CN113209361B - 一种生物材料复合水凝胶伤口敷料及其制备方法 - Google Patents
一种生物材料复合水凝胶伤口敷料及其制备方法 Download PDFInfo
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- CN113209361B CN113209361B CN202110354321.1A CN202110354321A CN113209361B CN 113209361 B CN113209361 B CN 113209361B CN 202110354321 A CN202110354321 A CN 202110354321A CN 113209361 B CN113209361 B CN 113209361B
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- wound dressing
- antibacterial
- hydrogel wound
- composite hydrogel
- polyvinylpyrrolidone
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Abstract
本发明属于医用敷料技术领域，尤其涉及一种生物材料复合水凝胶伤口敷料及其制备方法，该辅料包括以下重量百分比含量的原料：角蛋白1‑10％、丝素蛋白0.01‑0.08％、聚乙烯吡咯烷酮3‑21％、聚乳酸1‑5％、聚乙二醇1‑6％、抗菌剂0.01‑0.06％、交联剂0.1‑0.6％和余量为蒸馏水；本发明凝胶敷料利于伤口的愈合，具有优异的吸水、保湿与透气性。
Description
技术领域
本发明属于医用敷料技术领域，尤其涉及一种生物材料复合水凝胶伤口敷料及其制备方法。
背景技术
皮肤是覆盖整个机体的最大和最外面的器官，主要功能是保护下层肌肉、骨骼、韧带和内部器官免受外部生物、化学、机械和物理因素的影响。
此外，皮肤还涉及感觉、温度调节、免疫监视、防止脱水和促成维生素D3的合成。然而，皮肤的结构和功能会受到创伤、烧伤、手术切口或疾病(如糖尿病)的影响。皮肤结构受损后形成伤口，必须尽快重建其结构和功能，以确保身体稳态。为此，首先要充分了解皮肤的正常生理学，其次应充分了解皮肤伤口的愈合过程。伤口可定义为由物理或热损伤或存在潜在的医学病症而导致皮肤或粘膜表皮连续性的缺陷或破坏。创伤出现后应尽量避免细菌感染的风险，因为伤口愈合是一种复杂、动态和多步骤的过程。
通常伤口愈合包括连续和重叠的四个阶段:(1)凝血和止血，针对出血控制和限制微生物在体内的传播；(2)炎症与止血同时发生，促炎细胞因子的释放和肿胀同时出现；(3)增殖，其中成纤维细胞迁移至受损区域，促进毛细血管生长、胶原合成和肉芽组织的形成，最终上皮细胞迁移；(4)重塑，新组织不断重塑，这将决定最终瘢痕的性质。选择适当的伤口敷料对不同类型伤口的愈合是必不可少的。
而目前的医用敷料对于伤口愈合速度慢，同时吸水、保湿性差，不利于伤口的愈合，无法满足人们的需求。
发明内容
本发明为了解决上述技术问题提供一种生物材料复合水凝胶伤口敷料及其制备方法。
本发明解决上述技术问题的技术方案如下：一种生物材料复合水凝胶伤口敷料，包括以下重量百分比含量的原料：角蛋白1-10％、丝素蛋白0.01-0.08％、聚乙烯吡咯烷酮3-21％、聚乳酸1-5％、聚乙二醇1-6％、抗菌剂0.01-0.06％、交联剂0.1-0.6％和余量为蒸馏水。
进一步，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成。
进一步，所述交联剂为氯化钙。
本发明还提供一种生物材料复合水凝胶伤口敷料的制备方法，包括以下步骤：
A、称取如下重量百分比含量的原料：角蛋白1-10％、丝素蛋白0.01-0.08％、聚乙烯吡咯烷酮3-21％、聚乳酸1-5％、聚乙二醇1-6％、抗菌剂0.01-0.06％、交联剂0.1-0.6％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，得到抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入交联剂搅拌反应得到水凝胶伤口敷料。
进一步，在步骤B中，所述聚乙烯吡咯烷酮溶液质量分数为20-40％。
进一步，在步骤D中，所述抗菌分散液质量分数为5-20％。
进一步，在步骤D中，所述超声分散时间为30-40min。
进一步，在步骤E中，所述搅拌反应时间为30-60min，搅拌反应温度为30-50℃。
进一步，在步骤E中，反应溶液pH值控制在6.5-7.1。
本发明的有益效果是：本发明敷料通过角蛋白、聚乙烯吡咯烷酮、聚乳酸与聚乙二醇形成三维网状水凝胶结构，具有高机械强度与生物相容性的丝素蛋白复合于水凝胶中，提高力学性能，同时抗菌剂加载到水凝胶中提高抑菌率，满足治疗与恢复伤口的效果，角蛋白具有优异的生物相容性，利于水凝胶的形成和促进伤口愈合，改善细胞的粘附性能，提供湿润的环境，吸收伤口渗液和气体交换，此外聚乙烯吡咯烷酮与氧化石墨烯具有较强的液体吸收能力，透气且屏蔽细菌，还可促进皮肤对抗菌剂的吸收，加速伤口的愈合，本发明凝胶敷料利于伤口的愈合，具有优异的吸水、保湿与透气性。
具体实施方式
以下对本发明的原理和特征进行描述，所举实例只用于解释本发明，并非用于限定本发明的范围。
实施例1
A、称取如下重量百分比含量的原料：角蛋白1％、丝素蛋白0.01％、聚乙烯吡咯烷酮3％、聚乳酸1％、聚乙二醇1％、抗菌剂0.01％、氯化钙0.1％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，得到质量分数为20％的聚乙烯吡咯烷酮溶液，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，超声分散时间为30min，得到质量分数为5％的抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入氯化钙搅拌反应30min，反应温度为30℃，pH值控制在6.5，得到水凝胶伤口敷料。
实施例2
A、称取如下重量百分比含量的原料：角蛋白5％、丝素蛋白0.04％、聚乙烯吡咯烷酮12％、聚乳酸3％、聚乙二醇3％、抗菌剂0.03％、氯化钙0.3％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，得到质量分数为30％的聚乙烯吡咯烷酮溶液，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，超声分散时间为35min，得到质量分数为12％的抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入氯化钙搅拌反应50min，反应温度为45℃，pH值控制在6.8，得到水凝胶伤口敷料。
实施例3
A、称取如下重量百分比含量的原料：角蛋白10％、丝素蛋白0.08％、聚乙烯吡咯烷酮21％、聚乳酸5％、聚乙二醇6％、抗菌剂0.06％、氯化钙0.6％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，得到质量分数为40％的聚乙烯吡咯烷酮溶液，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，超声分散时间为40min，得到质量分数为20％的抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入氯化钙搅拌反应60min，反应温度为50℃，pH值控制在7.1，得到水凝胶伤口敷料。
对比例1
A、称取如下重量百分比含量的原料：角蛋白1％、聚乙烯吡咯烷酮3％、聚乳酸1％、聚乙二醇1％、抗菌剂0.01％、氯化钙0.1％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，得到质量分数为20％的聚乙烯吡咯烷酮溶液，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，超声分散时间为30min，得到质量分数为5％的抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入氯化钙搅拌反应30min，反应温度为30℃，pH值控制在6.5，得到水凝胶伤口敷料。
对比例2
A、称取如下重量百分比含量的原料：角蛋白5％、丝素蛋白0.04％、聚乙烯吡咯烷酮12％、聚乳酸3％、聚乙二醇3％、氯化钙0.3％和余量为蒸馏水；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，得到质量分数为30％的聚乙烯吡咯烷酮溶液，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、向步骤C得到的分散液中加入氯化钙搅拌反应50min，反应温度为45℃，pH值控制在6.8，得到水凝胶伤口敷料。
对比例3
A、称取如下重量百分比含量的原料：角蛋白0.5％、丝素蛋白0.005％、聚乙烯吡咯烷酮2％、聚乳酸0.5％、聚乙二醇0.3％、抗菌剂0.01％、氯化钙0.1％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，得到质量分数为20％的聚乙烯吡咯烷酮溶液，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，超声分散时间为30min，得到质量分数为5-20％的抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入氯化钙搅拌反应30min，反应温度为30℃，pH值控制在6.5，得到水凝胶伤口敷料。
对比例4
A、称取如下重量百分比含量的原料：角蛋白12％、丝素蛋白0.1％、聚乙烯吡咯烷酮25％、聚乳酸6％、聚乙二醇10％、抗菌剂0.06％、氯化钙0.6％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，得到质量分数为40％的聚乙烯吡咯烷酮溶液，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，超声分散时间为40min，得到质量分数为20％的抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入氯化钙搅拌反应60min，反应温度为50℃，pH值控制在7.1，得到水凝胶伤口敷料。
试验例1
力学性能试验，具体如下所示：
按照GB/T1040-92《塑料拉伸试验方法》，将凝胶敷料在电子万能试验机上进行拉伸性能测试，试验在室温下进行，拉伸速率为50mm/min。
弹性模量计算公式：E＝σ/ε，E为弹性模量(MPa)；σ为弹性变形阶段试样的应力(MPa)；ε为弹性变形阶段试样的应变。
拉伸强度计算公式：σ1＝p/(b×d)，σ1为试样的拉伸强度(MPa)；p为最大拉伸载荷(N)；b为试样宽度(mm)，d为试样厚度(mm)。
其具体的测试结果如下表1：
表1
| 试验例 | 弹性模量(MPa) | 拉伸强度(MPa) |
| 实施例1 | 0.783 | 0.695 |
| 实施例2 | 0.675 | 0.821 |
| 实施例3 | 0.684 | 0.764 |
| 对比例1 | 0.423 | 0.312 |
通过将实施例1、实施例2、实施例3与对比例1试验可知，丝素蛋白可有效的提高凝胶敷料的弹性模量与拉伸强度，使敷料使用时间更长，不易损坏。
试验例2
抗菌性试验，具体如下所示：
1、实验菌株的培养
复苏金黄色葡萄球菌和铜绿假单胞菌两种试验菌的冻干株，接种于普通肉汤培养基琼脂的平板上，在有氧条件下37℃培养24小时，必须经过形态学的鉴定为纯培养物，留作备用试验菌。
2、菌落数试验
参照中华人民共和国轻工业行业标准ISO 22196-2007和QB/T 2591-2003及日本抗菌制品技术协会的标准要求，拟用贴膜法对试验敷料进行检测，监测试验敷料的24小时抑菌率，同时参照GB 4789.2-2010的标准对试验后的菌落总数进行测定计数。将标准的菌株金黄色葡萄球菌、铜绿假单胞菌传代培养，混悬在TSB培养液中，运用麦氏比浊法分别将其配置成浓度为1.5×108CFU/mL的菌液，并依次做2次10倍递增稀释。之后分别取试验用菌液200μL滴加在放置于灭菌平皿的实施例1-3敷料、对比例2-4敷料、海藻酸钙敷料样品、空白对照，即直接滴在灭菌平皿上，每个样品做5个平行。灭菌镊子夹起已消毒的聚乙烯覆盖薄膜分别覆盖实施例1-3敷料、对比例2敷料、海藻酸钙敷料样品、空白对照上，铺平，使菌均匀接触样品，保证样品覆盖膜部位所铺的菌浓度不变。然后将灭菌培养皿放置在37℃、相对湿度大于90％条件下培养24小时，24小时后取出培养的样品，分别加入洗脱液20mL，反复洗实施例1-3敷料、对比例2-4敷料、海藻酸钙敷料样品、空白对照上的覆盖膜(用镊子夹起薄膜冲洗)，旋涡振荡1分钟充分摇匀后，取200μL接种于营养琼脂培养基(NA)中，在37℃下培养24小时后计算菌落数CFU，以上试验重复两次，试验结果取平均值。
所有试验数据都使用SPSS13.0软件包来做统计学的处理，数据用均数±标准差表示
各组间抗菌效果的统计方法是采用单因素的方差分析，组间两两比较采用的是SNK法，方差不齐的采用Dunnett T3法，P＜0.05表示有显著性差异。
按照中华人民共和国轻工业行业标准QB/T 2591-2003产品的抑菌及杀菌性能与稳定性试验方法评定试验敷料的抗菌性能，如计算出抑菌率是50-90％，提示试验敷料有抑菌作用，若抑菌率＞90％则提示试验样品有抗菌作用，若抑菌率＞99％，则提示试验样品具备强抗菌作用，计算公式为：抑菌率(％)＝(A-B)/A×100％；其中A代表是对照样品平均回收菌落，B代表的是试验样品平均回收菌数，具体计算结果如下表2所示：
表2
通过表2可知，本发明制备得到的敷料对金黄色葡萄球菌及铜绿假单胞菌抑菌率在99％以上，对比例2由于未加入抗菌剂，使抑菌率显著下降，同时对比例3-4的用量不满足要求，使得抑菌率下降但优于对比例2与海藻酸钙敷料样品，因此对于伤口的抑菌效果显著，同时提供局部的治疗。
试验例3
透气性、吸水和保湿性试验，具体如下所示：
吸水性是评价敷料好坏的一个重要指标，良好的敷料需要具有较强的吸收渗出液的能力，保持创面的清洁环境，防止二度感染。本试验在室温下采用吸水性能测试来测定敷料试样的吸液性。具体步骤如下：将干燥的敷料薄膜试样切成4cm×4cm的膜片，精确测量干重，每个样品浸在30mL去离子水中，室温下培养，相同间隔，溶胀的薄膜从水中取出，用滤纸吸去表面水分并测湿重。含水率和吸水率计算公式如下：
含水率(％)＝(w1-w0)/w1×100％
吸水率Q＝(w1-w0)/w1
式中；w1为试样的湿重，单位g；
w0为试样的干重，单位为g
透气性能测试，自制截面积为10cm2，高为3cm的透气杯。量取30mL蒸馏水透湿杯中，使溶液面与样品之间留有空隙，将样品剪成合适大小，放于容器上方，用上盖夹紧样品，为防止漏气，用生胶带将上盖和下盖固定住，在边缘涂上少量凡士林确保良好的密封性，称量并记录容器样品和液体的质量w1，将容器、液体和样品的质量w2，精准到0.0001g，透过率按照以下公式计算:
X＝((w1-w2)×1000×24)/Tg·m-2·d-1
具体如下表3所示：
表3
| 组别 | 吸水率Q/倍 | 保湿性RH％ | 透气性g·m<sup>-2</sup>·24h |
| 实施例1 | 10.32 | 21.36 | 1654.36 |
| 实施例2 | 9.34 | 24.15 | 1716.21 |
| 实施例3 | 9.68 | 20.34 | 1845.12 |
| 对比例3 | 3.25 | 5.36 | 865.32 |
| 对比例4 | 2.87 | 4.12 | 846.12 |
通过表3可知，聚乙烯吡咯烷酮带有亲水基团，容易与水结合成亲水凝胶氧化石墨烯具有亲水性以及静电斥力作用，同时含有羟基、羧基等极性基团，对极性溶液有较好的亲和力，从而聚乙烯吡咯烷酮与氧化石墨烯的相互协同作用促进吸水性与保湿性，同时聚乙烯吡咯烷酮含量较高时，聚乙烯吡咯烷酮环上的-C＝O与水分子结合变弱，使得吸水、保湿性能降低，反之聚乙烯吡咯烷酮含量较低时，与水结合达到一定的饱和状态，其吸水与保湿性稳定，同时氧化石墨烯为多孔结构，有效的促进透气性能，但各组分含量提高将堵塞该多孔结构，从而影响透气性，因此本发明的敷料在吸水率、保湿性与透气性优于对比例3与对比例4。
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (8)
1.一种生物材料复合水凝胶伤口敷料，其特征在于，
包括以下重量百分比含量的原料：角蛋白1-10％、丝素蛋白0.01-0.08％、聚乙烯吡咯烷酮3-21％、聚乳酸1-5％、聚乙二醇1-6％、抗菌剂0.01-0.06％、交联剂0.1-0.6％和余量为蒸馏水；
所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成。
2.根据权利要求1所述一种生物材料复合水凝胶伤口敷料，其特征在于，
所述交联剂为氯化钙。
3.一种生物材料复合水凝胶伤口敷料的制备方法，其特征在于，
包括以下步骤：
A、称取如下重量百分比含量的原料：角蛋白1-10％、丝素蛋白0.01-0.08％、聚乙烯吡咯烷酮3-21％、聚乳酸1-5％、聚乙二醇1-6％、抗菌剂0.01-0.06％、交联剂0.1-0.6％和余量为蒸馏水，所述抗菌剂为纳米银、氧化石墨烯与季铵化聚乙烯亚胺按质量比1:1:1组成；
B、将聚乙烯吡咯烷酮溶解于蒸馏水中，然后再加入聚乳酸与聚乙二醇混匀，得到混合液，备用；
C、向步骤B中的混合液中加入角蛋白与丝素蛋白分散均匀，得到分散液，备用；
D、将抗菌剂超声分散于蒸馏水中，得到抗菌分散液；
E、将步骤C得到的分散液与步骤D得到的抗菌分散液混匀，再加入交联剂搅拌反应得到水凝胶伤口敷料。
4.根据权利要求3所述一种生物材料复合水凝胶伤口敷料的制备方法，其特征在于，
在步骤B中，所述聚乙烯吡咯烷酮溶液质量分数为20-40％。
5.根据权利要求3所述一种生物材料复合水凝胶伤口敷料的制备方法，其特征在于，
在步骤D中，所述抗菌分散液质量分数为5-20％。
6.根据权利要求3所述一种生物材料复合水凝胶伤口敷料的制备方法，其特征在于，
在步骤D中，所述超声分散时间为30-40min。
7.根据权利要求3所述一种生物材料复合水凝胶伤口敷料的制备方法，其特征在于，
在步骤E中，所述搅拌反应时间为30-60min，搅拌反应温度为30-50℃。
8.根据权利要求3所述一种生物材料复合水凝胶伤口敷料的制备方法，其特征在于，
在步骤E中，反应溶液pH值控制在6.5-7.1。
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