針對糖尿病足潰瘍設計並製作一種抗菌且能促進傷口癒合的甲殼素複合式水凝膠之研究;Design and Fabrication of an Antimicrobial-Wound Healing-Enhanced Chitosan Complex Hydrogel for Diabetic Foot Ulcer Treatment.

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題名:	針對糖尿病足潰瘍設計並製作一種抗菌且能促進傷口癒合的甲殼素複合式水凝膠之研究;Design and Fabrication of an Antimicrobial-Wound Healing-Enhanced Chitosan Complex Hydrogel for Diabetic Foot Ulcer Treatment.
作者:	洪翊玲;Hong, Yi-Ling
貢獻者:	生物醫學工程研究所
關鍵詞:	第二型糖尿病;糖尿病足潰瘍;殼聚醣;銀離子;表皮生長因子;傷口癒合;Type 2 diabetes mellitus;Diabetic foot ulcer;Chitosan;Silver ions;Epidermal growth factor;Wound heling
日期:	2019-10-31
上傳時間:	2020-01-07 14:26:03 (UTC+8)
出版者:	國立中央大學
摘要:	糖尿病中最常見的是第二型糖尿病，屬於一種慢性疾病，它占糖尿病患者總人口數的 90-95%。通常會有較長的潛伏期，第二型糖尿病患者主要原因是自身的血液循環較差、新陳代謝異常和炎症時期較長等各種原因，導致傷口癒合緩慢，嚴重者會影響到患者的生活品質或導致死亡。而本研究中開發一種包含銀離子和包裹表皮生長因子 (EGF)之奈米粒子的殼聚醣水膠 (CPCHG)，此水膠可以提供抗菌能力也因表皮生長因子的存在幫助傷口癒合。本研究目前先以牛血清白蛋白(BSA)作為模擬蛋白代替表皮生長因子。奈米載體的製備方式以聚陰離子三聚磷酸鹽作為交聯劑與殼聚醣進行交聯製備包覆 BSA 的殼聚醣奈米載體(CTBNPs)。經過 DLS 分析，CTBNPs 的粒徑大小與表面電荷分別為 265±26 nm 和 30.02±1.42 mV；CTBNPs 在 48 小時的釋放率為 89% 但在 CPCHG 的 BSA 釋放約為 70%。在銀離子釋放分析中，銀離子在 48 小時中 4°C 和 37°C 的釋放量為 5.11 mg/L 及 6.53 mg/L。此外，在細菌抑制中，使用銀離子濃度為 24 mM 時，對於金黃色葡萄球菌有顯著抑制的效果；在細胞毒性試驗中，細胞的存活率也可達到 90%。生長曲線試驗中，將模擬蛋白(BSA)更改成表皮生長因子(EGF)製成的殼聚醣水膠(CPEHG)。使用不同濃度表皮生長因子(EGF)去測試 NIH-3T3，結果表明當 EGF 的濃度來到了 60 μg/mL 可讓細胞生長且不受銀離子的影響。選定水膠(CPEHG)最終配方為 60μg/ mL EGF 和 24 mM 銀離子濃度。在動II 物實驗中，我們使用 Sprague Dawley（SD）大鼠並注射鏈脲佐菌素觀察 2 周確定為糖尿病大鼠後在大鼠背上製造傷口並且使用敷料治療觀察傷口癒合情況。結果表明在各組實驗組中 CPEHG 水膠的傷口癒合速度最快、炎症細胞較少和有較多的膠原蛋白沉積，未來可用於治療糖尿病足傷口敷料。;Type II diabetes mellitus (T2DM) is the most common form of diabetes, accounting for 90-95% of diabetics, and is a chronic disease often preceded by a long asymptomatic period. Most of T2 diabetics suffer with delay wound healing resulted from poor blood circulation, prolonged inflammation and lack of growth signal due to disordered metabolism, and it may severely affect patients’ life quality and even cause death. To overcome the aforementioned issues, a synthetic chitosan-based hydrogel containing silver ions and growth factor-entrapped nanoparticles that may provide both antibacterial and would healing-enhanced functionalities were developed in this study. As being the preliminary investigation, the bovine serum albumin (BSA) was employed as the model protein molecule to substitute the practical growth factor protein in this study. Furthermore, the BSA-encapsulated chitosan nanoparticles were fabricated by using the polyanion tripolyphosphate as the coacervation crosslink agent. Through the DLS analysis, the size and surface charge of the BSA-loaded nanoparticles(CTBNPs) were 265 ± 26 nm and 30.02 ± 1.42 mV, respectively. The encapsulation efficiency and the loading rate of the BSA in the nanoparticles were 74± 8.62% and 1.23 ± 0.55%, respectively. Also, the release rate of BSA from the nanoparticles was 89% within 4 h, while that from the synthetic hydrogel was about 70% within the same time course. The nanoparticles were successful insides the chitosan hydrogel. From hydrogel property test, BSA releases were 70% . When the silver ions released in 48 hours, the 4 and 37 degrees release were 5.11 and 6.53 mg/L. Moreover, the bacterial inhibit study, when used 24 mM silver ions concentration the bacterial inhibit was significant. The hydrogel toxicity for the cell. In the cell study, we used 24 mM silver ions concentration the cell viability came to 90 %. In the growth curve study, we decide used epidermal growth factor with different concentration. However, when EGF came to 60 μg/mL cell weren′t affected by silver ions. We decided that the final formulation of the hydrogel was 60 μg/mL EGF and 24 mM silver ion concentration.In animal experiments, We used Sprague Dawley (SD) rats and injected with streptozotocin to observe wound healing on rats′ backs after 2 weeks of diagnosis as diabetic rats and to observe wound healing using dressings. The results showed that the wound healing rate of CPEHG water gel in each group was accelerated, inflammatory cell implantation and complication of collagen deposition, which could be used to treat diabetic foot wound dressing in the future
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