CN106581768B - 一种用于心肌修复的组织工程支架的制备方法 - Google Patents
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Abstract
一种用于心肌修复的组织工程支架的制备方法。向氧化石墨烯水溶液中加入壳聚糖粉末，搅拌均匀，得到乳浊液；逐滴向乳浊液中滴加冰醋酸，边滴加边搅拌，直至溶液澄清透明，得到粘稠的溶液；持续搅拌粘稠的溶液12～48小时后，缓慢倒入模具；将装有溶液的模具置于‑80℃温度条件下冷冻24小时以上，然后将模具置于预冻4小时以上真空冷冻干燥机中，冷冻干燥过夜，得到组织工程支架。本发明制备出的支架能够有利于细胞之间传导电信号，这是因为氧化石墨烯具有很强的导电能力，能够为细胞之间电信号传导提供介质，应用于心肌组织工程更加有利于心肌细胞间的信号传导；本发明的制备方法不采用有细胞毒性的交联剂，制备出的支架材料无细胞毒性。
Description
技术领域
本发明涉及一种适用于心肌组织工程的为心肌细胞生长及心肌细胞之间相互作用提供依托的壳聚糖为主要材料的组织工程支架材料的制备方法。
背景技术
目前，人造支架已经广泛应用于不同的组织重建工程，支架的主要功能是模仿天然细胞外基质，为细胞生长、分化提供一个微环境，同时，支架也能够携载生长因子，为细胞特定方向的增殖或者分化提供相应的刺激因素，为细胞与细胞之间、细胞与外基质之间有一个相互接触，相互作用的平台。
当前的心肌组织工程支架多数只考虑了细胞和细胞之间的化学信号传递，而忽略了心肌细胞与心肌细胞、心肌细胞与成纤维细胞、心肌细胞与内皮细胞之间的电信号传导，并且多数采用有细胞毒性的交联剂进行交联。
发明内容
本发明要解决的技术问题是提供一种为心肌细胞、心肌成纤维细胞、内皮细胞之间化学信号传导、电信号传导提供依托的心肌修复支架的制备方法。
本发明的技术方案：
一种用于心肌修复的组织工程支架的制备方法，步骤如下：
向氧化石墨烯水溶液中加入壳聚糖粉末，使得氧化石墨烯与壳聚糖的质量比为0.01％～0.1％，搅拌均匀，得到乳浊液；逐滴向乳浊液中滴加冰醋酸，边滴加边搅拌，直至溶液澄清透明，得到粘稠的溶液，使得滴加的冰醋酸和乳浊液中水的体积比为0.2/100～2/100；持续搅拌粘稠的溶液12～48小时后，缓慢倒入模具；将装有溶液的模具置于-80℃温度条件下冷冻24小时以上，然后将模具置于预冻4小时以上真空冷冻干燥机中，冷冻干燥过夜，得到组织工程支架。
所述的持续搅拌粘稠的溶液12～48小时后过程，在紫外灯照射条件下进行。
本发明的有益效果：本发明制备出的支架能够有利于细胞之间传导电信号，这是因为氧化石墨烯具有很强的导电能力，能够为细胞之间电信号传导提供介质，应用于心肌组织工程更加有利于心肌细胞间的信号传导；本发明的制备方法不采用有细胞毒性的交联剂，制备出的支架材料无细胞毒性。
具体实施方式
以下结合技术方案，进一步说明本发明的具体实施方式。
实施例1：
取5mg/L的氧化石墨烯水溶液50mL，向其中加入1g壳聚糖粉末，搅拌成乳浊液，由于壳聚糖不溶于水，向混合的乳浊液中逐滴加入800微升冰醋酸，边加边搅拌，待乳浊液逐渐澄清透明，置于磁力搅拌器，搅拌24小时，将溶液倒至模具中，置于-80℃冰箱冷冻过夜，24小时之后，用真空冷冻干燥机冷冻干燥得到支架材料。
实施例2：
采用15mg/L的氧化石墨烯水溶液50mL，向其中加入1g壳聚糖粉末，搅拌成乳浊液，向混合的乳浊液中逐滴加入300微升冰醋酸，边加边搅拌，待乳浊液逐渐澄清透明，置于磁力搅拌器，搅拌16小时，将溶液倒至模具中，置于-80℃冰箱冷冻过夜，24小时之后，用真空冷冻干燥机冷冻干燥24小时得到支架材料。
本发明方法的磁力搅拌过程是处于紫外灯照射下进行，经过紫外灯持续照射，可以营造一种无菌环境，同时减少支架材料内部的微生物。
本发明优选包括在制备壳聚糖/氧化石墨烯复合支架时不采用交联剂。
Claims (2)
1.一种用于心肌修复的组织工程支架的制备方法，其特征在于，步骤如下：
向氧化石墨烯水溶液中加入壳聚糖粉末，使得氧化石墨烯与壳聚糖的质量比为0.01％～0.1％，搅拌均匀，得到乳浊液；逐滴向乳浊液中滴加冰醋酸，边滴加边搅拌，直至溶液澄清透明，得到粘稠的溶液，使得滴加的冰醋酸和乳浊液中水的体积比为0.2/100～2/100；持续搅拌粘稠的溶液12～48小时后，缓慢倒入模具；将装有溶液的模具置于-80℃温度条件下冷冻24小时以上，然后将模具置于预冻4小时以上真空冷冻干燥机中，冷冻干燥过夜，得到组织工程支架。
2.根据权利要求1所述的制备方法，其特征在于，所述的持续搅拌粘稠的溶液12～48小时后过程，在紫外灯照射条件下进行。
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| CN103191464A (zh) * | 2013-04-07 | 2013-07-10 | 西南交通大学 | 一种抗菌多孔氧化石墨烯/壳聚糖复合支架的制备方法 |
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| JP2016158680A (ja) * | 2015-02-27 | 2016-09-05 | 国立大学法人北海道大学 | 酸化グラフェンを用いた骨再生用又は皮膚再生用スキャフォールド |
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| CN104922734A (zh) * | 2015-05-21 | 2015-09-23 | 东南大学 | 促进心肌修复的可注射壳聚糖复合水凝胶及其制备方法 |
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