源自不同真菌之幾丁質材料結合血小板破膜釋出液對於慢性傷口癒合之探討

Study of chitins isolated from fungi combined with platelet lysate on chronic wound healing

李漢楨(Han-Jen Li)

指導教授 ： 蘇慶華

臺北醫學大學/口腔醫學院/生醫材料暨工程研究所/碩士(2009年)

https://doi.org/10.6831/TMU.2009.00135

摘要

本實驗探討兩種真菌來源幾丁質包括SACCHACHITIN與RHIZOCHITIN是否可促進糖尿病慢性傷口的癒合，並評估其成效。實驗分別透過高糖細胞模型以及化學藥劑Streptozotocin(STZ)誘導糖尿病動物模型探討傷口癒合機制。細胞增生實驗中，Hs68纖維母細胞株先以高糖培養液(110 mM glucose)培養24小時，再分別加入不同濃度的SACCHACHITIN與RHIZOCHITIN(100, 250, 500 μg/ml )，於7天後以MTS assay分析其細胞存活率。結果顯示，在高糖環境下，SACCHACHITIN與RHIZOCHITIN皆能提高細胞的存活率。以in vitro scratch assay觀察細胞遷移的情形。結果顯示，在高糖環境下，SACCHACHITIN與RHIZOCHITIN亦能促進細胞移行的能力。動物實驗中，於STZ誘導型糖尿病Wistar Han IGS公鼠背部製作全層皮膚傷口(4 cm2)，分別以SACCHACHITIN與RHIZOCHITIN薄膜處理之，分於3、7、14、21天觀察傷口面積的變化。傷口面積試驗顯示，SACCHACHITIN與RHIZOCHITIN皆能促進糖尿病傷口的癒合。實驗進一步於SACCHACHITIN與RHIZOCHITIN中添加血小板破膜後富有生長因子之GF18，製成新的複合創傷敷料，評估其對於糖尿病導致慢性傷口癒合之成效。傷口面積試驗顯示，SACCHACHITIN結合GF18複合敷料與RHIZOCHITIN結合GF18複合敷料更具促進癒合之效果。將取下傷口樣本以酵素連結免疫法(Enzyme-linked immunosorbent assay, ELISA)分析其Transforming growth factor-beta1(TGF-β1)的分泌量，結果顯示，SACCHACHITIN、 RHIZOCHITIN、GF18及新的複合材料皆能在不同時期增加TGF-β1的分泌，且與傷口面積變化的趨勢相似，顯示敷料可能透過調控傷口部位生長因子的含量，達到促進傷口癒合的效果。

關鍵字

SACCHACHITIN ； RHIZOCHITIN ； GF18 ； 糖尿病 ； 傷口癒合

並列摘要

In the present study, SACCHACHITIN and RHIZOCHITIN were used to evaluate the effectiveness in accelerating chronic wound healing, by employing a cell line in vitro model and a chemical-induced diabetic rat in vivo model. For In vitro, Hs68 fibroblast cells were cultured in high-glucose medium (110 mM glucose) for 24 hours and then SACCHACHITIN and RHIZOCHITIN (100, 250, and 500 μg/ml) were supplemented as suspension to the medium. After 7 days, we observed both SACCHACHITIN and RHIZOCHITIN resumed cell viability and cell mobility of Hs68 cell in high-glucose condition. In in vivo model, full-thickness skin wounds (4 cm2) were created on the dorsum of wistar diabetic rats. The cutaneous wounds were treated with membrane of SACCHACHITIN or RHIZOCHITIN, and the area of wounds was analysed at 3, 7, 14, 21 days. The data in vivo indicated both SACCHACHITIN and RHIZOCHITIN induced wound healing, significantly. Furthermore, new wound dressings were designed to combine with GF18, a mixture of human platelet growth factor, with SACCHACHITIN or RHIZOCHITIN, and the effects of the novel wound dressings on diabetic wound healing were evaluated. The data in vivo indicated the addition of GF18 optimized the recovery effect of SACCHACHITIN and RHIZOCHITIN. Transforming growth factor-beta1(TGF-β1) was assayed by ELISA in the samples of wound area. As a result, SACCHACHITIN, RHIZOCHITIN, GF18 and new wound dressings all significantly increased the production of TGF-β1 in different stages of the healing process.

並列關鍵字

SACCHACHITIN ； RHIZOCHITIN ； GF18 ； Diabetes ； Wound healing

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