• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.89-89
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• 2003

본 연구에서는 1.7kb 및 3.1kb bovine $\beta$-casein promoter의 유전자 발현 조절능력을 알아보기 위해 1 7kb 및 3.1kb bovine $\beta$-casein promoter에 human Type II Collagen 유전자를 연결해서 DNA microinjection으로 형질전환생쥐를 생산하였다. 총 8마리의 founder생쥐(1.7kb collagen : 5마리, 3.1kb collagen 3마리)를 생산하였고 이 founder생쥐와 wild type 생쥐를 mating시켜서 $F_1 및 F_2$ 새끼를 얻었다. $F_1 및 F_2$새끼들에서 human Type II collagen 유전자의 transmission rate는 약 50%로 Mendel의 법칙에 따라 분리되어 안정적으로 유전자가 염색체에 정착되어 있음을 확인하였다. 이들 $F_1 및 F_2$새끼 중 암컷들을 임신시켜 분만 후 5-10 일경에 유선조직을 포함하여 여러 조직으로부터 RNA를 추출하여 Northern blotting 및 RT-PCR 방법을 이용하여 Type II collagen mRNA의 발현을 분석하였다. 유선에서의 발현은 1 7 kb 및 3.1 kb line별로 각각 1 line씩 발현되지 않았고, 그 외 line에서는 모두 발현되는 것으로 확인되었다. 유선에서의 Type II collagen mRNA 발형양은 1.7 kb 및 3.1 kb bovine $\beta$-casein promoter사이에서는 큰 차이를 나타내지 않았으나 1.7 kb promoter 형질전환생쥐의 경우 유선 이외 조직에서도 발현되는 양상을 나타내었고, 3.1kb promoter line에서는 유선특이적으로 발현시키는 양상을 나타내었다. 그러므로 bovine $\beta$-casein promoter의 1.7 kb와 3.1 kb 사이에 유선특이적 발현을 유도하는 조절부위가 있을 것으로 추정된다.

• BMB Reports
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• v.36 no.2
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• pp.154-158
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• 2003

Collagen is a family of proteins which consists of several genetically distinct molecular species and is intimately involved in tissue organization, function, differentiation and development. The purpose of this study was to investigate the concentration of different hydroxyproline (Hyp) fractions viz., total, free, peptide-bound, protein-bound, soluble- and insoluble-collagen hydroxyproline (Hyp) in various bovine tissues. Results showed that liver had the highest concentration of free Hyp followed by kidney, brain, spleen, lungs, muscle and heart. Liver also had the highest concentration of peptide-bound collagen Hyp followed by kidney, heart, spleen, lungs, brain and muscle. The concentration of protein-bound collagen Hyp was highest in the liver, followed by kidney, spleen, lungs, muscle, brain and heart. Total Hyp was highest in the liver, followed by kidney, spleen, brain, heart, muscle and lungs. Liver also had significantly high concentration of collagen as compared to other tissues examined (P<0.001). Spleen had the significantly higher concentration of soluble-collagen Hyp when compared to other tissues (P<0.001). This was followed by heart, muscle, lungs, brain, kidney and liver. Heart had the highest concentration of insoluble-collagen Hyp followed by lungs, kidney, liver, muscle, spleen and brain. The variation among the insoluble-collagen Hyp concentration of heart and muscle, spleen and brain was significant (P<0.001). We speculate that these differences could be due to the variation in turn over of rate of collagen metabolism in this species.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.91-109
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• 2001

The purpose of this study is to evaluate histologically the resorption and tissue response of various resorbale membranes used for guided tissue regneration procedures, using a subcutaneous model on the dorsal surface of the rat. In this study, 12 Sprague-Dawley male rats(mean BW 150gm) were used and the commercially available materials included dense collagen membrane, freeze-dried bovine dura mater loos collagen membrane, PLA/PLGA membrane. Animals were sacrificed at 3, 6 and 8 weeks after implantation of various resorbable membranes. Specimens were prepared with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows: 1. Resorption : Loose collagen membrane group was resorbed most rapidly. Dense collagen membrane group and freeze-dried bovine dura mater group were rarely resorbed. 2. Inflammatory reactions : PLA/PLGA membrane group showed persistent and severe inflammatory reactions for 3 to 8 weeks. Moderate inflammatory reactions and the ectopic formation of calcified material were observed in dense collagen membrane group. Freeze-dried bovine dura mater group and loose collagen membrane group showed mild inflammatory reactions 3. In PLA/PLGA membrane group, multinucleated giant cells by foreign body reactions were observed. In conclusion, the resorption of freeze-dried bovine dura mater didn't happen for 3-6weeks, which showed the best bio-compatibility. Therefore, freeze-dried bovine dura mater was considered proper resorbable membrane for guided tissue regeneration.

• The Journal of the Korean dental association
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• v.51 no.12
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• pp.650-657
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• 2013

The aim of this study was to achieve healing of Peri-implantitis defects and hard tissue augmentation using a bovine-derived bone mineral on the defect site. Two patients were treated with the surgical approach. With a full muco-periosteal flap elevation, the implant surfaces were exposed and granulation tissue removed around the implant and between the threads. Each surface of the contaminated implant was prepared with the air-abrasive device(PerioFlow$^{(R)}$) for decontamination. Bovine-derived bone mineral(Bio-Oss collagen$^{(R)}$) was then used to fill the defects and muco-periosteal flaps sutured to achieve transmucosal healing. Radiographs and clinical photographs were taken before and after 6 months of healing and an estimate of bone fill was assessed. Within the limits of the present case report, a surgical approach in treatment of peri-implantitis defects using a collagen form of bovine bone mineral was visited. Although limited, the two cases showed the stability and biocompatibility of a bovine-derived bone mineral and effectiveness of air-abrasive device(PerioFlow$^{(R)}$) as a decontamination method.

• Archives of Plastic Surgery
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• v.32 no.4
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• pp.529-532
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• 2005

Expanding cells ex-vivo is very important in tissue-engineering. Culture medium is usually supplemented with fetal bovine serum(FBS) in most of the experiments. However, cells grown in bovine serum media may posses the possibilities of disseminating bovine diseases and/or stimulating the patient's immune reactions. To overcome these problems, autologous or homologous serum should be used instead of the FBS. The purpose of this study is to compare cell proliferation and collagen synthesis depending on the kind of sera mixed on media and to provide a guideline on applying established experimental data to clinical cases. Human dermal fibroblasts were obtained from four patients. Five thousand cells per well in 96-well plates were incubated DMEM/F-12 Nutrient with varying serum mixture; 10% autologous serum, 10% homologous serum, and 10% FBS. Five days after incubation fibroblast proliferation and collagen production were determined by MTT assay and CICP enzyme immunoassay. The mean cell number were; $3.95{\times}10^4/well$, $2.97{\times}10^4/well$ and $2.30{\times}10^4/well$, respectively. The average amounts of collagen synthesized were; 238.13 ng/ml, 204.88 ng/ml, and 163.88 ng/ml in each. These results show that the use of human serum mixture may contribute to, not only preventing disseminated infection of bovine diseases. but also increase cell proliferation and collagen synthesis without simulating the patient's immune reactions.

• Journal of Periodontal and Implant Science
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• v.40 no.5
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• pp.220-226
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• 2010

Purpose: This study was performed to evaluate the periodontal wound healing effect of particulate equine bone mineral on canine alveolar bone defects. Methods: Twelve adult male beagle dogs were used as study subjects. The mandibular second and fourth premolars were extracted prior to the experimental surgery, and the extraction sites were allowed to heal for 8 weeks. After periodontal probing, two-walled defects were created at the mesial and distal sides of the mandibular third premolars bilaterally, and the defects were filled with equine particulate bone with collagen membrane or bovine particulate bone with collagen membrane, or collagen membrane alone. The defects without any treatment served as negative controls. After probing depth measurement, animals were sacrificed at 10, 16, and 24 post-surgery weeks for micro-computed tomographic and histomorphometric analysis. Results: The equine particulate bone-inserted group showed significantly decreased values of probing depth and first bone contact compared to the negative control and collagen membrane alone groups at weeks 10, 16, and 24 (P<0.05). There were no significant differences in the new cementum length, newly-formed bone area, or newly-formed bone volume between equine particulate bone- and bovine particulate bone-inserted groups, both of which showed significantly increased values compared to the negative control and collagen membrane alone groups (P<0.05). Conclusions: Equine particulate bone showed significant differences in probing depth, first bone contact, new cementum length, newly formed bone area, and bone volume fraction values when compared to the negative control and collagen membrane alone groups. There were no significant differences between equine and bovine particulate bone substitutes in these parameters; therefore, we can conclude that equine particulate bone is equivalent to bovine bone for periodontal regeneration.

• Journal of Acupuncture Research
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• v.26 no.6
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• pp.51-65
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• 2009

Objectives : This study was built to investigate the effect of Trachelospermi Caulis herbal-acupuncture on the Collagen-induced arthritis(CIA) in rats. Methods : Arthritis was induced by intradermal injection of Bovine type II collagen solution into base of tail. Experimental group were divided into 5 groups ; Normal(N) group, Control(C) group, Trachelospermi Caulis high(TH) group, Trachelospermi Caulis low(TL) group, Saline(S) group(n=7 for each group). Normal group was had no management. Control group was injected with Bovine type II collagen solution and taken no treatment. Trachelospermi Caulis high group was injected with Bovine type II collagen solution and taken high-intensity(10mg/kg) herbal-acupuncture treatment on $ST_{36}$. Trachelospermi Caulis low group was injected with Bovine type II collagen solution and taken low-intensity(5mg/kg) herbal-acupuncture treatment on $ST_{36}$. Saline group was injected with Bovine type II collagen solution and taken saline injection on $ST_{36}$. Body weight, paw edema volume and ankle joint thickness were measured during experimental day. On the last experimental day, we analyzed WBC count, TNF-$\alpha$ & IL-$1{\beta}$ concentration, c-fos immunohistochemistry and NADPH-d histochemistry for evaluating the effect of Trachelospermi Caulis herbal-acupuncture. Results : The results were as follows ; 1. In the change of paw edema volume, TH group only has significant difference compared with C group. 2. In the change of ankle joint thickness, TH group only has significant difference compared with C group. 3. In WBC count of serum, TH, TL groups have significant decrease compared with C group. 4. In TNF-$\alpha$ concentration of effusion, TH, TL groups have significant decrease compared with C group. and TH group has significant decrease compared with TL group. 5. In IL-$1{\beta}$ concentration of effusion, TH, TL groups have significant decrease compared with C group. and TH group has significant decrease compared with TL group. 6. In c-fos positive neurons of S1S2(cortex) region, TH, TL, S groups have significant decrease compared with C group. 7. In NADPH-d positive neurons of CPu(caudate putamen) region, TH, TL groups have significant decrease compared with C group. 8. In NADPH-d positive neurons of Tfp(transverse fibers of pons) region, TH, TL, S groups have significant decrease compared with C group. and TH group has significant decrease compared with S group. Conclusions : According to above results, we hope that Trachelospermi Caulis herbal-acupuncture may have the effect that decreases progression and development of CIA. And it can be suggested that Trachelospermi Caulis herbal-acupuncture may reduce the expression of c-fos and NOS.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.314-318
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• 2012

Most types of collagen used for biomedical applications, such as cell therapy and tissue engineering, are derived from animal tissues. Therefore, special precautions must be taken during the production of these proteins in order to assure against the possibility of the products transmitting infectious diseases to the recipients. The ability to remove and/or inactivate known and potential viral contaminants during the manufacturing process is an ever-increasingly important parameter in assessing the safety of biomedical products. The purpose of this study was to evaluate the efficacies of the 70% ethanol treatment and pepsin treatment at pH 2.0 for the inactivation of bovine viruses during the manufacture of collagen type I from bovine hides. A variety of experimental model viruses for bovine viruses including bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), bovine parainfluenza 3 virus (BPIV-3), and bovine parvovirus (BPV), were chosen for the evaluation of viral inactivation efficacy. BHV, BVDV, BPIV-3, and BPV were effectively inactivated to undetectable levels within 1 h of 70% ethanol treatment for 24 h, with log reduction factors of ${\geq}5.58$, ${\geq}5.32$, ${\geq}5.11$, and ${\geq}3.42$, respectively. BHV, BVDV, BPIV-3, and BPV were also effectively inactivated to undetectable levels within 5 days of pepsin treatment for 14 days, with the log reduction factors of ${\geq}7.08$, ${\geq}6.60$, ${\geq}5.60$, and ${\geq}3.59$, respectively. The cumulative virus reduction factors of BHV, BVDV, BPIV-3, and BPV were ${\geq}12.66$, ${\geq}11.92$, ${\geq}10.71$, and ${\geq}7.01$. These results indicate that the production process for collagen type I from bovine hides has a sufficient virus-reducing capacity to achieve a high margin of virus safety.

• The Journal of the Korean dental association
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• v.51 no.12
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• pp.643-649
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• 2013

The aim of this study was to achieve the healing of peri-implantitis defects and the hard tissue regeneration using the augmentation of a xenograft on defect site. Two patients were treated with the surgical approach. With a full muco-periosteal flap elevation, the implant surfaces were exposed and taken the debridement of granulation tissue around the abutment. Each surface of the abutments was prepared with the air-abrasive device (PerioFlow$^{(R)}$) for decontamination. Bovine-derived bone mineral (Bio-Oss collagen$^{(R)}$) was then used to fill the defects, and no membrane was placed on the grafting site. Radiographs and clinical photo was taken to compare from baseline status. Within the limits of the present case, this case shows the significance of the surgical treatment of peri-implantitis. And this also verifies the stability of bovine-derived bone mineral and effectiveness of Air-abrasive device (PerioFlow$^{(R)}$).

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.89-89
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• 2003