CN113637183A - 改性石墨烯负载纳米银/聚乙烯醇抗菌水凝胶及其制备方法 - Google Patents
改性石墨烯负载纳米银/聚乙烯醇抗菌水凝胶及其制备方法 Download PDFInfo
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- CN113637183A CN113637183A CN202110940788.4A CN202110940788A CN113637183A CN 113637183 A CN113637183 A CN 113637183A CN 202110940788 A CN202110940788 A CN 202110940788A CN 113637183 A CN113637183 A CN 113637183A
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical class [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 86
- 239000004372 Polyvinyl alcohol Substances 0.000 title claims abstract description 62
- 229920002451 polyvinyl alcohol Polymers 0.000 title claims abstract description 62
- 239000000017 hydrogel Substances 0.000 title claims abstract description 58
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 51
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 229910021389 graphene Inorganic materials 0.000 claims abstract description 27
- 239000004721 Polyphenylene oxide Substances 0.000 claims abstract description 20
- 150000001412 amines Chemical class 0.000 claims abstract description 20
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- 239000006185 dispersion Substances 0.000 claims abstract description 18
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000000034 method Methods 0.000 claims abstract description 14
- 229910001961 silver nitrate Inorganic materials 0.000 claims abstract description 8
- 239000012279 sodium borohydride Substances 0.000 claims abstract description 5
- 229910000033 sodium borohydride Inorganic materials 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 51
- 238000003756 stirring Methods 0.000 claims description 29
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 239000000203 mixture Substances 0.000 claims description 20
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 claims description 15
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
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- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 10
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 10
- DLYUQMMRRRQYAE-UHFFFAOYSA-N tetraphosphorus decaoxide Chemical compound O1P(O2)(=O)OP3(=O)OP1(=O)OP2(=O)O3 DLYUQMMRRRQYAE-UHFFFAOYSA-N 0.000 claims description 10
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- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 claims description 5
- 235000010344 sodium nitrate Nutrition 0.000 claims description 5
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- 238000000967 suction filtration Methods 0.000 claims description 5
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- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
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- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
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Abstract
本发明公开了一种改性石墨烯负载纳米银/聚乙烯醇抗菌水凝胶及其制备方法，属于生物材料技术领域。复合水凝胶由聚乙烯醇溶液、化学改性石墨烯、纳米银制成。采用改进Hummers法制备氧化石墨烯，用聚醚胺D230对其进行改性，利用改性后氧化石墨烯上的活性位点，静电吸附硝酸银后，加入硼氢化钠还原剂同时还原氧化石墨烯和硝酸银，制得聚醚胺D230改性石墨烯负载纳米银分散液，加入一定浓度的聚乙烯醇溶液，通过循环冻融法使聚乙烯醇中的羟基通过氢键进行物理交联，即得冷冻‑解冻法制备的改性石墨烯负载纳米银/聚乙烯醇抗菌水凝胶。本发明所得水凝胶具有良好的生物相容性，可应用于人造皮肤、抗菌敷料及伤口绷带等领域，在组织工程中具有潜在的应用价值。
Description
技术领域
本发明涉及一种抗菌水凝胶，特别涉及一种改性石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的制备方法。
背景技术
当今，随着生物材料和医疗器械的迅速发展，医疗感染给临床医生带来了严重的问题，这些生物材料和医疗设备，包括关节植入物、伤口敷料、导管、心脏起搏器和隐形眼镜，带来了与植入相关的感染，因此迫切需要固有的抗菌生物材料和医疗设备。在各类抗菌材料中，水凝胶具有亲水性、较强的保水能力、遇水膨胀、耐酸性、较强的对环境的敏感度和优良的生物相容性等优点。由于其可降解且具有较好的柔韧性，在医药领域尤其是抗菌领域引起了广泛的关注。纳米银具备强大抑菌能力，并且具备广谱杀菌的能力。和抗生素不同的是，其不会产生耐药性且稳定性较好，杀菌结束后可持续杀菌。这些优异的作用使纳米银在抗菌杀菌领域发挥出重要的作用。氧化石墨烯(GO)是石墨烯的氧化产物，其表面具有较大的比表面积和含氧基团，性质活泼。GO由于极性高，耐热性有限，利用聚醚胺D230改性氧化石墨烯可以有效提高耐热性，同时将氧化石墨烯引入水凝胶三维网络结构中可以显著提高水凝胶的力学性能。
因此，如何将改性氧化石墨烯、纳米银和聚乙烯醇组合制备出抗菌性能较好的材料，是本领域技术人员急需解决的技术问题。
现有技术中，例如中国专利CN 107854721 B公开了一种抗菌水凝胶的制备方法，该水凝胶以重氮树脂和羧化壳聚糖为原料制备。但是重氮树脂低毒，对人体有害，制备的水凝胶形成的抑菌圈较小，抗菌性能较差；中国专利CN 106432755 B公开了一种种羧甲基壳聚糖/氧化石墨烯/聚丙烯酰胺复合水凝胶的制备方法，但是其引入了N,N,N',N'-四甲基乙二胺作为催化剂，导致制得的抗菌水凝胶的生物相容性较差，对皮肤具有一定的刺激性。
因此，如何提供一种性能较优的改性氧化石墨烯、纳米银和聚乙烯醇组合制备出具有较好抗菌性能的材料是急需解决的问题。
发明内容
本发明的目的在于克服现有技术的不足，提供了一种改性石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶及其制备方法，以解决现有技术中制备的抗菌型水凝胶力学性能差、生物相容性不高等问题。
本发明是通过以下技术方案实现的：
本发明提供一种改性石墨烯负载纳米银/聚乙烯醇抗菌水凝胶，所述水凝胶的组成按重量组分计包括：80-100份聚乙烯醇、0.4-0.8份氧化石墨烯、5-8份聚醚胺D230和0.1-0.4份纳米银。
本发明提供一种上述水凝胶的制备方法，包括如下步骤：
(3)氧化石墨烯溶液的制备：冰浴条件下，将4.5ml浓硫酸，1.5g过硫酸钾粉末，1.5g五氧化二磷粉末置于250ml锥形瓶中，加入3g的石墨粉，升高温度保持6h，自然冷却至室温后，稀释、抽滤、洗涤至中性，室温下自然干燥，获得预处理样品；取1g预处理样品及0.5g硝酸钠加入23ml的浓硫酸溶液中，3h内缓慢滴加3g高锰酸钾，持续冰水浴搅拌0.5h，连续搅拌反应2h，再移至38℃反应0.5h，升高温度至95℃反应0.5h，高温反应后加入60ml去离子水继续缓慢搅拌，加入15ml 30％过氧化氢溶液，反应15min后加入40ml 1mol/L盐酸溶液，放置沉降，低速离心下层沉淀，洗涤后放置于冷冻干燥机持续干燥1天，制备得氧化石墨烯粉末；
(4)聚醚胺D230改性氧化石墨烯的制备：在80℃条件下，称取0.1g氧化石墨烯粉末加入到200mL去离子水中进行机械搅拌，超声处理并于30min时加入10-15g的聚醚胺D230，制得聚醚胺D230改性氧化石墨烯溶液；
(3)聚醚胺D230改性氧化石墨烯/聚乙烯醇分散液的制备：于恒温水浴锅中配置浓度为0.1-0.5g/ml的聚乙烯醇溶液，在90℃条件下，将1ml聚醚胺D230改性的氧化石墨烯溶液加入到20ml聚乙烯醇溶液中，搅拌10min，超声分散30min，得到聚醚胺D230改性的氧化石墨烯分散液；
(4)水凝胶的制备：在60℃的恒温水浴锅中，向步骤(3)所得分散液加入1-4ml的0.2mol/L硝酸银溶液，加入0.2mL 0.3mol/L的氢氧化钠溶液，搅拌10min，加入0.5ml0.1mol/L的硼氢化钠溶液，在N2气氛下超声处理30min，倒入玻璃模具中，冷冻24h后解冻2h，重复冷冻-解冻多次即得所述水凝胶。
进一步，步骤(4)中冷冻温度为-20±2℃，解冻温度为20±2℃。
进一步，步骤(4)中重复冷冻-解冻次数为3次。
本发明还提供一种上述制备方法制得的水凝胶在人造皮肤、抗菌敷料及伤口绷带等领域中的应用。
与现有技术相比，本发明的有益效果是：
本发明利用硝酸银作为银源，采用原位还原银离子制备氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶，操作简单且具有较高的条件可控性。其次，利用氧化石墨烯结构中的含氧基团与聚乙烯醇链中羟基相互作用形成氢键，增强水凝胶的机械性能。在氧化石墨烯上原位还原纳米银，优化水凝胶的抗菌性能，相较于在聚乙烯醇中加入戊二醛等交联剂来说，本发明采用的方法更加环保安全，利用聚醚胺D230改性氧化石墨烯可以有效提高耐热性。
本发明利用冷冻-解冻的方法，多次循环使得聚乙烯醇中的羟基之间形成氢键，相互交联得到力学性能优异的氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶。相较于普通聚乙烯醇基抗菌水凝胶，加入氧化石墨烯负载纳米银显著提高了抗菌性能及力学性能，且纳米银具有广谱抗菌的特点。采用冻融法制备，方便操作，具有良好的生物相容性，在人造皮肤、韧带以及作为伤口绷带敷料等领域具有广阔的前景。
具体实施方式
下面，举实施例说明本发明，但是，本发明并不限于下述的实施例。
根据下述实施例，可以更好地理解本发明。然而，本领域的技术人员容易理解，实施例所描述的内容仅用于说明本发明，而不应当也不会限制权利要求书中所详述的本发明。
空白对照
所述水凝胶其原料按重量份包括：聚乙烯醇含量为80-100份。
聚乙烯醇水凝胶的制备：在90℃的恒温水浴锅中，配置质量分数13％的聚乙烯醇溶液，超声处理30min除气泡后，倒入玻璃模具中，在-20±2℃中冷冻24h，在20±2℃解冻2h，重复冷冻-解冻3次，即得所述聚乙烯醇基水凝胶。
实施例1
一种聚醚胺D230改性氧化石墨烯/聚乙烯醇抗菌水凝胶的制备方法，包括如下步骤：
所述复合水凝胶其原料按重量份包括：聚乙烯醇含量为80-100份，氧化石墨烯0.4-0.8份，聚醚胺D230 5-8份。
1、氧化石墨烯水溶液的制备：冰浴条件下，将3.0g石墨粉、4.5ml浓硫酸，1.5g过硫酸钾粉末，1.5g五氧化二磷粉末置于250ml锥形瓶中混合，升高温度至80℃保持6h。自然冷却至室温后，稀释，抽滤。洗涤至中性，室温下自然干燥，获得预处理样品；取1g预处理样品及0.5g硝酸钠加入23ml的浓硫酸溶液中，3h内缓慢滴加3g高锰酸钾，持续冰水浴搅拌0.5h，连续搅拌反应2h，再移至38℃反应0.5h，升高温度至95℃反应0.5h，高温反应后加入60ml去离子水继续缓慢搅拌，加入15ml 30％过氧化氢溶液，直至混合体系变为棕黄色，反应15min后加入40ml mol/L盐酸溶液，静置沉降，低速离心下层沉淀，离心洗涤至混合液至pH值为5～7，放置于冷冻干燥机持续干燥1天，制备得氧化石墨烯粉末。
2、聚醚胺D230改性氧化石墨烯的制备：在80℃条件下，称取0.1g氧化石墨烯粉末加入到200mL去离子水中进行机械搅拌，超声处理并于30min时加入10g聚醚胺D230，制得聚醚胺D230改性氧化石墨烯溶液；
3、聚醚胺D230改性氧化石墨烯/聚乙烯醇分散液的制备：在90℃的恒温水浴锅中，配置质量分数13％的聚乙烯醇溶液。降低温度至85℃，取1ml聚醚胺D230改性氧化石墨烯溶液加入20ml配置所得聚乙烯醇溶液中，搅拌10min，超声分散30min得到聚醚胺D230改性氧化石墨烯/聚乙烯醇分散液。
4、聚醚胺D230改性石墨烯聚乙烯醇基抗菌水凝胶的制备：将上述制备的分散液在N2气氛下，超声处理30min除气泡后，倒入玻璃模具中，在-20±2℃中冷冻24h，在20±2℃解冻2h，重复冷冻-解冻3次，即得目标水凝胶。
实施例2
一种聚醚胺D230改性石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的制备方法，包括如下步骤：
1、氧化石墨烯水溶液的制备：冰浴条件下，将3.0g石墨粉、4.5ml浓硫酸，1.5g过硫酸钾粉末，1.5g五氧化二磷粉末置于250ml锥形瓶中混合，升高温度至80℃保持6h。自然冷却至室温后，稀释，抽滤。洗涤至中性，室温下自然干燥，获得预处理样品；取1g预处理样品及0.5g硝酸钠加入23ml的浓硫酸溶液中，3h内缓慢滴加3g高锰酸钾，持续冰水浴搅拌0.5h，连续搅拌反应2h，再移至38℃反应0.5h，升高温度至95℃反应0.5h，高温反应后加入60ml去离子水继续缓慢搅拌，加入15ml 30％过氧化氢溶液，直至混合体系变为棕黄色，反应15min后加入40ml 1mol/L盐酸溶液，静置沉降，低速离心下层沉淀，离心洗涤至混合液至pH值为5～7，放置于冷冻干燥机持续干燥1天，制备得氧化石墨烯粉末。
2、聚醚胺D230改性氧化石墨烯的制备：在80℃条件下，称取0.1g氧化石墨烯粉末加入到200mL去离子水中进行机械搅拌，超声处理并于30min时加入12g的聚醚胺D230，制得聚醚胺D230改性氧化石墨烯溶液；
3、聚醚胺D230改性氧化石墨烯/聚乙烯醇分散液的制备：在90℃的恒温水浴锅中，配置质量分数13％的聚乙烯醇溶液。降低温度至85℃，取2ml步骤3所得聚醚胺D230改性氧化石墨烯溶液加入20ml聚乙烯醇溶液中，搅拌10min，超声分散30min得到聚醚胺D230改性氧化石墨烯/聚乙烯醇分散液。
4、聚醚胺D230改性石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的制备：降低温度到60℃，称取1.5ml 0.2mol/L的硝酸银溶液加入步骤3所得分散液，加入0.2mL 0.3mol/L的氢氧化钠溶液，搅拌10min，加入0.5ml 0.1mol/L的硼氢化钠溶液，将上述制备的改性氧化石墨烯/聚乙烯醇分散液在N2气氛下，超声处理30min除气泡后，倒入玻璃模具中，在-20±2℃中冷冻24h，在20±2℃解冻2h，重复冷冻-解冻3次，即得目标水凝胶。
实施例3
一种聚醚胺D230改性石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的制备方法，包括如下步骤：
1、氧化石墨烯水溶液的制备：冰浴条件下，将3.0g石墨粉、4.5ml浓硫酸，1.5g过硫酸钾粉末，1.5g五氧化二磷粉末置于250ml锥形瓶中混合，升高温度至80℃保持6h。自然冷却至室温后，稀释，抽滤。洗涤至中性，室温下自然干燥，获得预处理样品；取1g预处理样品及0.5g硝酸钠加入23ml的浓硫酸溶液中，3h内缓慢滴加3g高锰酸钾，持续冰水浴搅拌0.5h，连续搅拌反应2h，再移至38℃反应0.5h，升高温度至95℃反应0.5h，高温反应后加入60ml去离子水继续缓慢搅拌，加入15ml 30％过氧化氢溶液，直至混合体系变为棕黄色，反应15min后加入40ml 1mol/L盐酸溶液，静置沉降，低速离心下层沉淀，离心洗涤至混合液至pH值为5～7，放置于冷冻干燥机持续干燥1天，制备得氧化石墨烯粉末。
2、聚醚胺D230改性氧化石墨烯的制备：在80℃条件下，称取0.1g氧化石墨烯粉末加入到200mL去离子水中进行机械搅拌，超声处理并于30min时加入15g的聚醚胺D230，制得聚醚胺D230改性氧化石墨烯溶液；
3、聚醚胺D230改性氧化石墨烯/聚乙烯醇分散液的制备：在90℃的恒温水浴锅中，配置质量分数13％的聚乙烯醇溶液。降低温度至85℃，取2.5ml步骤2所得聚醚胺D230改性氧化石墨烯溶液加入20ml配置所得聚乙烯醇溶液中，搅拌10min，超声分散30min得到氧化石墨烯/聚乙烯醇分散液。
4、聚醚胺D230改性石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的制备：降低温度到60℃，称取1.5ml 0.2mol/L的硝酸银溶液加入步骤3所得分散液中，加入0.2mL0.3mol/L的氢氧化钠溶液，搅拌10min，加入0.5ml 0.1mol/L的硼氢化钠溶液，将上述制备的改性氧化石墨烯/聚乙烯醇分散液在N2气氛下，超声处理30min除气泡后，倒入玻璃模具中，在-20±2℃中冷冻24h，在20±2℃解冻2h，重复冷冻-解冻3次，即得目标水凝胶。
将空白对照、实施例1、实施例2和实施例3制备的水凝胶进行抗菌试验，具体结果见表1。
改性氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的抗菌率
表1
抗菌试验表明，氧化石墨烯自身具有一定抑菌性，但抑菌性能不强，负载纳米银后抑菌性能显著提升。可见，利用本发明制备的抗菌水凝胶产品，抗菌性能较好。
改性氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的拉伸应力应变
| 拉伸应力/MPa | 应变/％ | |
| 实施例一 | 195.78 | 0.100 |
| 实施例二 | 211.30 | 0.104 |
| 实施例三 | 189.35 | 0.109 |
| 空白对照 | 237.17 | 0.063 |
表2
将得到的圆柱型水凝胶切割为尺寸60mm×2mm×2mm的试样，使用Instron5967型拉伸试验机进行拉伸性能测试。由实验数据可知，改性的氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶与聚乙烯醇水凝胶相比，断裂伸长率提高了约60％，力学性能得到了提升。
通过红细胞的溶血实验来探究生物相容性，在红细胞溶血实验中，在红细胞中加入所测的样品，如果样品的生物相容性较差，就会使得红细胞破裂并释放出血红蛋白，通过测定其的释放量，可以计算出样品的溶血率，从而评价材料的生物相容性。以红细胞的溶血实验来评价改性氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的生物相容性。
改性氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶的生物相容性
| 溶血率/％ | |
| 实施例一 | 0.25 |
| 实施例二 | 0.38 |
| 实施例三 | 0.44 |
表3
与阳性对照组(非离子型表面活性剂即0.1％TritonX-100处理过的红细胞)相比，改性氧化石墨烯负载纳米银/聚乙烯醇抗菌水凝胶溶血率均低于10％，说明改性氧化石墨烯负载纳米银/聚乙烯醇基抗菌水凝胶具有良好的生物相容性。
综上表明，我们提供了一种采用改进Hummers法制备氧化石墨烯，然后用聚醚胺D230对其进行改性，再在改性的氧化石墨烯的活性位点上，原位还原得到纳米银离子。通过冷冻-解冻法使聚乙烯醇上的羟基通过氢键相互交联，制备即得氧化石墨烯负载纳米银/聚乙烯醇基抗菌性水凝胶该水凝胶。该抗菌性水凝胶不仅具有较强的力学性能，还具有优异的抗菌性及良好的生物相容性。有望后期用于人造皮肤、韧带以及作为伤口绷带敷料等领域。
Claims (5)
1.一种改性石墨烯负载纳米银/聚乙烯醇抗菌水凝胶，其特征在于，所述水凝胶的组成按重量组分计包括：80-100份聚乙烯醇、0.4-0.8份氧化石墨烯、5-8份聚醚胺D230和0.1-0.4份纳米银。
2.权利要求1所述水凝胶的制备方法，其特征在于，包括如下步骤：
(1)氧化石墨烯溶液的制备：冰浴条件下，将4.5ml浓硫酸，1.5g过硫酸钾粉末，1.5g五氧化二磷粉末置于250ml锥形瓶中，加入3g的石墨粉，升高温度保持6h，自然冷却至室温后，稀释、抽滤、洗涤至中性，室温下自然干燥，获得预处理样品；取1g预处理样品及0.5g硝酸钠加入23ml的浓硫酸溶液中，3h内缓慢滴加3g高锰酸钾，持续冰水浴搅拌0.5h，连续搅拌反应2h，再移至38℃反应0.5h，升高温度至95℃反应0.5h，高温反应后加入60ml去离子水继续缓慢搅拌，加入15ml 30％过氧化氢溶液，反应15min后加入40ml 1mol/L盐酸溶液，放置沉降，低速离心下层沉淀，洗涤后放置于冷冻干燥机持续干燥1天，制备得氧化石墨烯粉末；
(2)聚醚胺D230改性氧化石墨烯的制备：在80℃条件下，称取0.1g氧化石墨烯粉末加入到200mL去离子水中进行机械搅拌，超声处理并于30min时加入10-15g的聚醚胺D230，制得聚醚胺D230改性氧化石墨烯溶液；
(3)聚醚胺D230改性氧化石墨烯/聚乙烯醇分散液的制备：于恒温水浴锅中配置浓度为0.1-0.5g/ml的聚乙烯醇溶液，在90℃条件下，将1-2.5ml聚醚胺D230改性的氧化石墨烯溶液加入到20ml聚乙烯醇溶液中，搅拌10min，超声分散30min，得到聚醚胺D230改性的氧化石墨烯分散液；
(4)水凝胶的制备：在60℃的恒温水浴锅中，向步骤(3)所得分散液加入1.5ml的0.2mol/L硝酸银溶液，加入0.2mL 0.3mol/L的氢氧化钠溶液，搅拌10min，加入0.5ml0.1mol/L的硼氢化钠溶液，在N2气氛下超声处理30min，倒入玻璃模具中，冷冻24h后解冻2h，重复冷冻-解冻多次即得所述水凝胶。
3.根据权利要求2所述的制备方法，其特征在于，步骤(4)中冷冻温度为-20±2℃，解冻温度为20±2℃。
4.根据权利要求3所述的制备方法，其特征在于，步骤(4)中重复冷冻-解冻次数为3次。
5.权利要求2-4任一所述制备方法制得的水凝胶在人造皮肤、抗菌敷料及伤口绷带等领域中的应用。
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