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A Dynamic Experimental Study of the Expression of iNOS in the Condylar Cartilage After Functional Mandibular Protrusion for the Rat

Author: ZhaoLiXia
Tutor: ZuoYanPing；ZuoBaoYong
School: Hebei Medical University
Course: Clinical Stomatology
Keywords: Functional appliance Inducible nitric oxide synthase (iNOS) Condylar cartilage Immunohistochemistry Mandibular Protrusion
CLC: R783.5
Type: Master's thesis
Year: 2010
Downloads: 28
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Abstract


Objective: The ultimate effect of functional appliance has been the focus of orthodontic workers, due to the different experimental methods, design ideas, as well as the patient's degree of cooperation with, the results are not the same. Academia exist two different points of view on the part of scholars believe that functional appliance through protrusive jaw muscle stress changes, thus contributing to the remodeling of the condyle, which affect the growth and development of the entire mandible improve Ⅱ malocclusion face ; otherwise, some scholars believe that functional appliance is just ahead of an advance the maximum growth of the lower jaw, and ultimately does not produce a significant role in bone. Normal cartilage cells can synthesize affect the generation of a small amount of NO mechanical stimulation of chondrocytes NO, iNOS is one of the main factors for the synthesis of NO. Therefore, the present study, the expression of iNOS in rat condylar cartilage study functional appliance process change, a preliminary understanding of iNOS and NO in condylar remodeling at the same time through the dynamic changes of iNOS observation of long-term stability of the functional appliance designed to provide experimental basis for clinical work. Methods: 60 4-week-old healthy SPrague Dawley (SD) male rats, weight about 90g, the conventional particles fed 7 days after random equivalent divided into two groups: the experimental group (S) and the control group (D), each 30. The removable maxillary inclined plane appliance 10-12 hours of experimental rats daytime wear homemade guide the lower jaw forward. The control group rats were not wearing the appliance for the natural growth of the group. All rats free access to water. Experimental rats free diet, control rats timed feeding, in order to ensure that the two groups of animals a considerable amount of weight gain. Experimental period of one week (1w), 2 weeks (2w), 3 weeks (3w), 4 weeks (4w), 5 weeks (5w), 6 weeks (6w), experimental period in each group were randomly selected rats 5 only were sacrificed, take the right condyle placed in 4% paraformaldehyde at 4 ℃ 24 hours, washed with distilled water at room temperature after 10% EDTA decalcification, after conventional treatment paraffin-embedded tissue blocks of condylar longitudinal slice . Paraffin sections were observed after HE staining condylar cartilage histological changes and measure the thickness of the the different experimental cycle condylar cartilage; Motic Med 6.0 digital medical image analysis system (Motic Med 6.0CMIAS) application by immunohistochemical staining to detect different experimental cycle condyle different parts of the cartilage iNOS expression changes. Application SPSS13.0 statistical software for all data processing, to analyze the changes in different time period condylar cartilage thickness and iNOS expression in condylar cartilage strength changes. Results: Histological observation to the naked eye: rat condylar anteroposterior diameter greater than the inner and outer diameters, the condyle anterior surface of a longer, after partial short slant. 1.1 control group, the histological features of the condylar cartilage microscope, a layer of fibrous tissue that articular disc surrounding the condylar cartilage surface corresponding slightly on part in the joint after-hours with cartilage tissue thickest condylar cartilage by backward before gradually thinning. The condylar cartilage below for trabecular bone. Along with the overall thickness of the condylar cartilage growth and development gradual thinning of to appear 増 changes associated with aging. Condylar cartilage has a unique layer localized sub-band, based on cell morphology can be divided into different the: ① fiber layer containing dense fibrous tissue fibroblasts ② germinal layer is composed of many dense small cell, this cell layer is more active visible mitotic phase (3) This layer of cells of the mature layer increases, irregular shape, irregular arrangement of the the nuclei mostly on one side of the cell, and an increase in cartilage mechanism. ④ transitional layer layer cell swelling, the nucleus is not obvious, significantly increased cartilage matrix, chondrocytes gradually transformed the area to the bone cells. 1.2 experimental group condylar cartilage histological features of functional appliance Protrusion in Rats jaw in the rear of the condylar cartilage thickness increased, the statistics from the first two weeks of the experiment, the middle and rear of the condylar cartilage compared with the control group learning differences, cartilage layer thickening performance increase in the number of germinal layer cell nucleus is large and obvious; the mature layer compared with the control group thickening, cell morphology similar to the control group. Condylar cartilage in the front of the thickness from 1 week to five weeks, compared with the control group, no statistically significant difference in the first six weeks of the cartilage thickness of the front of the control group compared to the thinning of the statistically significant. Observation of the animals found dismantled appliance worn after a period of time with the appliance, the individual rat mandibular habitual protrusive there anterior teeth crossbite. 2 immunohistochemistry results iNOS positive staining was brown, mainly located in the cytoplasm. Positive cells coloring levels significantly higher than background levels, blank control (PBS instead of primary antibody) group no staining particles. iNOS expression in rat condylar cartilage layer the mature layer shallow chondrocytes highest expression. iNOS expression intensity in the the condylar cartilage rear strongest expression for the front, middle, followed by the weakest. 2.1 condylar cartilage the front of iNOS expression in the control group and the experimental group on the front of the expression of the same trends with rear from 2 weeks to 4 weeks iNOS expression pairwise comparison significant difference. Protrusive mandibular Rats the expression level of the front portion is also enhanced, and from 1 to 6 weeks as compared with the control group always has a significant difference. 2.2 condylar cartilage Central iNOS expression in the expression of the control group and the experimental group in central trend, the same rear. The pairwise comparison control group from 2 to 4 weeks iNOS expression significant difference. Protrusion in Rats jaw iNOS expression in one week, two weeks, and the control group showed significant differences, 3 weeks to 4 weeks compared with the control group, no significant differences. 2.3 condylar cartilage rear iNOS expression in the positive control group of iNOS expression intensity gradually decreased, showing 増 age-related changes, from 2 weeks to 5 weeks iNOS expression pairwise comparison was statistically significant; iNOS in the experimental group The expression levels were higher than the control, the expression of the same trend with the control group, there are significant differences compared with control group from 1 to 5 weeks expression of iNOS. 6th week of iNOS expression levels compared with control group was no significant difference. Conclusion: Growing Rat condylar cartilage with the observation time goes to appear 増 age-related changes, gradual thinning of cartilage thickness; functional appliance condylar cartilage showed remodeling of the treatment of early stage cartilage thickness was significantly thicker and with the progress of the experiment gradually thinning. The growth in the developing rat condylar cartilage under normal physiological state iNOS expression decreased gradually with the growth and development, suggesting that iNOS participate in the the condyle normal growth and development process. 3 rats after wearing the the guide jaw forward functional appliance condylar cartilage iNOS expression increased gradually reduced with the experimental period, the trend is consistent with the control group, stabilized, suggesting that in the experimental late party After correction, should pay attention to maintain the effect.

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