• Title/Summary/Keyword: collagen gel

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Tissue engineering of dental pulp on type I collagen

  • Lee, Gwang-Hee;Huh, Sung-Yoon;Park, Sang-Hyuk
    • Restorative Dentistry and Endodontics
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    • v.29 no.4
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    • pp.370-377
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    • 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.

Effect of Exogenous Collagen on Re-epithelialization of Skin Wound in Rabbits

  • Jeon, Jin-Seok
    • Biomedical Science Letters
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    • v.7 no.1
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    • pp.17-25
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    • 2001
  • This study was performed to assess the effects of exogenous collagen gel for the re-epithelialization of partial thickness skin wound healing in rabbits. Adult male rabbits (New Zealand White Rabbit) 1.5~2 kg, were used for experimental animals. Skin wounds (1.5$\times$2 cm length) were created bilaterally on the flank of 10 rabbits and then treated a periods of 9 days. Wounds on the experimental site were treated with exogenous collagen gel as well as fabric material gauze dressing. Control site wounds were covered with fabric material gauze dressing alone. Histological findings indicated that the epithelial migration of the experimental site of rabbits was far more rapid than that in the other control wound sites. Moreover, exogenous collagen gel provided a moist environment to keep wound clean, and facilitate keratinocyte proliferation. The wound dressed with exogenous collagen gel demonstrated a significant increase in the healing rate and re-epithelialization.

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Microbial Transglutaminase Modifies Gel Properties of Porcine Collagen

  • Erwanto, Y.;Kawahara, S.;Katayama, K.;Takenoyama, S.;Fujino, H.;Yamauchi, K.;Morishita, T.;Kai, Y.;Watanabe, S.;Muguruma, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.2
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    • pp.269-276
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    • 2003
  • We studied the gel properties of porcine collagen with microbial transglutaminase (MTGase) as a catalyst. A creep meter was used to measure the mechanical properties of gel. The results showed samples with high concentration of MTGase gelled faster than those with a low concentration of MTGase. The gel strength increased with incubation time and the peaks of breaking strength for 0.1, 0.2 and 0.5% MTGase were obtained at 40, 20 and 10 min incubation time, respectively. According to SDS-PAGE, the MTGase was successfully created a collagen polymer with an increase in molecular weight, whereas no change in formation was shown without MTGase. The sample with 0.5% MTGase began to polymerize after 10 or 20 min incubation at $50^{\circ}C$, and complete polymerization occurred after 40-60 min incubation. Scanning electron microscopic analysis revealed that the gel of porcine collagen in the presence of MTGase produced an extremely well cross-linked network. The differential scanning calorimetric analysis showed the peak thermal transition of porcine collagen gel was at $36^{\circ}C$, and that with MTGase no peak was detected during heating from 20 to $120^{\circ}C$. The melting point of porcine collagen gel could be controlled by MTGase concentration, incubation temperature and protein concentration. Knowledge of the structural and physicochemical properties of porcine collagen gel catalyzed with MTGase could facilitate their use in food products.

Contraction Behavior of Collagen Gel and Fibroblats Activity in Dermal Equivalent Model

  • Yang, Eun-Kyung;Lee, Doo-Hoon;Park, Sue-Nie;Choe, Tae-Boo;Park, Jung-Keug
    • Journal of Microbiology and Biotechnology
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    • v.7 no.4
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    • pp.267-271
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    • 1997
  • We developed a dermal equivalent (DE) which was engineered using human dermal fibroblasts and a matrix of collagen gel. The in vitro construction of the DE was accomplished by casting a porcine collagen type I solution plus concentrated medium with isolated and cultured fibroblasts. These constructs were attached to culture dishes or left floating in culture medium. Contraction of attached gels results in decreased gel thickness without a change in gel diameter, and contraction of floating gels results in decreased gel thickness and diameter. After contraction, there was no increase in cell number in floating gels, but cells in attached gels began to increase after about 4 days of the lag phase in cell growth curve. At this lag phase, addition of fibroblast growth factor (FGF) at a concentration of $0.1{\mu}$/ml promoted cell proliferation in the attached collagen gels, but no effect in floating gels. These results indicate that the method of contraction had an influence on the extracellular matrix (ECM) organization, and this influenced not only cell growth but also fibroblast responsiveness to FGF. This suggests that attached collagen gel is more suitable as a dermal equivalent than the floating gel. And the final contracted area of attached gel is much larger than that of the floating gel since floating gel is contracted in all directions but attached gel is contracted only vertically.

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Comparison of Single and Sandwich Collagen Gel on the Survival and Metabolism of Rat Hepatocytes Primary Cell Culture (쥐 간세포 일차배양 세포의 생존능과 대사능에 단층과 복층 콜라젠 젤이 미치는 영향의 비교)

  • 정미경;이혜경
    • KSBB Journal
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    • v.11 no.4
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    • pp.453-461
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    • 1996
  • We compared the effects of two different systems of collagen matrix protein application on the survival and the biological functions of cultured primary hepatocytes. The rat liver primary hepatocytes were grown for approximately 40 days in vitro either on single collagen gel or between collagen sandwich gels. The morphological changes were observed for this culture period. While the hepatocytes grown on single gel began to die around at 7 days of culture, the cells grown between collagen gels still maintained their viability and began to die after 15 days. As markers for liver hepatic functions, we determined the biochemical activities of hepatocytes such as the secretions of albumin, fibronectin, fibrinogen, urea, and the reduction of secreted ammonia. We found that the rat hepatocytes cultured between collagen gels maintained fairly good biochemical functions than the hepatocytes cultured on single gel did. Therefore, the application of an extracellular matrix protein, collagen, in sandwich form was confirmed as a better choice for maintaining the functional hepatocytes culture for long term in vitro.

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Preparation of Living Skin Equivalent by using the Contracted Collagen Lattice and Cultured Human Keratinocytes (수축된 콜라겐 격자와 배양된 각질형성세포를 이용한 피부 대용물질의 제조에 관한 연구)

  • Park, Jae-Gyeong;Jo, Geum-Cheol;Park, Ho-Cheol
    • Journal of Biomedical Engineering Research
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    • v.14 no.1
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    • pp.51-62
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    • 1993
  • An experimental study was performed for the preparation of living skin-equivalent by the using collagen gel contraction with human fibroblasts as neodermls and cultured human keratinocytes as neoderm is . The results were as follows ; 1) The rate of collagen gel contraction was dependent on the number of fibroblasts into the lattice and collagen contraction was progressed according to the increment of the number of the cells. 2) The rate of collagen gel contraction was progressed according to the decrement of the contraction of the collagen. 3) The rate of gel contraction was progressed according to the increment of serum concentration in the fixed concentration of the fibroblasts and collagen. 4) The lattice contraction was decreased according to the increment of the population doublings of the fibroblasts. 5) Macroscopically, the artificial dermis was gray white in color and tissue-like consistency and elas- ticity. 6) Microscopically, three dimensionally contracted artificial dermis showed more dense fibroblasts and its newly formed collagen fibrils in the matrix than one dimensionally contracted one. 7) Finally prepared skin-equivalent showed good attachment of living stratified keratinocytes to the dermal equivalent microscopically. It has been proposed that newly formed skin-equivalent is suitable for the graft of extensively and deeply burned patients. Shortening of the manufacturing period of skin-equivalent and development of conservation technique as a readily usable state are to be solved for our ongoing works.

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Effects of Propolis on 808 nm Laser Induced Wounds

  • Shin, Jang-In;Rhee, Chung-Ku;Lee, Seung-Wan;Ahn, Jin-Chul
    • Biomedical Science Letters
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    • v.15 no.3
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    • pp.241-247
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    • 2009
  • Wound healing is a complex and highly organized biological response to injury that results in the loss of tissue integrity. Our particular interest was propolis, traditional used as an antimicrobial and an anti-inflammatory agent. The purpose of this study was to see the effects of propolis on healing of the laser induced wounds and the level of collagen formation. 808 nm laser (20 J) was irradiated on the back of rats. Irradiated wounds were divided into gel control and 6 experimental groups (3 and 5% of different three kinds of propolis). Gel type of propolis (supplied by Seoul Propolis Co., Daejeon, Korea) in water based were applied on the laser wound once daily for 14 days. One fourth of rats were sacrificed on $3^{rd}$, $7^{th}$, $10^{th}$, and $14^{th}$ day and each wound was evaluated for degree of wound healing and the level of collagen formation. Healing of wound was evaluated by measuring and comparing the width and depth of the wounds. The levels of collagen were increased in 3 and 5% gel groups comparing to control on $3^{rd}$ day and the collagen level were increased more on $7^{th}$ day. Wounds of the experimental groups showed better healing in width comparing to the healing of the control. The results of this study demonstrated that propolis accelerated healing of wounds induced by laser irradiation and good collagen formation.

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Effect of Demineralized Bone Particle Gel Penetrated into Poly(lactic-co-glycolic acid) Scaffold on the Regeneration of Chondrocyte: In Vivo Experiment (PLGA 다공성 지지체에 함침시킨 DBP젤의 연골재생 효과: In Vivo 실험)

  • Lee, Yun Mi;Shim, Cho Rok;Lee, Yujung;Kim, Ha Neul;Jo, Sun A;Song, Jeong Eun;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.36 no.6
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    • pp.789-794
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    • 2012
  • Poly(lactic-co-glycolic acid) (PLGA) has been most widely used due to its advantages such as good biodegradability, controllable rate of degradation and metabolizable degradation products. We manufactured composite scaffolds of PLGA scaffold penetrated DBP gel (PLGA/DBP gel) by a simple method, solvent casting/salt leaching prep of PLGA scaffolds and subsequent soaking in DBP gel. Chondrocytes were seeded on the PLGA/DBP gel. The mechanical strength of scaffold, histology (H&E, Safranin-O, Alcian-blue) and immunohistochemistry (collagen type I, collagen type II) were performed to elucidate in vitro and in vivo cartilage-specific extracellular matrices. It was better to keep the characteristic of chondrocytes in the PLGA/DBP gel scaffolds than that PLGA scaffolds. This study suggests that PLGA/DBP gel scaffold may serve as a potential cell delivery vehicle and a structural basis for in vivo tissue engineered cartilage.

Analysis of Low Molecular Weight Collagen by Gel Permeation Chromatography

  • Yoo, Hee-Jin;Kim, Duck-Hyun;Park, Su-Jin;Cho, Kun
    • Mass Spectrometry Letters
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    • v.12 no.3
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    • pp.81-84
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    • 2021
  • Collagen, which accounts for one-third of human protein, is reduced due to human aging, and much attention is focused on making collagen into food to prevent such aging. Gel permeation chromatography with Reflective Index (RI) detection (GPC/RI) was chosen as the most suitable instrument to confirm molecular weight distribution, and we explored the use of this technique for analysis of collagen peptide molecular sizes and distributions. Data reliability was verified by matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometric analysis. The data were considered meaningful for comparative analysis of molecular weight distribution patterns.

Biological effects of a root conditioning agent for dentin surface modification in vitro

  • Lee, Jue-Yeon;Seol, Yang-Jo;Park, Jang-Ryul;Park, Yoon-Jeong;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.40 no.6
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    • pp.257-264
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    • 2010
  • Purpose: Connective tissue reattachment to periodontally damaged root surfaces is one of the most important goals of periodontal therapy. The aim of this study was to develop a root conditioning agent that can demineralize and detoxify the infected root surface. Methods: Dentin slices obtained from human teeth were treated with a novel root planing agent for 2 minutes and then washed with phosphate-buffered saline. Smear layer removal and type I collagen exposure were observed by scanning electron microscopy (SEM) and type I collagen immunostaining, respectively. Cell attachment and lipopolysaccharides (LPS) removal demonstrated the efficiency of the root conditioning agent. Results: SEM revealed that the smear layer was entirely removed and the dentinal tubules were opened by the experimental gel. Type I collagen was exposed on the surfaces of the dentin slices treated by the experimental gel, which were compared with dentin treated with other root planing agents. Dentin slices treated with the experimental gel showed the highest number of attached fibroblasts and flattened cell morphology. The agar diffusion assay demonstrated that the experimental gel also has effective antimicrobial activity. Escherichia coli LPS were effectively removed from well plates by the experimental gel. Conclusions: These results demonstrated that this experimental gel is a useful tool for root conditioning of infected root surfaces and can also be applied for detoxification of ailing implant surface threads.