根據中華民國老年醫學會統計，台灣65歲以上男女罹患骨質疏鬆比率分別為13%及24%，造成個人及社會莫大成本耗損，因此許多的研究無不致力於尋找骨質疏鬆之治療或延緩發生的方法。本研究係建立一個評估中草藥或健康食品療效之方法並探討中藥骨碎補對於雞胚胎長骨發育之影響，作為研究治療或延緩骨質疏鬆發生之動物模式。將受精的雞蛋分成3組，於 99.8 oF狀態下孵育8天後每天以25 ml 濃度60μM之檸檬酸鋁 (n = 24，簡稱鋁組) 、0.7% NaCl (n = 14) 及骨碎補水萃取物 (n = 24，簡稱骨碎補組)分別注射至受精雞蛋之氣室後，繼續孵育。於連續注射之第4天及第8天，分別自3組受精雞蛋中取出雞胚胎，觀察其外型及發育情形並進行骨組織切片、染色及顯微鏡之觀察。結果發現經第4天之鋁組雞胚胎，其外觀上有明顯出血的情形、器官外露的現象，且胚胎之身長顯著地較控制組及骨碎補組為低 (p <0.05)，另外，其長骨發現有彎曲的現象，且第8天之長骨較第4天之長骨具有更顯著地彎曲。於控制組與骨碎補組，無論在外觀、胚胎身長及胚胎之長骨均無顯著的差異性。第4天骨碎補組之雞胚胎長骨經過切片及HE染色後，以顯微鏡觀察，發現其軟骨細胞（chondrocytes）多數已骨化形成骨小樑 (trabecular bone)。根據文獻記載骨面積量與骨徑寬與骨質量有關，因此以Image Pro-Lite軟體計算各組之軟骨細胞、骨面積量及骨徑寬，以便推論其形成之骨質量，結果發現第4天骨碎補組之骨徑寬較第8天控制組高 (p <0.05)，且二組之骨面積量相似，顯示骨碎補水萃取物促使第4天雞胚胎之長骨提早骨化達到控制組第8天之骨徑寬，暗示骨碎補水萃物具有促進鷄胚胎長骨骨化之現象。反之，鋁組無論第4天及8天皆發現大量未骨化之軟骨細胞，且其骨面積量及骨徑寬亦明顯低於控制組及骨碎補組 (p <0.05)，同時明顯地觀察到鋁破壞鷄胚胎長骨骨化之現象。本實驗利用鷄胚胎作為實驗動物模式，主要原因為此模式不會受到飼養食物的影響。此外，雞胚胎長骨之骨化作用於孵育後的第8天開始，因此給予樣品後，可由此階段觀察其對骨質生長發育的影響，實驗結果發現雞胚胎可作為一初期評估中草藥療效之具有潛力之實驗動物模式，且骨碎補水萃取物有促使雞胚胎長骨提早骨化之現象，而鋁則抑制長骨之骨化作用。; In recent years, there has been a lot of effort devoted to the study of the biological functions of Chinese herbs. Gusuibu is a traditional Chinese medicine commonly used for treating or preventing bone or joint diseases. The chick embryo has been used as the experimental animal model to examine the toxicity of aluminum. We applied this model to evaluate the effects of Gusuibu and aluminum on the status of long bone ossification of chick embryos. Embryos were treated on day 8 with 0.7% NaCl, 60 mM aluminum citrate, or the crude water extract of Gusuibu by instilling each solution separately into the air sacs. Aluminum citrate-treated embryos were found to have a significant inhibition on the growth and development of chick embryos, a persistent mid-diaphysis malformation of the tibia, and a significantly shorter tibia length on day 12-incubation. The growth and development of chick embryos treated with the crude water extract of Gusuibu and their reticular structures of trabecular bone were similar to those of the control group. In the embryos treated with Gusuibu for 4 day, ossification was initiated earlier and cortical bone thickness was wider than those of the control group. These results suggest that Gusuibu improves ossification, while aluminum inhibits the growth, development and bone ossification of chick embryos. We demonstrated that the chick embryo was useful and can be applied to evaluate the effects of functional foods on bone ossification and bone formation.
