電漿接枝中藥之傷口照護材的抗菌研究

The Study of Antibacterial Activity of Wound Care Products with Chinese Medicine Using Plasma Grafting Technology

徐翊庭(Yi-Ting Shu)

指導教授 ： 翁清松

中原大學/工學院/生物醫學工程研究所/博士(2017年)

https://doi.org/10.6840/cycu201700151

摘要

本研究以天然無毒含中藥敷材的方向進行開發，主要乃因中藥之使用相較於合成抗菌物較無副作用、無危害性等。在中藥的篩選是以古典文獻記載能促進傷口修復之功效（如抗菌、抗發炎、美白等）及高使用率為依循選用六種中藥（白芷、白朮、當歸、梔子、紫草及何首烏）進行不同重量之原藥材及中藥萃取後的抗菌效果比較，藉此初步選取適當之萃取物濃度，以作為後續細胞毒性及電漿技術實驗之中藥添加依據。
實驗之抗菌敷材是以聚酯（PET）短纖製成的針軋不織布為基材，再運用射頻電漿進行基材的表面活化處理，以接枝共聚合方式進行兩階段表面改質以製成含中藥之基材。其中，電漿接枝所採用之中藥，是依中藥的萃取率、藥材泛用性、敷材接枝率及藥材以往在皮膚美白效果的應用性等因素考量下，選擇了白朮及當歸來進行其抗菌敷材的製作。另外，經電漿處理後之基材亦會透過相關的物化性檢測（如：接觸角及FTIR等）、抗菌性及細胞毒性檢測來確認其基材經接枝後的效果，由此探討開發抗菌傷口照護材之可行性。
實驗結果顯示，原藥材抗菌試驗中，其抗菌效果並未非常顯著，整體上六種原藥材要添加至1000 mg才能呈現抑菌性；但若中藥經萃取後的抗菌則是呈現全面提升的趨勢，100 mg中藥萃取物就能顯現抑菌性。另外，中藥萃取物調配液濃度為1 mg/ml且細胞反應作用達48hr後，其促進細胞增生效果為最佳。抗菌敷材部份，是以2 mg/ml之中藥萃取物調配液濃度來進行中藥接枝之敷材製作，在PET不織布經電漿及丙烯酸（AAc）單體處理後，含白朮及當歸的敷材之接枝率分別為51.24%及82.36%，均顯著高於未經電漿處理的敷材。
在敷材的接觸角分析結果顯示PET表面經電漿處理後之接觸角均明顯變小。且透過SEM圖初步印證電漿處理對疏水性材料在表面改質上的有效性，可使PET不織布達到根本性的親水改質；透過ATR-FTIR分析，亦可看出中藥萃取物被成功地接枝到PET不織布基材上，且顯示出白朮水萃取物中主要存在的白朮多醣和類黃酮之吸收峰，以及當歸水萃取物中主要含有的當歸多醣和阿魏酸等親水性極性成分；另外，敷材的抗菌活性結果顯示，經過電漿及AAc處理後對金黃色葡萄球菌和大腸桿菌有更顯著的抗菌活性。以上數據表明了電漿表面改質技術是有助於提高含中藥萃取物之敷材的可加工性和抗菌效果；最後，由L929纖維母細胞之細胞毒性分析結果，亦可得知本研究所開發的白朮及當歸萃取物之傷口照護材（抗菌敷材）均被評定為0級，則代表不具細胞毒性反應。因此，本研究之加工方法與科學化驗證之結果，可提供作為未來天然植物抗菌敷材及衍生產品的開發方向與應用之參考。

關鍵字

傷口照護材 ； 市場分析 ； 中藥萃取 ； 電漿接枝 ； 抗菌 ； 細胞毒性

並列摘要

This study was done to develop a natural and non-toxic antibacterial dressing containing extract of traditional Chinese medicines because of its less side effects and harmfulness compared with synthetic antibiotics. In the screening of herbal medicines in this study, six Chinese medicines (Radix Angelicae Dahuricae, Rhizome Atractylodes macrocephalav, Radix Angelicae sinensis, Fructus Gardeniae, Radix Arnebiae and Radix Polygoni Multiflori) were chosen according to their wound repair efficacy (e.g. anti-bacterial, anti-inflammatory, skin-whitening, etc.) and the utilization rate recorded in the classical literatures. The antibacterial acitivities of original medicines and their extracts were evaluated and compared at different weights to determine the appropriate concentrate of extract of Chinese medicines in the subsequent cytotoxicity and plasma experiments.
In addition, the needle-punched nonwoven fabrics made by the polyethylene terephthalate (PET) staple fibers were used as the substrates of antibacterial dressing, and treated with radio frequency plasma-induced grafted technology to carry out the surface activation, and then grafted with the Chinese herbal extracts to fabricate the antibacterial herbal dressing by two-stage surface modification method. Rhizome Atractylodes macrocephala (RAM) and Radix Angelicae sinensis (RAS) were eventually chosen to use in antibacterial dressing according to some factors, such as their results in extraction yield and grafting rate of dressing, the utilization frequency, and the skin whitening efficacy in our past research. The effectiveness of gratfing of Chinese extracts onto PET substrates after plasma treatment were investigated through the assays of physical and chemical properites (contact angle, FTIR, etc.), antibacterial activities and cytotoxicity to evaluate the feasibility of development of antibacterial wound care products with Chinese herb extract.
The antibacterial test results showed that the antibacterial effect of the original medicines was not very significant, and the six original medicines must be reached in dosage of 1000 mg that would be present their antibacterial effect. However, their antibacterial effect can be improved signicantly after extraction, and the effective dosage can be reduced to 100 mg. The cell proliferation was the best at 1 mg/ml of deployment concentration of Chinese herb extract for 48hr. In the part of the preparation of antibacterial dressing, 2 mg/ml of deployment concentration of Chinese herb extract was applied in the grafting onto substrate. In the assessment results of effectiveness of herbal dressings, the grafting rate of RAM and RAS extracts onto PET nonwovens after plasma and acrylic acid treatment were 51.24 % and 82.36 %, respectively, which were significantly higher than those without plasma treatment.
The result of water contact angle showed that the contact angle of surface of PET nonwovens reduced significantly after plasma treatment, which confirm the validity of plasma treatment in the surface modification for hydrophobic materials and result in the hydrophobicity improvement of surface characteristics of PET nonwovens from hydrophobic to hydrophilic. The successful grafting was also confirmed from SEM and ATR-FTIR results by the appearance of new peaks associated with the presence of Atractylodes macrocephala polysaccharides and flavonoids in RAM extract and Angelica sinensis polysaccharides and ferulic acid in RAS extract. In addition, the plasma treatment further increased the broad-spectrum antibacterial effects of PET nonwoven dressings containing RAM and RAS extracts. Finally, the L929 cytotoxicity result indicated that these antibacterial dressings with RAM and RAS extracts are both non-cytotoxic and encourage cells to grow in a single layer with good cellular attachment. Therefore, the results of this research method and scientific verification of this study could be used as a reference for the future development and application of natural plant antibacterial dressing and derivative products.

並列關鍵字

wound care products ； market analysis ； Chinese herb extract ； plasma-induced grafting ； antibacterial ； cytotoxicity

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