論文名稱:

以組織免疫染色法觀察後牙缺失之大白鼠顳顎關節盤中一、三類膠原蛋白之變化

 

Immunohistochemical observation of type Ⅰ Ⅲ collagen in temporomandibular joint discs of rats after posterior teeth loss

研究生:

張治平  Zhang, Zhi-Ping

 

(以作者名查詢陽明大學館藏系統)

 

(以作者名查詢全國圖書書目資訊網)

指導教授:

許明倫  Hsu, Ming-Lun

        學校名稱:

國立陽明大學

          學年度:

86

摘要:

顳顎關節為咀嚼系統中之重要環結。其功能有別於身體上其他關節,而其構造自亦異於其他關節。顳顎關節之構造包含關節窩、關節盤、探頭、關節囊及其相鄰之肌肉、骨骼及韌帶等組織。由於顳顎關節之運動型式不同於身體上其他關節因此其關節盤受力之方式,於下顎運動時所扮演之角色,亦有別於身體上其他關節之關節盤。關節盤之構造主要為第IIll類膠原蛋白彼此糾結成緻密網狀結構。不同形式之膠原蛋白含不同之多胜肱鍵(polypeptide chain)而有不同之功能,體內各組織亦因其所含不同膠原蛋白之型式和濃度而其不同之功能。

 

  本實驗之目的為探討拔除後牙是否造成顳顎關節盤中第IIll類膠原蛋白之變化。研究中原以十二隻大白公鼠(Sprague-Dawley malerat)為樣本,隨機分為三組,其中四隻為單側拔牙組,另四隻為雙側拔牙組,于二十週大時拔牙,其他四隻為對照組。而對照組中一隻大白鼠,于拔牙後二十一週時死亡,故對照組為三隻。手術前,使用乙醚作深度麻醉。單側組拔除左側上、下顎所有臼齒;雙側組拔除上顎左右兩側所有臼齒;對照組不作拔牙處理。拔牙三十週後,犧牲老鼠取出顳顎關節盤,經急速冷凍後包埋,使用冷凍切片機切片,每個切片厚度 8μm。最後再使用第IIII類膠原蛋白抗體等試劑作組織免疫染色。每個關節盤各切二片,一片染第I類膠原蛋自,另一月染第III類膠原蛋白。染色之深淺即代表膠原蛋白含量之多寡,將觀察到之結果,依染色之深淺分"1""2""3",三個等級,以對照組最常見到之染色深淺為中等染色(2),比中等染色顏色深者,也就是濃染的顏色為"3",比中等染色顏色淺者也就是淡染的顏色為"1"。結果發現對照組與實驗組間各個體出現濃染、中等染色與淡染之機會不一,並無一定之趨勢可循,但仍可獲兩點、結論:(1)IIII類膠原蛋白在關節盤中,均以前側出現濃染之機會為多,此可能因受外翼狀肌拉力所致。(2)IIII類膠原蛋白之變化則以單側拔牙組較為明顯,可能由於單側拔牙後咀嚼模式改以非拔牙側為主之單側咀嚼模式所致。

   
 

  Temporomandibular joint (TMJ) is an important segment in the masticatory system. Both it's function and structure are different from another joints of the body. TMJ consists ofcondyle, disc, glenoid fossa, capsule, and the adjacent muscles, ligaments, and bones. The role of the TMJ disc in the mandibular movement and the force directing on the TMJ disc are different from another joints of the body. Disc mainly composes of type I and type III collagens. Collagen fibers in disc twist one another to form a dense reticular structure. Because different type of collagen fiber contains different polypeptide chain, so each type of collagen possesses it's own unique character. Different type of tissue in the body has different function due to the composition of different collagen.

 

  The purpose of this experiment was to study the influence of type I and III collagens in temporomandibular joint discs of the rats after posterior teeth loss. Twelve 20 weeks adult male SD (Sprague-Dawley) rats were used as the experimental samples. Rats were randomly divided into three groups: Unilateral extraction group, left upper and lower molars were removed. Bilateral extraction group, upper bilateral molars were extracted. Control group consisted of rats without extraction. But one rat of control group expired after twenty-one week. So the amout of control group reduced to three. After thirty weeks rats were sacrificed and bilateral temporomandibular joint discs were surgically removed and prepared for immunohistochemical study.

 

  Two slides were made from each disc, one for staining of type I collagen another for type III collagen.The staining time was unified, 2 minutes. The more collagen concentration existed in the disc, the deeper staining color to be observed. The results were divided into three categories according to the deepness of staining color: "2" (standard color) represents the most common color observed in the control group, "3" is the color which is deeper than standard color which "1" is the color which is lighter than standard color. Four locations from anterior, posterior, medial, and lateral parts of each disc were observed under light microscope (200X). The results showed that the percentage of standard color, deep color and light color to be observed in each slide is not the same, there was no special trend could be followed. But the results suggested that: (l)Deep color could be noticed in the anterior part of most slides both in type I & III collagens, this could be due to the pulling force of lateral pterygoid muscle. (2)Variation of type I & III collagens are more prominent in the unilateral extraction group, this could be influenced by changing of the chewing pattern to be unilateral (non-extraction site) chewing.