• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.38 no.6
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• pp.321-325
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• 2012

Collagen is widely used for regenerative therapy and pharmaceutical applications as one of the most useful scaffolds. Collagen is the most abundant protein in vertebrates and the natural substrate of various types of animal cells. Bone and dentin are mineralized tissues and almost similar in chemical components. They consist of collagen (18%), non-collagenous proteins (2%), hydroxyapatite (70%) and body fluid (10%) in weight volume. Pepsin-digested, type I collagen (atelocollagen) and heat-denatured collagen (gelatin) are basic collagenous materials for medical use. Demineralized dentin matrix (DDM) and demineralized bone matrix (DBM) belong to acid-insoluble group, and vital tooth-derived DDM is a unique dentin material including cementum and growth factors. In this review, collagen-based materials will be introduced and discussed for bone regenerative surgery.

• Journal of Ginseng Research
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• v.22 no.3
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• pp.216-221
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• 1998

We examined the anti-invasive activity of ginsenosides Rhl, Rha on the highly metastatic HT1080 human fibrosarcoma cell line. In vitro invasion assay showed ginsenoside Rhr reduced tumor cell invasion through a reconstituted basement membrane in a transwell chamber more than ginsenoside Rh1. Significant down-regulation of matrix metalloproteinase-9 (MMP-9) by ginsenosides Rh, and Rh2 was detected by Northern blot analysis. However, the expression of MMP-2 was not affected by Rh, and Rhr. The expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) was increased by Rhl after 0.5, 1 or 3 day-treatment but reduced after 6 day-treatment. However, the expression of TIMP-2 was not changed by treatment with Rh2. Plasminogen activator inhibitor (PAI) and urokinase-type plasmlnogen activator (uPA) were not changed by treatment with Rh1 and Rh2 for 3 and 6 days. Quantitative gelatin-based zymography confirmed a markedly reduced expression of MMP-9 but MMP-2 after treatments with ginsenosides Rhl and Rha. These results suggest that down-regulation of MMP-9 contributes to the anti-invasive activity of ginsenosides Rhl and Rhr in the HT1080 cells.

• KSBB Journal
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• v.8 no.4
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• pp.397-404
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• 1993

Two strains were isolated from the vinegar of Korean traditional fermented rice wine and the vine gar of fermented persimmon, respectively. These strains, designated as KM and BPV, were identified as the genus Acetobacter with respect to morphological, physiological, and biochemical characteristics. The Isolates oxidized ethanol to acetate and over-oxidized acetate or lactate to CO2 and H2O. They were positive in catalase test, while being negative in oxidase, gelatin liquefaction, VP test, H2O production and indole formation tests. No ${\gamma}$-pyrones ware produced from glucose and fructose. KM was tolerant of 11% ethanol while BPV was relatively sensitive to ethanol at a higher concentration than 5%. The guanine-plus-cytosine contents of the DNA of KM and BPV strains were 53.8 and 56.6 mol%, respectively. The cellular fatty acid compositions contained in these isolates were saturated straightchain C14:0 and C16:0,, and unsaturated straight-chain C18:1. Major ubiquinone system of KM was Q-9, but that of BPV was Q-10. In morphophysiological and biochemical aspects, KM strain was similar to Acetobacter pasteurianus. However, BPV strain was different from other Acetobacter type strains.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.68.3-69
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• 2003

Matrix metalloproteinases (MMPs) play an important role in tumor invasion and metastasis by extracellular matrix degradation. To analyze the effect of DA-125, a anthracyclin derivative, on the invasion or metastasis of cancer cells the expression of matrix metalloproteases (MMPs) was investigated in human fibrosarcoma HTl080 cells by RT-PCR or gelatin zymographic methods. As result, DA-125 suppressed the expression of MMP-2 and 9 as well as tissue inhibitor of metalloproteinase-1 (TIMP-1) TIMP-2 and MT1-MMP with a time- and dose-dependent manner. Inaddition, DA-125 inhibited cancer cell migration and colony formation, and also exhibited the inhibitory activities of invasion and motility with a matrigel and type I collagen assay. (omitted)

• 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.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.245-252
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• 2006

A bacterial strain, identified as Staphylococcus aureus JJ-11, producing collagenase was isolated out of 40 persons having skin troubles. S. aureus JJ-11 produced collagenase optimally in the media containing 1.5%(w/v) gelatin, 1%(w/v) yeast extract, 0.4%(w/v) $K_2HPO_4$, 0.005%(w/v) $NiSO_4{\cdot}6H_2O$ at $37^{\circ}C$ for 18 hrs. The collagenase produced by Staphylococcus aureus JJ-11 was purified at 6.66-folds purity through application of chromatography with Amberlite IRA-900 and Sephacryl S-300 HR columns. The molecular weight of the partially purified enzyme was estimated to be 62 kDa by SDS-PAGE. The protein exhibited optimum enzymatic activity at pH 7.0, and showed a stable activity at pH 4-8. The optimum temperature for collagenase was at $37^{\circ}C$, and activity was maintained upto $40^{\circ}C$. The enzyme activity was slightly elevated in the presence of divalents such as, $Fe^{2+},\;Co^{2+}\;and\;Ba^{2+}$ However, the activity was inhibited in the presence of $Sr^{2+}\;or\;Hg^{2+}$. The inhibition of activity by O-phenanthroline and EDTA suggested that the enzyme may contain metal which is required for activity. The enzyme showed the highest activity when insoluble collagen (type I) was, used as a substrate.

• Journal of Life Science
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• v.13 no.3
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• pp.308-313
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• 2003

The collagenase gene from Vibrio parahaemolyticus 04 was subcloned into an expression vector pET-29b. The recombinant collagenase was expressed in Escherichia coli BL2l(DE3) and partially purified by Hi-Trap affinity and Sephacryl S-100 size exclusion chromatographies. The recombinant enzyme was purified by 43.7-fold and the yield was 73%. SDS-PAGE revealed that the molecular weight of the enzyme was approximately 35 kDa. Substrate specificity study of the enzyme displayed that the enzyme showed the highest activity with the type I collagen and the synthetic peptide, Z-GPGGPA, indicating that the enzyme was indeed a collagenase. The enzyme showed broad pH optimum around pH 6-12 and was stable between pH 5.5 and 11.5. The optimum temperature for the type I collagen degradation was $35^{\circ}C$. The thermostability measurement of the enzyme indicated that the enzyme was stable up to $55^{\circ}C$, but the activity was diminished quickly above $60^{\circ}C.$

• Journal of Periodontal and Implant Science
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• v.29 no.2
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• pp.311-326
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• 1999

This study was investigated to observe the effect of Treponema denticola(TDC) and Treponema lecithinolyticum(TLC) on cultured human periodontal ligament cells. Several experiments were performed including MTT test for the inhibition effect of cell proliferation, LDH test for the cytotoxicity , gelatin zymography for the gelatinase activation and observation of cell morphology change using the phase-contrast microscopy. The results were as follows. 1. The effect of concentration on cell proliferation with time showed an inhibitory effect at high concentration $(150{\mu}g/well)$ for TLC and at low concentration( $9.4{\mu}gwell$ ) for TDC. 2. The effect of time on cell proliferation with concentration showed an inhibitory effect at $150{\mu}g/well$ on 2-day incubation for TLC and at $9.4{\mu}g/well$ on 2-day incubation for TDC. 3. The effect of heat-treated TDC and TLC on the inhibition of cell proliferation showed the difference in the heat-treated group compared to the non-heat treated group for TDC, whereas no difference was found for TLC. 4. The morphological changes which were observed from the phase-contrast microscopy showed the difference in the test group compared to the control group. The loss of spindle-like appearance, cell-to-cell detachment and inhibition of cell proliferation were observed. 5. There was no difference of the cytotoxicity effect between the test group and the control group in the LDH test. 6. The active form of progelatinase A with molecular weight 72kDa was activated in both TDC and TLC on the gelatin zymography. Regarding to the above results, TDC and TLC have an effect on periodontal ligament cells by playing an inhibitory role in cell proliferation and appears to activate progelatinase A which degrades type IV collagen.

• Food Science and Biotechnology
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• v.15 no.5
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• pp.721-727
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• 2006

This study was carried out to investigate the effect of hydrocolloids on the changes in physical properties of a model ice cream. The model ice cream contained water, sugar, skin milk powder, com oil, and 4 different hydrocolloid stabilizers (gelatin, pectin, hydroxyethylstarch, locust bean gum), was manufactured in a batch type freezer. The following physical characteristics of ice cream were examined: flow behavior, overrun, air cell size, ice crystal size, and melt resistance. With regard to flow behavior, all of aged mixes had a lower apparent viscosity relative to the mix before aging, and ice cream mix containing locust bean gum had the highest viscosity. Air cell size was observed to range from 20 to $38\;{\mu}m$, and ice cream with locust bean gum showed the largest size. There was an inverse correlation between overrun and air cell size. The ice crystal sizes of all samples ranged from 25 to $35\;{\mu}m$. Ice cream with added pectin contained the smallest ice crystal size, which was significantly difference from other stabilizers (p<0.05), and resulted in superior melt resistance with increased melting time compared to other samples.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.89-92
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• 2001

Aspergillus niger has been used as a host system to express many heterologous proteins. It has various advantages over other expression systems in that it is a small eukaryotic GRAS (Generally Recognized aS Safe) organism with a capacity of secreting large amount of foreign proteins. However, it has been known that the presence of an abundant protease is a limiting factor to express a heterologous protein. The proteases deficient mutants of A. niger were obtained using UV -mutagenesis. A total of 1 ${\times}$ $10^5$ spores were irradiated with 10-20% survival dose of UV, 600J/M2 at 280nm, and the resulting spores were screened on the casein -gelatin plates. Ten putative protease deficient mutants were further analyzed on the starch plates to differentiate the pro from the secretory mutant. An endogenous extracellular enzyme, glucose oxidase, was also examined to confirm that the mutant phenotype was due to the proteases deficiency rather than the mutation in the secretory pathway. The reduced proteolytic activity was measured using SDS-fibrin zymography gel, casein degradation assay, and bio-activity of a supplemented hGM -CSF (human Granulocyte-Macrophage Colony Stimulating Factor). Comparing with the wild type strain, less than 30 % of proteolytic activity was observed in the culture filtrate of the protease deficient mutant (pro -20) without any notable changes in cell growth and secretion.