• 作者： 鄭景侯連團; 李正?; 謝易修
• 作者服務機構： 國立台灣大學醫學院牙醫學系暨牙醫科學研究所; 國立台灣大學醫學院生化學科
• 中文摘要： 齒槽骨、牙本質和牙骨質是口腔的鈣化組織。它們所含的非膠原蛋白雖然少，但已有報告提出該等蛋白在骨誘導、組織鈣化的過程中可能扮演重要的角色；同時對某些結締組織細胞的生物活性也有調節作用。然而不同的非膠原蛋白成份的生物功能和作用機轉則仍不清楚。本文主要的目的乃在分析比較這三種鈣化組織非膠原蛋白成份之異同。我們由健康的人在拔除第三大臼齒時取得齒槽骨和牙齒，經切成lmm薄片後，牙齒切片在立體顯微鏡下，把牙骨質和牙本質分開，三種組織依序以0.5M acetic acid， 4M Guanidine∕0.5M EDTA，和250 units∕ml bacterial collagenase三種萃取液萃取之，所萃取的蛋白質以單向和雙向SDS-PAGE分析之。由冷凍乾燥所得之產物可知，三種萃取液對組織的萃取效果各有不同。骨中含有較多的mineral-associated protein，牙骨質中則分佈較多的collagen-associated protein。10％ SDS-PAGE之分析發現，acetic acid和Guanidine∕EDTA所萃取的成份有許多類似，但是和collagenase所萃取的成份則差異很大。雙向SDS-PAGE之分析則得知，有許多特異蛋白點分佈在齒槽骨（MW=61，55，40，35，34，33kD，而在MW 10-29 kD，p1 5.6-6.4間也有多個點），牙本質（MW=59， 54，35， 28， 25， 24， 21 kD）和牙骨質（MW＝71，64-65，58，55，52，50，47，43，40，31，19 kD）中。這個結果顯示齒槽骨，牙本質和牙骨質雖然在形態和構造上有些相似，但成份的分佈上有所差異，同時又各有其特異蛋白點之存在。確認這些不同組織中的特異蛋白，將有助於我們進一步去瞭解它們在組識中的功能以及生化特性。
• 英文摘要： Matrix proteins of bone, dentin and cementum have been shown to play a role in bone induction duringthe mineralization process, and in regulating the activities of several types of mesenchymal cells. Whether thesebiological functions are mediated through the same mechanism or whether there is specific modulation in eachbiological process is still open to speculation. The purpose of this pilot investigation was to compare the non-collagenous proteins among these tissues. Bone and teeth, obtained from clinically healthy subjects, were sec-tioned into 1 mm thick pieces. With the aid of a dissecting microscope, cementum and dentin were separatedand collected. Tissue specimens were extracted sequentially in three steps by solutions containing 0.5 M aceticacid, 4 M Guanidine/0.5 M EDTA, and 250 units/ml bacterial collagenase, respectively. Proteins extractedwere dialyzed, lyophilized and then further analyzed by both 10% SDS-PAGE (1-D) and two-dimensional (2-D)SDS-PAGE. Comparison showed that the three extraction buffers had relatively different extraction capacitieswithin and among each tissue. The components extracted by acetic acid and Guanidine/EDTA were similar,but seemed different from that extracted by bacterial collagenase as shown by 10% SDS-PAGE. Two-dimensionalSDS-PAGE further characterized numerous distinct protein spots from bone (MW of 61, 55, 40, 35, 34, 33kD and eleven distinct spots which showed MW between 10 and 29 kD, pl range of 5.6-6.4), dentin (MW of59, 54, 35, 28, 25, 24, 21 kD), and cementum (MW of 71, 64-65, 58, 55, 52, 50, 47, 43, 40, 31, 19 kD). Theseresults implied that, although bone, dentin and cementum show morphological and structural similarities, manyproteins were present specifically in each tissue. The distribution of mineral-associated and collagen-associatedproteins in these tissues were also different. Further studies will focus on isolation and characterization ofthese tissue-specific proteins and will delineate their biological functions.
• 中文關鍵字： non-collagenous protein; alveolar bone; dentin; cementum; two-dimensional gel electrophoresis.
• 英文關鍵字： --