• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.323-327
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• 1995

This experiment was carried out to observe the origin and developmental pattern of somatic embryos of Oenanthe javanica ($B^{L}.$) DC. The experiment included observation of embryogenic cells and their development stages by light microscope, transmission electron microscope and scanning electron microscope. The embryogenic cells, which were smaller than non-embryogenic cells in size with expanded nucleus and dense cytoplasm. When stained with hematoxylin, the embryogenic cells were readily distinguished from the non-embryogenic cells of which cell walls were stained with safranin. It was observed at somatic embryos developed from single cells on the epidermis of developing embryos or in the surface or inside of embryogenic clumps by segmentation pattern. Observation with a transmission electron microscope revealed that the embryogenic cells had dense cytoplasm expanded nucleus, small vacuoles, large amyloplasts containing starch grains, and abundant organelles including lipid bodies. Under a scanning electron microscope, embryogenic callus was shown to consist of very smaller cells than non-embryogenic cells in an orderly arrangement and covered with a net-like structure, while the non-embryogenic callus consisted of large cells, irregular in size and arrangement, and covered with a gelatin-like material.

• Analytical Science and Technology
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• v.28 no.1
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• pp.72-77
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• 2015

To study the efficacy of extract powder of Angelica gigas in preventing and treating degeneration of the articular cartilage in rats with monosodium iodoacetate (MIA)-induced osteoarthritis, A total of 30 six-week-old Sprague-Dawley rats were randomly divided into normal control group, untreated group and Angelica gigas treated group, with 10 rats in each group. During the treatment period, body weight were measured in each four days interval from starting date. The rat were sacrificed at the end of 3rd week after daily administration of Angelica gigas and then rat tibia articular cartilage was removed. In articular cartilages, glycosaminoglycan (GAG) amount increased by MIA treatment were reduced while proteoglycan (PG) amount decreased by MIA treatment were fairly recovered by Angelica gigas treatment, respectively. The content of TNF-a was also slightly reduced sections of the cartilage were stained with safranin-0 were also partially recovered by Angelica gigas treatment. By HPLC analysis, the content of main compounds decursin and decursinol angelate was analyzed as $10.5{\pm}0.2%$ of total extracts.

• Journal of Korean Medicine Rehabilitation
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• v.25 no.2
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• pp.51-64
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• 2015

Objectives This study was carried out to investigate the effects of Gyejigabuja-tang extracts on the Monosodium iodoacetate (MIA) induced osteoarthritis in rats. Methods Osteoarthritis was induced by injection of MIA into knee joint cavity of rats. Rats are divided into 4 groups (normal, control, positive comparison group, GBT group, each n=5). Normal group was injected by normal saline into knee joint cavity only. Control group was induced for osteoarthritis by MIA and oral medicated with distilled water. Positive comparison group was injected with MIA and taken Joins tablet 25 mg/kg. GBT group was injected with MIA and taken Gyejigabuja-tang extracts 300 mg/kg. Positive comparison group and GBT group were oral medicated for each substance once a day for 4 weeks. ALT, AST and creatinine were evaluated for hepatotoxicity and nephrotoxicity. Hind paw weight bearing ability was examined and inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$), $PGE_2$, $LTB_4$, osteocalcin and deoxypyridinoline (DPD) within serum were analysed. Knee joint structures were observed by Hematoxylin & Eosin (H&E), Safranin-O staining method. Results 1. Function of liver and kidney was not affected. 2. Hind paw weight bearing ability was significantly improved. 3. IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ in experimental group were significantly decreased compared with control group. 4. $PGE_2$, $LTB_4$, Osteocalcin and DPD in experimental group were decreased compared with control group. 5. In histopathologic observation, injury on synovial membrane and cartilage of experimental group was lesser than control group (H&E, Safranin-O staining). Conclusions Based on these results, it can be suggested that Gyejigabuja-tang has anti-inflammation effects on the MIA-induced osteoarthritis in rats.

• The Journal of Korean Physical Therapy
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• v.26 no.6
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• pp.418-424
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• 2014

Purpose: This study was performed to investigate the effect of joint mobilization on pain relief and cartilage repair in an induced osteoarthritis rat model by analyzing the expression of heat shock protein 70 in articular cartilage. Methods: MIA was injected into SD rats to induce osteoarthritis. These rats were divided into 4 groups: control group (n=30), no further treatment after the MIA injection ; experimental group I(n=30), performed swimming exercise after the MIA injection experimental group II (n=30), underwent joint mobilization after the MIA injection and experimental group III (n=30), performed swimming exercise and underwent joint mobilization after the MIA injection. For the histologic and pathophysiologic evaluation, safranin-O staining and for the immunohistochemical evaluation, the expression of HSP 70 in articular cartilage was analyzed 1, 7, 14, and 21 days after the MIA injection. Results: The inflammatory response and loss of tissue declined in experimental groups I and II over time, whereas the greatest decreases were noted in experimental group III. In the articular cartilage, low expression of HSP 70 was observed in every group on day 1, whereas HSP 70 expression was elevated on days 7 and 14 in experimental groups II and III. After 21 days, experimental group II displayed the strongest positive reaction, whereas HSP 70 was higher in experimental group III at this time point compared to that after 14 days. Conclusion: Our results showed that swimming exercise and joint mobilization had positive effects on pain relief and histologic and functional recovery in an induced osteoarthritis rat model.

• Journal of Korean Medicine Rehabilitation
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• v.33 no.3
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• pp.17-32
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• 2023

Objectives The purpose of this study was to investigate the antioxidant, anti-inflammatory and cartilage regeneration effects of Euiiin-tang water extract (EIIT) in the treatment of monosodium iodoacetate (MIA)-induced osteoarthritis in rats. Methods Animal models were divided into five groups. The normal group didn't do any treatments causing osteoarthritis. The control group was orally administerd distilled water instead of the drug, the positive control group used indomethacin 5 mg/kg, the EIIT 100 group used EIIT 100 mg/kg and the EIIT 200 group used EIIT 200 mg/kg, and seven rats were placed per group. We administered drug to rats for 2 weeks and analyzed oxidative stress-related proteins in joint tissue. Inflammation mediators and inflammatory cytokines induced by the activity of inflammation-related proteins were analyzed. In addition, the expression of anti-inflammatory cytokines and collagen-related factors were analyzed, and H&E staining and Safranin-O staining were performed to see the effect on histopathological changes. Results 1) Oxidative stress-related proteins were significantly reduced. 2) Inflammationrelated proteins, inflammatory mediators and inflammatory cytokines were significantly reduced. 3) Anti-inflammatory cytokines were significantly increased. 4) Collagen proteolysis factors significantly decreased, and collagen degradation inhibitory factor was significantly increased. 5) EIIT administration significantly reduced cartilage degeneration and deformation in H&E staining, and reduced proteoglycan destruction in Safranin-O staining. Conclusions From the above experimental results, it judges that Euiiin-tang has antioxidant, anti-inflammatory, and cartilage regeneration effects on osteoarthritis in rats induced by MIA.

• Journal of Oral Medicine and Pain
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• v.36 no.4
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• pp.261-271
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• 2011

Osteoarthritis in patients with temporomandibular disorders(TMDs) induces pain, limitation of mouth opening, occlusal problems, and most commonly affects their life quality. Control method and progressive process of osteoarthritis are being extensively researched. The researchers focus on histologic changes, synovial changes, muscular and ligamental changes and observed reaction to pain. Therefore most of them developed the animal model for osteoarthritis in TMD patients. In this study, we applied several methods which induces osteoarthritis of temporomandibular joint(TMJ) in rats or mice. For locally induce osteoarthritis in TMJ, Monosodium iodoacetate(MIA) or interleukin-$1{\alpha}$(IL-$1{\alpha}$) were injected into TMJ joint space for 5 or 3 weeks. Other groups are chosen for osteoarthritis under systemic control including hormonal changes and aging. To observe cellular change, increased collagen, degenerative bony destruction and distribution of proteoglycans (PGs), safranin-O staining and Masson's trichrome staining were used.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.528-533
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• 2009

In the current study, the mesenchymal stem cells (MSCs) isolated and propagated from the human umbilical cord blood (UCB) were tested for their capabilities of differentiation into chondrocytes in vitro. The mesenchymal progenitor cells (MPCs) collected from UCB were cultured in a low glucose DMEM medium with 10% FBS, L-glutamine and antibiotics. The human MSC colonies were positively stained by PAS reaction. When the immunophenotypes of surface antigens on the MSCs were analyzed by fluorescence-activated cell sorter (FACS) analysis, these cells expressed positively MSC-related antigens of CD 29, CD44, CD 90 and CD105, whereas they did not express antigens of CD14, CD31, CD34, CD45, CD133 and HLA-DR. Following induction these MSCs into chondrocytes in the chondrogenic differentiation medium for 3 weeks or more, the cells were stained positively with safranin O. We clearly confirmed that human MSCs were successfully differentiated into chondrocytes by RT-PCR and immunofluorescent stain of type-II collagen protein. These data also indicate that the isolation, proliferation and differentiation of the hUCB-derived MSCs in vitro can be used for elucidating the mechanisms involved in chondrogenesis. Moreover this differentiation technique can be applied to developing cell-based tissue regeneration or repair damaged tissues.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.5
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• pp.404-416
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• 2006

Purpose : This study was to clarify the changes in mandibular condyle after unilateral mandibular distraction osteogenesis throughout histological changes and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2). Materials & Methods : Intraoral distractors were placed via submandibular incision in 8 dogs. Two unoperated animals served as controls. Distraction was performed five days after osteotomy as a rate of 0.5 mm twice per day for 10 days. Two animals were sacrificed on 7, 14, 28, and 56 days after completion of distraction, respectively. Ipsilateral condyles were harvested and processed for histological and immunohistochemical examinations. Results : The condyle cartilage is separated into four layers: fibrous layer, proliferative layer, hypertrophic layer, and calcified layer. At 7 days and 14 days after distraction, the condylar cartilage showed the decreased thickness of the articular cartilage and reduced cellularity. At 28 days after distraction, there was an increase in cellularity of fibrous, proliferative, and hypertrophic layer. However, it demonstrated reduced cellularity compared to the control. At 56 days of after distraction, the articular cartilage was an almost normal histologic structure. Positive Safranin-O staining, indicative of sulfated proteoglycans, was examined in the condylar cartilge of nonloaded control. At 7 days and 14 days after distraction, the sulfated proteoglycans is almost completely depleted from the noncalcified part of the condylar cartilage. At 28 days after distraction, there was an increase in Safranin-O staining intensity. However, the staining intensity of the experimental condyle was weaker than that of the control. At 56 days of after distraction, the condylar cartilage showed almost normal Safranin-O staining pattern. In control condyle, MMP-2 immunostaining was seen in fibrous, proliferative, and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in fibrous and proliferative layer. At 7 days and 14 days after distraction, strong MMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, MMP-2 immunostaining was seen in the fibrous and hypertrophic layer of condylar cartilage, however, their immunoactivity was reduced. At 56 days after distraction, MMP-2 immunoreactivity showed almost normal immunostaining pattern. In control condyle, TIMP-2 immunostaining was primarily seen in fibrous and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in proliferative layer. At 7 days after distraction, very weak TIMP-2 immunoreactivity appeared in fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 14 days after distraction, weak TIMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, TIMP-2 immunoreactivity was increased in the fibrous and hypertrophic layer of condylar cartilage. At 56 days after completion of distraction, TIMP-2 immunoreactivity showed almost normal immunostaining pattern. Conclusions : The results show that short-term outcome of physiologic distraction osteogenesis may lead to degenerative changes in the condylar cartilage. These alterations in the condylar cartilage may be considered as a pressure-related degeneration of the cartilage tissue. However, the long-term results suggest that the condylar cartilage display repair activity after mandibular distraction osteogenesis.

• Polymer(Korea)
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• v.36 no.6
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• pp.789-794
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• 2012

Poly(lactic-co-glycolic acid) (PLGA) has been most widely used due to its advantages such as good biodegradability, controllable rate of degradation and metabolizable degradation products. We manufactured composite scaffolds of PLGA scaffold penetrated DBP gel (PLGA/DBP gel) by a simple method, solvent casting/salt leaching prep of PLGA scaffolds and subsequent soaking in DBP gel. Chondrocytes were seeded on the PLGA/DBP gel. The mechanical strength of scaffold, histology (H&E, Safranin-O, Alcian-blue) and immunohistochemistry (collagen type I, collagen type II) were performed to elucidate in vitro and in vivo cartilage-specific extracellular matrices. It was better to keep the characteristic of chondrocytes in the PLGA/DBP gel scaffolds than that PLGA scaffolds. This study suggests that PLGA/DBP gel scaffold may serve as a potential cell delivery vehicle and a structural basis for in vivo tissue engineered cartilage.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.4
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• pp.273-278
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• 2015

The purpose of this study was to evaluate the in-vitro efficacy of PDT using red light emitting diode (LED) with Radachlorin for biofilm inhibition of clinical Candida albicans isolates. The suspensions containing C. albicans at $9{\times}10^8CFU/mL$ were prepared on yeast nitrogen base containing 5% glucose. The biofilm formation was grown for 3 h after seeding suspensions each 100 ul on a 96-well plate and then supernatant was discarded. Each well was treated with $0.39{\mu}g/mL$ from $50{\mu}g/mL$ concentrations of Radachlorin on adherent biofilm. After a 30-minute incubation, light was irradiated for 30, 60, or 90 minutes using the following light source of wavelength 630 nm LED, at energy densities of 14, 29, and $43J/cm^2$. Afterwards, all supernatant was removed and dried. Adherent cells were stained with safranin O and dried. The cell viability was measured using a microplate reader at 490 nm. Also, a fluorescent signal on C. albicans was observed by saturation of a photosensitizer. In conclusion, a significant inhibition of 72.5% was observed to C. albicans on biofilm at the Radachlorin dose of $50{\mu}g/mL$ with 630 nm LED. The Photosensitizer (Radachlorin) was adequate at 30 minuttes for C. albicans. Overall, the results showed that inhibition of biofilm formation was Radachlorine dose-dependent. The results suggest that PDT, using Radachlorin with 630 nm LED, is able to decrease biofilm formation of C. albicans.