• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권3호
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• pp.228-238
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• 2005

Purpose: Several cryoprotectants are in use to help the survival of cells during cryopreservation of bone in maxillofacial region. Among them, $Me_2SO$(dimethyl sulfoxide), EG(ethylene glycol), sucrose were used for experimentally created defects with accompanying cryopreserved bone graft in the rabbit model. The aim of this study is to analyze the effect of above mentioned agents on bone formation using histologic and histomorphometrical methods, thus to provide experimental support for clinical application of these agents. Materials and methods: Nine rabbits were used as experimental animals. Surgical defects were created on the distal femoral heads and mesial tibial heads of each animal using trephine drill(5mm diameter and 5mm length). The harvested bones were cryopreserved in $-80^{\circ}C$ refrigerator for one week. The defects were filled with cryopreserved bone with cryoprotectants as experimental groups and cryopreserved bone without cryoprotectant as control. Then, the animals were sacrificed at 1, 2, and 3 weeks after surgery. With Goldner's modified Masson trichrome staining and semiautomatic image analysis system, we observed the change of the cells and bone formation. Results: After bone graft, bone formation and active remodeling process were examined in all experimental groups and the control. But the intensity of such activities of the control were somewhat weaker than that of the experiments. Especially $Me_2SO$+sucrose group was the best in bone formation and bone remodeling. $Me_2SO$ group was more than that of EG group in bone fomation. Sucrose seems to be helpful in survival of the bone cell. Histologic findings showed superior bony quantity and quality in experimental groups than that in control. Conclusions: The data from this study provides the basis for future studies for evaluating the effect of cryoprotectants in the cryopreservation of bone and clinical study for predictable use of these agents.

• 대한두개안면성형외과학회지
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• 제18권3호
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• pp.155-161
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• 2017

Background: Because of the relatively similar size of organs to human and the physiological and structural similarities, the use of porcine as xenograft donors is progressing very actively. In this study, we analyzed the characteristics of porcine ear cartilage and evaluated its suitability as graft material in reconstructive and cosmetic surgery. Methods: The auricular cartilage was harvested from two pigs, and subjected to histological examination by immunohistochemical staining. To determine the collagen content, samples were treated with collagenase and weight changes were measured. After sterilization by irradiation, the samples were grafted into rats and stained with Hematoxylin and Eosin and Masson Trichrome to observe inflammation and xenograft rejection. Results: In IHC staining, extracellular matrices were mainly stained with type II collagen (20.69%), keratin sulfate (10.20%), chondroitin sulfate (2.62%), and hyaluronic acid (0.84%). After collagenase treatment, the weight decreased by 68.3%, indicating that about 70% of the porcine ear cartilage was composed of collagen. Upon xenograft of the sterilized cartilages in rats, inflammatory cells were observed for up to 2 months. However, they gradually decreased, and inflammation and reject-response were rarely observed at 5 months. Conclusion: The porcine ear cartilage was covered with perichondrium and cellular constituents were found to be composed of chondrocytes and chondroblasts. In addition, the extracellular matrices were mainly composed of collagen. Upon xenograft of irradiated cartilage into rats, there was no specific inflammatory reaction around the transplanted cartilage. These findings suggest that porcine ear cartilage could be a useful alternative implant material for human cosmetic surgery.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제21권1호
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• pp.1-12
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• 1999

The purpose of this study was to evaluate the healing process of the calvarial defect filled with hydroxylapatite(HA) and $TGF-{\beta}$ in Rat. 72 Sprague-Dawly rats were divided into 3 groups, control and two experimental groups. Bony defect were artificially prepared in the calvaria of all 72 rats and followed by implantation of HA (experimental group of 24 rats) and HA+$TGF-{\beta}$(another experimental group of 24 rats) into the defects. Sequential sacrifice was performed at 1, 2, 4, 6, 8, 12 weeks of experiment. Obtained specimen was stained with Hematoxylin and Eosin, Masson's Trichrome and Immunohistochemistry. The results were as follows, 1. Granulation tissue was prominent on control group in 1 and 2 weeks. Bony defects were filled with dense fibrous tissue through the whole experimental period and osteoinduction could not be observed in all groups. 2. Inflammatory cell infiltration was prominent on control group in 1 and 2 weeks and osteoclastic activity was high in HA implanted experimental group at 1 and 2 weeks. 3. Inflammatory cell infiltration was less and maturation of fibrous tissue could be found on HA+$TGF-{\beta}$ implanted experimental group at 1 and 2 weeks. 4. Osteoconduction activity was high in HA+$TGF-{\beta}$ implanted experimental group at 2 and 4 weeks but there was no difference after 6 weeks among 3 groups. 5. In grafted site of HA+$TGF-{\beta}$ implanted group, osteonectin expression was slightly increased from 1 week to 6 weeks. In the host site, it was increased from 1 to 4weeks. 6. In grafted site of HA+$TGF-{\beta}$ implanted group, osteocalcin expression was high at 4 weeks. In the host site, we could find the difference among 3 groups. From above results, the HA with mixture of $TGF-{\beta}$ has the potentiality of promoting bone formation in the bony defect area in the rat.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제13권3호
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• pp.241-251
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• 1991

During the condylar shaving procedure, the articular soft tissue cover can be removed. Author compaired the histological healing process of the articular soft tissue cover between the preservative and unpreservative group group with 45 New Zealand rabbits(Average wt. : about 2.5kg). In unpreservative group, the usual high condylar shave with the removal of soft tissue cover was performed. In the preservative group, the underlying bone, replaced in its original position and sutured. The animals were sacrified 1, 2, 3, 4, 6 weeks interval after operation. The specimens were fixed in 10% neutral formalin and decalcified, paraffin embedded and stained by Hematoxylin & Eosin, and Masson's trichrome. The obtained results were as follows. 1. The condyles of the both group were covered with an articular sop tissue layer. 2. The cartilage cells in subarticular layer has regular continuous patterns in the preservative group but frequently interrupted in the unpreservative group. 3. The incision made in the posterior part of the articular surface for the elevation of the articular soft tissue frequently caused a deformity such as the interruption of the subarticular layer of cartilage. 4. By the above findings, the preservation of articular sop tissue cover may be the effective operation method on concept of bone remodelling.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제22권1호
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• pp.33-42
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• 2000

The purpose of this study was to compare the wound healing process after skin incision using scalpel, $CO_2$ laser and pulsed Nd:YAG laser in rats. After skin on the back was incised 3 cm long, rats were sacrificed at 1, 3, 7, 14, 21 and 28 days. Macroscopic, histologic and immunohistochemical examinations using the collagen type IV and the CD34 antibodies which are necessary to the forming process of new capillary were performed. Results obtained were as follows ; Macroscopically the initial wound healing of the laser group was about $1{\sim}2$ weeks slower than that of the scalpel group. There weren't however any remarkable differences in all groups in 4 weeks after incision. By histologic finding, acute inflammatory cells were more prominent during the initial wound healing in the scalpel group than in the other groups. Epithelialization started in the order of scalpel, $CO_2$ and Nd:YAG laser group after skin incision. By the Masson's trichrome stain, collagen synthesis in the Nd:YAG laser group was more slowly initiated than in the other groups. But it was completed at the $3{\sim}4$ weeks in all groups. Immunohistochemically, collagen type IV and CD34 expression were markedly increased at 2 weeks in the scalpel and $CO_2$ laser group. Meanwhile, in the Nd:YAG laser group, these reactions were observed later tan the other groups. Collagen type IV and CD34 expression were decreased in all groups after 4 weeks. These results suggest that $CO_2$ and Nd:YAG laser showed similar healing process compared with scalpel and a potential substitute for scalpel in skin incision.

• 치과방사선
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• 제12권1호
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• pp.105-113
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• 1982

Because of the development of rampant caries, osteomyelitis and osteoradionecrosis that occur after radiation therapy of oral cancers, extraction of teeth at or near the malignant lesion has been done in the past. Few, however, have studied the radiation effect on the healing of extraction wounds. This study is concerned with the effect of Co-60 irradiation on the healing process of extraction wounds in rats. Fifty six, male, Spraque-Dawley rats are used. The right first molar of the mandible is extracted from all animals. They are divided into three experimental groups of 14 each and a control group of 14. Three experimental groups are irradiated respectively with 200 rad, 400 rad and 600 rad and a pair of rats in each group are killed on days 1, 3, 5, 7, 14, 21 and 28 after irradiation. Two animals from the control group are killed on the day when the experimental rats are killed. The irradiated hemimandibles are fixed in 10% neutral formalin, decalcified in 5% trichloroacetic acid, embedded in paraffin and sectioned. The sections are stained in hematoxylin and eosin, van Gieson, Masson's trichrome or silver nitrate. Results show that in general radiation effects on healing extraction wounds are dose dependent; i.e., the higher is the dose, the greater is the histologic changes observed: 1. Irradiation tends to retard blood clot organization and epithelial regeneration. 2. An increase in the number of giant cells and osteoclasts is noted after irradiation. 3. Formation of regenerating connective tissues around and within the extraction site is com- promised, and a clear reduction of primitive mesenchymal type connective cells is noted. 4. The healing process begins along the lateral aspect of the extraction socket in the control, while irregular histologic appearances of the brabecular pattern is present in the experimental rats.

• 한방재활의학과학회지
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• 제20권2호
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• pp.17-34
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• 2010

Objectives : This study was carried out to know the effects of Gwanjul8-bang(here in after reffered to GJ8) on the inhibition of arthritis induced by collagen on DBA/1J mice. Methods : For this purpose, GJ8 was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, hepatotoxicity, arthritis index, value of immunocyte in draining lymph node(DLN) and paw joint, cytokine in serum were measured in vivo. Results : 1. The arthritis index was decreased significantly. 2. In total cell counts of DLN and paw joint, the cells in DLN increased significantly while there was a significant decrease in paw joint. 3. In lymph nodes, $CD19^+$, $CD3^+$, $CD4^+/CD25^+$ cells increased significantly, and $B220^+/CD23^+$ cells decreased significantly. 4. In joints, $CD3^+$, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly. 5. The levels of $TNF-{\alpha}$, IL-6, IL-17, MCP-1 and vascular endothelial growth factor(VEGF) in serum was significantly decreased compared with control. 6. Anti-collagen II in serum was significantly decreased compared with control. 7. The degree of arthritis induced damage of joint of GJ8 group is slight compared with control group in histopathologic observation(hematoxylin & eosin(H&E), Masson-Trichrome(MT) staining). Conclusions : Comparison of the results for this study showed that GJ8 had immunomodulatory effects. So we expect that GJ8 should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease. Therefore there seems to be many clinical research needed in the future.

• Journal of Periodontal and Implant Science
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• 제30권1호
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• pp.91-104
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• 2000

Many synthetic bone materials have been studied for their potential of regenerative effects in periodontal tissue. Safflower seeds have been traditionally used as a drug for the treatment of fracture and blood stasis in oriental medicines. The purpose of this study was to assess and compare the osseous responses in rat calvarial defects between bone substitutes such as calcium carbonate and bovine-derived hydroxyapatite and feeding of safflower seeds. The calvarial defects were made with 8 mm trephine bur in 24 Sprague-Dawley rats. Two graft materials were implanted in each experimental groups, whereas the control and safflower seed feeding groups were sutured without any other treatment. And then the rats of safflower seed feeding group were supplied with 3 g/day of safflower seeds. Each group was sacrificed at 4 weeks and 8 weeks. To study a histopathology related to bone healing and regeneration, Goldner's Masson Trichrome stain was done at each weeks. The tissue response was evaluated under light microscope. There were more osteoblastic activity, new bone formation, dense bony connective tissues in bovine-derived hydroxyapatite group compared to other groups at 8 weeks. The osseous defect area of safflower seed feeding group was filled with prominent fibrous tissues, where less inflammatory infiltration and new capillary proliferation. In the early phase of bone healing, safflower seed feeding reduces the inflammatory response and promotes the proliferation of connective tissue. These results suggest that natural bovine-derived HA and safflower seed feeding could enhance the regenerative potential in periodontal defects.

• Journal of Ginseng Research
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• 제41권3호
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• pp.316-325
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• 2017

Background: Ginseng essence (GE) is a formulation comprising four medicinal and edible herbs including ginseng (Panax ginseng), American ginseng (Panax quinquefolius), lotus seed (Nelumbo nucifera), and lily bulb (Lilium longiflorum). This study was aimed at investigating the hepatoprotective effect of GE against carbon tetrachloride ($CCl_4$)-induced liver injury in rats. Methods: We treated Wistar rats daily with low, medium, and high [0.625 g/kg body weight (bw), 1.25 g/kg bw, and 3.125 g/kg bw, respectively] doses of GE for 9 wk. After the 1st wk of treatment, rats were administered 20% $CCl_4$ (1.5 mL/kg bw) two times a week to induce liver damage until the treatment ended. Results: Serum biochemical analysis indicated that GE ameliorated the elevation of aspartate aminotransferase and alanine aminotransferase and albumin decline in $CCl_4$-treated rats. Moreover, $CCl_4$-induced accumulation of hepatic total cholesterol and triglyceride was inhibited. The hepatoprotective effects of GE involved enhancing the hepatic antioxidant defense system including glutathione, glutathione peroxidase, glutathione reductase, glutathione S-transferase, superoxide dismutase, and catalase. In addition, histological analysis using hematoxylin and eosin and Masson's trichrome staining showed that GE inhibited $CCl_4$-induced hepatic inflammation and fibrosis. Furthermore, immunohistochemical staining of alpha-smooth muscle actin indicated that $CCl_4$-triggered activation of hepatic stellate cells was reduced. Conclusion: These findings demonstrate that GE improves $CCl_4$-induced liver inflammation and fibrosis by attenuating oxidative stress. Therefore, GE could be a promising hepatoprotective herbal formulation for future development of phytotherapy.

• Korean Journal of Acupuncture
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• 제24권3호
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• pp.117-137
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• 2007

Objectives & Methods : The purpose of this study is to observe the effects of Gentianae macrophyllase Radix herbal-acupuncture solution(GR-HAS) at Joksamni(ST36) on Collagen-induced arthritis(CIA) in DBA/1J mice. The author performed several experimental items to analyze arthritis evaluation, change of weight, spleen size and adhesion rate, change of cytokine level, IgG, IgM and anti-collagen II , change of immunocyte count, histological change of CIA mouse joint related with RA. Results : 1. In the GR-HA group, the arthritis index, the incidence of arthritis and joint edema were significantly decreased. 2. In GR-HA group, there were no weight loss and similarly maintained as normal group. Spleen size, adhesion rate and the edema and transformation of knee joint were low and similarly maintained as normal group. 3. In the GR-HA, cytokine level, IgG, IgM and anti-collagen II were significantly decreased. 4. In the GR-HA, the change of immunocyte count were similarly maintained as normal group. 5. In the histological changes of the CIA mouse joint, the cartilage destruction and synovial cell proliferation were decreased in the GR-HA(Hematoxylin and eosin stain). The collagen fiber expressions in the GR-HA were similar with that of the normal group(Masson's Trichrome stain) Conclusions : These results suggest that GR-HA at ST36 has an effect to control immune reaction and suppress inflammation, synovial cell proliferation and cartilage destruction in rheumatoid arthritis.