• Title/Summary/Keyword: toluidine blue

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REMODELING PROCESS OF THE STREPTOZOTOCIN-INDUCED DIABETIC RAT'S RESECTED CONDYLE (Streptozotocin유도 당뇨병이 백서하악 절단과두 치유에 미치는 영향의 실험적 연구)

  • Kim Won-Cheol;Hwang Eui-Hwan;Lee Sang-Rae;Hong Jung-Pyo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.24 no.2
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    • pp.263-274
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    • 1994
  • The purpose of this study was to investigate the remodeling process of the streptozotocin-induced diabetic rat's resected condyle. This experiment was performed with male Sprague-Dawly strain rats weighing approximately 250 gm, which were rendered diabetic by an intravenous injection of streptozotocin(70㎎/㎏ body weight). After condylectomy, experimental rats were serially terminated on the 1st week, the 2nd week, the 3rd week, and the 4th week. The following termination, the mandibles were dissected out to make specimens. Each mandibular condyle was radiographed with Hitex HA-80(Hitex Co., Ltd. Japan). In addition to radiographic observation, the mandibular condyles, further decalcified and embedded in paraffin, were sectioned and stained with Hematoxylin and Eosin, Toluidine blue and Masson's trichrome. They were observed with a light microscope and a polarizing microscope. The results were as follows. 1. Soft X-ray radiograms revealed proliferation of bone after 1 week in both groups. Irregularly repaired bones and dense trabeculae were clearly observed in experimental group. 2. The resected condyles were repaired by intramembraneous and endochondral bone formation in both groups. 3. Bone tissue repair was initiated from the adjacent margin of resected bone, and cartilaginous tissues were observed at the top of repaired bone in both groups. 4. The number of osteoblasts of experimental group was small, compared with control group. Each osteoblast was small and flat. The thin trabeculae were irregularly formed. 5. Collagens of bone were gradually matured in both groups, but the degree of maturation was lower in experimental group. 6. Fibrous tissues covered the upper parts of repaired bone were densely arranged in the both groups. Conclusively, atrophied osteoblasts, immature collagen of bone, and thin and irregular trabeculae which were characterized in the diabetes experimental group showed diabetes disturbed osteoblastic function and caused disturbance of remodeling process of bone.

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Acer tegmentosum Maxim Prevents Bone Loss by Inhibiting Osteoclastogenesis and Promoting Osteoblast Mineralization in Ovariectomized Mice

  • Oh, Tae Woo;Park, Kwang-Il;Do, Hyun Ju;Kim, Kyungho;Yang, Hye Jin;Cho, Won Kyung;Ma, Jin Yeul
    • Natural Product Sciences
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    • v.26 no.1
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    • pp.83-89
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    • 2020
  • Osteoporosis is a worldwide disease leading to significant economic and societal burdens globally. Osteoporosis is caused by unbalanced bone remodeling between the rate of osteoclast bone resorption and osteoblast bone formation. Acer tegmentosum Maxim (AT) is a traditional herbal medicine containing multiple biological activities such as anti-oxidant and anti-inflammatory purposes. However, its role in osteoporosis has not been fully studied. Therefore, we investigated whether AT has a potent inhibitory effect on osteoporosis and its mechanism through a systemic evaluation in ovariectomized (OVX) mice. OVX mice were orally administrated with the AT at doses of 50, 100, and 200 mg/kg for 10 weeks. Histological images and histomorphometry analyses were performed by H&E and Toluidine blue satin, and the expression levels of receptor activator for nuclear factor-kB ligand (RANKL), nuclear factor of activated T cells cytoplasm 1 (NFATc1), c-Fos, and matrix metalloproteinase 9 (MMP9) related to the osteoclast differentiation were investigated using immunohistochemical analysis. Administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Further, administration of AT increased periosteal bone formation in a dose-dependent manner. Meanwhile, AT inhibited not only the expression of NFATc1 and c-Fos, which are two major regulators of osteoclastogenesis but also reduced bone resorbed encoding expression of MMP9 and RANKL. Our results indicated that administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Also AT has the bone protective effect through the suppression of osteoclast and promotion of osteoblast, suggesting that it could be a preventive and therapeutic candidate for anti-osteoporosis.

INFLUENCE OF CIGARETTE SMOKE INHALATION ON DENTAL IMPLANT OSSEOINTEGRATION IN THE RAT (백서에 식립한 치과용 임플란트의 골유착에 흡연이 미치는 영향)

  • Kim, Se-Ho;Kim, Soung-Min;Kim, Ji-Hyuck;Park, Young-Wook;Park, Chan-Jin;Jung, Ki-Myoung;Lee, Suk-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.4
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    • pp.291-299
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    • 2005
  • Objectives : It is well known that cigarette smoking is harzardous to the osseointegration of dental implant, due to the impaired wound healing accompanied by reduced alveolar bone density. The aim of this study is to evaluate the influence of cigarette smoke on the implant osseointegration by the time factor consideration. Materials and methods : Twenty-four male Spraque-Dawley rats (8 weeks, weighting 200 to 250g) were used in this study. In the experimental group, 13 rats were exposed to cigarette smoke, 8 minutes per day during 6 weeks, and 12 rats in the control group were not exposed at any time. RBM (Resorbed blasting media) surfaced implant (diameter 3.3mm, length 5.0mm, AVANA Co., Korea) was placed in the right femur of each rat. Each implant with surrounding bone was prepared with microtome (cutting band 0.2mm$^{(R)}$, EXAKT Co., Germany) after 1 day, 2, 4, 8 and 12 weeks and stained with toluidine blue (1%). Another clinical investigation of each implant was also done at each evaluation time. Results : Clinical investigation around implant fixture showed that there were no significant differences between the control and experimental group. Microscopic observation around implant fixture showed that there were significant differences between the control and experimental group at the initial stage after implant fixture installation. Experimental group showed a decreased bone to implant contact within 4 weeks compared to control group, but showed similar characteristics after 4 weeks. Conclusion : Smoking inhalation effect on the dental implant showed the impaired wound healing by vasoconstriction and decreased intramedullary blood flow at initial stage of osseointegration. This experimental results can be clinically useful to the implant surgery of smoking patients.

Correlation between Infiltrations of Tumor-associated Macrophages, Mast Cells, and Dendritic Cells with Clinicopathologic Factors in Advanced Gastric Cancer (진행성 위암에서 종양 연관성 대식세포, 비만세포, 가지세포의 침윤과 임상-병리학적 인자와의 연관성)

  • Lee, Seung-Bum;Chi, Kyong-Chon
    • Journal of Gastric Cancer
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    • v.5 no.3 s.19
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    • pp.206-212
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    • 2005
  • Purpose: Angiogenesis has a critical role in tumor proliferation, invasion, and metastasis. In gastric cancer, tumor-associated macrophages and mast cells produce angiogenic factors such as VEGF, that inhibit the functional maturation of dendritic cells. The aim of this study is to identify tumor-associated macrophages, mast cells, dendritic cell infiltrations, and microvessel densities (MVD) to investigate the relationship between them and the prognosis for gastric-cancer patients. Materials and Methods: The subjects were 79 patients selected from those who had undergone a curative gastric resection for stomach cancer. With them, Immune-histochemical staining was done using CD34 for the MVD, CD68 antigen for macrophages, and S-100 protein for dendritic cells, and toluidine blue staining was done for mast cells. Results: Macrophage infiltration showed a statistically significant positive correlation with histologic differentiation and a negative correlation with invasion depth, nodal metastasis, and stage. S-100 (+) dendritic cells and mast cells had no significant correlations with histologic differentiation, invasion depth, nodal metastasis, distant metastasis, stage, and MVD. As survival, no statistically significant differences were seen between the variables. Conclusion: Tumor-associated macrophages should be evaluated as possible prognostic markers in gastric-cancer patients.

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FORMATION OF BASEMENT MEMBRANE AND STRATIFICATION OF RABBIT ORAL KERATINOCYTES CULTURED ON HUMAN ACELLULAR DERMAL MATRIX (인간 무세포성 진피기질 위에 배양한 가토 구강각화상피세포의 중충화와 기저막 형성에 관한 연구)

  • Kim, Yong-Deok;Ahn, Kang-Min;Yum, Hak-Yeol;Chung, Hun-Jong;Kim, Soung-Min;Jang, Jeong-Won;Sung, Mi-Ae;Park, Hee-Jung;Hwang, Soon-Jung;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.6
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    • pp.510-522
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    • 2005
  • To assess the clinical applicability of bio-artificial mucosa which was made with autologous oral keratinocytes and human acellular dermal matrix, the formation of basement membrane and stratification of oral keratinocytes were evaluated. Six New Zealand white rabbits (around 2kg in weight) were anesthetized and its buccal mucosa was harvested (1.0 $\times$ 0.5cm size). Oral keratinicytes were extracted and cultured primarily with the feeder layer of pretreated NIH J2 3T3 fibroblast. These confluent cells were innoculated on the human acellular dermal matrix and cultured in multiple layer by air-rafting method. After 3, 5, 7, 10, 14 days of culture, each cultured bio-artificial mucosa was investigated the number of epthelial layer of by H&E stain and toluidine blue stain. The immuhohistochemical methods were used to evaluate the cell division capacity, the formation of basement membrane, and it's property of specific cells (PCNA, cytokeratin 14, laminin). Transmission electromicroscopy was used for the attachment between cells and matrix with the number of hemidesmosome. In result, the numbers of layer of stratified growth of oral keratinocyte cultured on the human acellular dermal matrix and the number of hemidesomal attachment between epithelial cells and human acellular dermal matrix were similar to the layers of normal oral mucosa after 10 days of culture. The cell division rate, basement membrane formation and proliferation rate increased as culture period increased. With these results, bio-artificial mucosa with autologous oral epithelial cells cultured on the acellular dermal matrix had clinically adaptable properties after 10 days' culture and this new bio-artificial mucosa model with relatively short culture time can be expected clinical applicability.

Effect of Persicae Semen for Atopic Dermatitis Skin Tissue and Regulate to Inflammation Mediator in Serum (도인(桃仁)의 아토피 피부염 모델 피부조직 및 혈청 내 염증매개물질 조절 효과)

  • Kim, Sangwoo;Hong, SooYeon;Kwon, Boguen;Kim, Myunghyun;Kim, Sang-bae;Jin, Dae-hwan;Choi, Woochan;Sohn, Youngjoo;Jung, Hyuk-Sang
    • The Korea Journal of Herbology
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    • v.35 no.4
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    • pp.51-60
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    • 2020
  • Objective : The objective of this study was to demonstrate the effect of Persicae Semen (PS) in DNCB-induced atopic dermatitis mouse and HaCaT cell. Methods : The BALB/c mice were divided into four groups. To develop atopic dermatitis, 200 ㎕ of 1 and 0.5% DNCB solution was put on the back of mice in the Control group, the PS-Low group and the PS-High group once a day. After application of DNCB, 200 ㎕ of the PS extract was also treated. The Normal group was given PBS. The mice dorsal skin was stained with Masson's trichrome, H&E, and toluidine blue to evaluate the thickness of the epidermis and dermis, infiltration of eosinophils and mast cells respectively. ELISA was applied to measure the serum level of IgE and IL-6. Toxicity of PS was measured by MTS assay in HaCaT cell. To investigate the effects of PS on HaCaT cells, cells were pre-treated with PS for 1h, and then stimulated with TNF-α and IFN-γ. After 24 hours, the expression of TARC was analyzed using RT-PCR. Results : PS not only significantly diminished the thickness of the epidermis and dermis, but also reduced the infiltration of eosinophil and mast cell in skin lesion. PS also reduced the serum IgE and IL-6 level which plated important roles in the atopic dermatitis. The expression of TARC was decreased significantly in TNF-α/IFN-γ stimulated HaCaT cell. Conclusion : These results suggest that PS may be effective in alleviating the atopic dermatitis induced by DNCB and inflammation by TNF-α/IFN-γ.

The Effects of BGG on Various Immunological Factors Related to Pathogenesis of Allergic Dermatitis in NC/Nga Mice Induced by Der-f (보음거풍지양탕(補陰祛風止痒湯)이 아토피 피부염을 유발(誘發)한 NC/Nga 생쥐의 면역(免疫) 조절작용(調節作用)에 미치는 영향(影響))

  • Bang, Chan-Kuk;Choi, Jeong-Jun;Eom, Dong-Myung;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.16 no.2
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    • pp.147-169
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    • 2007
  • To evaluate the therapeutic effects of BGG on atopic dermatitis, we investigated the composition of immune cells of lymph node, PBMC and skin of Dermatophagoides farinae-induced NC/Nga mice. The levels of immunoglobulins in serum were analyzed at the protein level and the amount of pathologic cytokines were investigated using CD3/CD28 stimulated splenocytes. The results are summarized below; 1. BGG showed no cytotoxic effect up to $200\;{\mu}g/m{\ell}$ on mLFC in vitro. 2. BGG showed no hepatotoxicity in vivo based on the levels of ALT and AST. 3. Atopic dermatitis was improved through naked eye examination. BGG reduced the skin clinical index from 2.9 to 1.3 (p<0.01). 4. H&E and toluidine blue staining of tissue biopsies revealed that BGG inhibited the infiltration of lymphocytes and mast cells to skin. 5. BGG reduced the number of CD19 positive B cells in PBMCs by 16% (p<0.01), whereas cells were increased by 26% (p<0.05) in lymph nodes. 6. BGG reduced the numbers of B220+/CD23+ cells by 15% (p<0.01) and 33% in PBMCs and lymph node, respectively. 7. BGG reduced the numbers of B220+/IgE+ cells in PBMCs and lymph node by 21% and 33% (p<0.01), respectively. 8. BGG suppressed the levels of IgE (13%, p<0.001) as well as IgM (34%, p<0.001), IgG2a (40%, p<0.001) and IgG2b (26%, p<0.05). 9. BGG reduced the levels of IL-4 and IFN-$\gamma$ by 7% (p<0.05) and 13% (p<0.001) in anti-CD3 and anti-CD28-activated splenocytes, respectively. 10. BGG considerably inhibited the production of TNF-$\alpha$ and IL-6 by 42% (p<0.01) and 15% in the serum, respectively. Based on the results above, we concluded that BGG has therapeutic effects on atopic dermatitis by regulating the differentiation of B cells and isotype switching of IgE. Further investigations on the molecular mechanisms of BGG on atopic dermatitis are anticipated.

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Morphometric study of the calculus and periodontal tissues adhered to the root surfaces in periodontitis (치주염이환 치아표면에 부착된 치석과 치주조직의 형태 계측학적 연구)

  • Kim, Chong-Kwan;Yi, Seung-Won
    • Journal of Periodontal and Implant Science
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    • v.27 no.3
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    • pp.621-631
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    • 1997
  • To verify the effect of subgingival calculus on the periodontal tissues in periodontitis and the effectiveness of supragingival scaling to remove the calculus, 30 teeth from healthy group (Probing pocket depth:$PPD{\leq}mm$: HP group), 15 teeth from moderate group ($4{\leq}PD<7mm$:MP group), 30 teeth from advanced group (PPD>7mm: AP group) were selected and supragingival scaling was performed before extraction of all experimental teeth. After careful extraction, the teeth were cleaned with saline and disclosed with toluidine blue and carefully examined the relationship and distance between the calculus attached on the root surface and periodontal tissues. As a result, it was; 1. The calculus was not discovered on the root surface of teeth in HP group, but was in MP and AP group, mostly on interproximal surface and furca area. The shape of the attached calculus was ovoid, trepazoid and polygonal and the calculus was distributed randomly over the root surface. 2. PPD was more than the distance between the gingival margin to the level of attached connective tissue in AP group rather than in HP and MP group. 3. The length of calculus was $2.7mm{\pm}.44mm$ in HP group and $4.1{\pm}.89in$ AP group. 4. The distance between the apical margin of calculus and the level of attached connective tissue was $2.4{\pm}.33mm$ in MP group and $3.4{\pm}.89mm$ in AP group. 5. The length of subgingival calculus was tended to increase in relation to the probing pocket depth. Therefore, it can be concluded, the calculus in periodontal pocket can not be removed completely with supragingival scaling. As the terminal part of calculus was far away with limited distance from the periodontal tissue, it can be said that the calculus was not a direct factor in destroying the periodontal tissue. In this study, the extent of the plaque was not verified but the location of calculus can be used in clinical practice for complete removal of calculus when the distance relation bewteen calculus and plaque will be known.

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Effects of Three Types of GagamBangpungtongseong-san(Except Talcum) on the Atopic Dermatitis in Mice (아토피피부염이 유발된 마우스에 대한 3종 가감방풍통성산(활석 제외)의 효과 비교)

  • Lim, Kyeong-Min;Song, Ji-Hoon;Kim, Jong-Han;Choi, Jung-Hwa;Jung, Min-Yeong;Park, Soo-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.33 no.3
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    • pp.1-26
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    • 2020
  • Objectives : This study was to examine the effects of 3 types of BTS which were excluded talcum only or replaced talcum to Lonicera japonicae Flos or Kochiae Fructus on the DNCB-induced atopic dermatitis in mice. Methods : In this study, Balb/c mice were divided into five groups: normal, control, GBT(BTS except talcum), GBTG(GBT added Lonicera japonicae Flos), and GBTJ(GBT added Kochiae Fructus). And the effects on atopic dermatitis were evaluated by weight change, ear's thickness and weight, thickness of dorsal skin, severity scale of dorsal skin, histopathologic findings of dorsal skin by H&E and toluidine blue stain, proliferation of splenocyte and thymocyte in vitro, proliferation of splenocyte in vivo, IL-4, TNF-α, IgE in serum. Results : There were no significantly changes in body weight and effect of ear's weight in GBT, GBTG, and GBTJ group. The thickness of ear of GBT and GBTJ group showed significant decrease. And the thickness of dorsal skin of GBTJ group significantly decreased compared to the control, GBT, and GBTG group. All the treated groups significantly decreased in severity scale, histopathologically reduced epidermal thickness, and mast cell infiltration. In vitro, all the treated groups increased in the proliferation rates of splenocyte. However, in vivo study, it showed a falling tendency and GBT group significantly decreased compared to control, GBTG, and GBTJ group. In vitro study, GBTG group significantly decreased in the proliferation rates of thymocyte. There was no IgE contents chnage in GBT, GBTG, and GBTJ groups but IL-4 and TNF-α contents were significantly decreased. Conclusions : GBT, GBTG, and GBTJ are expected to improve symptoms of atopic dermatitis and further studies are needed for development of BTS's transformation.

Degenerative Changes of the Rat Dorsal Root Ganglion (DRG) Cells Following a Tight Spinal Nerve Ligation (랫드 척수신경 결찰에 따른 척수신경절세포의 퇴행성변화)

  • Kim, Yi-Suk;Jo, Seung-Mook
    • Applied Microscopy
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    • v.39 no.3
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    • pp.261-266
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    • 2009
  • This study aim to disclose a possible mechanism for the neuronal cell death induced by peripheral nerve injury following a spinal nerve ligation (SNL) as a neuropathic pain model. Male Sprague-Dawley rats (270~290 g) were used for this study. Pain threshold was evaluated for their response to mechanical (von Frey hairs) stimuli 1, 3, and 7 days after a tight ligation of L5 ventral ramus. In control group, the small ganglion cells were strongly stained with routine toluidine blue (TB), whereas the large ganglion cells showed a little bit weak stainity. Each large ganglion cell is surrounded by perineuronal satellite cells. In experimental groups, small ganglion cells showing apparent degenerative changes increased on 1 day, and showed a peak in degenerative cell number at 3 days group, and decreased gradually at 7 days group. We also found a small number of large-sized ganglion cells showing mild degenerative changes. However their satellite cells ware relatively intact with no typical findings throughout this experiment. Under the electron microscope, small ganglion cells showed various stage and typical features of the dark degeneration including mitochondrial swelling.