As the first stage of development of an artificial skin, fibroblasts were cultured in the collagen matrices to make a living dermal equivalent. Mouse embryonic fibroblasts were incorporated into a collagen matrices on plastic dishes containing concentrated DMEM culture media supplemented with sodium bicarbonate, hepes, antibiotics and fetal bovine serum. As the growth stimulation components, glycosaminoglycans were added: hyaluronic acid, chondroitin sulfate, heparin, chitosan were incorporated into the media at a concentration of either 1% or 5% w/w/ to collagen in order to investigate the effect on development of dermal equivalent. After the few days of incubation, gel matrics were contracted and firm dermal equivalent were formed. And the keratinocytes were cultured on top of dermal equivalent and make a three dimensional artificial skin tissue.
Purpose: Collagen is the principal structural biomolecule in cartilage extracellular matrix, which makes it a logical target for cartilage engineering. In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four types of scaffolds(n=4) were inserted. The types included DHT cross-linked(Group 1), DHT and EDC cross- linked(Group 2), CS added DHT cross-linked(Group 3), and CS added DHT and EDC cross-linked(Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated respectively 4, 8, and 12 weeks after implantation. Results: The largest quantity of regenerated cartilage was found in DHT cross-linked groups 1 and 3 in the 8th week and then decreased in the 12th week, while calcification increased. Calcification was observed from the 8th week and the area increased in the 12th week. Group 4 was treated with EDC cross-linking and CS, and the matrix did not degrade in the 12th week. Cartilage-specific type II collagen mRNA expression increased with time in all groups. Conclusion: CS did not increase chondrogenesis in all groups. EDC cross-linking may prevent chondrocyte infiltration from the perichondrium into the collagen scaffold.
Kim, Hyunji;Son, Daegu;Choi, Tae Hyun;Jung, Samhyun;Kwon, Sunyoung;Kim, Junhyung;Han, Kihwan
Archives of Plastic Surgery
/
v.40
no.1
/
pp.11-18
/
2013
Background To minimize the inflammatory reaction and improve healing, a new modified dermal substitute composed of an atelocollagen, chondroitin-6-sulfate, and amniotic membrane (AM) was applied to full-thickness skin defects in a pig. Atelocollagen was extracted from bovine skin, and two modified dermal substitutes were generated according to the cross-linking type. Methods The AM-collagen dermal substitutes were characterized and compared with currently used dermal substitutes in a pig skin defect model. There were five experimental groups: dehydrothermal (DHT) cross-linking atelocollagen with the AM on the top (AM-DHT), DHT and chemical cross-linking atelocollagen with the AM on the top (AM-DHT/chemical), Terudermis, Integra, and AlloDerm. After $3{\times}3cm$ full-thickness skin defects on the back of a pig were created, each dermal substitutes dermal substitutes was randomly grafted on the defects. Two weeks after grafting, autologous partial-thickness skin was over-grafted on the neodermis. The take rate of the dermal substitutes, skin, and histological sections were all assessed at 1, 2, and 4 weeks postoperatively. Results More rapid healing and a higher take rate were evident in the AM-DHT and Terudermis groups. Histological examination revealed fewer inflammatory cells and more fibroblast hyperplasia in these two groups. Four weeks after surgery, the amount of newly formed collagen was significantly more appropriate in the AM-DHT group. Conclusions These observations provide supporting evidence that a newly developed amniotic-collagen dermal substitute may inhibit inflammatory reactions and promote wound healing.
Objectives: Capsaicin (CAP) is the chief pungent principle found in the hot red peppers and the chili peppers that have long been used as spices, food additives and drugs. This study investigated the anticancer potential of CAP through its ability to modify extracellular matrix components and proteases during mice lung carcinogenesis. Methods: Swiss albino mice were treated with benzo(a) pyrene (50 mg/kg body weight dissolved in olive oil) orally twice a week for four successive weeks to induce lung cancer at the end of $14^{th}$ week. CAP was administrated (10 mg/kg body weight dissolved in olive oil) intraperitoneally. Extracellular matrix components were assayed; Masson's trichome staining of lung tissues was performed. Western blot analyses of matrix metalloproteases 2 and 9 were also carried out. Results: In comparison with the control animals, animals in which benzo(a)pyrene had induced lung cancer showed significant increases in extracellular matrix components such as collagen (hydroxy proline), elastin, uronic acid and hexosamine and in glycosaminoglycans such as hyaluronate, chondroitin sulfate, keratan sulfate and dermatan sulfate. The above alterations in extracellular matrix components were effectively counteracted in benzo(a)pyrene along with CAP supplemented animals when compared to benzo(a) pyrene alone supplemented animals. The results of Masson's trichome staining for collagen and of, immunoblotting analyses of matrix metalloproteases 2 and 9 further supported the biochemical findings. Conclusion: The apparent potential of CAP in modulating extracellular matrix components and proteases suggests that CAP plays a chemomodulatory and anti-cancer role working against experimentally induced lung carcinogenesis.
The purpose of this study was to evaluate 1) in vivo, the expression of chondroitin 4-sulfate (CH-4S), a structural element of glycosaminoglycans(GAGs), in periodontal tissue during the experimental movement of rat incisors, by labelled streptavidine biotin immunohistochemical staining for CH-4S, 2) In vitro, the expression of CH-4S in cultured human periodontal ligament(PDL) cells supplemented with 10ng/ml of $TGF-{\beta}_1$, 20ng/ml of PDGF-BB, 1ng/ml $TNF-\alpha$, or $1{\mu}g/ml$ LPS by western blot analysis. The results of this study were as follows ; 1. The expression of CH-4S was stronger in pulp, PDL, osteoblasts, osteoclasts and osteocytes in experimental group than in control group, but was rare in dentin, and cementum of experimental groups, regardless of the duration of force application, which was not different from that of control group. 2. In experimental group, the expression of CH-4S in pulp began to increase at 1 day after force application and got to the highest degree at 7 days. After 14 days, the expression in CH-4S immunoreactivity was decreased, and became similar to that of control group at 28 days. 3. The expression of CH-4S in PDL was noted in adjacent to alveolar bone. PDL showed higher intensity of immunolabelling after 1 day of orthodontic tooth movement. And the expression was more stronger in the tension side than that of pressure side of PDL at 1 day, but more stronger in the pressure side than that of tension side of PDL at 4 days. After 7 days, a decrease in CH-4S expression was observed. 4. The expression of CH-4S in alveolar bone got to the highest degree at 4 days, and At 7 days, a decrease in CH-4S expression was observed. 5. PDGF-BB notably raised the expression of CH-4S in the PDL cells at 3 days of cultivation 6. The expression of CH-4S of PDL cells was decreased with the application of $TNF-\alpha$ at 1 day. 7. Admixture of $TGF-{\beta}_1$ and PDGF-BB got more expression of CH-4S in PDL as compared to only $TGF-{\beta}1$ or PDGF-BB. A similar decrease of the expression of CH-4S was observed in the case of application of LPS or $TNF-\alpha$.
This study was conducted to investigate the effect of maturity scores [2 (bull), 2 (steer), 3-9 (cow)] and the number of extractions (up to 4 times) on the chemical properties of water extract from Hanwoo shank bones (arm, fore shank, round and hind shank). The turbidity, meat color (CIE L value), collagen, protein, caloric and chondroitin sulfate contents of samples were observed. The turbidity and lightness were higher for water extract of Hanwoo shank bones with a maturity score of 2 (bull and steer) than maturity scores of 3-9 (cow) (p<0.05). The turbidity and lightness of water extract from shank bones of all Hanwoo maturity scores significantly increased with the 1st and 2nd extractions, but significantly decreased with 3rd and 4th extractions (p<0.05). The collagen and protein contents were highest for water extract from Hanwoo shank bones of maturity score 2 (bull and steer) (p<0.05). The caloric and chondroitin sulfate contents were higher for water extract from Hanwoo shank bones of maturity score 2 (bull and steer) than maturity scores of 3-9 (cow) (p<0.05). As the number of extractions increased, the chondroitin sulfate content significantly decreased (p<0.05). Based on these results, differences correlating with maturity scores were found only with collagen and protein contents. Therefore, further studies should be considered to address whether different maturity scores affect the price of shank bones in the meat industry.
Moon, Sang Ha;Kim, Jaecheol;Hwang, Keum Taek;Cha, Yong Bum
Korean Journal of Food Science and Technology
/
v.47
no.6
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pp.725-732
/
2015
This study was conducted to compare the physicochemical and sensory characteristics of beef-bone broths prepared under atmospheric pressure (BBAP) and overpressure (BBOP). Beef-bone was boiled in water (bone/water=1:2, w/w) for 1, 2, 4, 6, and 12 h under atmospheric pressure and overpressure ($121^{\circ}C$, 1.25 atm). The BBOP broth samples were found to contain significantly higher amounts of solid, crude protein, crude ash, collagen, and chondroitin sulfate than the BBAP broth samples for a given boiling time (p<0.05). In addition, the Ca and Mg contents in the BBAP samples were higher than those in the BBOP samples, whereas the P, Na, and K contents were higher in BBOP than BBAP. The L value of BBAP was also significantly higher than that of BBOP (p<0.05). Further, as the boiling time increased, turbidity increased in the BBAP samples. In the quantitative descriptive analysis, the BBOP samples exhibited stronger brownness, transparency, meaty off-odor, meaty off-flavor, and sulfuric odor than the BBAP samples.
Ha, Soon-Ok;Park, In-Hye;Lee, Yong-Seok;Kim, Jae-Kyu;Chung, Soo-Yeol;Choi, Yong-Lark
Journal of Life Science
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v.18
no.7
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pp.958-962
/
2008
To get the nutritional data of hot water extract from imported kangaroo tails, research was done about contents of collagen, chondroitin sulfate, protein, fat, mineral ions, and conjugated linoleic acid (CLA) in tails. Collagen content of sulfuric acid digested sample was way higher at bones than meats in both kangaroo tail and cow tail. Comparing kangaroo tail and cow tail, meat of kangaroo tail have 1.7 times higher collagen content than that of cow tail. Content of collagen in bone parts of kangaroo tail was also higher 1.2 times than that of cow tail. Meat sample of kangaroo tail (liquid extraction) have 1.3 times higher content of muco-polysaccharide than that of cow tail. In the born part, kangaroo tail was 2.4 times higher than cow tail in its content of muco-polysaccharide. The CLA content of the kangaroo tail showed the content that was higher than 0.9% of the cow tail for 4.9% and showed about 5.3 times high ratios. Especially in kangaroo tail, a band with high content of CLA was found between C18:1 and C18:2.
This study was carried out to investigate the effect of heparin, CSA and PC12 on sperm motility and acrosome reaction in bovine fresh and frozen semen which were washed and incubated in mTALP, and also the effect of heparin-pretreatment on motility and acrosome reaction in mTALP, and also the effect of heparin-pretreatment on motility and acrosome reaction of sperm treated with PC12, and the results obtained were as follows : 1. When fresh sperm was once washed and then incubated for 15 minutes in mTALP containing heparin 1, heparin 2, CSA and PC12, the percent of motile sperm of PC12 was significantly lower than that of control, heparin 1, heparin 2 and CSA. But the percent of acrosomereacted sperm of PC12 was signifciantly higher than that of control, heparin 1, heparin 2, and CSA. 2. When frozen sperm was once washed and then incubated for 15 minutes in mTALP containing heparin 1, heparin 2, CSA and PS12, there was no significant difference in the percent of motile sperm among treatments, but the percent of acrosome-reacted sperm of PC12 was signifciantly higher than that of heparin 2, and there was no significant difference in the percent of acrosome-reacted sperm among control, heparin and CSA. 3. When fresh sperm was twice washed and then incubated for 15 minutes in mTALP containing heparin and PC12, there was no significant differrence in the percent of motile sperm among treatments, but the percent of acrosome-reacted sperm of PC12 was significantly higher than that of control and heparin. When the sperm was incubated for 120 minutes, the percent of motile sperm of PC12 was significantly lower than that of control and heparin, but the percent of acrosome-reacted sperm of PC12 was significantly higher than that of control and heparin. 4. When fresh sperm was twice washed and preincubated in mTALP containing heparin for 0, 15, 120, and 240 minutes, and then incubated with PC12 for 15 minutes, there was no significant difference in the perce수 of motile sperm among treatments, but the percent of acrosome-reacted sperm of 120 and 240 minutes was significantly higher than that of 0 and 15 minutes.
Kim, Eun Young;Noh, Eun Hyung;Noh, Eun Ji;Park, Min Jee;Park, Hyo Young;Lee, Dong Sun;Riu, Key Zung;Park, Se Pill
Asian-Australasian Journal of Animal Sciences
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v.26
no.2
/
pp.178-188
/
2013
The glycosaminoglycans (GAGs) present in the female reproductive tract promote sperm capacitation. When bovine sperm were exposed to 10 ${\mu}g/ml$ of one of four GAGs (Chondroitin sulfate, CS; Dermatan sulfate, DS; Hyaluronic acid, HA; Heparin, HP) for 5 h, the total motility (TM), straight-line velocity (VSL), and curvilinear velocity (VCL) were higher in the HP- or HA-treated sperm, relative to control and CS- or DS-treated sperm. HP and HA treatments increased the levels of capacitated and acrosome-reacted sperm over time, compared to other treatment groups (p<0.05). In addition, sperm exposed to HP or HA for 1 h before IVF exhibited significantly improved fertilizing ability, as assessed by 2 pronucleus (PN) formation and cleavage rates at d 2. Exposure to these GAGs also enhanced in vitro embryo development rates and embryo quality, and increased the ICM and total blastocyst cell numbers at d 8 after IVF (p<0.05). A real-time PCR analysis showed that the expression levels of pluripotency (Oct 4), cell growth (Glut 5), and anti-apoptosis (Bax inhibitor) genes were significantly higher in embryos derived from HA- or HP-treated sperm than in control or other treatment groups, while pro-apoptotic gene expression (caspase-3) was significantly lower in all GAG treatment groups (p<0.05). These results demonstrated that exposure of bovine sperm to HP or HA positively correlates with in vitro fertilizing ability, in vitro embryo developmental potential, and embryonic gene expression.
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