多孔性膠原蛋白與泡沫型膠原蛋白於體內外組織再生之研究

Tissue regeneration for porous and foamy collagen in vivo and in vitro

梁修銘(Shiu-Ming Liang)

指導教授 ： 姚少淩

元智大學/工程學院/化學工程與材料科學學系/碩士(2012年)

摘要

膠原蛋白具有良好生物相容性及生物可降解性，因此成為一個理想之天然高分子生醫材料，目前膠原蛋白在組織工程修復應用之型態主要分為水膠與多孔性結構，其中水膠型態膠原蛋白由於無孔洞之結構，較不利於組織的再生；多孔性膠原蛋白可提供良好的組織生長環境，但卻有降解過快之問題。因此針對上述之問題，工業技術研究院提供以高壓氣體與膠原蛋白混合發泡技術，開發出泡沫型態之多孔結構膠原蛋白。
本研究主要探討泡沫型膠原蛋白與傳統水膠型態和多孔性膠原蛋白對組織再生之影響，多孔性膠原蛋白是經由-20℃冷凍後，再以冷凍乾燥法製備成型。在體外測試方面包含分析孔徑大小之差異及比較不同型態對降解速率之影響。在體內測試方面，將三種不同型態之膠原蛋白植入至大鼠皮下，以組織切片染色評估植入後三、七及十四天組織生長之情形。
由掃描式電子顯微鏡觀察結果得知，泡沫型膠原蛋白與多孔性膠原蛋白之孔徑大小分別為166.5 ± 24.2 μm與151.9 ± 26.1 μm，其孔洞大小並無顯著差異(P>0.3)，而在酵素降解測試之結果顯示，多孔性膠原蛋白於六小時後完全被分解，水膠型態與泡沫型膠原蛋白仍有91%與72% 膠原蛋白未被降解。在老鼠皮下植入三天後取樣觀察顯示，發現三種型態之膠原蛋白其周圍組織有些免疫細胞；植入後七天，泡沫型膠原蛋白與多孔性膠原蛋白之外圍孔洞位置均有纖維母細胞生長進入；植入後十四天，泡沫型膠原蛋白與多孔性膠原蛋白孔洞位置均有纖維母細胞生長進入，且有組織再生之情形，但纖維母細胞大部分被阻擋在水膠型態膠原蛋白周圍。
綜合上述實驗結果，泡沫型膠原蛋白具有均勻的孔洞結構與較慢降解之特性，可使組織漸次取代材料，以達到組織再生之目的，因此泡沫型膠原蛋白為組織工程提供一新的研究方向。

關鍵字

多孔性膠原蛋白 ； 泡沫型膠原蛋白 ； 組織再生

並列摘要

Collagen has a biocompatible and biodegradable material, and has become an ideal natural polymer biomaterial. Currently gel-like and porous collagens are focusing in tissue engineering. Nevertheless the gel-like collagen has a non-porous structure, and is no beneficial to tissue engineering application. Although porous collagen has a structure beneficial to tissue regeneration, it degrades too fast. The porous structure of foamy collagen has been developed using high pressure and gel-like collagen mixed foaming technology provided by industrial technology research institute (ITRI).
This research was to compare the feasibility of tissue regeneration of the foamy collagen with that of conventional gel-like and porous collagen. Porous collagen was fabricated by lyophilization after storing at -20℃. Analysis of the pore size of three types of collagen was done by SEM. These three types of collagen on the other procedure was degraded by enzyme in virto. Foamy collagen is compared in terms of degradation rate by enzyme with gel-like and porous collagen. In the case of in vivo test, gel-like, foamy and porous collagen were implanted of subcutaneously in SD rat and the tissue regeneration was observed after 3, 7 and 14 days.
Measurements of the pore size of foamy (166.5 ± 24.2 μm) and porous collagen (151.9 ± 26.1 μm) have no significant difference in the pore size (p>0.05). With the regard of enzyme test, the porous collagen has been completely degraded while gel-like and foamy collagen remain 91.5% and 72.1%. 3 days after implantation, there were immune cells observed in the periphery of the three types of collagen. 7 days after implantation, fibroblast ingrowth was observed in the periphery of foamy and porous collagen. 14 days after implantation, tissue regeneration and fibroblast ingrowth were observed in the pores of foamy and porous collagen but almost in the periphery of gel-like collagen.
As a conclusion of this study foamy collagen has a uniform porous structure and degradation rate is highly beneficial to tissue regeneration. Thus, the foamy collagen has become a new research direction for tissue regeneration in the future.

並列關鍵字

porous collagen ； foamy collagen ； tissue regeneration

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