1. Synthesis and in vivo evaluation of a novel hydroxyapatite/ collagen–alginate as a bone filler and a drug delivery carrier of bone morphogenetic protein 指導教授 : 林鴻儒 博士 學生 : 徐楓茜 日期 :98/12/15 Materials Science and Engineering C 24 (2004) 341–347 Shinichi Sotome, Toshimasa Uemura, Masanori Kikuchic, Jiani Chen, Soichiro Itoh, Junzo Tanaka, Tetsuya Tateishi, Kenichi Shinomiya*

2. 前言 硬骨具有良好的再生能力，但需要有適當 的治療方式，較嚴重患者通常是使用自身 移植，例如：骨癌。 自身移植通常都使用腸骨或小腿骨，但是 通常都會造成捐贈部位疼痛且數量又有限。 基於這些原因，在骨組織有顯著的研究， 像是骨組織成長因素或間葉幹細胞的發展 來取代自身移植。

3. 實驗步驟 - 實驗步驟 - HAp/col-alginate HAp/col powder 0.5g3ml 0.9% NaCl 35 μ l of 5M CaCO 3 mixture 1.5ml 3% alginate HAp/col-alginate mixture D-gluconic acid lactone Injected into a mold 45min, room temperture HAp/col-alginate was incubated in100 Mm CaCl 2 30min Cut to 2*2*3 and 2*2*5

4. 實驗步驟 - Implantation into bone holes Wistar rats(10weeks, 280-320g) Femur by a lateral approach, and a 3mm hole using electric drill The drill hole rinsed with 0.9% NaCl solution The implant was placed into the hole The fascia and skin were sutured

5. 實驗步驟 - Ectopic bone formation model HAp/col-alginate rh-BMP2 (0, 4, 20, 100 μ g/ml) sample Atelo-collagen rh-BMP2 (0, 4, 20, 100 μ g/ml) sample Implanted into the bilateral side of dorsal muscle (5weeks, male, Wistar rat)

6. 結果討論 Fig. 1. Photograph of HAp/Col–alginate. (A) Dry implant. (B) Wet implant.

7. Fig. 2. Implant site 2 weeks after implantation. (A, B) HAp/Col– alginate. [(B) Higher magnification of rectangular area in panel A.] (C) Alginate. (D) Porous HA. Black arrowheads indicate implant. White arrowheads indicate newly formed bone. The borders of HAp/Col– alginate and newly formed bone were not clear and bone tissue invaded the implant.

8. Fig. 3. Implant site of HAp/Col–alginate 4 weeks after implantation. (B) Higher magnification of rectangular area of panel A. Tissue invasion and bone formation around the HAp/Col– alginate implants increased.

9. Fig. 4. Implant site 8 weeks after implantation. (A–C) HAp/Col alginate. (D) Alginate. (E) Porous HA. (A, B) Fragmentation of HAp/Col– alginate and increase of tissue invasion and bone formation were observed. (C) TRAP-positive multinuclear cells attached to the implant were observed. (White arrowheads indicate TRAP-positive cells.) (D) Tissue invasion of the implants was not observed except at tears. (E) Most of the pores were not filled with bone tissue.

10. Fig. 5. Ectopic bone formation induced by BMP2 (100 g/ml), 5 weeks after implantation. (A–C) HAp/Col–alginate with BMP. (A) HE section of the whole implant. (B) Immunostaining section with antiosteocalcin. (C) Higher magnification. Bone formation extended throughout almost the entire implant and bone marrow-like tissue was also observed. White arrows indicate bone marrow-like tissue. (D) Collagen sponge with BMP. The area of bone formation was less than that of HAp/Col– alginate and occupied only a part of the implant. Black arrowheads indicate soft tissue which invaded the remnant of the collagen sponge. White arrowheads indicate bone.

11. 結論 由組織切片圖可看出， HAp/Col-alginate 可 作為藥物載體且順利地使骨增生。 由組織切片圖可看出， HAp/Col-alginate 可 作為人造骨材料