• Toxicological Research
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• v.17 no.3
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• pp.187-194
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• 2001

The oxidative stress causes the cell damage and death and thereby, stimulates membrane lipid peroxidation. In this study, the correlation between the lipid peroxidation product and the parameter of liver fibrosis (cirrhosis) was investigated in cholestasis induced rats. The Sprague-Dawley rats were divided into 3 groups (sham: sham operation, BDL/S-I and BDL/S-II : bile duct ligation/scission) and were observed for 2 or 4 weeks. After observation period, the organs were weighed and the ratio of organ weight/body weight was calculated. Sera and liver tissue were used for the measurement of malondealdehyde (MDA), parameter of clinical biochemistry, total collagen content and the staining. The ratio of organ weight/body weight in BDL/S-I and BDL/S-II was significantly increased compared to sham operated group. Serological parameters (Alanine transaminase, Aspartate transaminase, Alkaline phosphatase and Total bilirubin) in BDL/S-I and BDL/S-II group were significantly higher than those in sham operated group. Concentration of MDA in BDL/S-I (261%) and BDL/S-II(790%) was significantly increased compared to MDA in sham operated group. And the content of hydroxyproline (hyp) in BDL/S-I and BDL/S-II group was significantly increased 2~4 times than in sham operated group. The good correlations between hyp in liver tissue and MDA in sera of sham operated group and all operated group were found (r=0.825). The significantly higher value of MDA, hyp and serological parameters in BDL/S-I and BDL/S-II group suggests the stimulation of lipid peroxidation and chronic liver damage. Especially the activation of lipid peroxidation and the stimulation of liver fibrosis was stronger in BDL/S-II group than in BDL/S-I group. The stronger fibrosis, portal-portal septum formation, the more massive bile duct proliferation in portal triads and stroma, and hepatocytes swelling were observed in liver tissue of and BDL/S-II group compared to BDL/S-I group. Conclusively, a good correlation between MDA as a lipid peroxidation marker and hyp as a liver fibrotic parameter could be connected with the process of liver fibrosis. Moreover, cholestasis condition may cause jaundice, activation of lipid peroxidation, and collagen accumulation in liver. Additionally, optimal observation period of bile duct obstruction for the screening of antioxidant and antifibrotic effect in rats would be four weeks.

• Applied Biological Chemistry
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• v.37 no.2
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• pp.85-93
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• 1994

The fish skin gelatin hydrolysates were produced using a continuous two-stage membrane (MWCO 10,000, MWCO 5,000) reactor, and molecular weights, amino acids and functional properties of the hydrolysates were investigated. The major molecular weights distribution of the major fractions were $8{\sim}10\;KDa$ and $4.5{\sim}6.5\;KDa$ in the 1st-step hydrolysates, $2{\sim}6\;KDa$ and $0.5{\sim}2\;KDa$ in the 2nd-step hydrolysates. Among the amino acids in the hydrolysates, glycine, proline, serine, alanine, hydroxyproline, glutamic acid and aspartic acid having sweet taste were responsible for $68{\sim}72%$ of the total amino acids. But valine, methionine, isoleucine, leucine, phenylalanine and histidine having a bitter taste were only $23{\sim}25%$ Taste evaluations show that the gelatin hydrolysates have a brothy and sweet taste, 2nd-step hydrolysate have more a favorable taste than 1st-step hydrolysate. The hydrolysates were completely soluble and clear over the entire pH range. Moisture sorption at intermediate water activities of the 2nd-step hydrolysate was much higher than the unmodified fish skin gelatin, but foaming and emulsification properties were poor. Buffer capacity of the 2nd-step hydrolysate was higher than the fish skin gelatin and 1st-step hydrolysate, while viscosities of the hydrolysates were lower than the fish skin gelatin.

• Polymer(Korea)
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• v.32 no.1
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• pp.26-30
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• 2008

This study was designed to investigate the effect of hybridization of synthetic/natural materials for annulus fibrosus (AF) tissue regeneration in vitro and in vivo. The synthetic/natural hybrid scaffolds were prepared using PLGA (poly (lactic-co-glycolic) acid), SIS (small intestinal submucosa) and DBP (demineralized bone particles). PLGA, PLGA/SIS(20%), PLGA/DBP(20%) and PLGA/SIS (10%)/DBP (10%) scaffold were manufactured by solvent casting/salt leaching method. Compressive strength was measured. Rabbit AF cells were isolated, cultured and seeded into experimental groups. Hydroxyproline production and DNA quantity of AP cells on each scaffold was measured at 2, 4 and 6 weeks after in vitro culture. Cell-scaffold composites were implanted subcutaneously into athymic mice. After 1,4 and 6 weeks postoperatively, specimens were taken and H&E, Safranin-O and type I collagen staining were carried out concerning formation of cartilagenous tissue. In vitro PLGA/SIS scaffold was evaluated for total collagen content (bydroryproline/DNA content) and PLGA scaffold was evaluated for compressive strength.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.1
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• pp.118-122
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• 2003

Oxidative stress and its consequent lipid peroxidation exert harmful effects, which have been currently involved in the generation of carbon tetrachloride (CCl₄)-induced fibrosis(cirrhosis). In this study, it was investigated whether dried extract of 田螺(Concha Cipangopaludinae; CC) has liver functional improvement, antioxidative and antifibrotic effect in rats those were induced liver fibrosis by CCl₄ administration. The female Sprague-Dawley rats were divided into 3 groups(Normal, AC, AC-CC) and were observed in 6 weeks. Except for normal group, liver fibrosis(cirrhosis) in rats were developed by CCl₄ administration(0.8 ㎖/rat/week). And the rats were treated with prepared CC(p. o. 2 ㎖/day/rat). At the time of sacrifice, the liver, kidney and spleen were weighed and the ratio of organ weight/body weight was calculated. The MDA, hyp and biochemical parameters(AST, ALT, ALP, t-bili) were measured in sera and liver tissue of rats. The strong yellow color of urine was observed in all CCl₄-treated group compared with normal group, but jaundice didn't appear in CCl₄-treated group. The mortality of CCl₄-treated group is very low(<13%) during 6 weeks of observation time. The ratio of liver/body as well as the weight of liver in CCl₄-treated rats significantly increased compared with that in normal group(p<0.001). The level of clinical parameters in sera of all liver fibrosis(cirrhosis) developed rats were significantly higher than that of normal group(p<0.001-0.05). Especially the value of BUN, ALP, t-bilirubin in AC-CC group showed 20.9%, 19.6%, 47.9% lower than that in AC group. The content of hyp in CCl₄-treated rats was significantly higher than normal group(p<0.001～<0.05), and showed 12.2% lower value in the AC-CC group than AC group(p<0.05). The production of lipid peroxidation(MDA) in sera and liver tissue significantly increased under the fibrotic(cirrhotic) condition(p<0.001～<0.05). Especially the MDA value of AC-CC group in sera significantly 46.5% decreased compared with that of AC group(p<0.05), and the MDA value of AC-CC in liver tissue showed 21.4% lower than that of AC group. Concha Cipangopaludinae can be improved hepatic function, and maybe have effect of liver protection, antioxidation and antifibrosis.

• Applied Biological Chemistry
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• v.36 no.4
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• pp.290-295
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• 1993

In order to utilize effectively fish skin from fish processing, characteristics of the yellowfin sole and dover sole skins were investigated. In the yellowfin sole, the crude protein content and yield of fish skin used for the preparation of gelatin were 22.3% and 11.3%, respectively and in the dover sole, 17.2% and 8.9%, respectively. In the yellowfin sole skin, the soluble and insoluble collagen occupied 66.1% and 33.9%, respectively and in the dover sole skin, 78.8% and 21.1%, respectively. No difference in the amino acid composition between soluble and insoluble collagen was detected. The sum of proline and hydroxyproline content in the collagen extracted from fish skin was lower than that of those from pork skin or bone. The molecular weight of the two major subunits from the soluble collagen in the yellowfin sole skin were found to be 143 KDa and 202 KDa. Those in the dover sole skin were 142 KDa and 207 KDa. The physico-chemical properties such as the melting point and gelling point of yellowfin sole skin gelatin were superior to those of dover sole skin gelatin.

• Journal of Life Science
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• v.8 no.5
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• pp.589-597
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• 1998

In order to effectively utilize marine processing by-product such as fish scale, chemical compositions for the scale were analyzed. The selected fishes were gray mullet, Mugil cephalus, living in the sea and carp, Cyprinus carpio in the fresh water, having a lot of scales among the fishes living in seawater and fresh water. And we also investigated the difference in the chemical compositions between gray mullet and carp, depending on both living circumstances. The major components of the scales were found to be crude ash and crude protein which were each about 49% for gray mullet and which were about 20% and 79% for carp, respectively, on the basis of dried scales. The proteins extracted from both scales proved to be collagen through amino acid compositions and SDS polyacrylamide gel electrophoretic patterm. Also this scale collagen was assumed to by Type I collagen because the migration rate of $\alpha$1 and $\alpha$2 subunit of the collagen were almost the same those as calf skin Type I collagen. Most of proteins from gray mullet was collagen, however, the collagen content in proteins from carp was estimated to be only about 53%, on the basis of the ratio of hydroxyproline to protein. The crude ashes of both scales identified to be hydroxyapatite through element compositions and X-ray diffraction analysis. In conclusion, both fishes in different living circumstances were almost similar to in the chemical compositions but chemical contents for crude ash and crude protein.

• The Korean Journal of Nuclear Medicine
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• v.24 no.2
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• pp.222-228
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• 1990

The development of histomorphometric and histodynamic investigations has permitted the description of a specific and complex osteopathy in hyperthyroidism. The increased bone turnover rate in hyperthyroid patients may be accompanied by a considerable bone loss. These features are associated with both inclosed osteoclastic bone resorption and increased osteoblastric bone formation, with an accelerated calcification rate. Conventional biochemical markers of bone metabolism, i.e. serum calcium and alkaline phosphatase and urinary hydroxyproline and calcium are normal in most patients with hyperthyroidism. However, the correlation between serum BGP and serum concentration of thyroid hormon suggests that serum BGP may be a sensitive marker of increased bone formation due to the hypersecretion of thyroid hormones. Any increase in bone turnover, whether focal or diffuse, will result in an increase in $^{99m}Tc-methylenediphosphonate$ uptake (MDP). The measurement of this uptake in hyperthyroid patients by bone provides a sensitive and objective means of quantifying skeletal metabolism. Using a standard shadow-shield whole-body monitor and radioimmunoassay kit, we have measured whole-body retention of $^{99m}Tc-MDP$ up to 24hr and concentration of serum Osteocalcin in 20 patients with hyperthyroidism and in 42 normals. The results were as follows; 1) The average of serum Osteocalcin level in 42 patients with normals was $9.90{\pm}4.87(ng/ml)$ and in 20 patients with hyperthyroidism was $19.54{\pm}5.7(ng/ml)$. Both the averages of serum Osteocalcin and 24hr $^{99m}Tc-MDP$ uptakes in hyperthyroid patients were higher than those in normals. 2) $^{99m}Tc-MDP$ uptakes in skeletal system increased in proportion to normal ageing after 40 yrs old in 42 patients with normals. The average of $^{99m}Tc-MDP$ uptakes in hyperthyroid patients were higher than those in normals without related ageing. 3) A significant relationships between the $^{99m}Tc-MDP$ uptakes and serum Osteocalcin level were peformed (r=0.55, $y=17.58+6.7\times$). From the above results we concluded that the measurement of serum Osteocalcin and 24hr $^{99m}Tc-MDP$ uptakes can be used for evaluation of bone turnover as a specific marker in hyperthyroid patients.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.8
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• pp.1390-1393
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• 2003

Measuring in vivo rate of bone collagen synthesis has so far been technically difficult and often subject to quite large errors. In the present study, bone collagen synthesis rate was measured using a precursor-product method, based on the exchange of $^2$$H_2O$ into amino acids. Mass isotopomer abundance in hydroxyproline from bone collagen was analyzed by gas chromatography/mass spectrometry. The $^2$$H_2O$ labeling protocol consisted of an initial intraperitoneal injection of 99.9% $^2$$H_2O$, to achieve approximately 2.5% body water enrichment followed by administration of 4% $^2$$H_2O$ in drinking water for 9 weeks. Body $^2$$H_2O$ enrichments were stable at 2.7 ∼ 3.0% over labeling Period. In growing rats, the fractional synthesis rate ( $k_{s}$) of bone collagen was 0.066 $\pm$ 0.049 w $k^{-1}$ . The unique features of stable $^2$$H_2O$ pools and label incorporation allowed the precursor-product approach to be used for measuring bone collagen synthesis rate..

• The Journal of Korean Academy of Prosthodontics
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• v.41 no.3
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• pp.325-341
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• 2003

Statement of problem: In cases where bony defects were present, guided bone regenerations have been performed to aid the placement of implants. Nowadays, the accepted concept is to isolate bone from soft tissue by using barrier membranes to allow room for generation of new bone. Nonresorbable membranes have been used extensively since the 1980's. However, this material has exhibited major shortcomings. To overcome these faults, efforts were made to develop resorbable membranes. Guided bone regenerations utilizing resorbable membranes were tried by a number of clinicians. $Bio-Gide^{(R)}$ is such a bioresorbable collagen that is easy to use and has shown fine clinical results. Purpose: The aim of this study was to evaluate the histological results of guided bone regenerations performed using resorbable collagen membrane($Bio-Gide^{(R)}$) with autogenous bone, bovine drived xenograft and combination of the two. Surface morphology and chemical composition was analyzed to understand the physical and chemical characteristics of bioresorbable collagen membrane and their effects on guided bone regeneration. Material and methods: Bioresorbable collagen membrane ($Bio-Gide^{(R)}$), Xenograft Bone(Bio-Oss), Two healthy, adult mongrel dogs were used. Results : 1. Bioresorbable collagen membrane is pure collagen containing large amounts of Glysine, Alanine, Proline and Hydroxyproline. 2. Bioresorbable collagen membrane is a membrane with collagen fibers arranged more loosely and porously compared to the inner surface of canine mucosa: This allows for easier attachment by bone-forming cells. Blood can seep into these spaces between fibers and form clots that help stabilize the membrane. The result is improved healing. 3. Bioresorbable collagen membrane has a bilayered structure: The side to come in contact with soft tissue is smooth and compact. This prevents soft tissue penetration into bony defects. As the side in contact with bone is rough and porous, it serves as a stabilizing structure for bone regeneration by allowing attachment of bone-forming cells. 4. Regardless of whether a membrane had been used or not, the group with autogenous bone and $Bio-Oss^{(R)}$ filling showed the greatest amount of bone fill inside a hole, followed by the group with autogenous bone filling, the group with blood and the group with $Bio-Oss^{(R)}$ Filling in order. 5. When a membrane was inserted, regardless of the type of bone substitute used, a lesser amount of resorption occurred compared to when a membrane was not inserted. 6. The border between bone substitute and surrounding bone was the most indistinct with the group with autogenous bone filling, followed by the group with autogenous bone and $Bio-Oss^{(R)}$ filling, the group with blood, and the group with $Bio-Oss^{(R)}$ filling. 7. Three months after surgery, $Bio-Gide^{(R)}$ and $Bio-Oss^{(R)}$ were distinguishable. Conclusion: The best results were obtained with the group with autogenous bone and $Bio-Oss^{(R)}$ filling used in conjunction with a membrane.

• Polymer(Korea)
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• v.32 no.6
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• pp.516-522
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• 2008

Biodegradable polymers have been used extensively as scaffolding materials to regenerate new tissues and the ingrowth of tissue have been reported to be dependent directly of the porosity, pore diameter, pore shape, and porous structure of the scaffold. In this study, porous poly (L-lactide-co-glycolide) (PLGA) scaffolds with five different pore sizes were fabricated to investigate the effect of pore sizes for AF tissue regeneration. Cellular viability and proliferation were assayed by MTT test. Hydroxyproline/DNA content of AF cells on each scaffold was measured. sGAG analyses were performed at each time point of 2 and 6 weeks. Scaffold seeded AF cells were implanted into the back of athymic nude mouse to observe the difference of formation of disc-like tissue depending on pore size in vivo. We confirmed that scaffold with $180{\sim}250{\mu}m$ pores displayed high cell viability in vitro and produced higher ECM than scaffold with other pore sizes in vivo.