• Title/Summary/Keyword: Collagen gel

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Extraction and characterization of pepsin-soluble collagen from different mantis shrimp species

  • Hiransuchalert, Rachanimuk;Oonwiset, Nakaweerada;Imarom, Yolrawee;Chindudsadeegul, Parinya;Laongmanee, Penchan;Arnupapboon, Sukchai
    • Fisheries and Aquatic Sciences
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    • v.24 no.12
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    • pp.406-414
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    • 2021
  • The objective of this study was to investigate the yield and characteristics of collagen protein extracted from the muscle of four different species of mantis shrimp: Miyakella nepa, Harpiosquilla harpax, Erugosquilla woodmasoni, and Odontodactylus cultrifer. Mantis shrimp muscle was extracted by using a pepsin-solubilization technique, with 0.5 M acetic acid and 5% pepsin enzyme. The highest collagen yield was from M. nepa muscle (0.478 ± 0.06%), which was significantly greater (p < 0.05) than that from H. harpax, O. cultrifer, and E. woodmasoni (0.313 ± 0.03%, 0.123 ± 0.02%, and 0.015 ± 0.00%, respectively). The freeze-dried collagen appeared as thin fibers, and formed an opaque film. The pepsin-soluble collagen (PSC) from four mantis shrimp species was analyzed by gel electrophoresis. The results showed that all species of mantis shrimp contained type I collagen, consisting of β, α1, and α2 subunits with average molecular weights of 250, 145, and 118 kDa, respectively. The study of the solubility of collagen showed that, for NaCl, collagen had the highest relative solubility in 2% NaCl (80.20 ± 4.95%). In contrast, the solubility decreased at higher NaCl concentrations. However, in terms of pH, collagen had the highest relative solubility at pH 3 (91.32 ± 5.14%), and its solubility decreased at higher pH. FT-IR spectroscopy was used to compare the collagen with a model compound. Five wavenumbers in the spectrum for model collagen were identified: Amide A (3,406-3,421 cm-1), amide B (2,916-2,940 cm-1), amide I (1,639-1,640 cm-1), amide II (1,539-1,570 cm-1), and amide III (1,234-1,250 cm-1).

Selection of Immobilization Material for Stabilization of Bioluminescence from Photobacterium phosphoreum (Bioluminescence 안정성을 위한 Photobacterium phosphoreum의 고정화 물질에 관한 연구)

  • Lee, Eun-Su;Kim, Hyeon-Suk;Jeon, Eok-Han
    • KSBB Journal
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    • v.14 no.4
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    • pp.403-407
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    • 1999
  • Various materials including sodium alginate, k-carragreenan, collagen and polyacrylamide were studied in order to maintain stability of bioluminescence of P. phosphoreum for the purpose of continuos monitoring of toxic subtances. Collagen and polycryamide were shown to be inadequate for immobilization of p. phosphoreum since the bioluminescence decreased when cells were mixed with such materials. In case of k-carrageenan, the bioluminescence was stable when compared with collagen and polyacryamide. However, the k-carrageenan was not suitable for immobilization of p. phosphoreum as cells could not be mixed with the material properly in temperature at which gel formation already occurred. P . phosphoreum must be treated at low temperature below that of gel formation since these are psychrophilic luminescent bacterial. When cells were immobilized on sodium alginate, the bioluminescence was stably maintained for 20 minutes.

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A Study on Growth of Human Testicular Tissue in 3-Dimensional Collagen Gel Tissue Culture (Collagen Gel을 이용한 사람의 고환 조직배양에 관한연구)

  • Lee, Choong-Hyun;Lee, Sang-Cheol;Lee, Sun-Joo;Sohn, Joon-Woong;Chang, Sung-Goo;Kim, Jin-Il;Chai, Soo-Eung
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.1
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    • pp.53-56
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    • 1993
  • A recently developed collagen gel culture technique has been applied to study on growth in tissue of human testicular tissue. Minimum Eagle's medium supplemented with amino acid, 10% Fetal Bovine Serum and 0.1mM non-essential amino acid are emploid. Tissue fragments on collagan gel are fixed at time intervals for the histologic findings of testis. The mature spermatids are maintained for 2 weeks and can be observed until four weeks. But the rate of glucose consumption is increased contrary to histologic findings.

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Preparation and quality characteristics of low molecular weight collagen treated with hydrolytic enzymes from Korean native chicken feet (효소를 이용한 저분자 토종 닭발 콜라겐의 제조 및 품질 특성)

  • Jeong, Gyeong A;Lee, Chang Joo
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.695-700
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    • 2021
  • The purpose of this study was to prepare low-molecular weight collagen using a commercial proteolytic enzyme (Protamex) from collagen extracted from feet of Korean native chicken and to investigate the quality characteristics of this collagen. The collagen content of Korean native chicken feet was 13.9 g/100 g, which was higher than the 6.21 g/100 g of general broilers. It was found that the content of low molecular weight collagen increased as the concentration of proteolytic enzymes and reaction time increased. In particular, reaction with 1% Protamex for 7 h resulted in 55.6% of low molecular weight (1,000-5,000 Da) collagen content, and the average molecular weight was 5,390 Da. Regarding the texture of the enzyme-treated collagen, the collagen with high molecular weight peptides decomposed into low molecular weight peptides, and the gel type could not be formed, whereas the sol type was maintained.

Simvastatin as a Modulator of Tissue Remodeling through Inhibition of Matrix Metalloproteinase (MMP) Release from Human Lung Fibroblasts

  • Ra, Ji-Eun;Lee, Ji-Kyoung;Kim, Hui-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.71 no.3
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    • pp.172-179
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    • 2011
  • Background: Statins can regulate the production of pro-inflammatory cytokines and inhibit MMP production or activation in a variety of types of cells. This study evaluated whether statins would inhibit MMP release from human lung fibroblasts, which play a major role in remodeling processes. Methods: This study, using an in-vitro model (three-dimensional collagen gel contraction system), evaluated the effect of cytokines (tumor necrosis factor-${\alpha}$, TNF-a and interleukin-$1{\beta}$, IL-1b) on the MMP release and MMP activation from human lung fibroblasts. Collagen degradation induced by cytokines and neutrophil elastase (NE) was evaluated by quantifying hydroxyproline. Results: In three-dimensional collagen gel cultures (3D cultures) where cytokines (TNF-a and IL-1b) can induce the production of MMPs by fibroblasts, it was found that simvastatin inhibited MMP release. In 3D cultures, cytokines together with NE induced collagen degradation and can lead to activation of the MMP, which was inhibited by simvastatin. Conclusion: Simvastatin may play a role in regulating human lung fibroblast functions in repair and remodeling processes by inhibiting MMP release and the conversion from the latent to the active form of MMP.

Effects of High Pressure/High Temperature Processing on the Recovery and Characteristics of Porcine Placenta Hydrolysates

  • Lee, Mi-Yeon;Choi, Ye-Chul;Chun, Ji-Yeon;Min, Sang-Gi;Hong, Geun-Pyo
    • Food Science of Animal Resources
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    • v.33 no.4
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    • pp.474-480
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    • 2013
  • This study was performed to investigate the effects of high pressure/high temperature (HPHT) treatment on the recovery efficiency and characteristics of porcine placenta hydrolysates. The placenta hydrolysates were characterized by solubility, free amino acid contents, gel electrophoresis, gel permeation chromatography (GPC) and amino acid composition. Placenta was treated at 37.5 MPa of pressure combined with various temperatures (150, 170, and $200^{\circ}C$) or various holding times (0, 30, and 60 min at $170^{\circ}C$). Insoluble raw placenta collagen was partially solubilized (> 60% solubility) by the HPHT treatment. Free amino group content of placenta collagen was increased from 0.1 mM/g collagen to > 0.3 mM/g collagen after HPHT treatment, reflecting partial hydrolysis of collagen. The molecular weight ($M_w$) distribution showed evidence of collagen hydrolysis by shifting of $M_w$ peaks toward low molecular weight when treated temperature or holding time was increased. Alanine (Ala), glycine (Gly), hydroxyproline (Hyp), and proline (Pro) contents increased after the HPHT treatments compared to a decrease in the others. In particular, the increase in Gly was obvious, followed by Hyp and Pro, reflecting that placenta hydrolysates were mainly composed of these amino acids. However, increasing temperature or holding time hardly affected the amino acid compositions. These results indicate that the HPHT treatment is advantageous to hydrolyze collagen derived from animal by-products.

A STUDY OF THE EFFECT OF CULTURED BONE MARROW STROMAL CELLS ON PERIPHERAL NERVE REGENERATION (체외 배양한 골수줄기세포를 이용한 말초신경재생에 관한 연구)

  • Choi, Byung-Ho;Zhu, Shi-Jiang;Jung, Jae-Hyung;Huh, Jin-Young;Lee, Seoung-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.6
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    • pp.492-495
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    • 2005
  • The role of cultured bone marrow stromal cells (BMSCs) in peripheral nerve regeneration was examined using an established rabbit peroneal nerve regeneration model. A 15-mm peroneal nerve defect was bridged with a vein filled with BMSCs $(1{\times}10^6)$, which had been embedded in collagen gel. On the contralateral side, the defect was bridged with a vein filled with collagen gel alone. When the regenerated tissue was examined 4, 8 and 12 weeks after grafting, the number and diameter of the myelinated fibers in the side with the BMSCs were significantly higher than in the control side without the BMSCs. This demonstrates the potential of using cultured BMSCs in peripheral nerve regeneration.

SHRINKAGE OF VITREOUS BODY CAUSED BY HYDROXYL RADICAL

  • Park, Myoung-Joo;Shimada, Takashi;Matuo, Yoichirou;Akiyama, Yoko;Izumi, Yoshinobu;Nishijima, Shigehiro
    • Journal of Radiation Protection and Research
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    • v.33 no.4
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    • pp.143-150
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    • 2008
  • In this study, we examined the effect of hydroxyl radical generated by $\gamma$-ray and UV irradiation on shrinkage of vitreous body. Change in gel ratio of vitreous body and change in the properties of its components (collagen, sodium hyaluronate) were analyzed. By comparing these results, the amount of hydroxyl radical, which induces the considerable shrinkage of vitreous body, was evaluated from theoretical calculation based on experimental condition and some reported kinetic parameters. It was concluded that the integrated amount of hydroxyl radical required to liquefy half of the vitreous body (Vitreous body gel ratio = 50%) was estimated as $140\;{\mu}molg^{-1}$ from $\gamma$-ray irradiation experiment. Also, from UV irradiation experiment result, it was confirmed that the effect of hydroxyl radical is larger than that of other reactive species. The causes of shrinkage of vitreous body are supposed as follows, 1) decrease in viscosity by cleavage of glycoside bond in sodium hyaluronate, 2) leaching of collagen from vitreous body and 3) leaching of crosslinked products and scission products of collagen.

Establishment of Embryo Culture System using Co-incubated Collagen Matrix Gel with Porcine Endometrial Cells (돼지 자궁내막 상피세포와 공동배양된 Collagen Matrix Gel을 이용한 체외수정란 배양체계 확립)

  • Lee, Sang-Hee;Han, Hye-In;Hwangbo, Yong;Lee, Seunghyung;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.39 no.3
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    • pp.49-57
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    • 2015
  • 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 $IL-1{\beta}$. 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 $IL-1{\beta}$ groups than EC without hCG and $IL-1{\beta}$. The VEGF protein was detected at co-incubated CM with EC, EC treated with hCG and $IL-1{\beta}$ 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 $IL-1{\beta}$ is beneficial and useful for enhancing the production of porcine blastocysts in vitro.

Characterization of Low-Molecular-Weight Collagen from Korean Native Chicken Feet Hydrolyzed Using Alcalase

  • Heedong Woo;Gyeong A Jeong;Hyunwook Choi;Chang Joo Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.656-661
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    • 2023
  • The aims of this study were to optimize the preparation of low-molecular-weight collagen using a proteolytic enzyme (alcalase) derived from the feet of Korean native chickens, and to characterize the process of collagen hydrolysis. Foreign bodies from chicken feet were removed using ultrasonication at 28 kHz with 1.36 kW for more than 25 min. The hydrolytic pattern and molecular weight distribution of enzyme-treated collagen from chicken feet were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high-performance liquid chromatography, respectively. Ideally, chicken feet should be treated at 100℃ for 8 h to obtain a high collagen content using hot water extraction. The collagen content of the chicken foot extract was 13.9 g/100 g, and the proportion of low-molecular-weight collagen increased with increasing proteolytic enzyme concentration and reaction time. When treated with 1% alcalase, the average molecular weight of collagen decreased rapidly to 4,929 Da within 5 h and thereafter decreased at a slower rate, reaching 4,916 Da after 7 h. Size exclusion chromatography revealed that low-molecular-weight collagen peptides of approximately 1,000-5,000 Da were obtained after hydrolysis with 1% alcalase for 1 h.