CN108778351A - 用于改进罗非鱼皮的方法及其用于覆盖皮肤损伤的用途 - Google Patents

用于改进罗非鱼皮的方法及其用于覆盖皮肤损伤的用途 Download PDF

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CN108778351A
CN108778351A CN201680051079.8A CN201680051079A CN108778351A CN 108778351 A CN108778351 A CN 108778351A CN 201680051079 A CN201680051079 A CN 201680051079A CN 108778351 A CN108778351 A CN 108778351A
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埃德马尔·马西埃尔·利马·儒尼奥尔
曼诺尔·鄂多立克·德·莫赖斯·菲略
马塞洛·何塞·博格斯·德·米兰达
尼尔森·萨尔托·皮科洛
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Saiala Federal University
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Abstract

本发明涉及罗非鱼皮的用途，该罗非鱼皮基于在5型和7型洁净室环境中浓度在50‑99％范围的甘油进行若干步骤的处理，并且在某些情况下，当微生物数量高时，有必要通过伽马辐照进行补充辐射消毒。该鱼皮可以用作诸如烧伤和急性或慢性伤口的皮肤损伤的闭合性生物敷料。

Description

用于改进罗非鱼皮的方法及其用于覆盖皮肤损伤的用途
技术领域
本发明涉及罗非鱼皮的用途，该罗非鱼皮基于在5型和7型洁净室环境中浓度在50-99％范围的甘油进行若干步骤的处理，并且在某些情况下，当微生物数量高时，有必要通过伽马辐照进行补充辐射灭菌。该鱼皮可用于烧伤和急性或慢性伤口，包括其在浅层或深层的二度烧伤造成的损伤表面上的应用，在急性伤口例如自体移植的供体皮肤区域或者慢性伤口中的应用，或在麻醉下进行简单外敷、外科清洁或外科清创的最后阶段以及经常出现在这类损伤中的自体植皮中的应用。
背景技术
目前，巴西的医学并不具备用于(动物来源的)临时异源皮肤外敷物的替代选择，以作为烧伤和伤口治疗的一部分。数十年来，在发达国家，尤其是美国，工业化猪皮被大规模地用于这一目的。考虑到高昂的成本和巴西的经济现状，将该产品进口到巴西并不具备商业可行性。
具体实施方式
本发明描述了罗非鱼皮在诸如烧伤、慢性伤口等皮肤损伤的表面处理中的用途，所述罗非鱼获自使用培养***(tanks-net)的养鱼场，通过灭菌过程，如下所述。
屠宰后，使用绿松石(工具)剥除罗非鱼皮(重量为800g至1000g之间)并将其在活水中进行清洗，以移除任何微量的血液和其它杂质，并将其置于预先冷却至4℃的无菌生理盐水(0.9％NaCl溶液)中进行最终清洗。随后，移除仍贴附于鱼皮上多余的肌肉，并将鱼皮切割成10.0cm×5.0cm的块并用生理盐水清洗，随后通过以下步骤进行灭菌。
第一步：将鱼皮置于包含2％葡萄糖酸氯己定(具有表面活性剂的溶液)的无菌容器中，并在其中保持30分钟。
第二步：在上一步操作后，使用无菌生理盐水清洗鱼皮并将其置于另一个包含另外的2％葡萄糖酸氯己定溶液的容器中，并在其中保持30分钟。
第三步：使用无菌生理盐水冲洗鱼皮并将其置于包含50％甘油、49％生理盐水和1％的具有青霉素、链霉素和fungisol的溶液的容器中，将其包装在含冰的恒温箱中用于运输(Jaguaribara-NPDNI)。
下述步骤在具有水平或垂直层流的无菌环境中在UFC的NPDM中进行。
第四步：24小时之前，应将鱼皮从前述溶液中取出，使用无菌生理盐水清洗，并置于无菌、密封的包含75％甘油、24％生理盐水和1％的具有青霉素、链霉素和fungisol的溶液的密闭容器中，在该容器中，将鱼皮在该溶液中按揉5分钟，并且在具有恒定转速为15转/分钟的搅拌器和温度为37℃的水浴(离心机)中保持3小时。
第五步：将鱼皮取出，再次用无菌生理盐水清洗，并置于另一个无菌且密封的包含99％甘油和1％的具有青霉素、链霉素和fungisol的溶液的容器中，将鱼皮在该溶液中按揉5分钟，并在转速为15转/分钟且温度为37℃的离心机中的水浴中另外保持3小时。
第六步：在使用甘油的上一步结束时，将鱼皮包装于无菌的双层塑料包封中并于4℃下储存以供日后使用，保存限期为最长达两年。
从天然鱼皮开始，即在第一步之前和上述六个步骤之中，对***、革兰氏阴性细菌和真菌进行七次微生物学测试。当用于细菌计数(即生物负载微生物分析或微生物限度测试)的生物负载水平在可接受范围内时，该鱼皮可供使用，其中所述生物负载微生物分析或微生物限度测试在加工和处理期间对需要控制微生物水平的药物和医疗产品上进行。
当使用鱼皮时，将鱼皮从包封中取出并且在不同的无菌生理盐水中清洗三次，每次清洗五分钟。在此步骤之后，鱼皮被切割成伤口的尺寸并作为封闭敷料使用。
若细菌计数高于10/3，应进行如下所述的补充灭菌步骤：
第七步：补充辐射灭菌，伽马-钴60，其中负荷剂量范围为15至50千戈瑞，取决于生物负载水平(微生物计数)。
通过粘附在创面上，罗非鱼皮在与这些损伤相互作用时会促进真皮基质的愈合和修复过程的加速(由于其组织结构中I型胶原蛋白的作用)，避免渗出液停留和体液流失，增强阻止细菌侵入的屏障，并缓解疼痛。

Claims (6)

1.一种用于改进罗非鱼皮的方法，其特征在于，所述方法通过以下步骤进行：
-使用绿松石剥除罗非鱼皮并在活水中冲洗，置于预先冷却至4℃的无菌生理盐水(0.9％NaCl溶液)中进行最终清洗，随后将多余的肌肉去除，并切割成10.0cm×5.0cm的块，用生理盐水清洗，随后按照以下步骤进行灭菌：
-将所述罗非鱼皮置于包含2％葡萄糖酸氯己定(具有表面活性剂的溶液)的无菌容器中，并在其中保持30分钟；
-使用无菌生理盐水清洗所述罗非鱼皮并将所述罗非鱼皮置于另一个包含另外的2％葡萄糖酸氯己定溶液的容器中，并在其中保持30分钟；
-使用无菌生理盐水溶液冲洗所述罗非鱼皮，并将所述罗非鱼皮置于包含50％甘油、49％生理盐水和1％的具有青霉素、链霉素和fungisol的溶液的容器中。
2.根据权利要求1所述的方法，其特征在于，下述步骤在具有水平或垂直层流的无菌环境中进行。
3.根据权利要求1或2所述的方法，其特征在于以下步骤：
-24小时之前，将所述罗非鱼皮从所述溶液中取出，使用无菌生理盐水清洗，并置于无菌且密封的包含75％甘油、24％生理盐水和1％的具有青霉素、链霉素和fungisol的溶液的容器中(在所述溶液中按揉所述罗非鱼皮5分钟)，并且在具有恒定转速为15转/分钟的搅拌器和温度为37℃的水浴(离心机)中保持3小时；
-将所述罗非鱼皮取出，再次用无菌生理盐水清洗，并置于另一个无菌且密封的包含99％甘油和1％的具有青霉素、链霉素和fungisol的溶液的容器中(在所述溶液中按揉所述罗非鱼皮5分钟)，并在转速为15转/分钟且温度为37℃的水浴(离心机)中另外保持3小时，并且在最后，将所述罗非鱼皮包装于无菌的双层塑料包封中并于4℃下储存以供日后使用。
4.根据权利要求1至3任一项所述的方法，其特征在于，若细菌计数高于10/3，应进行补充灭菌步骤。
5.根据权利要求4所述的方法，其特征在于，其是补充辐射灭菌步骤，伽马-钴60，其中负荷剂量范围为15至50千戈瑞，取决于微生物计数的生物负载水平。
6.根据前述权利要求所述的罗非鱼皮的用途，其特征在于，所述罗非鱼皮作为诸如烧伤和急性或慢性伤口的皮肤损伤的封闭敷料施用。
CN201680051079.8A 2015-09-03 2016-09-02 用于改进罗非鱼皮的方法及其用于覆盖皮肤损伤的用途 Pending CN108778351A (zh)

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US11612675B1 (en) 2018-06-28 2023-03-28 The Regents Of The University Of California Fish skin biologic bandage
BR102018073515A2 (pt) * 2018-11-14 2020-05-26 Edmar Maciel Lima Junior processo de obtenção de matriz extracelular de pele de tilápia (oreochromis niloticus) e uso da matriz extracelular de tilápia
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