聚乙烯醇-聚乳酸-磷酸三鈣化學交聯細胞支架的特性應用於組織工程

Characterization of cross-linked Poly(vinyl alcohol)- Polylactic Acid-Tricalcium phosphate 3Dscaffold for tissue engineering

黃佳軍(Chia-Chun Huang)

指導教授 ： 謝文權

義守大學/醫學院/生物科技學系/碩士(2017年)

摘要

本實驗的研究目的是利用聚乙烯醇、聚乳酸和磷酸三鈣經由化學交聯的方式製作出具有多孔性、吸水性能、生物相容性高、結構穩定的3D細胞支架。化學交聯的PVA-PLA-Ca3(PO4)2 3D細胞支架由熱差分析儀的實驗可得知，此三種材料可透過化學交聯的方式形成共聚合物。在吸水性測試中可以達到700%；在抗壓測試中可得知隨著材料中PLA的增多會使的3D細胞支架的抗壓強度增加，最高可達到9x10-3MPa。
利用電子顯微鏡的觀察，可以發現利用一定的轉速來產生氣泡可以製作出大量的孔隙，在模擬人體內的酵素分解試驗中，3D細胞支架在各種酵素的分解下，在21天內平均的分解率約在20-25%的分解效果。
在體外細胞培養實驗中，利用3D支架培養細胞在3天後可達2.3 x105，再利用能量散射光譜儀分析，更能夠證明此細胞支架具有生物可相容性；在動物實驗中，把PVA-PLA-Ca3(PO4)2 D細胞支架縫入大鼠的表皮下觀察一個月進行抽血檢測，在組織切片觀察中並未發現細胞發炎；在抽血檢驗中白血球的數量並沒有增多，因此證明支架在大鼠體內並沒有產生排斥反應。由上結果得知，此細胞支架材料是可以作為再生醫療之應用。

關鍵字

聚乙烯醇 ； 聚乳酸 ； 磷酸三鈣 ； 化學交聯 ； 3D細胞支架 ； 細胞培養

並列摘要

Polylactic Acid were grafted to poly(vinyl alcohol) (PVA) to form a porous scaffold. The grafted PVA- Polylactic Acid 3D scaffold was then observed by Differential scanning calorimetry(DSC). Grafting increased the water absorbency of the scaffold by 700%. Scanning electron microscopic (SEM) observations of the material revealed that the 3D scaffold was highly porous when formed using a homogenizer at 100 rpm.
Compression testing revealed that, increasing the amount of Polylactic Acid and adding increased the strength of the 3Dscaffold to 9x10-3 MPa. Over 21 days, the 3D scaffold was degraded by various enzymes with a weight loss of up to 20-25%.In the animal experiment, the PVA-PLA-Ca2(PO4)2cell stent was sutured into the epidermis of the rats for one month . In the tissue section observation, we did not find cell inflammation. In the blood test, we observation the number of white blood cells did not increase, In vitro tests revealed favorable cell proliferation and growth in a 3D scaffold.

並列關鍵字

poly(vinyl alcohol) ； Polylactic Acid ； Tricalcium phosphate

參考文獻

[1] Y.W. Wang, Q. Wu, and G.Q. Chen, Attachment, “proliferation and differentiation ofosteoblasts on random biopolyesterpoly (3-hydroxybutyrate-co-3-hydroxyl- hexanoate) scaffolds” Biomaterials 25:669-675(2004).

[2] J. Jiang, N. Kojima, T. Kinoshita, A. Miyajima, W. Yan, Y. Sakai, “Cultivation and inductionof fetal liver cells in poly-L-lactic acid scaffold” Materials Science and Engineering C24:361-363(2004).

[3] Z. Li, H.R. Ramay, K.D. Hauch, D. Xiao, and M. Zhang, “Chitosan–alginate hybrid scaffolds for bone tissue engineering” Biomaterials 26:3919-3928(2005).

[4] Wen-Chuan Hsieh, Wan-Nan Uang Chen, Shih-Jung Yang, Hsin-Hao Chou, “ 3D tissue culture and fermentation of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) from Burkholderiacepacia D1” Journal of the Taiwan Institute of Chemical Engineers, 57: 250-255(2011).

[5] Chih-FengKuo, Nina Tsao, Hsin-Hao Chou, Yi-Ling Liu, Wen-Chuan Hsieh, “Release of FITC-BSA from poly(L-lactic acid) microspheres analysis using flow cytometry” Colloids & Surfaces B : Biointerfaces, 89:271-276(2012).