為克服人體移植骨的取得困難與避免疾病傳染之高危險性，特比照自溶及抗原去除後同種移植骨(AAA bone) 之製備，進行實驗評估運用異種骨移植之可能性，本實驗第一階段是在40隻SD老鼠腹直肌中植入0.5*0.5公分人或豬骨之AAA bone後分別在 3,7,14,21,28天以免疫組織化學染色法(IHC)分析骨誘導蛋白BMP-2，BMP-4，BMP-7在誘導過程中所扮演的角色並在第28天作H&E染色分析骨再生情形。第二階段則在是6隻狗之前肢分別植入人、豬、狗之AAA bone後，每隔3周以X光觀察並在第40周後取出以肉眼及骨密度儀器分析骨頭生長之情況以評估同、異種骨移植的差異性。 在老鼠腹直肌中的誘導表現顯示無論人或豬骨均在植入骨頭附近的肌肉有BMPs的聚集，其中 BMP-2/-4在植入早期3-7天即有明顯的表現，而BMP-7則在晚期14-21天才有較明顯表現。但植入的骨頭本身卻沒有BMPs的表現。第28天之H&E染色則顯示人或豬骨均有血管侵入及骨新生作用。 狗前肢移植實驗則顯示植入後第3周所有的狗前肢都恢復了重力承受(weight-bearing)功能。以肉眼、X光及骨密度儀器檢查發現同種(狗骨AAA bone)植入狗前肢是完全癒合與正常骨一致。植入豬AAA bone的其中一隻在第20周左右因植入的骨頭被吸收而宣告失敗，但另一隻經過四十周後外觀上則發現植骨部分被軟組織取代，以X光及骨密度儀器檢查發現是有一部分的骨頭被吸收，但骨頭截斷面處已經有骨生長但密度較正常對照組低。植入人骨AAA bone的其中一隻狗在第15周左右因感染造成骨髓炎而失敗。另一隻經過四十周後外觀上發現無特殊異常，以X光及骨密度儀器檢查發現癒合良好，且有明顯骨增生現象而整體密度與正常對照組相差不大。 本研究之初步結果顯示AAA bone之骨誘導乃源發於周邊組織之內生性BMPs聚集而AAA bone則有如人工魚礁提供間葉細胞轉化時的鷹架附著效果，更證實經過自溶與抗原去除處理後之異種移植骨(異種AAA bone)是具有臨床運用的可能性。; Either in the Oral and Maxillofacial Surgery or Orthopaedic Surgery, bone graft is still the best resolution of bone defect. However, even now in clinic use, no substitute of autograft bone could be satisfied with both mechanical strength and potential regeneration. In 1965，Marshall Urist made the key discovery that demineralized bone induced heterotopic bone formation when implanted subcutaneously or intramuscularly in animals. As a spark of revolution of bone regeneration changed the traditional theory of osteoconduction into osteoinduction. Subsequent purification studies of these bone inductive proteins from bovine and swine resulted in classification of many members of bone morphogenetic proteins(BMPs). AAA bone(Autolyzed antigen-extracted allogeneic bone graft) is the bone which preserves BMPs and demineralized bone structure matrix. Then BMPs can release from the bone structure scaffold and exert their inductive potential. The BMPs which belong to transforming growth factor-βsuperfamily have shown to transduce signals through heteromeric trans-menbrane serine-threonine kinase receptors and transform mesenchymal cells into chondrocytes and osteoblasts and ossification. At present, most studies still focus on allogeneic transplantation. But our research wants to compare the xenogeneic osteoinduction characterization . Because transmission disease among the human has become critical problem during transplantation. Although xenogeneic transplantation frequently fails due to immune response through calssⅠ(CD8+) T cytotoxic lymphocytes response, the AAA bone which extract the major antigen can decrease the graft to be destroyed by host immune system can satisfy clinical application. We therefore believe that the study of xenogeneic osteoinduction of autolyzed and antigen-extracted human and swine bone---A comparative characterization in rat and canine model will be also a great contribution to bone transplantation in the near future.
