CN107998439B - 一种功能性食用菌来源复合敷料的制备方法 - Google Patents

一种功能性食用菌来源复合敷料的制备方法 Download PDF

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CN107998439B
CN107998439B CN201711313907.3A CN201711313907A CN107998439B CN 107998439 B CN107998439 B CN 107998439B CN 201711313907 A CN201711313907 A CN 201711313907A CN 107998439 B CN107998439 B CN 107998439B
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程抒劼
徐辉
曹崇江
郭子君
朱震菡
王星
胡凯宇
陈贵堂
王岁楼
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China Pharmaceutical University
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Abstract

本发明公开了一种功能性食用菌来源复合敷料的制备方法，属于医疗器械产品技术领域。由如下步骤制备而成：将食用菌菌根干燥后磨粉，过筛，得到菌根粉末，密封保存备用；将菌根粉末进行脱色；将冷冻干燥至恒重的脱色菌根粉末分别于有机溶剂和碱液中浸泡后，进行微波处理；待冷却后进行离心，沉淀用水反复冲洗至无醇味，分散在去离子水中形成悬浮液；收集粒径在一定范围内的粉末，与纳米银颗粒以及金花葵精油均匀混合后，制成多孔膜；冷冻干燥即得食用菌来源复合敷料。制得的敷料能够有效促进小鼠和大鼠受损皮肤的修复与再生，该产品应用于皮肤损伤护理，进一步提高食用菌工厂化生产的附加值，增加企业利润，具有很好的经济价值和社会效应。

Description

一种功能性食用菌来源复合敷料的制备方法
技术领域
本发明涉及一种具有促进创伤愈合功效的新型食用菌来源复合敷料的制备方法,属于医疗器械产品技术领域。
背景技术
我国是食用菌生产大国，食用菌工厂化生产过程中产生的废弃菌根大多就地堆置，或直接施入田中，这造成了农业有机资源的巨大浪费。菌根来源于食用菌的子实体，和药材相比安全性更高，并且其中含有大量多糖、有机酸、果胶、酶等生物活性物质，具有高值化利用的潜力。
皮肤是人体的重要器官，它起着控制体温、防止感染及体液流失、免疫及传感的作用。由于创伤、擦伤、皮肤溃烂和烧伤等原因，可能导致皮肤的大范围伤害，严重创伤甚至可以威胁生命或导致伤残。
创伤愈合过程主要包括血小板凝集、血液凝固、纤维蛋白形成、炎性反应发生、角质细胞增殖和再生、受损组织修复和重塑。目前对创伤愈合的促愈措施主要有皮肤移植、敷料敷盖、细胞因子外用和基因治疗等。其中皮肤移植是修复大面积软组织缺损最有效的方法，但无论自体或异体皮肤移植均受到供体来源的限制。此外，异体和人工合成皮肤代用品的移植，尚未突破免疫排斥反应的瓶颈问题；传统敷料敷盖仅能够在一定程度上促进组织修复，且促愈效果不显著；细胞因子外用则存在半衰期短和治疗费用昂贵等问题；基因治疗虽然克服了细胞生长因子应用所存在的一些不足，但是由于基因转染效率低下以及转染表达的可调控性差等，技术难题难以解决，尚无法大规模应用。
本发明首次以天然来源的食用菌废弃菌根为原料，在此基础上添加具有抗菌作用的纳米银颗粒和具有抗炎、镇痛、生肌、淡化疤痕作用的金花葵精油，制备出了一种新型复合敷料。该敷料能够快速吸收创面渗出液，有效促进角质细胞再生、迁移以及肉芽组织的形成，并通过温和的急性炎性反应吸引大量的多核细胞和巨噬细胞以清除组织碎片和血凝块，从而促进创伤的修复和愈合。该产品可以变废为宝，广泛应用于皮肤损伤的后续护理，进一步提高食用菌工厂化生产的附加值，增加企业利润。
发明内容
本发明的目的是解决现有技术中食用菌废弃菌根浪费以及市面上可用于皮肤损伤护理的经济有效的医疗产品缺乏的问题，制备出一种能够加速创伤愈合的新型食用菌来源复合敷料，从而变废为宝，提高食用菌工厂化生产的附加值，增加企业利润。
本发明的技术方案
一种功能性食用菌来源复合敷料，按照下述步骤制备而成：
(1)将食用菌菌根干燥后磨粉，过筛，得到菌根粉末，密封保存备用；
(2)将菌根粉末进行脱色。
(3)将冷冻干燥至恒重的脱色菌根粉末分别于有机溶剂和碱液中浸泡后，进行微波处理。
(4)待冷却后进行离心，沉淀用水反复冲洗至无醇味，分散在去离子水中形成悬浮液。
(5)收集粒径在一定范围内的粉末，与纳米银颗粒以及金花葵精油均匀混合后，制成多孔膜。
(6)将多孔膜冷冻干燥至恒重，即得功能性食用菌来源复合敷料，保存于-20℃。
步骤(1)中，所述食用菌选自平菇、香菇、草菇、杏鲍菇、金针菇、白玉菇、蟹味菇或双胞蘑菇中的一种或两种以上任意比例的混合物。
步骤(1)中，所述干燥温度为70℃，时间为15h。
步骤(1)中，所述过筛为过60目筛。
步骤(2)中，所述脱色试剂为0.1％次氯酸钠。
步骤(3)中，所述有机溶剂为80％乙醇溶液，浸泡时间为2.5h。
步骤(3)中，所述碱液为50％NaOH溶液，浸泡时间为10min。
步骤(3)中，所述微波处理的功率为462W，处理时间20min。
步骤(4)中，所述粉末的粒径小于100微米，粉末与纳米银颗粒以及金花葵精油的比例为100：5：1。
步骤(5)中，所述多孔膜的厚度为0.1mm-0.2mm。
本发明的有益效果
本发明采用废弃食用菌菌根作为主要原料，在此基础上添加具有抗菌作用的纳米银颗粒和具有抗炎、镇痛、生肌、淡化疤痕作用的金花葵精油，制备出的食用菌来源复合敷料能够有效促进小鼠和大鼠受损皮肤的修复与再生，并且效果好于云南白药对照组。该医疗产品的开发可以变废为宝，应用于皮肤损伤护理，进一步提高食用菌工厂化生产的附加值，增加企业利润，具有很好的经济价值和社会效应。
附图说明
图1为实施例1的功能性食用菌来源复合敷料对小鼠体表伤口愈合效果的时间关系图；
图2为实施例1的功能性食用菌来源复合敷料对大鼠体表伤口愈合效果的时间关系图；
图3为实施例1的功能性食用菌来源复合敷料与云南白药对大鼠体表伤口愈合效果的对比图；
具体实施方式
下面结合具体实施例对本发明作进一步说明：
实施例1
将金针菇菌根70℃干燥15h后磨粉，过60目筛，得到菌根粉末，密封保存备用；用0.1％次氯酸钠使菌根粉末褪色，并用去离子水反复洗涤去除任何残留的次氯酸盐；称取2g冷冻干燥至恒重的脱色菌根粉末于50ml烧杯中，加入20ml 80％乙醇在90℃条件下浸泡2.5小时；离心除去乙醇，加入30ml 50％NaOH溶液，搅拌均匀后，浸泡10min。(注：所有溶液体积根据粉末重量等体积放大)；将烧杯放入微波炉内，杯口罩保鲜膜以尽可能减少反应过程中反应液水分的蒸发，在微波功率462W下处理20min；微波反应结束后，取出烧杯，待冷却后进行离心，沉淀用水反复冲洗至中性，分散在去离子水中形成悬浮液；使用滤纸过滤悬浮液，收集粒径小于100微米的粉末，与纳米银颗粒以及金花葵精油以100：5：1的比例均匀混合后，制成厚度为0.2mm的多孔膜；将多孔膜冷冻干燥至恒重，即得食用菌来源复合敷料，产物记为TG05，保存于-20℃。
实施例2
将金针菇菌根70℃干燥15h后磨粉，过60目筛，得到菌根粉末，密封保存备用；用0.1％次氯酸钠使菌根粉末褪色，并用去离子水反复洗涤去除任何残留的次氯酸盐；称取2g冷冻干燥至恒重的脱色菌根粉末于50ml烧杯中，加入20ml 80％乙醇在90℃条件下浸泡2.5小时；离心除去乙醇，加入30ml 50％NaOH溶液，搅拌均匀后，浸泡10min。(注：所有溶液体积根据粉末重量等体积放大)；将烧杯放入微波炉内，杯口罩保鲜膜以尽可能减少反应过程中反应液水分的蒸发，在微波功率462W下处理20min；微波反应结束后，取出烧杯，待冷却后进行离心，沉淀用水反复冲洗至中性，分散在去离子水中形成悬浮液；使用滤纸过滤悬浮液，收集粒径小于100微米的粉末，与纳米银颗粒以及金花葵精油以100：5：1的比例均匀混合后，制成厚度为0.2mm的多孔膜；将多孔膜冷冻干燥至恒重，即得食用菌来源复合敷料，产物记为TG06，保存于-20℃。
实验测试：
将实施例1制得的食用菌来源复合敷料分别进行小鼠和大鼠体表伤口愈合效果的研究，并且与市购云南白药创可贴进行效果对比，方法和结果如下：
1.食用菌来源敷料对小鼠体表伤口愈合效果的影响测定
(1)建立小鼠体表创伤模型:将6只雄性小鼠适应性喂养3天后，分别进行编号并称量体重，腹腔注射20％乌拉坦试剂(5mL/100g)进行麻醉，待昏迷后用剃须刀去除小鼠的背部毛发并用碘伏消毒。在脊柱两侧，距离脊柱0.5cm、耳朵后方2.5cm的两个大小相同的面积为1cm2的镜像区域(1.0cm*1.0cm)进行标记，并切除全层皮肤，止血消毒后备用。
(2)给药:用已灭菌的敷料覆盖每只小鼠背部的右侧创面作为实验组，用无菌纱布覆盖每只小鼠背部的左侧创面作为对照组，处理完毕后用绷带加以包扎固定。分别在术后隔天用游标卡尺对伤口面积进行测量、记录、拍照以及换药，直至伤口完全愈合。实验期间小鼠每日自由进食、饮水。由图1可以看出与对照相比，敷料TG05能够有效促进小鼠体表伤口的愈合。
2.食用菌来源敷料对大鼠体表伤口愈合效果的影响测定
(1)建立大鼠体表创伤模型:将9只雄性大鼠适应性喂养3天后，分别进行编号并称量体重，腹腔注射20％乌拉坦试剂(5mL/100g)进行麻醉，待昏迷后用剃须刀去除大鼠的背部毛发并用碘伏消毒。在脊柱两侧，距离脊柱1cm、耳朵后方4.5cm的两个大小相同的面积为1cm2的镜像区域(1.0cm*1.0cm)进行标记，并切除全层皮肤，止血消毒后备用。
(2)给药:用已灭菌的敷料覆盖每只大鼠背部的右侧创面作为实验组，用无菌纱布覆盖每只大鼠背部的左侧创面作为对照组，处理完毕后用绷带加以包扎固定。分别在术后隔天用游标卡尺对伤口面积进行测量、记录、拍照以及换药，直至伤口完全愈合。实验期间大鼠每日自由进食、饮水。由图2可以看出与对照相比，敷料TG05能够有效促进大鼠体表伤口的愈合。
3.食用菌来源敷料与云南白药对大鼠体表伤口愈合效果的对比测定
用已灭菌的敷料TG05覆盖每只大鼠背部的右侧创面作为实验组，用市售云南白药创口贴覆盖每只大鼠背部的左侧创面作为对照组，其余实验步骤同上，结果见图3。由图3可以看出，敷料TG05对大鼠体表伤口愈合的促进效果优于云南白药对照组。

Claims (1)

1.一种功能性食用菌来源复合敷料的制备方法，其特征在于按照下述步骤制备而成：
(1)将食用菌菌根干燥后磨粉，过筛，得到菌根粉末，密封保存备用；
(2)将菌根粉末进行脱色；
(3)将冷冻干燥至恒重的脱色菌根粉末分别于有机溶剂和碱液中浸泡后，浸泡时间为2.5h；
进行微波处理；
(4)待冷却后进行离心，沉淀用水反复冲洗至无醇味，分散在去离子水中形成悬浮液；
(5)收集粒径在一定范围内的粉末，与纳米银颗粒以及金花葵精油均匀混合后，制成多孔膜；
(6)将多孔膜冷冻干燥至恒重，即得食用菌来源复合敷料，保存于-20℃；
步骤(1)中，所述食用菌为金针菇；
步骤(1)中，所述干燥温度为70℃，时间为15h；
步骤(1)中，所述过筛为过60目筛；
步骤(2)中，所述脱色试剂为0.1％次氯酸钠；
步骤(3)中，所述有机溶剂为80％乙醇溶液，所述碱液为50％NaOH溶液，浸泡时间为10min；
步骤(3)中，所述微波处理的功率为462W，处理时间20min；
步骤(5)中，所述粉末的粒径小于100微米；步骤(5)中，所述粉末与纳米银颗粒以及金花葵精油的比例为100：5：1；
步骤(5)中，所述多孔膜的厚度为0.1mm-0.2mm。
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