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    Please use this identifier to cite or link to this item: http://tkuir.lib.tku.edu.tw:8080/dspace/handle/987654321/87909


    Title: 幾丁聚醣磷酸鈣複合材料之製備及作為引導骨再生薄膜之評估
    Other Titles: Preparation of chitosan/calcium phosphate composite materials useful for the guided bone regeneration
    Authors: 戴宏穎;Tai, Hung-Yin
    Contributors: 淡江大學化學工程與材料工程學系博士班
    董崇民;Don, Trong-Ming
    Keywords: 幾丁聚醣;磷酸鈣;引導骨再生;Chitosan;calcium phosphate;guided bone regeneration;asymmetric composite membrane
    Date: 2012
    Issue Date: 2013-04-13 11:51:33 (UTC+8)
    Abstract: 本研究利用幾丁聚醣作為引導骨再生膜的主材料,幾丁聚醣具有良好細胞相容性、無毒性、生物可分解性、成膜性以及價格便宜等特性,加上本身具有抑菌性,可防止傷口細菌感染。而為了加速引導骨再生,另外在幾丁聚醣中加入不同結晶型態之磷酸鈣混合成複合材料,藉由磷酸鈣不斷釋放出鈣離子來引導骨性細胞吸附及加速骨組織的再生,同時亦藉著磷酸鈣來改善幾丁聚醣的機械性質。本研究乃利用逆乳化法製備缺鈣之氫氧基磷灰石(CDHA),然後經800°C與1000°C高溫煆燒後,可分別轉換成結晶性較高且同時含有缺鈣氫氧基磷灰石及磷酸三鈣(TCP)的雙相磷酸鈣(BCP)與磷酸三鈣(TCP),將不同的磷酸鈣(CDHA, BCP, TCP)引入幾丁聚醣之中,並以冷凍乾燥法製備成多孔結構薄膜(chitosan-P, CDHA/chitosan-P, BCP/chitosan-P, TCP/chitosan-P),適當的孔洞尺寸與孔隙度,可以讓細胞或組織的增生能力提高,但使其不易侵入薄膜而破壞薄膜結構。實驗發現,骨母細胞較易貼附於含有CDHA結晶相態的CDHA/chitosan-P與BCP/chitosan-P孔洞薄膜上,而骨母細胞在具有TCP相態的BCP/chitosan-P與TCP/chitosan-P孔洞薄膜上則有較強的細胞增生能力,最終骨母細胞分化與礦化時,CDHA/chitosan-P與 BCP/chitosan-P孔洞薄膜中的CDHA結晶相態又可引導骨母細胞分化形成新生骨礦物,總結來說,冷凍乾燥法製備之薄膜具有的多孔結構,可讓牙齦纖維母細胞在薄膜的一側生長且無不良反應,而對另一側骨缺損的部份,又有加強其骨細胞再生的能力。其中又以BCP/chitosan-P因同時擁有CDHA與TCP兩結晶相態,讓BCP/chitosan-P具備與骨母細胞貼附相關的骨親合性,同時還具有加速骨母細胞增生,與增強骨母細胞分化礦化的能力,因此在引導骨再生材料上,BCP/chitosan-P確實擁有其應用發展的空間。將幾丁聚醣溶液直接冷凍乾燥可製備得到單純孔洞型態之幾丁聚醣薄膜,其材料較為柔軟,操作性較不佳。為了這個缺失,本研究在幾丁聚醣孔洞膜的一側加上一層生物可分解聚羥基丁酯(PHB)緻密層,將PHB薄膜電漿接枝聚丙烯酸,並以聚丙烯酸為架橋固定幾丁聚醣,直接將幾丁聚醣冷凍乾燥製備出非對稱型幾丁聚醣/PHB複合薄膜,此種薄膜不論在乾燥或濕潤的狀態下,材料皆具有優異的起始模數,最終,在動物實驗方面,本研究比較不同磷酸鈣(CDHA, BCP與TCP)與幾丁聚醣製備之多孔膜,將材料貼附在大鼠頭蓋骨缺損位置21天後,可以發現BCP/chitosan-P薄膜覆蓋在骨缺損部位後,可以在21天內將骨缺損恢復將近58%,大幅的加速骨修復之速度。本研究所製備之添加BCP顆粒之幾丁聚醣孔洞薄膜確實具有在引導骨組織再生膜的應用發展之空間。
    To fulfill the properties of barrier membrane useful for guided bone tissue rgegenration in the treatment of periodontitis, a simple process containing oven-drying and lyophilization was proposed in this study to produce composite membrane from biodegradable chitosan (CS) and fnctional calcium phosphate. Calcium deficient hydroxyapatite (CDHA) having an average particle size of 45 nm was synthesized by reverse emulsion method. It was converted to the respective biphasic calcium phosphate (BCP, 226 nm) and ß-tricalcium phosphate (TCP, 450 nm) by calcination at 800°C and 1000°C, and the BCP consisted of 92% TCP and 8% CDHA. Subsequently, chitosan was mixed with calcium phosphates to prepare CDHA/chitosan, BCP/chitosan, and TCP/chitosan membranes. The lyophilization at -20°C then gave the rest of material an interconnected pore structure with high porosity (91~95%) and suitable pore size (102 to 147 μm) which could reveal the biocompatibility and prevention of migration of gingival fibroblast. Furthermore, the composite membrane promoted the permeability and adhesiveness to bone cells as demonstrated by the in-vitro cell culture of primary osteoblast. The CDHA and BCP particles separately added into chitosan could further increase the cell attachment and differentiation of osteoblast, and then the BCP and TCP particles separately added into chitosan could increase great cell proliferation of osteoblast. This is beneficial for maintaining a secluded space for the bone regeneration as well as to prevent the invasion of other tissues. On the other hand, asymmetric chitosan/PHB composite membrane was produce to promote the wet operation of barrier membrane. The dense PHB membrane grafted poly(acrylic acid) by plasma induced polymerization, then chitosan was immobilized by reacting with poly(acrylic acid) and lyophilized. The asymmetric membrane (chitosan/PHB) with a skin PHB layer had initial modulus value almost 50 times that of the symmetric porous membrane (chitosan-P). Finally, five different samples of a control, chitosan, CDHA/chitosan, BCP/chitosan, and TCP/chitosan porous membrane were assessed as periodontal barrier membranes for the calvarial critical size bone defects in SD rats. Histological and histomorphometric analyses revealed that BCP/chitosan resulted in the most effective bone regeneration compared to other samples with rgenerated bone area of 57% covering the bone defect area. The BCP/chitosan has the potential to be used as the barrier membrane for guided bone regeneration.
    Appears in Collections:[化學工程與材料工程學系暨研究所] 學位論文

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