• Journal of Chest Surgery
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• 제46권1호
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• pp.1-13
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• 2013

Background: Glutaraldehyde (GA) is a widely used cross-linking agent for improving mechanical properties and resistance to enzymatic degradation of collagenous tissue, but it has several drawbacks such as calcification and cytotoxicity. The aim of this study was to find the alternative effective cross-linking methods to GA. Materials and Methods: Bovine pericardium was processed with GA with ethanol+octanol and glycine detoxification, and polyethylene glycol (PG) space filler, dimethyl 3,3'-dithiobispropionimidate (DTBP), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) treatment, and the physical fixation of ultraviolet irradiation were done. The biologic material properties of variously treated pericardial tissues were assessed by biochemical, mechanical and histological tests. Treated pericardial tissues were also implanted subcutaneously or intramuscularly into the rabbit for 10 weeks to assess the xenoreactive antibody response of immunoglobulin G and M, their anti-calcification effect. Results: The biochemical and mechanical properties of EDC fixed pericardial tissues were comparable to the GA fixed tissue. The cytotoxicity was lowest in space filler treated GA fixed group. In rabbit subcutaneous or intramuscular implantation models, decellularization, space filler, EDC treatment group showed significantly lower calcium content than GA only and DTBP treatment group (p<0.05, analysis of variance). The titer of anti $Gal{\alpha}1-3Gal{\beta}1$-4GlcNAc-R antibodies did not change in the postimplantation serial enzyme-linked immunosorbent assay. Hematoxylin and eosin and von Kossa staining showed that decellularization, space filler, EDC, and ultraviolet treatment had less inflammatory cell infiltration and calcium deposits. Conclusion: The decellularization process, PG filler, and EDC treatments are good alternative cross-linking methods compared to GA only fixation and primary amine of DTBP treatment for cardiovascular xenograft preservation in terms of the collagen cross-linking stability and in vivo anti-calcification effects.

• 한국수산과학회지
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• 제25권5호
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• pp.432-442
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• 1992

동남참게, Eriocheir japonicus의 난황단백전구체 합성부위를 면역조직화학적인 방법으로 구명하기 위하여, 난황단백질을 정제하고, 이 단백질에 대한 항혈청을 제작하여 난황단백질에 대한 면역학적인 성질을 밝힌 후, 난황단백전구체의 항체가 존재하는 항 난황단백혈청을 사용하여 PAP(peroxidase-antiperoxidase)법으로 생식소, 간췌장, 근육조직을 대상으로 면역조직화학 반응을 시켰다. 성숙한 암, 수혈청에 대한 면역전기영동과 Ouch-terlony 면역확산 실험에 의하여 암혈청내에는 암 특이성혈청단백질이 있음이 밝혀졌다. 암, 수혈청에 대한 항혈청과 난황형성중인 암혈청에 대한 조난황단백추출액에 대하여 면역전기영동과 Ouchte-rlony 면역확산 실험을 행한 결과, 암 특이성혈청 단백질은 난황단백전구체임이 밝혀졌다. 조난황단백추출액으로 부터 DEAE 이온교환 chro-matography와 sepharose CL-4B gel 여과법에 의하여 난황단백질을 정제하였으며, 이 난황단백질의 분자량을 sepharose CL-4B gel 여과법에 의하여 추정한 결과 분자량은 약 245,000 dalton이었다. 또한 난황단백질은 면역학적인 분석에 의하여 난황 단백질에 대한 항혈청에는 난황단백전구체에 대한 항체가 존재하고 있음이 증명되었다. 난황단백질에 대한 토끼 항혈청을 이용하여 PAP법에 의한 면역조직화학반응을 시킨 결과, 난황형성중인 난모세포와 성숙한 암참게의 간췌장은 양성반응이 나타는데 반하여 난소내의 여포세포층과 난황형성전 난모세포, 근육층 그리고 성숙한 수참게의 간췌장은 전혀 양성반응이 나타나지 않았다. 따라서 본 종의 난황단백전구체의 합성부위는 간췌장임을 알 수 있다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제31권6호
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• pp.469-477
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• 2009

육안적 검사결과 실험군과 대조군 모두에서 특별한 염증 소견이나 창상 이개 없이 반원 모양의 골형성이 관찰되었다. 조직학적으로 3주째에 대조군, 실험 1군, 실험2군, 실험 3군 모두 이식골 주위 및 티타늄 반구 내면을 따라 신생 골형성이 관찰되었다. 조직학적으로 6주째에 모든 군에서 3주째에 비하여 신생골 면적의 증가 및 성숙 소견이 관찰되었고, 실험2군에서는 부분적으로 이식골이 흡수되면서 신생골이 형성되는 것이 관찰된 반면, 실험 3군에서는 이식골의 흡수 소견이 관찰되지 않았다. 조직형태계측학적으로 3주, 6주 모두 자가골에서 가장 많은 신생골 형성이 나타났고, 신생골 면적 비교시 자가골, 이종골, 합성골 순으로 크게 나타났고, 각 군간의 통계학적으로 유의한 차이는 없었다(p>0.05). 본 연구결과 골유도 재생술시 골형성 능력은 자가골이 가장 좋지만, 자가골 채취가 불가능할 경우, 적절한 차폐막을 사용한 합성골과 이종골 복합 이식방식도 좋은 대체제가 될 것으로 생각된다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제38권
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• pp.16.1-16.9
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• 2016

Background: The association of biomaterial combined with repair factor-like platelet-rich plasma (PRP) has prospective values. Bovine-derived xenograft has been identified as an osteoconductive and biocompatible grafting material that provides osseointegration ability. PRP has become a valuable adjunctive agent to promote healing in a lot of dental and oral surgery procedures. However, there are controversies with respect to the regenerative capacity of PRP and the real benefits of its use in bone grafts. The purpose of this study was to assess the influence of PRP combined with xenograft for the repair of peri-implant bone defects. Methods: Twelve rabbits were used in this study, and the experimental surgery with implant installation was performed simultaneously. Autologous PRP was prepared before the surgical procedure. An intrabony defect (7.0 mm in diameter and 3.0 mm deep) was created in the tibia of each rabbit; then, 24 titanium dental implants (3.0 mm in diameter and 8.5 mm long) were inserted into these osteotomy sites. Thus, a standardized gap (4.0 mm) was established between the surrounding bony walls and the implant surface. The gaps were treated with either xenograft alone (control group) or xenograft combined with PRP (experimental group). After healing for 1, 2, 3, 4, 5, and 6 weeks, the rabbits were sacrificed with an overdose of KCl solution. Two rabbits were killed at each time, and the samples including dental implants and surrounding bone were collected and processed for histological analysis. Results: More newly formed bone and a better bone healing process were observed in control group. The histomorphometric analysis revealed that the mean percentage of bone-to-implant contact in the control group was significantly higher than that of the experimental group (25.23 vs. 8.16 %; P < 0.05, independent-simple t test, analysis of variance [ANOVA]). Conclusions: The results indicate that in the addition of PRP to bovine-derived xenograft in the repair of bone defects around the implant, PRP may delay peri-implant bone healing.

• Toxicological Research
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• 제31권1호
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• pp.49-59
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• 2015

Eye is a highly vascularised organ. There are chances that a foreign substance can enter the systemic circulation through the eye and cause oxidative stress and evoke immune response. Here the eyes of rabbits were exposed, for a period of 7 days, to 5 known ocular irritants: Cetyl pyridinium chloride (CPC), sodium salicylate (SS), imidazole (IMI), acetaminophen (ACT) and nicotinamide (NIC). The eyes were scored according to the draize scoring. Blood collected from the treated rabbit were analyzed for haematological and biochemical parameters. After sacrifice, histological analysis of the eye and analysis of pro-inflammatory biomarkers ($IL-1{\alpha}$, $IL-1{\beta}$, IL-8 and $TNF-{\alpha}$) in the cornea using ELISA was carried out. Spleen was collected and the proliferation capacities of spleenocytes were analyzed. Liver and brain were collected and assessed for oxidative stress. The eye irritation potential of the chemicals was evident from the redness and swelling of the conjunctiva and cornea. Histopathological analysis and ELISA assay showed signs of inflammation in the eye. However, the haematological and biochemical parameters showed no change. Spleenocyte proliferations showed only slight alterations which were not significant. Also oxidative stress in the brain and liver were negligible. In conclusion, chemicals which cause ocular irritation and inflammation did not show any systemic side-effects in the present scenario.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권5호
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• pp.391-399
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• 2004

Distraction osteogenesis has been thought to be promising technique for replacing bone graft in maxilla and mandible. The purpose of this study was to investigate the expression of osteonectin on distraction osteogenesis. Sixteen rabbits were used for this experiment. Osteotomy was performed between premolar and mental foramen. On the experimental group, distraction device was connected to the respective bone segments. On the control group, bone segments were fixed using plate and screws after osteotomy. Distraction was carried out at the rate of 0.7mm per day to obtain a 4.9mm elongation on the experimental group. After 3 days, 7 days, 14 days, and 28 days two rabbits of each group were sacrificed. The results obtained from this study were as follow : Experimental group was observed that the gaps between the distracted bone edges were occupied by new bone. Expression of Osteonectin were detected throughout the experiment in both groups and Expression of Osteonectin were markedly increased during distraction and consolidation period in experimental group than control group. From these results, it could be stated that distraction was shown to improve and accelerate bone formation and mechanical stress like distraction has considerable effects on osteonectin.

• 대한한방내과학회지
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• 제29권2호
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• pp.318-333
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• 2008

Objectives : In this study, the author tried to examine whether Cheogjogupye-tang (淸燥救肺湯, CGPT) and Yieum-jeon (理陰煎, YEJ) significantly affect in vitro and in vivo mucin secretion, MUC5AC gene expression in airway epithelial cells and contractility of isolated tracheal smooth muscle of rabbit. Materials and Methods : For in vitro experiment, confluent hamster tracheal surface epithelial (HTSE) cells were chased for 30 minutes in the presence of CGPT and YEJ to assess the effects of the agents on mucin secretion by enzyme-linked immunosorbent assay (ELISA), with removal of oriental herbal medicine extract from each agent-treated sample by centrifuge microfilter. Also, the effects of the agents on TNF-alpha or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Possible cytotoxicities of the agent were assessed by examining both LDH release from HTSE cells and the rate of survival and proliferation of NCI-H292 cells. For in vivo experiment, hypersecretion of airway mucin and goblet cell hyperplasia was induced by exposure of rats to $SO_2$ over 3 weeks. Effects of CGPT and YEJ orally administered for 1 week on in vivo mucin secretion from tracheal goblet cells of rats and hyperplasia of goblet cells were assessed using ELISA and histological analysis after staining the epithelial tissue with alcian blue, respectively. Also, the effects of CGPT and YEJ on contractility of isolated tracheal smooth muscle were investigated. Results : (1) CGPT significantly inhibited in vitro mucin secretion from cultured HTSE cells. However, YEJ did not affect in vitro mucin secretion; (2) CGPT and YEJ did not affect hypersecretion of in vivo mucin and hyperplasia of tracheal goblet cells; (3) CGPT and YEJ slightly increased the expression levels of TNF-alpha or EGF-induced MUC5AC gene in NCI-H292 cells; (4) CGPT and YEJ inhibited acetylcholine-induced contraction of isolated tracheal smooth muscle of rabbit; (5) CGPT and YEJ did not affect LDH release from HTSE cells and the survival and proliferation of NCI-H292 cells. Conclusion : The results from the present study suggest that CGPT and YEJ mainly affect the expression of mucin gene rather than secretion of mucin and do not show remarkable cytotoxicity to respiratory epithelial cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제33권6호
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• pp.459-466
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• 2011

Purpose: This study evaluated the capability of bone formation with silk fibroin/nano-hydroxyapatite/corn starch composite scaffold as a bone defect replacement matrix when grafted in a calvarial bone defect of rabbits $in$ $vivo$. Methods: Ten New Zealand white rabbits were used for this study and bilateral round-shaped defects were formed in the parietal bone (diameter: 8.0 mm). The silk fibroin 10% nano-hydroxyapatite/30% corn starch/60% composite scaffold was grafted into the right parietal bone (experimental group). The left side (control group) was grafted with a nano-hydroxyapatite (30%)/corn starch (70%) scaffold. The animals were sacrificed at 4 weeks and 8 weeks. A micro-computerized tomography (${\mu}CT$) of each specimen was taken. Subsequently, the specimens were decalcified and stained with Masson's trichrome for histological and histomorphometric analysis. Results: The average ${\mu}CT$ and histomorphometric measures of bone formation were higher in the control group than in the experimental group at 4 weeks and 8 weeks after surgery though not statistically significant ($P$ >0.05). Conclusion: The rabbit calvarial defect was not successfully repaired by silk fibroin/nano-hydroxyapatite/corn starch composite scaffold and may have been due to an inflammatory reaction caused by silk powder. In the future, the development of composite bone graft material based on various components should be performed with caution.

• Asian-Australasian Journal of Animal Sciences
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• 제33권1호
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• pp.166-176
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• 2020

Objective: An experiment was conducted to determine the effects of L-arginine (L-Arg) and N-carbamoylglutamic acid (NCG) on the growth, metabolism, immunity and community of cecal bacterial flora of weanling and young rabbits. Methods: Eighteen normal-grade male weanling Japanese White rabbits (JWR) were selected and randomly divided into 6 groups with or without L-Arg and NCG supplementation. The whole feeding process was divided into weanling stage (day 37 to 65) and young stage (day 66 to 85). The effects of L-Arg and NCG on the growth, metabolism, immunity and development of the ileum and jejunum were compared via nutrient metabolism experiments and histological assessment. The different communities of cecal bacterial flora affected by L-Arg and NCG were assessed using high-throughput sequencing technology and bioinformatics analysis. Results: The addition of L-Arg and NCG enhanced the growth of weanling and young rabbit by increasing the nitrogen metabolism, protein efficiency ratio, and biological value, as well as feed intake and daily weight gain. Both L-Arg and NCG increased the concentration of immunoglobulin A (IgA), IgM, and IgG. NCG was superior to L-Arg in promoting intestinal villus development by increasing villus height, villus height/crypt depth index, and reducing the crypt depth. The effects of L-Arg and NCG on the cecal bacterial flora were mainly concentrated in different genera, including Parabacteroides, Roseburia, dgA-11_gut_group, Alistipes, Bacteroides, and Ruminococcaceae_UCG-005. These bacteria function mainly in amino acid transport and metabolism, energy production and conversion, lipid transport and metabolism, recombination and repair, cell cycle control, cell division, and cell motility. Conclusion: L-Arg and NCG can promote the growth and immunity of weanling and young JWR, as well as effecting the jejunum and ileum villi. L-Arg and NCG have different effects in the promotion of nutrient utilization, relieving inflammation and enhancing adaptability through regulating microbial community.

• Archives of Plastic Surgery
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• 제33권5호
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• pp.592-600
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• 2006

Purpose: Numerous materials, both autologous and nonautologous, have been used for augmentation of sunken areas, but they have their own limitations. The purpose of this study is to determine the histologic response and volume change of the xenogenic collagen-based scaffold($Terudermis^{(R)}$) to the transfer into a subcutaneous soft tissue location in vivo rabbit model. Methods: Eighteen New Zealand white rabbits were used. Three $1.2{\times}1.2cm$ sized subcutaneous pockets were created on the dorsal surface of each ear. $1{\times}1cm$ sized collagen matrix($Terudermis^{(R)}$) and autologous dermal graft were implanted into each pocket. Full thickness of ear was harvested in 3 days, 1, 2, 4 weeks, 3, 6 months after implantation. Results: Histological analysis of implants demonstrated progressive neovascularization, fibroblast infilteration, neocollagen bundle synthesis and organization, and few foreign body reaction. The thickness of the collagen matrix in 3 days after the operation was 87.69% of the thickness of the collagen matrix in wet state. Then it decreased to 30.17% in 6 months after the operation. The rate of decrease was similar at all points at the same time compared with autologous dermal graft. Conclusion: Our experimental study suggests that $Terudermis^{(R)}$ could be a safe material as an implant for permanent augmentation in subcutaneous tissue. However the choice of graft for augmentation should be remained to the clinical situations.