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Development of nanobiomaterials for peripheral nerve repair
|關鍵字:||PVA/chitosan conduit;聚乙烯醇/幾丁聚醣導管;nerve regeneration;PCL-PEG diblock copolymer;microphase separation;nanotopography;cell response;inflammation;platelet activation;cell migration;sliding angle;polyurethane;nanocomposite;biocompatibility;gold nanoparticles;tissue reaction.;神經再生;聚己內酯-聚乙烯醇雙團聯式共聚合物;微相分離;奈米地形學;細胞反應;發炎;血小板活化;細胞遷移;滑動角度;聚胺酯;奈米複合材料;生物相容性;金奈米粒子;組織反應||出版社:||化學工程學系所||引用:||Chapter.1  Noble J, Munro CA, Prasad VSSV, et al. Analysis of upper and lower extremity peripheral nerve injuries in a population of patients with multiple injuries. J Trauma 1998; 45:116-122.  Kelsey JL, Praemer A, Nelson L, et al. Upper Extremity Disorders. Frequency, Impact, and Cost. New York, NY, Churchill Livingstone, 1997.  Mohanna PN, Young RC, Wiberg M, et al. A composite poly-hydroxybutyrate-glial growth factor conduit for long nerve gap repairs. J Anat 2003; 203:553-565.  Lee GW, Collins ED, Kupfer DM, Weeks PM. 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本論文第二部分研究一奈米相分離的生物可降解高分子的細胞效應，使用聚己內酯-聚乙烯醇雙團聯式共聚合物表面，調查奈米小島對纖維母細胞及內皮細胞貼附及增生的影響。結果顯示在雙團聯式共聚合物中增加聚乙烯醇鏈段會改善表面親水性，而細菌的附著性會被抑制。PCL-PEG 23:77表面具有奈米形態，內皮細胞、血小板及單核球在PCL-PEG 77:23表面貼附的數目被抑制，而在PCL-PEG 23:77會增加。然而在PCL-PEG 23:77表面上，血小板及單核球活化的數目是減少的。如同血小板及單核球活化程度降低，PCL-PEG 23:77 對細胞具有較好的反應。另一方面，調查奈米小島對於不同種類細胞如纖維母細胞及內皮細胞移動行為之影響，其中基質的水滑動角被量測，以曠時攝影系統結合光學顯微鏡調查細胞移動速度，發現較快的細胞移動速率與基質具有較小的滑動角有關
本論文第三部份研究奈米複合材料生物效應，製備不同形式的聚胺酯-金奈米粒子之複合材料，其中含43.5 ppm金的聚醚型聚胺酯的複合材料顯示，相對於聚胺酯或是不同含量金之聚醚型聚胺酯複合材料，具有增加細胞增生、減少血小板及單核球活化及較少的細菌附著，良好的生物相容性與複合材料表面形態變化有關。在含43.5 ppm金的聚醚型聚胺酯複合材料中發現氧化降解作用被抑制，氧化穩定性的增加與奈米複合材料較高的自由基清除能力有關。另一方面，針對聚酯型聚胺酯-金的奈米複合材料之型態、熱性質、生物相容性、氧化降解及自由基清除能力進行調查，當聚酯型聚胺酯奈米複合材料中含有定量的金奈米粒子(43.5至65 ppm)顯示理想的熱性質及生物性質。藉由原子力顯微鏡確認，在含有43.5 ppm或65 ppm金奈米粒子之聚酯型聚胺酯奈米複合材料中顯出不同的表面形態，相對於聚酯型聚胺酯或是不同濃度金的聚酯型聚胺酯複合材料，顯示出具有增加細胞增生、減少血小板及單核球活化及較少的細菌附著。聚胺酯及其奈米複合材料對氧化降解作用都具有抵抗性，但奈米複合材料有較大的自由基清除能力。植入老鼠19天後，奈米複合材料顯示有較小的纖維包膜厚度，表示具有低度的組織反應。奈米複合材料僅含有非常低量之金奈米粒子(43.5至65 ppm)，但生物相容性的確顯著提高，歸因於表面形態的顯著變化及自由基清除能力的結果。
In the first part, a method to prepare hydrogel nerve conduit based on chitosan and PVA was developed. The materials was characterized and the in vitro biocompatibility was investigated. The effect on repair of peripheral nerve injury in short time was be evaluated in animal studis. The functional assessment and histological analyse demonstrated that chitosan and chitosan/PVA 70 /30 conduits were better choices for nerve regeneration.
In the second part, PCL/ PEG diblock copolymers was used as model surfaces to examine the effect of nanoislands on attachment, proliferation and migration of fibroblasts, and endothelial cells. Our results showed that surface hydrophilicity improved with the increased PEG segments in diblock copolymers and that bacteria adhesion was inhibited by increased PEG contents. PCL-PEG 23:77 showed nanotopography on the surface. The number of adhered endothelial cells, platelets and monocytes on diblock copolymer surfaces was inhibited in PCL-PEG 77:23 and enhanced in PCL-PEG 23:77. Nevertheless, the platelet and monocyte activation on PCL-PEG 23:77 was reduced. PCL-PEG 23:77 had better cellular response as well as lower degree of platelet and monocyte activation. Moreover, the effect of nanoislands on migration of different cells including fibroblasts and endothelial cells was investigated. The water sliding angle of the substrates was measured. The cell migration rate was examined under a real-time optical microscope. It was found that a greater cell migration rate correlated with the smaller sliding angle of the substrate.
In the third part, two types of polyurethane-gold nanocomposites were prepared. The poly(ether)urethane-gold nanocomposite at 43.5 ppm of gold showed enhanced cellular proliferation, reduced platelet and monocyte activation and much less bacterial adhesion, relative to PU alone or nanocomposites at the other Au contents, in general. This better biocompatibility was associated with the surface morphological change in the presence of Au. The oxidative degradation in nanocomposite at 43.5 ppm of gold was also inhibited. The increased oxidative stability corresponded to the greater free radical scavenging ability of the nanocomposites. Moreover, the morphology, thermal properties, biocompatibility, oxidative degradation and free radical scavenging ability of the poly(ester)urethane-gold nanocomposites were characterized in vitro. The nanocomposite containing a certain amount (43.5-65 ppm) of gold demonstrated the optimal thermal and biological properties. The nanocomposites at 43.5 ppm or 65 ppm of gold exhibited a different surface morphology confirmed by the AFM. They also showed enhanced cellular proliferation, reduced platelet and monocyte activation and much less bacterial adhesion, relative to PU alone or nanocomposites at the other Au contents, in general. This better biocompatibility was associated with the surface morphological change in the presence of Au. PU and nanocomposites were all resistant to oxidative degradation. The nanocomposites had greater free radical scavenging ability. After 19 days of rat implantation, the nanocomposites also showed lower tissue reaction indicated by the smaller fibrous capsule thickness. The significant enhancement of biocompatibility in the nanocomposites in such low Au contents (43.5-65 ppm) appeared to be a result of the extensively modified surface morphology as well as the free radical scavenging effect in the presence of Au.
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