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標題: | 褐藻醣膠/幾丁聚醣薄膜與氧化石墨烯/幾丁聚醣薄膜於抗沾粘應用 Fucoidan/Chitosan Film and Graphene oxide/Chitosan Film in Prevention of Post-surgical Adhesion |
作者: | Guan-Wei Chen 陳冠維 |
指導教授: | 黃義侑 |
關鍵字: | 術後沾粘,抗沾黏薄膜,幾丁聚醣,褐藻醣膠,氧化石墨烯, post-surgical adhesion,anti-adhesion barrier,chitosan,fucoidan,graphene oxide, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | 沾粘仍然是手術後一大隱憂,病患可能會因為手術後續處理不慎,或因個人體質關係,導致腸道阻塞、肚子痛,甚至因卵巢沾粘形成不孕症。然而抗沾粘薄膜仍然有使用限制,例如需在無血情況下使用,容易粘附在手術手套上,加上其單價高昂,因此我們選用廣泛應用在生醫上的幾丁聚醣為基材,搭配褐藻醣膠和氧化石墨烯,製成褐藻醣膠/幾丁聚醣薄膜以及氧化石墨烯/幾丁聚醣薄膜以應用在抗沾粘材料上。
幾丁聚醣可促進傷口癒合、抗菌性質,搭配褐藻醣膠的抗發炎、抗血管新生的特性,在玻尿酸薄膜中加入褐藻醣膠可降低沾粘,與氧化石墨烯的抗菌性、生物相容性互相結合,製成薄膜。因為組織來自於細胞,如果細胞不貼附在材料上,就不會有組織沾粘的問題。把基材的幾丁聚醣與褐藻醣膠以1:1的比例混合平舖在幾丁聚醣基材上,以抓住更多的褐藻醣膠;另一方面,氧化石墨烯製備後經傅立葉轉換遠紅外線光譜儀(FTIR)定性測出羰基與羥基,尺寸達10微米;褐藻醣膠可以增加硫基,這些官能基可降低NIH-3T3細胞對材料的貼附力,然而加入這些材料,表面的粗糙度經由也會上升至320nm左右,親水性增加,接觸角降低約30-400,可以幫助材料更加貼附在受傷組織表面。薄膜的生物相容性皆達90%,細胞貼附性深受官能基與粗糙度影響,透過光學顯微鏡、螢光顯微鏡、掃描式電子顯微鏡可以觀察褐藻醣膠與氧化石墨烯的減少細胞貼附的能力。動物實驗中發現盲腸附近沒有殘留的薄膜,表示在傷口修復過程中,材料已完全分解。解剖後進行抗沾粘評估,觀察到純幾丁聚醣薄膜的抗沾粘分數些微下降至2.3分,比控制組3分來得低,而褐藻醣膠與氧化石墨烯的加入更加使抗沾粘分數下降至0-0.3分,H&E染色更加看清細部的沾粘層的消失,故這兩種材料運用在術後沾粘是有幫助的。 Post-surgical adhesion remains such a severe problem that patients suffer complications such as bowl obstruction, chronic abdominal pain and infertility. However, the usage of the relative commercial products is limited such as the bloodless condition during surgeon. The price is high due to the value of the materials. For reducing the expenses of the anti-adhesion products, we are trying to use chitosan film, fucoidan/chitosan film and graphene oxide/chitosan film in anti-adhesion application. The standing point we are concerned about is cell anti-adhesion properties of these materials. Chitosan accelerates wound healing process and possesses anti-bacterial property. Fucoidan added in the hyaluronic acid film lower the adhesion grade with anti-inflammation, anti-angiogenesis properties. Graphene oxide owns anti-bacterial property and biocompatibility with limited content. We combine fucoidan with little chitosan content to catch more water-soluble fucoidan and let the mixture coat on chitosan film. On the other hand, graphene oxide we prepared is qualitatively proved by Fourier transform infrared spectroscopy (FTIR) to get carbonyl, carboxyl and hydroxyl group. The size of graphene oxide is 10μm observed by transmission electron microscopy (TEM). The addition of the fucoidan and graphene oxide is used to decrease the affinity of NIH-3T3 fibroblasts based on the sulfur functional group and chiefly carboxyl group respectively. Following from that, the surface roughness evaluated by atomic force microscopy (AFM) with addition of second material is approximately 320nm, which is twice as rough as the pure chitosan film. The contact angle test with 30-40 degree reduction also tells the additional materials raise the hydrophilicity of film surfaces, which assists the film adhesion to the wound area during the abdominal surgeries. Cell viability by MTT assay test are all getting up to 90%, which would not be an issue in each kind of film, even with limited graphene oxide content. NIH-3T3 fibroblasts tend to inhibit the linkage from chitosan film with fucoidan or graphene oxide rather than pure chitosan film as a result of the interaction of surface roughness and functional groups through MTS assay tests and cell image by optical microscopy (OM), fluorescence microscopy (FM) and scanning electron microscopy (SEM). Animal study evaluation shows that chitosan film lower the adhesion grade from 3 to 2.3. The fucoidan/chitosan film and graphene oxide/chitosan film are obviously decreased the tissue adhesion score to 0-0.3. Additionally, residues of films cannot be found near the cecum wound, which means the biodegradability of the film within the wound healing process. H&E stain reveals the adhesion layer disappears when adding fucoidan or graphene oxide in chitosan film. These results firmly suggested that the fucoidan/chitosan film and graphene oxide/chitosan film are favorable biomaterials in application of post-surgical adhesion. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61959 |
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顯示於系所單位: | 醫學工程學研究所 |
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