010
  • We present here the effect of extracellular matrix (ECM) on the proliferation and physiology of HL-1 cardiac cells. HL-1 cell is from AT-1 mouse atrial cardiomyocyte tumor lineage. HL-1 cell can be serially passaged, yet they maintain the ability to contract which is a promising character of HL-1 cell for the cell based biosensors. HL-1 cells grow up on the ECM which can affect on the attachment and growth of HL-1. In this paper, we discuss HL-1 cell-ECM interactions with three different ECMs and non-treated surface. HL-1 cells are grown for 4 days after seeding then observed their attachment. Also they were immunostained by hoechst and EthD-1 for proliferation, phalloidin for Factin, and DAPI for nuclei. Fibronectin was revealed as the proper ECM material for HL-1 cell culture. This study can provide basic information for understanding the cell-ECM interactions and growth of HL-1 cells.

α-Asarone Modulates Activity of Matrix Metalloproteinase as well as Antioxidant Activity (α-Asarone이 항산화 활성 및 기질금속단백질 분해효소 활성 조절에 미치는 영향)

  • Park, Hye-Jung;Kim, Moon-Moo
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1000-1006
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    • 2015
  • α-Asarone is the main component of Acorus gramineus, which is a widely used oriental traditional medicine. A. gramineus is known to have a variety of medicinal effects, such as anti-gastric ulcer, antiallergy and antioxidant activity. It is also known to inhibit the release of histamine. However, the mechanism of its action remains unclear in humans. In this study, the effects of α-asarone on matrix metalloproteinase (MMP) and its antioxidant effect in a cell-free system were examined in HT1080 cells. In an MTT assay, the effect of α-asarone on cell viability showed no cytotoxicity below 16 μM. In an antioxidant assay, α-asarone increased reducing power in a dose-dependent manner but not the scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. In addition, α-asarone exhibited the protective effect against DNA oxidation induced by hydroxyl radicals produced by the Fenton reaction. Furthermore, in a gelatin disk assay, α-asarone enhanced collagenase activity. It also increased the activities of MMP-2 and MMP-9 stimulated by phorbol 12-myristate 13-acetate (PMA) in a gelatin zymography. On the other hand, the activity of MMP-9 stimulated by phenazine methosulfate (PMS) but not that of MMP-2 was increased in the presence of α-asarone. These findings suggest that α-asarone could be a candidate for the prevention and treatment of pathological diseases related to oxidative stress and MMPs.

Secretory Differentiation of Hamster Tracheal Epithelial Cells Increases Activation of Matrix Metalloproteinase-2

  • Shin, Chan-Young;Lee, Woo-Jong;Park, Kyu-Hwan;Ryu, Jae-Ryun;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • v.12 no.1
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    • pp.1-8
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    • 2004
  • In chronic airway inflammatory diseases such as asthma and chronic bronchitis, it has been suggested that matrix metalloproteinases secreted from infiltrating neutrophil contribute the pathogenesis of the disease and have been a focus of intense investigation. We report here that hamster tracheal surface epithelial goblet cells (HTSE cells) produce matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2). Matrix metalloproteinase activities were investigated using [$^3H$]collagen-digestion assay and gelatin zymography. The subtype of matrix metalloproteinases expressed from HTSE cells was MMP-2 (gelatinase A), which was determined by Western blot with various subtype selective anti-matrix metalloproteinase antibodies. The MMP-2 and TIMP-2 cDNAs from HTSE cells were partially cloned by RT-PCR and they reveal more than 90% of sequence homology with those from human, rat and mouse. The collagenolytic activity was increased with the secretory differentiation of the HTSE cell and it was found that zymogen activation was responsible for the increased MMP-2 activity in HTSE cells. The results from the present study suggest that the metaplastic secretory differentiation of airway goblet cells may affect chronic airway inflammatory process by augmenting the zymogen activation of MMP-2.

A study on separation and characterization of matrix metalloproteinase-9 inhibitors from natural plants (천연 식물 추출물에서 Matrix Metalloproteinase-9 활성 억제제의 분리 및 특성화에 관한 연구)

  • Hur, Yong-Chul;Park, Sung-Woo;Kim, Tai-Jin
    • Analytical Science and Technology
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    • v.18 no.3
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    • pp.188-193
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    • 2005
  • Three different oriental natural plants (Angelicae Dahuricae Radix, Sanguisorba Officinalis L, Euonymus alatus) were extracted with 70% methanol under refluxing for 4 hr in order to investigate their inhibitory effect on Matrix Metalloproteinase-9 (MMP-9) by a modified gelatin zymography, where only euonymus alatus showed the inhibitory effect on the activity of Matrix MMP-9. The fraction was collected by using the mixture of ethyl acetate and hexane on silica gel column. Seven portions were obtained and three fractions of them (first, third and forth) showed inhibitory effect on the zymography. To verify the effect of these substances on cells, human hepatoma, Hep3B cells as a cancer model, and Chang liver cells as a normal model were selected. In order to examine the cell viability, $1{\mu}g/mL$ of each extract was treated on cells. Most of the methanol soluble fractions showed negligible toxicity on human liver cell line.

Analysis of Matrix Metalloproteinase-9 Expression in Renal Cell Carcinoma (신세포암종에서 Matrix Metalloproteinase-9 발현의 분석)

  • Kim, Ji-Yoon;Park, Tong-Choon
    • Journal of Yeungnam Medical Science
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    • v.23 no.1
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    • pp.82-89
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    • 2006
  • Background: Matrix metalloproteinases (MMPs) are involved in the degradation of the extracellular matrix, which is an important step in tumor invasion and metastasis. A positive correlation between the expression of MMP-9 and aggressive behavior of renal cell carcinomas (RCCs) has been reported. MMP-9 expression in RCCs and adjacent normal kidney tissues were examined in this study. Materials and Methods: Twenty-five patients pathologically diagnosed as clear cell RCCs, from specimens obtained at radical nephrectomy, between May 2003 and December 2004 were enrolled in this study. MMP-9 activity was estimated using gelatin zymography, and quantified using a laser densitometer. The results were compared with clinicopathological characteristics. Results: The expression of MMP-9 was significantly elevated in the RCC compared with non-tumor kidney specimens (p<0.01). The levels of MMP-9 expression in the RCC patients with large tumors (>4 cm) or vascular invasion were significantly higher than in those without these clinical manifestations (p<0.01). There were also significant differences in the expression of MMP-9 among T stages (p<0.01). The tissue MMP-9 level was the highest in nuclear grade 4, but there was no statistical significance between the histological grades (p=0.17). Conclusions: These results suggest that enhanced MMP-9 expression contributes to carcinogenesis and tumor progression in the later stages of RCC.

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Possible Role of Matrix Metalloproteinase in Osteolytic Intracranial Meningiomas

  • Moon, Hyung-Sik;Jung, Shin;Jung, Tae-Young;Cao, Van Thang;Moon, Kyung-Sub;Kim, In-Young
    • Journal of Korean Neurosurgical Society
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    • v.47 no.1
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    • pp.11-16
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    • 2010
  • Objective: Abnormalities of the bone are frequently encountered in patients with meningioma, and hyperostosis and endostosis are common bone alterations in these tumors. Extensive bony destruction is very unusual in patients with meningioma. We report six cases of intracranial meningioma associated with an osteolytic lesion of the skull and discuss the underlying mechanisms that may be responsible for bone destruction in patients with meningioma. Methods: Six patients were classified into three groups, severe, moderate and mild, according to the degree of osteolytic bony destruction. The tumor was classified as intracranial or extracranial, depending on its location. We investigated the potential role of matrix metalloproteinase (MMP) in meningioma-associated osteolysis. The levels of MMP expression were determined by gelatin zymography, reverse transcription-quantitative PCR analysis (RT-PCR) and immunohistochemical analysis. Results: Complete surgical removal of the lesion was performed in each patient. Histological examination revealed benign meningioma in four cases, and two cases of atypical meningioma. Patients did not have a poor prognosis except one case of recurred atypical meningioma. Gelatin zymography and RT-PCR detected high levels of MMP-2 in almost all extracranial masses in comparison with the intracranial masses and MMP9 in two. There was no difference in the severity of bone destruction. Immunohistochemical analysis revealed MMP-2 expression in the vicinity of the bone destruction, and a few MMP-9-positive stainings were observed. Conclusion: Osteolysis of the skull in patients with meningiomas might not be indicative of malignant pathological features and poor prognosis. Invasion to the extracranial portion and osteolysis might be associated with MMP-2 expression in meningioma.

TIMP-2 Overexpression by Retrovirus Effectively Inhibits Invasive Phenotype - A Gene Therapy Approach

  • Ahn, Seong-Min;Yeowon Sohn;Kim, Yun-Soo;Aree Moon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.106-106
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    • 2001
  • Matrix metalloproteases (MMPs) 는 다양한 세포에서 전이와 침윤성에 중요한 역할을 한다. MMP의 내인성 저해제인 tissue inhibitor of motalloprotease-2 (TIMP-2) 는 MMP-2에 높은 특이성을 지닌다. MMP-2와 TIMP-2사이의 불균형은 침윤성과 전이와 같은 병리학적 과정과 관계되는 extracellular matrix (ECM)의 퇴화를 일으킨다. TIMPs는 분비되는 분자이기 때문에 특정한 암의 유전자 치료에 사용될 가능성을 지닌다. 본 연구에서는 MMP-2가 H-ras에 의해 유도된 침윤성에 책임지는 것으로 보여지는 H-ras MCF10A 세포에 TIMP-2 유전자를 함유하는 retrovirus를 이용하여 연구하였다. TIMP-2 유전자를 함유하는 재조합 retrovirus는 PG13 세포를 infection 시키는데 사용되었다. H-ras MCF10A 세포는 PGl3 세포의 conditioned media로 처리되었을 때, gelatin zymography에서 MMP-2의 분비가 농도의존적으로 저해되었다. 또한 retrovirus에 의한 TIMP-2의 과잉 발현은 농도의존적으로 H-ras MCF10A 세포의 침윤성과 이동성을 상당히 감소시킨다. 이와 같은 실험 결과는 TIMP-2가 H-ras MCF10A 세포에서 MMP-2 분비와 세포의 침윤성, 이동성을 감소시키는 역할을 지닌다는 것과 TIMP-2 유전자를 함유하는 retrovirus가 효과적으로 MMP-2 분비, 세포 침윤성, 세포 이동성을 감소시켰다는 것을 보여 준다. 이는 암의 예방과 치료를 위한 유전자 치료법의 적용에 상당한 가능성을 제시한다.

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Differentiation of the Fetal Rat Pulmonary Epithelial Cells in Organotypic Culture (기관형 배양에서 흰쥐 태자 폐상피세포의 분화)

  • 홍혜남;조운복
    • The Korean Journal of Zoology
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    • v.35 no.3
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    • pp.295-307
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    • 1992
  • In order to study the differentiation of the epithelial cells during the development of fetal rat lung tissue, histological changeB in organotypic culture and in vivo were examined. Light microscopy and scanning electron microscopy were used to analvre the histological change in rat lung from the 15th nary of gestation to the 111th nary after birth. In organotypic culture system, the pulmonary epithelial cell differentiation was studied by scanning electron microscopy. The results obtained from this study were as follows. 1. During deveiopment of lung, the glandular stage lasted from the Isth day to the lsth naut of gestation; the canalicular stage from the 17th nay to the 19th naut of gestation; the saccuiar stage from 20th nary to the birth. Alveolar stage was observed at the 3rd nary of postnatal rat lung. 2. In organotvpic culture of fetal rat lung cells organized alveolar-like structures resembling those of in uiuo state were observed on the gelatin matrix. In contrast with in vivo state, fetal lung cells formed group of type ll pneumocytes predominently along the contours of the matrix. These cells have large apical surface, short microvilli and secreted materials which may be sunactant. These results suggested that an orsanotypic culture retaining epithelial- -mesenchvmal relationships is appropriate culture model to study the pulmonary epithelial cell (especially type ll pneumocvte) differentation.

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