海藻酸鈉/透明質酸/幾丁聚醣複合敷料之製備技術及其生物適應性評估

Technology and Biological Adaptation Assessments of Sodium Alginate / Hyaluronic Acid / Chitosan Composite Dressing

吳松濂(Song-lien Wu)

指導教授 ： 樓靜文

中臺科技大學/健康科學院/醫學工程暨材料研究所/碩士(2010年)

摘要

具有生物特性多醣體的生醫材料被廣泛應用於眾多醫療與手術當中。本研究選用幾丁聚醣(Chitosan,Cs)、海藻酸鈉(Alginate,Alg)以及透明質酸(Hyaluronic Acid，HA)等多醣體生醫材料,複合聚乳酸(Polylactic Acid,PLA)纖維與脫脂棉以製備醫用敷料並探討其相關物性及生物相容性。
在實驗製程方面可分為兩大部分,第一部分為製備敷料使用的不織布基布,其中變化PLA纖維與脫脂棉混合比例,再經喂棉,開棉,給棉,梳棉,棉網舖疊基重,軋針密度等製程。接著對其進行抗拉伸強力,抗撕裂強力,吸水性,透氣度,水氣透過率,柔軟度等物性測試及分析,藉以尋求敷料基布之最佳化製程參數。
第二部分則為進行複合敷材之抗菌層與膨潤層製備,抗菌層選用幾丁聚醣為材料,再以噴霧方式塗佈於前述最佳化之基布,並對複合基布進行抑菌測試。膨潤層係使用氯化鈣溶液交聯之海藻酸鈉/透明質酸混合溶液,以冷凍乾燥法製備成多孔狀薄膜,之後進行細胞培養,測試不同透明質酸濃度對細胞之影響,尋求最佳化透明質酸濃度薄膜。接續對其進行膨潤度,重量損失率,接觸角及掃描式電子顯微鏡(SEM)觀察形態等物性測試及分析,且測試不同交聯濃度對膠體之影響。最後將兼具抗菌層與膨潤層之功能性複合敷料進行動物傷口癒合之生物適應性實驗及其應用評估。
將PLA纖維與脫脂棉纖維混合後,經由不織布基布製程,可製備出具有一定強力之基布,最佳化混棉比例為8：2,幾丁聚醣複合基布經交聯後之海藻酸鈉/透明質酸薄膜具有良好之結構穩定性,在測試中浸泡於磷酸緩衝溶液(PBS)的環境下96小時,損失率其重量皆能保持40 %以下。進行金黃色葡萄球菌抑菌試驗時,以幾丁聚醣濃度3 wt%,噴塗量0.154 g/cm2有最佳化之抑菌效果,本論文所製作之海藻酸鈉/透明質酸/幾丁聚醣複合PLA/脫脂棉不織布基布之水氣透過率約為2039 ± 178.8 g/m2day,符合文獻中的水氣透過率範圍。本研究使用L929纖維母細胞進行MTT細胞活性測試觀察得到HA濃度為3mg/ml具有最佳細胞活性。將海藻酸鈉/透明質酸薄膜與細胞共培養24 h,結果顯示以氯化鈣添加濃度0.15 %所製備之海藻酸鈉/透明質酸薄膜有最佳之細胞貼附性。最後藉由動物體外試驗結果觀察,海藻酸鈉/透明質酸/幾丁聚醣複合PLA/脫脂棉不織布基布複合敷料確實具有促進傷口癒合效果。

關鍵字

複合敷料 ； 幾丁聚醣 ； PLA ； 海藻酸鈉 ； 透明質酸

並列摘要

Abstract
Polysaccharide is the most prevalent material in the surgery. Chitosan, Sodium alginate and Hyaluronic Acid were compound with Polylactic acid fiber and absorbent cotton into a composite dressing whose biological and mechanical property were studied.
Two phases were included in this study. The optimum manufacturing parameter of the base fabric was earned in the first phase. PLA and absorbent cotton were mixed in different ratios before the mixture were processed in the following procedures: feeding, opening, channeling, carding, lamination and needle punching. Tensile strength, tear strength, moisture absorption, air permeability, vapor permeability, and flexibility of the base fabrics involving various ratios of PLA and absorbent cotton were explored.
The anti-bacterial layer and swelling layer were prepared in the second phase. Chitosan, the dominant anti-bacterial material, was spray-coated upon the base fabric chose on the basic of the result in the first phase. An anti-bacterial test was carried out on this Chitosan-coated layer. The swelling layer was prepared by crosslink-ing the sodium alginate/Hyaluronic acid solution with calcium chloride. Next, the sodium alginate/Hyaluronic acid/calcium chloride was freezing dried into a porous membrane on which cells were later cultured upon for effects of Hyaluronic acid concentrations on cell vitality. After the best Hyaluronic acid concentration was earned, swelling rate, weight loss rate, far-infrared ray, contact angle and SEM of this sodium/Hyaluronic acid/calcium chloride membrane were also examined for effects of cross-link concentrations on the membrane’s mechanical property. Finally, the optimum anti-bacterial layer and the swelling layer were compounded with the base fabric, so a composite dressing was ready for further animal tests and application evaluations.
When the PLA and absorbent cotton were mixed and processed, a base fabric with promising strength was yielded. The best mixing ratio was 8:2 (PLA: absorbent cotton). The Alg/HA membrane is firmly structured after Chitosan was cross-linked. After the Chitosan layer was immersed in the PBS for 96 hours, its average weight loss rate was under 40 %. The anti-bacterial test on staphylococcus aureus exhibited that coating the 3 wt% Chitosan in 0.154 g/cm2 brings out the best anti-bacterial effect. The average moisture permeability of this sodium alginate/ Hyaluronic acid/PLA/ absorbent cotton composite dressing was 2039±178.8 g/m2 day. According to previous studies, the result was acceptable.
The MTT cell vitality test was conducted using L929. The excellent cell survival rate was found in the HA concentration of 3 mg/ml. The Alg/HA membrane was cultured with L929 for 24 hours. The result indicated that the 0.15 % calcium chloride achieved sound cell adhesive effect. Last but not least, the animal test also confirmed that this sodium alginate/Hyaluronic acid/Chitosan/PLA/ absorbent cotton composite dressing accelerated healing.

並列關鍵字

Hyaluronic acid ； Sodium alginate ； composite dressing ； Chitosan ； PLA fiber

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