本文選題：下頜功能前伸 + 髁突軟骨��； 參考：《河北醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：髁突軟骨在整個生命過程中始終處于改建狀態(tài),改建除了受到機體自身遺傳、體液、神經(jīng)等因素控制之外,其局部所受的各種應(yīng)力刺激也起著重要作用。髁突軟骨細胞能夠感受各種應(yīng)力刺激，在生物機械力的作用下，髁突軟骨內(nèi)會產(chǎn)生生物力學(xué)信號的轉(zhuǎn)換，表現(xiàn)出細胞增殖、分化和程序性細胞死亡等一系列的反應(yīng)。然而,髁突軟骨受到機械應(yīng)力作用后,體內(nèi)軟骨細胞如何應(yīng)答機械應(yīng)力刺激,并將刺激信號轉(zhuǎn)導(dǎo)進入細胞內(nèi),從而調(diào)節(jié)髁突骨改建的細胞機制仍未能得到清楚的認識。近年來發(fā)現(xiàn)c-fos基因參與髁突骨改建的信號轉(zhuǎn)導(dǎo)和調(diào)控過程,并受到學(xué)者的關(guān)注。c-fos屬于細胞癌基因中的核內(nèi)蛋白類,集中分布于細胞核內(nèi),細胞處于靜止期時不表達,它的表達一般與細胞生長、增殖同步出現(xiàn)。原癌基因的主要功能是調(diào)節(jié)細胞生長、增殖和分化,有研究發(fā)現(xiàn)該基因與軟骨細胞增殖以及骨形成有著密切的關(guān)系。 本實驗采用免疫組織化學(xué)染色的方法檢測大鼠下頜功能前伸后髁突軟骨中c-fos的表達及其變化規(guī)律，探討細胞內(nèi)c-fos介導(dǎo)功能矯治前伸下頜后髁突骨改建的機制，為臨床上功能矯治提供實驗依據(jù)。 方法：選用4周齡健康雄性Sprague-Dawley(SD)大鼠70只，體重約80克，隨機等量分為實驗組和對照組，組內(nèi)根據(jù)實驗周期（1、3、7、14、21、28和35天）分為7組（每組5只），共14組。適應(yīng)性飼養(yǎng)一周后，實驗組利用3M玻璃離子將自制下頜前伸斜面導(dǎo)板矯治器粘固于大鼠上切牙，通過鏈狀橡皮圈經(jīng)鼻上頜復(fù)合與兩側(cè)口外弓連接，使得斜面導(dǎo)板獲得穩(wěn)定的固位，將大鼠四肢用醫(yī)用橡皮膏緊緊包裹有效地防止大鼠前爪對裝置的破壞及對面部組織的傷害，矯治器24h戴用；對照組不做任何實驗操作。所有動物均以軟食飼養(yǎng)，自由飲水，同環(huán)境下飼養(yǎng)至實驗結(jié)束。實驗后1、3、7、14、21、28、35天分別取材，常規(guī)固定，脫鈣，脫水，包埋。石蠟切片后進行常規(guī)HE染色觀察髁突組織學(xué)變化；免疫組化方法（二步法）檢測FOS蛋白的表達，采用ZKPACS-G型中科恒業(yè)細胞圖象分析系統(tǒng)進行免疫組化染色評分，以IHS值表示染色強度。所有數(shù)據(jù)結(jié)果均應(yīng)用SPSS13.0統(tǒng)計軟件進行處理，，統(tǒng)計分析時實驗組與對照組組間比較采用t檢驗，組內(nèi)比較采用單因素方差分析，以P＜0.05為有統(tǒng)計學(xué)差異。檢驗水準α=0.05。 結(jié)果： 1大體觀察 實驗組大鼠下頜處于適度前伸狀態(tài)，上下頜前牙呈對刃關(guān)系，摘除矯治器后，下頜不能后退。而對照組大鼠前牙覆蓋約3mm。 2髁突形態(tài)學(xué)變化 對照組隨著觀測時間的延長，髁突軟骨呈現(xiàn)增齡性變化，表現(xiàn)為軟骨厚度變薄，其中軟骨中部和后部變化更為顯著；實驗組自實驗第3天起,髁突軟骨各層組織變厚,軟骨細胞數(shù)量增多,髁突中后份有明顯變化,而前份變化不明顯。之后到21天時，軟骨軟骨各層逐漸穩(wěn)定，移行層較對照組仍明顯增厚，并可見處于末期的軟骨細胞，成骨細胞以及大量原始骨小梁及發(fā)育不全的骨髓腔。 3FOS蛋白表達水平及分布 空白對照（PBS代替一抗）組未見陽性著色顆粒。對照組髁突軟骨中僅肥大層少量細胞FOS蛋白表達陽性，且一直到第21天都維持在較低的表達水平，第21天以后表達逐漸下降到極低的水平，組間比較差異無統(tǒng)計學(xué)意義（P0.05）;實驗組第3天時,髁突軟骨中FOS蛋白表達明顯增強,IHS=2.2±1.10,與對照組IHS=1.0±0.70顯著差異（P＞0.05）,第7天時，實驗組IHS=6.0±1.22陽性表達持續(xù)增強，與對照組IHS=1.0±0.70比較有明顯差異（P 0.05）；第14天，實驗組IHS=6.0±1.58,與第7天基本維持在同一表達水平，第21天,實驗組IHS=3.6±1.67，大部分樣本陽性表達細胞開始減少,但與對照組相比仍有顯著差異（P0.05）；第28天，實驗組IHS=2.4±1.14，對照組IHS=0.8±0.45，差別仍具有統(tǒng)計學(xué)意義；第35天，髁突軟骨FOS蛋白表達回落至極低水平，實驗組IHS=0.6±0.55，與對照組IHS=0.6±0.55相比無統(tǒng)計學(xué)差異。 結(jié)論： 1c-fos參與了大鼠下頜前伸后髁突軟骨改建過程的調(diào)節(jié)。 2c-fos在髁突軟骨細胞增殖和分化中起到重要作用。 3c-fos介導(dǎo)了大鼠髁突軟骨骨改建過程中機械外力-生物化學(xué)信號的轉(zhuǎn)導(dǎo)，在生物信號由細胞核外轉(zhuǎn)入核內(nèi)的過程中起到重要的作用。
[Abstract]:Objective: the condylar cartilage has been rebuilt throughout the life process. In addition to the control of the body's own inheritance, body fluid and nerve, the local stress stimulation also plays an important role. The condyle cartilage cells can feel a variety of stress spines. Under the effect of biological mechanical force, the condylar cartilage will be within the condylar cartilage The transformation of biomechanical signals shows a series of responses to cell proliferation, differentiation and programmed cell death. However, after the condylar cartilage is subjected to mechanical stress, how the cartilage cells in the body respond to mechanical stress stimulation and transduce the stimulus signal into the cell, thereby regulating the cell mechanism of the remodeling of the condyle bone still failed. In recent years, the c-fos gene has been found to be involved in the signal transduction and regulation of the reconstruction of the condylar bone, and the attention of the scholars is concerned that.C-fos belongs to the intracellular protein of the cell carcinoma gene, which is concentrated in the nucleus and the cell is not expressed at the stationary phase. Its expression is in synchro with cell growth and proliferation. Its main function is to regulate cell growth, proliferation and differentiation. Some studies have found that this gene is closely related to chondrocyte proliferation and bone formation.
In this experiment, immunohistochemical staining was used to detect the expression of c-fos in the mandibular condylar cartilage and the changes in the mandibular protrusion in rats. The mechanism of c-fos mediated function in the reconstruction of the posterior condyle of the mandibular protrusion was explored to provide the experimental basis for the clinical functional correction.
Methods: 70 healthy male Sprague-Dawley (SD) rats of 4 weeks old, weighing about 80 grams, were divided into the experimental group and the control group randomly. The group was divided into 7 groups according to the experimental period (1,3,7,14,21,28 and 35 days) and 14 groups. After a week of adaptation, the experimental group used the 3M glass ion to stick the self-made mandibular protrusive inclined plane guide appliance. The rat upper incisor was fixed on the upper incisor by the chain like rubber ring through the nasal maxillary compound and the two side pantograph, making the oblique guide plate stable. The rat limbs were tightly wrapped with medical rubber cream to effectively prevent the damage of the rat's front claw and the injury of the facial tissue. The orthodontic 24h was worn and the control group did not do any experimental operation. All animals were fed with soft food, free drinking water, and kept in the same environment to the end of the experiment. After 1,3,7,14,21,28,35 days, the animals were harvested, routinely fixed, decalcified, dehydrated and embedded. After paraffin section, routine HE staining was used to observe the histological changes of the condyle; the immunohistochemical method (two step method) was used to detect the expression of FOS protein, and ZKPACS-G medium was used. The industrial cell image analysis system was used to perform immunohistochemical staining score with IHS value. All data were processed with SPSS13.0 software. The experimental group was compared with the control group by t test, and the single factor variance analysis was used in the group. The statistical difference between the group and the P < 0.05 was statistically significant. The test level was alpha =0.0. Five
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本文編號：1815955