聚己內酯與聚麩胺酸團聯共聚物與明膠複合敷材搭載兒茶素於傷口癒合之應用研究

傷口癒合過程中適當的發炎反應，可調節免疫系統抵禦外來物質的入侵，但反之則會引起細胞壞死。本研究目的是製備天然與合成高分子複合傷口敷材，藉由合成聚己內酯與分子量8k、15k、28k、35k之聚麩胺酸，形成團聯共聚物，並利用黏度法、傅立葉轉換紅外線光譜儀、核磁共振儀與凝膠滲透層析儀鑑定化學結構與分子量。利用體外傷口癒合試驗，可知聚麩胺酸團聯共聚物可促進纖維母細胞移動與增生的能力為控制組(單純細胞培養)的116 %，敷材以10 %明膠為主混合聚己內酯與聚麩胺酸團聯共聚物，經由0.7 % 梔子素交聯形成半互穿型網狀分子結構，經由冷凍乾燥法成形。掃描式電子顯微鏡觀察敷材表面為光滑表面，截面可看到約50-100 μm的孔洞，在純水中具有436- 529 %的膨潤度，另一方面，利用溶劑萃取綠茶兒茶素，經液相層析定量兒茶素中主要成分為表没食子兒茶素没食子酸酯(EGCG)，每克兒茶素含0.974 克EGCG，以自由基分子，1，1-二苯基-2-三硝基苯肼進行體外抗氧化試驗，結果顯示兒茶素在10分鐘之內清除95 % 自由基，半抑制劑量為10.3 mg/L，與EGCG標準品EC50 = 10.9 mg/L相同擁有很高的抗氧化能力，利用脂多醣刺激巨噬細胞評估抗兒茶素抗發炎之能力，在濃度22.9 μg/mL的兒茶素下，不論先加入兒茶素後刺激巨噬細胞，或是先刺激巨噬細胞後加入兒茶素均能減緩巨噬細胞產生發炎物質。最後經大鼠傷口癒合實驗，可得知明膠混合聚麩胺酸團聯共聚物敷材能夠使大鼠傷口14天內達91 %的傷口癒合率，進一步經由觀察組織切片內表皮層厚度與免疫細胞的情形，可得知兒茶素具有幫助傷口處上皮組織重建與減少傷口處發炎細胞數目的情形。

關鍵字

明膠；傷口癒合；聚己內酯與聚麩胺酸團聯共聚物；兒茶素

並列摘要

Wound healing is a complex physiological process and requires dressing that exhibited the following properties: maintain moist environment at the wound/dressing interface and absorbs excess exudates without leakage to the surface of the dressing and non-toxic. Hence, this study aims to prepare a biocompatible wound dressing material that exhibits the above properties. The material is compose of a natural and synthetic polymer composite by linking polyethylene glycol-poly(ε-caprolactone), PEG-PCL and, with poly(γ-glutamic acid), PGA with different molecular weights on both ends of the block copolymers. The material was characterized by solution viscometry, fourier transform infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, and gel permeation chromatography. The study is composed of two parts: first is the preparation of the wound dressing material. The second part is the in vivo investigation of the wound healing of the dressing and anti-inflammatory property of catechin extracted from green tea. In vitro wound healing experiment showed that PGA-PCL-PEG-PCL-PGA block copolymers can promote fibroblast migration and proliferation at 116 % ability which exceeded that of the control cells. Using gelatin crosslinked with 0.7% genipin to mix with the block copolymers forming a semi-interpenetrating network, IPN structure through the lyophilization method to form the final dressing. SEM micrographs showed that the dressing cross section was porous with pore sizes in the range of 50-100 μm. The wound dressing swelling ratio was 436 - 529 % and can maintain moist environment at wound/dressing interface for 7 days. Catechin extracted from green tea was composed of 97.4 % epigallocatechin gallate (EGCG) assayed using High-performance liquid chromatography, HPLC. The catechin showed an anti-oxidant capacity because it can remove 95 % of the DPPH radicals within 10 minutes with 50 % effective inhibition concentration (EC50) of 10.3 mg/L. In vitro anti-inflammatory experiment showed that catechin exhibited an anti-inflammatory property. Results showed that a concentration of 22.9 μg/mL of catechin can slow down the LPS stimulation macrophages to produce nitric oxide. The dressing material was used as skin patch on excised rat skin for in vivo wound healing ability of the dressing. Results showed that gelatin based dressing exhibited a wound recovery of 91 % within 14 days. The catechin was loaded to the gelatin-based dressing for anti-inflammatory in vitro study. Histological results showed that gelatin based dressing loaded with catechin can reduce wound inflammation and promote epithelial tissue reconstruction.

並列關鍵字

wound healing ； catechin ； gelatin ； poly(ε-caprolactone)-poly(γ-glutamic acid) block copolymer

參考文獻

71. 何怡儂，促進綠茶萃取物生體可利用率之纖維素經皮遞藥系統，中原大學碩士論文，民國98年。

87. 邱鉅樺，明膠與聚己內酯與聚麩胺酸團聯共聚物複合支架於小腸組織工程之研究，中原大學碩士論文；民國100年。

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聚己內酯與聚麩胺酸團聯共聚物與明膠複合敷材搭載兒茶素於傷口癒合之應用研究

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