• Journal of Food Hygiene and Safety
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• v.9 no.3
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• pp.123-131
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• 1994

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

• The korean journal of orthodontics
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• v.22 no.2 s.37
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• pp.427-447
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• 1992

The purpose of this study was to evaluate the effect of the lateral displacement on the mandibular condylar growth in the rabbit. The experimental animals were twenty White NewZealand rabbits of 4-week old. Ten of them was used as control group, and experimental animal was composed of remaining ten. Laterodeviation appliance was made of cast base metal and appliance was cemented with resin in permanent fashion. Experimental group were sacrificed at 1, 2, 4, 6, 8 weeks form beginning of the experiment. Both of temporomandibular joint were prepared for histologic study. The conclusions are: 1. In control group, there was slight increase of proliferative zone and hypertrophic zone at 2-week control animal and slight reduction at 4-week. 6-week and 8-week control animal were similiar to 1-week control animal. 2. In right mandibular condyle of experimental group, 2-week experimental animal showed marked increase of proliferative zone and hypertrophic zone at posterior surface of condylar head. In 8-week experimental animal marked increase at anterior surface of anticular surface is observed. 3. In left mandibular condyle of experimental group, proliferative zone and hypertrophic zone were reduced at 1-week experimental animal and slight increase at 2-week. Proliferative zone and hypertrophic zone were reduced at 4-week experimental animal and were slightly increased at 6 week. 4. After 8 weeks, right and left condyle were not different in experimental group. The condylar cartilage was stabilized 8 weeks after the experiment. No marked traumatic change was seen, but minute focal bleeding was seen at articular cavity in 1-week, 2-week and 4-week experimental animal. 6-week and 8-week experimental animal did not show bleeding tendency in articular cavity.

• Archives of Plastic Surgery
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• v.42 no.2
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• pp.150-158
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• 2015

Background Fat is widely used in soft tissue augmentation. Nevertheless, it has an unpredictably high resorption rate. Clinically, external expansion with negative pressure is used to increase fat graft survival. In this study, fat graft recipient sites were preconditioned by external application of negative pressure in order to test for improvements in vascularity and fat graft survival. Methods Negative pressure was applied randomly to either the left or right dorsal ear of 20 New Zealand male white rabbits at a pressure of -125 mm Hg. The negative pressure was removed one week after the skin perfusion was measured. The skin flap at each ear was elevated, and 1 g of fat was grafted above the dorsal perichondrium. After one week, the fat weight, microvessel density, mature vessel density of the skin and fat, and amount of glycerol released were measured. Three months after the grafting, the same measurements were performed, with the exception of glycerol release. Results The fat survival rate of the experimental group ($75.4%{\pm}3.9%$) was higher than that of the control group ($53.1%{\pm}4.3%$) (P<0.001). Skin perfusion was higher in the experimental group. The glycerol release in the experimental group was significantly higher than in the control. The microvessel density of the skin and fat was significantly higher in the experimental group. Three months after the grafting, the skin and fat mature vessel density was significantly higher in the experimental groups. Conclusions Negative pressure prior to fat grafting increased the vascularity of the recipient site, and, accordingly, enhanced fat graft survival.

• Archives of Plastic Surgery
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• v.41 no.6
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• pp.647-653
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• 2014

Background Administration of growth factors has been associated with increased viability of composite grafts greater than 1-cm in diameter. Platelet-rich plasma (PRP) contains many of the growth factors studied. In this study, we evaluate the effect of PRP injection on composite graft viability and the proper time for injection. Methods A total of 24 New Zealand White rabbits were divided into four groups. Autologous PRP was injected into the recipient sites three days before grafting in group 1, on the day of grafting in group 2, and three days after grafting in group 3. Group 4 served as control without PRP administration. Auricular composite grafts of 3-cm diameter were harvested and grafted back into place after being rotated 180 degrees. Median graft viability and microvessel density were evaluated at day 21 of graft via macroscopic photographs and immunofluorescent staining, respectively. Results The median graft survival rate was 97.8% in group 1, 69.2% in group 2, 55.7% in group 3, and 40.8% in the control group. The median vessel counts were 34 (per ${\times}200$ HPF) in group 1, 24.5 in group 2, 19.5 in group 3, and 10.5 in the control group. Conclusions This study demonstrates that PRP administration is associated with increased composite graft viability. All experimental groups showed a significantly higher survival rate and microvessel density, compared with the control group. Pre-administration of PRP was followed by the highest graft survival rate and revascularization. PRP treatments are minimally invasive, fast, easily applicable, and inexpensive, and offer a potential clinical pathway to larger composite grafts.

• Archives of Plastic Surgery
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• v.32 no.4
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• pp.521-528
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• 2005

The purpose of this research is to find out the degree of cartilage regeneration by inserting the atelo-collagen scaffold obtained from dermis of a calf on cartilage defect site. Dissection underneath the perichondrium by the periosteal elevator on both side of ears of six New Zealand white rabbits were made to expose the cartilage, leaving pairs of circular holes 3, 6, 9 mm width with punches. One hole was left for a control, and on the other hole atelo-collagen scaffold of the same size was transplanted. In postoperative 1, 2, 4 weeks, the tissues were dyed. The length of long axis of neocartilage was measured through an optical microscope with a 0.1 mm graduation at original magnification, ${\times}40$. In the first and second week, both group showed no sign of cartilage regeneration. In the fourth week, regeneration on marginal portions was observed on all groups and the average values of length of long axis of neocartilage according to defect size were as follows: In the cases with 3mm defect, it was $0.85{\pm}0.30mm$ in the control group, and $1.85{\pm}0.38mm$ in the graft group; in the cases with 6 mm defect, $1.33{\pm}0.58mm$ in the control group, and $2.25{\pm}0.46mm$ in the graft group; and in the cases with 9 mm defect, $2.33{\pm}0.77mm$ in the control group, and $4.47{\pm}1.39mm$ in the graft group. This means that the collagen scaffold has an influence on the regeneration of neocartilage. But the relative ratio of the length of neocartilage to cartilage defect size was not significant in the statistics.

• Archives of Plastic Surgery
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• v.33 no.5
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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.

• Journal of Biomedical Engineering Research
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• v.29 no.2
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• pp.159-163
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• 2008

The goal of this study is to investigate the effect and potential of three-dimensional Co-culture of BMSCs (bone marrow stromal Cells) and NP (nucleus pulposus) Cells on the differentiation of BMSCs into NP-like Cells. The NP Cells and BMSCs were isolated and cultured from New Zealand White rabbits. The isolated NP Cells and BMSCs were prepared in different alginate beads. Those two types of beads were separated by a track-etched membrane of $3\;{\mu}m$ pore in a 6-well culture plate. No growth factors were used. In addition to these, NP and BMSC were cultured in the beads independently for control. The number of Cells in Co-culturing system was half of those in two control groups. Proliferation and production of glycosaminoglycan (GAG) were evaluated along with histological observation. The GAG production rate(GAG contents/Cell) of Co-cultured BMSCs were much higher than that of BMSCs cultured alone. The total amounts of GAG produced by BMSCs in Co-culturing system were larger than those produced by BMSCs in control group and were comparable with those produced by NP alone even the number of each Cell was half of BMSCs in Co-culturing system. This study showed the potential of differentiation of BMSCs into NP-like Cells through three-dimensional Co-culture system even without any chemical agents.

• Journal of Korean Neurosurgical Society
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• v.55 no.5
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• pp.237-243
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• 2014

Objective : Symptomatic disc degeneration develops from inflammatory reactions in the annulus fibrosus (AF). Although inflammatory mediators during annular inflammation have been studied, the roles of matrix metalloproteinases (MMPs) and their inhibitors have not been fully elucidated. In this study, we evaluated the production of MMPs and tissue inhibitors of metalloproteinase (TIMPs) during annular inflammation using an in vitro co-culture system. We also examined the effect of notochordal cells on annular inflammation. Methods : Human AF (hAF) pellet was co-cultured for 48 hours with phorbol myristate acetate-stimulated macrophage-like THP-1 cells. hAF pellet and conditioned media (CM) from co-cultured cells were assayed for MMPs, TIMPs, and insulin-like growth factor (IGF)-1 levels using real-time reverse-transcriptase polymerase chain reaction and enzyem-linked immunosorbent assay. To evaluate whether notochordal cells affected MMPs or TIMPs production on annular inflammation, hAF co-cultured with notochordal cells from adult New Zealand White rabbits, were assayed. Results : MMP-1, -3, -9; and TIMP-1 levels were significantly increased in CM of hAF co-cultured with macrophage-like cells compared with hAF alone, whereas TIMP-2 and IGF-1 levels were significantly decreased (p<0.05). After macrophage exposure, hAF produced significantly more MMP-1 and -3 and less TIMP-1 and -2. Interleukin-$1{\beta}$ stimulation enhanced MMP-1 and -3 levels, and significantly diminished TIMP-2 levels. Co-culturing with rabbit notochordal cells did not significantly influence MMPs and TIMPs production or COL1A2 gene expression. Conclusion : Our results indicate that macrophage-like cells evoke annular degeneration through the regulation of major degradative enzymes and their inhibitors, produced by hAF, suggesting that the selective regulation of these enzymes provides future targets for symptomatic disc degeneration therapy.

• Journal of Korean Neurosurgical Society
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• v.48 no.1
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• pp.1-7
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• 2010

Objective : Notochordal cells in the intervertebral disc interact with nucleus pulposus (NP) cells and support the maintenance of disc homeostasis by regulation of matrix production. However, the influence of notochordal cells has not been evaluated in the annulus fibrosus (AF), which is the primary pain generator in the disc. We hypothesized that the notochordal cell has the capacity to modulate inflammatory mediators secreted by AF cells secondary to stimulation. Methods : Notochordal and AF cells were isolated from adult New Zealand white rabbits. AF pellets were cultured with notochordal cell clusters or in notochordal cell-conditioned media (NCCM) for 24 or 48 hours with proinflammatory cytokines at varying concentrations. Gene expression in AF pellets were assayed for nitric oxide synthase (iNOS), cyclo-oxygenase (COX)-2, and interleukin (IL)-6 by real time reverse transcriptase polymerase chain reaction (RT-PCR). Results : AF pellet in NCCM significantly decreased the iNOS and COX-2 messenger ribonucleic acid (mRNA) levels compared to AF pellets alone and AF pellets with notochordal cells (p < 0.05). AF pellet resulted in dose-dependent iNOS and COX-2 expression in response to IL-$1{\beta}$, stimulation, demonstrating that 1 ng/ml for 24 hours yielded a maximal response. AF pellet in NCCM significantly decreased the expression of iNOS and COX-2 in response to 1ng/ml IL-$1{\beta}$, stimulation at 24 hours (p < 0.05). There was no difference in IL-6 expression compared to AF pellets alone or AF pellets with notochordal cell clusters. Conclusion : We conclude that soluble factors from notochordal cells mitigate the gene expression of inflammatory mediators in stimulated AF, as expected after annular injury, suggesting that notochordal cells could serve as a novel therapeutic approach in symptomatic disc development.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.35 no.1
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• pp.7-12
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• 2009

Purpose: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. Material & Methods: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. Results: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores ($20-40{\mu}m$). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. Conclusion: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.