• Title/Summary/Keyword: paraffin-embedded specimens

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Effects of Electrical Stimulation on the Nucleolar Organizer Region in Keratinocyte of Wounded Skin in Rat (전기자극이 흰쥐 창상 표피세포의 핵소체형성부위에 미치는 영향)

  • Lee Jae-Hyoung;Jekal Seung-Joo;Park Seung-Teack
    • The Journal of Korean Physical Therapy
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    • v.12 no.1
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    • pp.1-7
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    • 2000
  • The purpose of this study was to determine the effect of microamperage electrical stimulation on the number of argyrophilic nucleolar organizer region (AgNOR) in rat skin. Twenty four male Sprague-Dawley rats were divided into electrical stimulation and control group. Bach animals hair on the back was removed. The electrical stimulation group received an positive rectangular positive electrical stimulation with $500{\mu}A$, while the control group was given the same treatment without electricity. The rats were sacrificed at 4 and 7 day of stimulation, respectively. The biopsy specimens were fixed in formalin, embedded in paraffin and stained with silver nitrate. The AgNOR were counted using a light microscope and computerized image analysis system and calculated as the mean number of AgNOR per nucleus in the epidermal keratinocyte. In control skin, the mean AgNOR count of epidermal keratinocyte at 4 and 7 day were 1.67 and 1.72, whereas electrical stimulated rat had mean AgNOR counts of 2.0 and 2.14, respectively. A Student's t-test showed a significantly higher mean AgNOR number at 4 ana 7 day in the electrical stimulated rats than control rats (p<0.05). The microamperage electric current stimulation increased the epidermal AgMOR expression in incisional wound skin. These results suggest that the microamperage electrical stimulation may promote migration and proliferative activity of epidermal keratinocyte in surgical wound.

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Expression of PDL-specific protein;PDLs22 on the developing mouse tooth and periodontium (발생중인 생쥐 치아 및 치주조직에서 치주인대-특이 단백질; PDLs22의 발현)

  • Park, Jung-Won;Park, Byung-Ki;Kim, Sang-Mok;Kim, Byung-Ock;Park, Joo-Cheol
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.1-12
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    • 2002
  • The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

Study on DNA Content and Ki-67 Antibody Expression by Means of Image Analyzer for the Benign and Malignant Lesions of the Larynx (후두 편평상피의 전암성 및 악성병변에서 화상분석기를 이용한 DNA 배수성검사와 Ki-67 항체 양성세포의 분석에 관한 연구)

  • 주형로;이선희;최종욱;김인선
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.89-89
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    • 1993
  • The laryngeal epithelial cell kinetics of 26 laryngeal lesions(invasive squamous cell carcinoma 14, epithelial hyperplasia 5, laryngeal nodule 7) were studied by immunehistochemical analysis with the monoclonal antibody Ki-67, which reacts with nuclear antigen in proliferating cells using paraffin embedded tissue. For DNA analysis, touch implint with fresh biopsy specimens were stained with feulgen and analyzed by image analyzer in 22 cases. 1) The proportion of Ki-67-positive cells were 32.65$\pm$ 11.59% in invasive squamous cell ca, 20.14$\pm$3.38% in epithelial hyperplasia lesion and 11.66$\pm$3.02% in laryngeal nodule. 2) DNA aneuploidy was found in 7 cases of 10(70%) invasive squamous cell carcinomas, 2 cases of 5(40%) epithelial hyperplasia lesions and all cases of laryngeal 3) Proliferation index(S phase+G2/M phase) show 23.42$\pm$11.33% in squamous cell carcinoma, 13.09$\pm$ 10.90% in epithelial hyperplasia lesion and 4.50$\pm$1.19% in laryngeal nodule. As the results, measuring the DNA content from touch imprint method together positivity of Ki-67 antibody from the microtissue during the laryngeal microscopic surgery, cell kinetics can be assessed as an effort of deciding the prognosis and provide a key to the management of precancerous lesions.

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Clinical Significance of Argyrophilic Nucleolar Organizer Regions(AgNORs) In Squamous Cell Carcinoma of the Lung (편평세포폐암에서 Argyrophilic Nucleolar Organizer Regions(AgNORs)의 임상적의의)

  • Han, Seung-Beom;Jeon, Young-June;Lee, Sang-Sook
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.513-521
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    • 1995
  • Background: Nucleolar organizer regions(NORs) are chromosomal segments encoding for ribosomal RNA and associated with argyrophilic nonhistone protein. Ribosomal RNA genes ultimately direct ribosome and protein synthesis, and it has been suggested the numbers of NORs detected in the cell may reflect nuclear and cellular activity. This study was performed to evaluate the applicability of AgNORs to the diagnosis of squamous cell carcinoma of the lung. Method: The one step silver methods(AgNORs) was used to stain NORs in the routinely processed, formalin fixed, paraffin embedded sections of 36 cases of squamous cell carcinoma of the lung obtained by surgical resection of primary tumor. In each specimen, 100 tumor cells and 100 normal cells adjacent to the tumor chosen at random were examined under an oil immersion lens at a magnification of ${\times}1000$. The mean number of AgNORs per nucleus was calculated for each specimen. Results: The mean number of AgNORs per nucleus(mAgNORs) of normal bronchial epithelium and squamous cell carcinoma of the lung was $1.74{\pm}0.25$ and $4.05{\pm}0.80$, respectively. The difference of mAgNOR between normal and tumor tissue was statistically significant(p<0.001). There was no statistical difference among tumors of different stages. The difference of mAgNOR between normal and tumor tissue was statistically significant in each TNM stage(p<0.05). Conclusion: Mean AgNOR count may be used as a useful marker for the differential diagnosis of benignancy and malignancy, and proliferative activity of the cell in squamous cell carcinoma of the lung. But there was no statistical difference in mean AgNOR count among tumors of different surgical stages. Further studies for the application of mAgNORs to the diagnosis of other histologic types and cytologic specimens of the lung cancer are needed.

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Thyrocyte HLA-DR Expression in Variety of Thyroid Diseases (갑상선 질환들에서 여포세포의 HLA-DR발현)

  • Chang Eun-Sook;Sohn Soo-Sang;Cho Seung-Che;Kim In-Ho;Kim Sang-Pyo;Kang Joong-Shin
    • Korean Journal of Head & Neck Oncology
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    • v.13 no.1
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    • pp.16-23
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    • 1997
  • Although normal thyroid epithelial cells do not constitutively express HLA-DR antigen, their expression in wide spread within thyroid glands obtained from the human with autoimmune thyroid disease and with many neoplastic thyroids. We have, therefore, studied immunohistochemically with regard to the expression of HLA-DR antigen of thyroidectomy specimens from 50 patients of various thyroid diseases with use of paraffin-embedded tissue. One or two sections from each case were stained with commercially available mouse monoclonal antibody for class II HLA-DR antigen(HLA-DR/Alpha, DAKO) and examined by semiquantitative counting system for thyrocytes, neoplastic thyrocytes and other cells expressing HLA-DR antigen. All patients with lymphocytic thyroiditis(2/2) and diffuse hyperplasia(Graves' disease)(5/5), most patients with Hashimoto's disease(9/ll) expressed HLA-DR antigens in thyrocyte with abundant HLA-DR expressing lymphocytic infiltrates with lymph follicle formation in its vicinity or adjacent to the lesion. Most patients with papillary carcinoma(9/1l) had HLA-DR antigen detected in malignant thyrocytes ; while follicular carcinoma(0/3) and follicular adenoma(0/5) did not have detactable HLA-DR immunoreactivity. Adenomatous goiter(3/7) had HLA-DR antigen detected focally in lesser than half cases. Conversely, in four papillary carcinomas and three adenomatous goiters, HLA-DR expression of thyrocytes was found in the absence of HLA-DR expressing lymphoid infiltrates. In such cases therefore other factors more than thyroid autoimmunity must be causative for HLA-DR immunoreactivity. The results of this study indicate as follows. 1) The expression of HLA-DR on thyrocytes involved in autoimmune reactions appeared to be secondary to cytokine release from associated lymphocytic infiltrates. 2) Thyrocytes in thyroid lesions with equal degrees of lymphocytic infiltration without HLA­DR expression exhibited no HLA-DR immunoreactivity. 3) In neoplastic thyrocytes, most papillary carcinoma(9/11) exhibited detactable HLA-DR expression, while follicular carcinoma/adenoma(0/3/0/5) exhibited no detactable HLA-DR immunoreactivity which suggest the existence of divergent mechanisms inducing and modulating HLA-DR expression of different types of neoplastic thyrocytes.

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HISTOLOGICAL CHANGES IN THE ELONGATED BONE AFFECTED BY OSTEODISTRACTION OF THE MANDIBLE IN THE DOG (성견 하악골 절단 후 기계적 골 견인에 의해 형성된 골 신장부에 대한 시기별 조직학적 변화)

  • Baek, Sun-Ho;Ahn, Byoung-Keun;Park, Young-Ju;Park, Hee-Geon;Park, Jun-Woo;Rhee, Gun-Joo;Lee, Yong-Chan;Cho, Byoung-Ouck
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.5
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    • pp.404-416
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    • 2001
  • Purpose : Traditionally, the treatement of choice has been a bone grafting procedure to increase the length of bone in case of actual length discrepancy. But, bone grafting procedure has many disadvantages, for example, graft resorption, donor site morbidity, and so on. So, many trials have been performed to avert the use of autogenous bone graft via introducing new materials or methods. And, one of those trials has been realized by the development of a technique inducing bone lengthening by osteotomy (or corticotomy) and slow gradual distraction of the osteotomized segments. This new technique of bone lengthening dates back to the early 20th century. But, the majority of information concerning the biology of new bone formation during bone lengthening and technical details of the procedure were produced by extensive clinical and experimental studies performed by Ilizarov, a Russian surgeon. According to Ilizarov, with adequate blood supply, preservation of periosteum, rigid fixation of the osteotomized segments, and proper rate and rhythm of distraction, intramembranous bone rapidly develops within the distraction gap in the limb lengthening procedure. In the limb lengthening, many orthopedic surgeons try to observe the biologic and clinical principles recommended by Ilizarov. In the oral and maxillofacial region, however, not a few studies must be performed to apply this surgical technique in the clinical cases. Besides, the mechanism of bone formation in the distraction gap is not clear, yet. The purpose of this experiment was to scrutinize serially the histological changes in the elongated bone affected by osteodistraction of the mandibular body in an adult canine model. In addition, it was performed to confirm the presence of specific region(s) which was important in the bone formation in the gap through the observation of the expression pattern of osteocalcin and osteonectin with the immunohistochemical examination. Materials and Methods : The experimental and control specimens were obtained from seven adult male mongrel dogs weighing over 20kg. The distractors were custom-made linear extraoral devices and bicortical fixation screws were 2.3mm in diameter, 50mm in total length, 15mm in screw length. The distractors were devised to produce a linear gap of 0.75mm between two bony segments every $360^{\circ}$ turn of the rotation rod of the device. The mandibular body of the right side of each animal was corticotomized perpendicular to the occlusal plane and then two bony segments were separated completely by careful manipulation of the segments with bone forceps. The left side of each animal was left intact. This side was served as control. At sixth day after osteotomy and fixation of the segments were performed, distraction of the segments was commenced with a rate of 1.1mm/day and a rhythm of two/day for ensuing 7 days. The animals were euthanized at the 16th. 29th, and 44th day after the osteotomy. The bony specimens were decalcified, embedded in paraffin, sectioned $5{\mu}m$ thick and stained with H&E. The prepared specimens were examined under the light microscope. And, immunohistochemical examinations using anti-osteocalcin antibody (OC1, Biodesign, USA) and anti-osteonectin antibody (Haematologic Technologies Inc., Essex, VT) to locate the expressions of osteocalcin and osteonectin, respectively, were performed. Results : 1. New bone was observed already at the 16th. day after osteotomy. This suggests that new bone formation in osteodistraction was commenced at an early stage of the regenerative process. But, radiologically and microscopically, bony union was not completed in the distraction gap at the 44th. day after osteotomy. Therefore, rigid fixation must be maintained between the bony fragments till the complete bony union is confirmed clinically rather than one month or so after the completion of distraction.

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Prognostic Value of Vascular Endothelial Growth Factor (VEGF) in Resected Non-Small Cell Lung Cancer (비소세포폐암의 예후인자로서 Vascular Endothelial Growth Factor(VEGF)의 의의)

  • Ko, Hyeck-Jae;Park, Jeong-Hyun;Shim, Hyeok;Yang, Sei-Hoon;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.6
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    • pp.676-685
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    • 2001
  • Background : Angiogenesis is an essential component of tumor growth and metastasis, and the vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors. Several solid tumors produce substantial amounts of VEGF, which stimulates proliferation and the migration of endothelial cells, thereby inducing neovasculization by a paracrine mechanism. To evaluate the prognostic roles of angiogenesis and VEGF expression in patients with non-small cell lung cancer, the relationship between VEGF expression in tumor tissues, the clinicopathologic features and the overall survival rate were analysed. Methods : Sixty-nine resected primary non-small cell lung cancer specimens were evaluated. The paraffin-embedded tumor tissues were stained by anti-VEGF polyclonal antibodies using an immunohistochemical method to assess VEGF expression. Results : In Forty-one patients (59%), the VEGF antigen was expressed weakly in their tumor tissue, whereas in twenty-eight patients (41%) the VEGF antigen was expressed strongly. The median survival time of the weak VEGF expression group was 24 months, and that of the strong VEGF expression group was 19 months. The three year-survival rates were 35%, 33%, respectively. The survival difference between both groups was not statistically significant. Conclusion : Although results were not statistically significant, the strong expression group tended to poorer prognosis than the weak expression group.

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Prognostic Value and Histologic Correlation of Sonography in Childhood Nephrotic Syndrome (신증후군 환아에서 신장초음파의 예후적 가치 및 조직검사와의 상관관계에 대한 연구)

  • Cheon Hae-Won;Yoo Kee-Hwan;Hong Young-Sook
    • Childhood Kidney Diseases
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    • v.2 no.1
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    • pp.26-33
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    • 1998
  • This retrospective study was designed to evaluate the prognostic value and histologic correlation of sonography in childhood nephrotic syndrome. Sixty-nine patients with proteinuria over 2g per day at the time of presentation who were treated at the Korea University Hospital were included in this review. They were 1 to 15 years old(mean age, 7.8 years) with 49 males and 20 females. In each patient an ultrasound examination was done using SPA 1000(Diasonics, C.A., U.S.A.) on admission. Tissue specimens were obtained from 46 patients. The paraffin-embedded specimens were reviewed with special reference to interstitial edema, interstitial fibrosis, tubular atrophy, global sclerosis or inflammatory cell infiltrates. Biopsy proven renal disease were minimal change disease(n=20), focal segmental glomerulosclerosis(n=7), membranous glomerulonephritis(n=2), membranoproliferative glomerulonephritis(n=1), $Henoch-Sch\"{o}nlein$ purpura nephritis(n=6), IgA nephropathy(n=5), poststreptococcal glomerulonephritis(n=2), systemic lupus erythematosus(n=1) and Alport syndrome(n=2). There was a significant relationship between increased cortical echogenicity and global sclerosis or tubular atrophy(P<0.05). But no significant relationship was found between increased cortical echogenicity and interstitial fibrosis, interstitial edema, or inflammatory cell infiltration. In biopsy-proven primary nephrotic syndrome(n=30), no significant relationship was found between the increased conical echogenicity and the interstitial edema, interstitial fibrosis, global sclerosis, tubular atrophy or inflammatory cell infiltration. But there was a significant relationship between increased cortical echogenicity and resistance to corticosteroid (P<0.05). These results suggest that increased cortical echogenicity may be due to tubular atrophy or global sclerosis in patients with proteinuria and may be an effective indicator of resistance to corticosteroid in primary nephrotic syndrome.(J Korean Soc of Pediatr Nephrol 2:26-33, 1998)

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Comparison of Proliferative Activity in Each Histological Subtypes of Benign and Atypical Intracranial Meningiomas by PCNA and Ki-67 Immunolabeling (양성 뇌수막종의 조직학적 아형 및 이형성 뇌수막종에서 PCNA와 Ki-67 표지지수의 비교)

  • Choi, Seung Jin;Chang, Eun Deok;Kwon, Seung Oh;Kye, Dae Kon;Park, Choon Keun;Lee, Sang Won;Kang, Joon Ki
    • Journal of Korean Neurosurgical Society
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    • v.29 no.9
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    • pp.1215-1221
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    • 2000
  • Objective : The clinical prognosis and biological behavior of atypical and especially malignant meningiomas are well known to be worse than benign meningioma, but the degree of biological aggressiveness in each classical subtypes of benign meningioma is controversy. This study was performed to see whether there is a difference in the proliferative activity between each different histological subtypes of benign meningioma as well as atypical meningioma. Methods : Paraffin-embedded surgical specimens of 27 meningiomas, including two recurrent tumors, were studied to evaluate proliferative activity by immunohistochemical method with monoclonal antibodies to proliferating cell nuclear antigen(PCNA) and MIB-1. The specimens consisted of 8 cases of meningothelial, 9 cases of transitional, 5 cases of fibroblastic subtypes and 5 cases of atypical meningiomas. Results : Mean PCNA labeling indices of meningothelial, transitional and fibroblastic meningiomas were $4.82{\pm}5.10%$, $9.01{\pm}4.25%$ and $5.66{\pm}5.32%$, but that of atypical meningiomas was $27.62{\pm}19.67%$, noting a higher value compared to all three subtypes of benign meningiomas. Mean Ki-67 labeling indices of the above 3 subtypes were $0.43{\pm}0.85%$, $0.44{\pm}1.08%$ and $0.24{\pm}0.18%$, and that of atypical meningiomas was also revealed to be of higher value ($0.84{\pm}0.59%$). PCNA and Ki-67 labeling indices were not statistically different between histological subtypes of benign meningioma(p>0.05), but the differences of both immunolabeling between benign and atypical meningiomas were statistically significant(p<0.05). Conclusion : Immunolabeling of PCNA and Ki-67 in intracranial meningiomas reveals no prognostic difference between meningothelial, transitional and fibroblastic subtypes in classical benign meningiomas by measuring expression of PCNA and Ki-67, but it seems to be helpful in differentiating benign and atypical meningioma, later showing more proliferative activity and biological aggressiveness.

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Experimental Study on the Effects of Ovariectomy and Estrogen on the Bone Pattern of Mandible in Rats (난소적출과 에스트로젠 투여가 백서의 하악골 구조에 미치는 영향)

  • Lee, Hyung-Soon;Hong, Sung-Gyu;Kim, Jong-Ghee
    • The korean journal of orthodontics
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    • v.29 no.1 s.72
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    • pp.83-94
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    • 1999
  • The purpose of this study was to evaluate the changes of cancellous and cortical bone and the effect of estrogen in ovariectomized rats. Fifty female rats, 250gm in body weight, were divided into three groups : ovariectomized group(OVE), ovariectomized and estrogen-injected group(OVE-EST), and sham operated and estrogen-injected group(EST). Bilateral ovariectomy was performed at the onset of the experiment. In OVE-EST group and EST group, estrogen was injected $50{\mu}g/kg$ B.W. every other days from 3 weeks after surgery to sacrifice. Each five rats were sacrificed after 5, 6, 7 weeks. One side of mandibular body was radiographed with a soft x-ray apparatus(Hitex Co., Ltd., Japan). Thereafter the obtained microradiographs were used for the morphometric analysis using a Image analyzer. The morphometric analysis was perforrmed for parameters such as total bone area, cortex bone area and medullary bone area. The other side of the mandibular bone was decalcified and embedded in paraffin as using a general method. The specimens were sectioned and stained with Mallory's anilline blue and observed light microscopically. The results were as follows. 1 In all groups, the proportion of cortex to total bone area was not significantly different. 2. In ovariectomized(OVE) group, the proportion of marrow cavity to medullary bone area increased significantly from 5 to 7 weeks(p<0.05). In ovariectomized and estrogen-injected(OVE-EST) group, it decreased significantly at 7 weeks, and in estrogen-injected(EST) group, it decreased significantly from 6 weeks(p<0.05). 3. Microradiogram and histopathologic findings revealed that marrow cavity was enlarged and osteoclasts were observed around irregular bone surface in OVE group. In OVE-EST group, the size of marrow cavity at 7 weeks was similar to that of control group. In EST group, as dense trabecular bone increased from 5 to 7 weeks, marrow cavity decreased.

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