Park, Jeong-Eun;Ham, Jun-Sang;Kim, Hey-Kyung;Lee, Chi-Ho;Kim, Dong-Wook;Seol, Kuk-Hwan;Oh, Mi-Hwa;Kim, Dong-Hun;Jang, Ae-Ra
Food Science of Animal Resources
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v.32
no.2
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pp.234-240
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2012
This study was conducted to examine the effects of low molecular weight gelatin hydrolysates (GH, less than 3kDa), extracted from pig skin collagen on the bone metabolism of ovariectomized (OVX) rats. The rats in the experimental groups were randomly segregated into six different treatment groups such as 1) NC, the normal rat fed AIN 93 diet (basal diet) only; 2) OC, the OVX rat fed the basal diet only; 3) GH 0.1, the OVX rat fed the basal diet with 0.1% GH; 4) GH 0.8, the OVX rat fed the basal diet with 0.8% GH; 5) G 0.1, the OVX rat fed the basal diet with 0.1% gelatin; 6) G 0.8, the OVX rat fed the basal diet with 0.8% gelatin. Body weight gain in the GH 0.1, GH 0.8, and G 0.8 was significantly higher than those in the NC and OC. Feed intake of the GH 0.1 and GH 0.8 was higher than that of the NC and OC, while no significant difference was found in feed efficiency ratio (FER). BMD of the GH 0.8 was higher than that of the OC. However, gelatin hydrolysates and gelatin resulted in higher BMC level compare to the OC. Serum HDL-cholesterol of rat fed GH and gelatin was higher than that of OC (p<0.05). LDL-C of the GH 0.1 and the GH 0.8 tended to be less than that of OC. Serum alkaline phosphatase (ALP) of the GH 0.1 was lower than that of the OC. The serum of GH 0.8 showed lower osteocalcin value than the OC (p<0.05). In addition, GOT and GPT levels significantly decreased in all treatment groups. These results indicated that gelatin hydrolysates from pig skin gelatin hydrolysates enhanced BMD and serum biochemical parameters related to bone metabolism. Therefore, the gelatin hydrolysates could be used as a beneficial material to improve bone health.
Periosteum and periosteum-derived progenitor cells have demonstrated the potential for stimulative applications in repairs of various musculoskeletal tissues. It has been found that the periosteum contains mesenchymal progenitor cells capable of differentiating into either osteoblasts or chondrocytes depending on the culture conditions. Anatomically, the periosteum is a heterogeneous multi-layered membrane, consisting of an inner cambium and an outer fibrous layer. The present study was designed to elucidate the cellular phenotypic characteristics of cambium and fibrous layer cells in vitro, and to assess whether structural integrity of the tendon in the bone tunnel can be improved by periosteal augmentation of the tendonbone interface. It was found the cells from each layer showed distinct phenotypic characteristics in a primary monolayer culture system. Specifically, the cambium cells demonstrated higher osteogenic characteristics (higher alkaline phosphatase and osteocalcin levels), as compared to the fibrous cells. Also in vivo animal model showed that a periosteal augmentation of a tendon graft could enhance the structural integrity of the tendon-bone interface, when the periosteum is placed between the tendon and bone interface with the cambium layer facing toward the bone. These findings suggest that extra care needs to be taken in order to identify and maintain the intrinsic phenotypes of the heterogeneous cell types within the periosteum. This will improve our understanding of periosteum in applications for musculoskeletal tissue repairs and tissue engineering.
Although considerable research has been done on the blood chemistry of domestic animals, little work has been made of the changes associated with age. Moreover, the records about physiology of the goat were not much available in Korea, and a comprehensive survey of the blood chemical values of the Korean native goat has not been made. The authors intended to biometric study on the hardness of bone of race horse and Jeju horse in Korea. The measurement of hardness of bone were performed in 272 race horses (Thoroughbred 91, Anglo-Arab 107, Arab 74) and in 109 Jeju horses by the caliper by Toryba's method. Some interesting facts obtained through this study were as follows: 1. There was not significant difference of bone hardness between male and female. The average of bone hardness by the Toryba's meteod were $23.07{\pm}1.01$ in race horse and $19.44{\pm}1.84$ in Jeju horse. 2. The grade of bone hardness of race horse were higher than those of Jeju horse(P<0.001). 3. The correlation coefficient between age and grade of bone hardness were r= +0.344 in Jeju horse, theme were statistically significant (P<0.01) and the regression equation was Y=0.29x+18.497.
This research was conducted to compare the effects of vitamin E (VE) when supplemented in either feed or water on the performance and meat quality of broilers. For a six-week feeding trial, a total of 330 broiler chicks were allotted to five treatments. The treatments were 1) 0 ppm VE, 2) 10 ppm VE in feed, 3) 20 ppm VE in feed, 4) 5 ppm VE in water and 5) 10 ppm VE in water. During the starter phase (0-3 weeks) chicks on non-supplemented groups grew slower (p<0.05) than the supplemented ones and the same trend was followed during the finisher (4-6 weeks) and overall period (0-6 weeks). The feed intake was significantly higher in feed supplemented groups as compared with water-supplemented groups and at higher levels as compared with lower levels of supplementation. The nutrient digestibility studies conducted after 15 and 35 days on the feeding trial showed that the digestibility of all nutrients was significantly (p<0.05) higher in supplemented groups than the non-supplemented one. The dressing percentage was higher in supplemented groups, when fed in feed and at higher levels when compared with their respective counterparts. Similar trends were noticed with respect to bone resistance. The calcium and phosphorus contents in tibia were also significantly (p<0.05) higher in supplemented, feed fed groups at higher levels than other groups. The TBARS values measured after 5 and 10 days of storage, which reflect the degree of oxidation, showed significantly lower levels in supplemented diets. The plasma and muscle vitamin E levels also showed a positive linear correlation with the levels supplemented both in feed and water. Overall it can be inferred that supplementation of VE was beneficial and there was not much difference observed when fed either in feed or water at the levels measured in the present study.
Palma, M.N.N.;Rocha, G.C.;Valadares Filho, S.C.;Detmann, E.
Asian-Australasian Journal of Animal Sciences
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v.28
no.11
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pp.1624-1628
/
2015
Rigorously standardized laboratory protocols are essential for meaningful comparison of data from multiple sites. Considering that interactions of minerals with organic matrices may vary depending on the material nature, there could be peculiar demands for each material with respect to digestion procedure. Acid digestion procedures were evaluated using different nitric to perchloric acid ratios and one- or two-step digestion to estimate the concentration of calcium, phosphorus, magnesium, and zinc in samples of carcass, bone, excreta, concentrate, forage, and feces. Six procedures were evaluated: ratio of nitric to perchloric acid at 2:1, 3:1, and 4:1 v/v in a one- or two-step digestion. There were no direct or interaction effects (p>0.01) of nitric to perchloric acid ratio or number of digestion steps on magnesium and zinc contents. Calcium and phosphorus contents presented a significant (p<0.01) interaction between sample type and nitric to perchloric acid ratio. Digestion solution of 2:1 v/v provided greater (p<0.01) recovery of calcium and phosphorus from bone samples than 3:1 and 4:1 v/v ratio. Different acid ratios did not affect (p>0.01) calcium or phosphorus contents in carcass, excreta, concentrate, forage, and feces. Number of digestion steps did not affect mineral content (p>0.01). Estimated concentration of calcium, phosphorus, magnesium, and zinc in carcass, excreta, concentrated, forage, and feces samples can be performed using digestion solution of nitric to perchloric acid 4:1 v/v in a one-step digestion. However, samples of bones demand a stronger digestion solution to analyze the mineral contents, which is represented by an increased proportion of perchloric acid, being recommended a digestion solution of nitric to perchloric acid 2:1 v/v in a one-step digestion.
Gu, Na-Yeon;Park, Mi Jeong;Lee, Jienny;Byeon, Jeong Su;Jeong, Da-Un;Cho, In-Soo;Cha, Sang-Ho
Korean Journal of Veterinary Research
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v.58
no.4
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pp.211-217
/
2018
Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-${\beta}1$ and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3D-TN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.
We tried to extract bone morphogenetic protein (BMP) from the freeze-dried bovine cortical bone (FBCB) for bone graft, which were defatted with chloroform-methanol for 20 days, freeze-dried at $-80^{\circ}C$ for 7 days and sterilized by ethylene oxide gas. Two kg of FBCB were pulverized in a wheel mill to $0.5-2.0mm^3$ cubic in size. The bone particles were demineralized in 0.6N HCI for 10 days at chloroform-methanol$4^{\circ}C$ and defatted with chloroform-methanol for 6 hours at room temperature, which was going to be defatting and demineralized cortical bone (DDM). For extracting BMP, DDM was agitated continuously through 72 hours with magnetic stirrer at $4^{\circ}C$ into 12 times of volume of 6 M guanidine hydrochloride (Gdn-HCl) solution containing proteinase inhibitors to protect BMP such as 2mM N-ethylaleimide, 1mM iodoacetic acid, 1mM phenylmethylsulfonyl fluoride and a sterilizer, 1mM sodium azide. The extraction procedure was repeated for three times. All extracted solution was centrifuged at 10,000 rpm for 30 min and then, the supernatant was dialyzed with 12 times of volume of deionized water at $4^{\circ}C$ for 24-72 hours, which cut off below 6,000-8,000 molecular weight. The dialyzed specimen contained crude-BMP was centrifuged, freeze-dried, and weighted. Through these processing, we could obtained $84.9\%$ as FBCB, $17.8\%$ as DDM and $0.71\%$ as crude-BMP from the wet cortical bone without cancellous bone, marrow and muscles. The crude-BMP were obtained $68.3\%$ from the first extraction, $29.6\%$ from secondary and $2.1\%$ from tertiary, respectively. It was suggested that high yield of crude-BMP migth be explained by three-time repetition of the extraction processing for crude-BMP with Gdn-Hcl sol.
Kim, Gi-Beom;Kim, Jae-Ryong;Ahn, Myun-Hwan;Seo, Jae-Sung
Journal of Yeungnam Medical Science
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v.24
no.2
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pp.262-274
/
2007
Purpose : Bone morphogenetic proteins (BMPs) play an important role in the formation of cartilage and bone, as well as regulating the growth of chondroblasts and osteoblasts. In this study, we investigated whether recombinant human BMP adenoviruses are available for ex vivo gene therapy, using human fibroblasts and human bone marrow stromal cells in an animal spinal fusion model. Materials and Methods : Human fibroblasts and human bone marrow stromal cells were transduced with recombinant BMP-2 adenovirus (AdBMP-2) or recombinant BMP-7 adenovirus (AdBMP-7), referred to as AdBMP-7/BMSC, AdBMP-2/BMSC, AdBMP-7/HuFb, and AdBMP-2/HuFb. We showed that each cell secreted active BMPs by alkaline phosphatase staining. Since AdBMP-2 or AdBMP-7 tranducing cells were injected into the paravertebral muscle of athymic nude mice, at 4 weeks and 7 weeks, we confirmed that new bone formation occurred by induction of spinal fusion on radiographs and histochemical staining. Results : In the region where the AdBMP-7/BMSC was injected, new bone formation was observed in all cases and spinal fusion was induced in two of these. AdBMP-2/BMSC induced bone formation and spinal fusion occurred among one of five. However, in the region where AdBMP/HuFb was injected, neither bone formation nor spinal fusion was observed. Conclusion : The osteoinductivity of AdBMP-7 was superior to that of AdBMP-2. In addition, the human bone marrow stromal cells were more efficient than the human fibroblasts for bone formation and spinal fusion. Therefore, the results of this study suggest that AdBMP-7/BMSC would be the most useful approach to ex vivo gene therapy for an animal spinal fusion model.
This study was aimed to clarify the histopathological changes in the experimental animal model subjcted to rigid fixation performed across the frontonasal sutrue in growing rabbits. Sixteen rabbits aged 6 weeks used. In experimental group(n=12), rigid fixation with miniplates and screws was performed across the frontonasal suture. Control group(n=4) was those with periosteal elevation only. Experimental animals were sacrificed on the 2nd, 4th, 8th, and 12th week after operation, and frontonasal suture area was excised for light microscopic and scanning electron microscopic examination. The results obtained were as follows : 1. In control groups, collagen fiber bundles ran in the midportion of bone sutrue and cambial layers were seen at bone surface. Sutural surfaces are beveled and external and internal bony projected portions were observed. 2. In experimental groups, distance of bone suture was decreased by new bone formation on the 2nd week, while increased by bone resorption at the miniplate applied area and bone formation in the adjacent bone on the 4th week. 3. In experimental groups, the original bone surface was almost resorbed and new bone formation was found on the 8th week. Regulary-run collagen fibers, smooth and dense bone surfaces were similar to the bone patterns of control groups on the 12th week. Above results suggest that bone formation is restricted where the miniplate is applied, while compensatory growth is appeared in the adjacent bont. It is considered that rigid fixation with miniplates and acrews results in a little disturbance of sutural growth of the craniofacial bone in infancy and children when applied for short duration.
Objective : The purpose of this study was to verify the appropriateness of ovariectomized rats as the osteoporosis animal model. Methods : Twelve female Sprague-Dawley rats underwent a sham operation (the sham group) or bilateral ovariectomy [the ovariectomy (OVX) group]. Eight weeks after operations, serum biochemical markers of bone turnover were analyzed; osteocalcin and alkaline phosphatase, which are sensitive biochemical markers of bone formation, and C-terminal telopeptide fragment of type I collagen C-terminus (CTX), which is a sensitive biochemical marker of bone resorption. Bone histomorphometric parameters and microarchitectural properties of 4th lumbar vertebrae were determined by micro-computed tomographic (CT) scan. Results : The OVX group showed on average 75.4% higher osteocalcin and 72.5% higher CTX levels than the sham group, indicating increased bone turnover. Micro-CT analysis showed significantly lower bone mineral density (BMD) (p=0.005) and cortical BMD (p=0.021) in the OVX group. Furthermore, the OVX group was found to have a significantly lower trabecular bone volume fraction (p=0.002). Conclusion : Our results showed that bone turnover was significantly increased and bone mass was significantly decreased 8 weeks after ovariectomy in rats. Thus, we propose that the ovariectomized rat model be considered a reproducible and reliable model of osteoporosis.
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