CN114230814B - 一种担载伤科黄水的水凝胶及其制备方法 - Google Patents
一种担载伤科黄水的水凝胶及其制备方法 Download PDFInfo
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- CN114230814B CN114230814B CN202111656270.4A CN202111656270A CN114230814B CN 114230814 B CN114230814 B CN 114230814B CN 202111656270 A CN202111656270 A CN 202111656270A CN 114230814 B CN114230814 B CN 114230814B
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- Medicinal Preparation (AREA)
Abstract
本发明提供了一种担载伤科黄水的水凝胶,由聚乙烯醇和溶剂经过冷冻交联法制备,所述溶剂包括10~100wt%的伤科黄水和余量的水。本发明以聚乙烯醇和伤科黄水为主要原料,经过冷冻交联法制备得到担载有伤科黄水有效成分的PVA水凝胶,其可以用作伤口敷料,能够提供湿性的愈合环境,吸收渗出液,防止积液形成;能够提供有效的微生物屏障和持续的、持久的抗菌抗炎活性;能够持续释放药物,提供持续治疗效果;使用时具有舒适性,贴附性,使用方便和可避免去除时创面损坏的优点。实验结果表明,本发明提供的水凝胶具有良好的缓释性。
Description
技术领域
本发明涉及医用材料技术领域,具体是一种担载伤科黄水的水凝胶及其制备方法。
背景技术
伤科黄水是佛山市中医院骨伤科的院内洗剂,由黄连、黄柏、栀子、紫草、薄荷、白矾共6味中药组成。在临床多采用湿敷的形式,用于治疗骨骼损伤以及软组织损伤,具有抗炎消肿、活血化瘀、祛腐生新的功效。现代药理研究发现,该制剂具有抗感染、抗炎镇痛、改善微循环等多种药理作用,在治疗骨伤科患者伴有软组织损伤和创伤感染中具有其突出的优势。目前伤科黄水在临床上一般通过将药水喷洒在医用棉纱上直接湿敷使用,药效短,需反复用药,操作频繁,工作量大;且易挥发干燥,容易与创面粘连,影响创面修复;一般的纱布覆盖使用时对伤口恢复程度观察不便。因此,采用现代新材料技术进行剂型改进非常必要。
水凝胶(Hydrogel)是一类极为亲水、且自身含水的三维网络结构凝胶。它在水中迅速溶胀并在此溶胀状态可以保持大量体积的水而不被溶解。近来,由于水凝胶含水量高,柔软且富有弹性,与人体组织在物理性能上非常相似,并具备良好的生物相容性,水凝胶在生物医学领域如药物控释系统、经皮缓释制剂、创伤敷料、接触镜等方面的应用越来越受到关注。
根据来源不同,水凝胶材料分为天然水凝胶材料和人工合成水凝胶材料。天然水凝胶材料主要有多糖类,例如壳聚糖、海藻酸钠、纤维素、淀粉等;和多肽类,例如胶原、明胶、聚氨基酸等。人工合成水凝胶材料包括聚甲基丙烯酸、聚乙烯醇(PVA)。
聚乙烯醇(PVA)水凝胶的主要原料为聚乙烯醇。聚乙烯醇分子中含有大量的羟基,正因为这些羟基的存在使聚乙烯醇具备水溶性及溶胀特性。在目前发现的高聚物中PVA是唯一具备水溶性的高分子物质。正是由于聚乙烯醇具备如此多的优点,其正作为一种绿色的材料被广泛地应用在不同领域的生产生活中,受到科研和实际生产的广泛。通过科学家们长时间的努力,聚乙烯醇类薄膜产业化的发展已经初具规模,国内外如日本合成化学公司、法国GREENSOL公司、美国W.T.P公司等己经生产此类材料。不单是国外,在国内如中国包装总公司科技部支持株洲工学院和广东肇庆方兴包装材料公司研发和生产了此类材料,并且通过检验,正在走向市场。
室温条件下,PVA水溶液呈液态状,此时PVA分子之间不发生任何交联,因此不具备任何力学性能,为了得到具有一定力学性能的水凝胶就需要进行特别的工艺加工。PVA水凝胶的制备方法可以分为三种,即物理交联法,化学交联法,辐照交联法。冷冻交联法是一种将PVA溶液经过反复低温冷冻、升温融化,在反复冷冻过程中,PVA分子会发生团聚,由于氢键及物理缠绕而形成水凝胶。目前,冷冻交联法制备的PVA水凝胶具有白色、不透明、触感类似于橡胶等特点,一般在矫形外科手术中用于修复或替代关节软骨。
发明内容
有鉴于此,本发明所要解决的技术问题在于提供一种担载伤科黄水的水凝胶,其用作伤口敷料时,能够释放伤科黄水主要成分,具有抗菌抗炎活性,而且能够吸收渗出液,防止形成积液。
本发明提供了一种担载伤科黄水的水凝胶,由聚乙烯醇和溶剂经过冷冻交联法制备,所述溶剂包括10~100wt%的伤科黄水和余量的水。
本发明以聚乙烯醇和伤科黄水为主要原料,经过冷冻交联法制备得到担载有伤科黄水有效成分的PVA水凝胶,具有缓释效果。本发明提供的PVA水凝胶可以用作伤口敷料,能够提供湿性的愈合环境,吸收渗出液,防止积液形成;能够提供有效的微生物屏障和持续的、持久的抗菌抗炎活性;能够持续释放药物,提供持续治疗效果;使用时具有舒适性,贴附性,使用方便和可避免去除时创面损坏的优点。
本发明以聚乙烯醇为主要原料,在一个实施例中,所述聚乙烯醇的醇解度为78%以上,即为78%-100%;在一个实施例中,所述聚乙烯醇的醇解度为88%~100%。在一个实施例中,所述聚乙烯醇的醇解度为99%-100%。
在一个实施例中,所述聚乙烯醇的分子量为2×104~3×105。在一个实施例中,所述聚乙烯醇的分子量为5×104~1.5×105。在一个实施例中,所述聚乙烯醇的分子量为7×104~9×104。
本发明以伤科黄水为药物成分和溶剂,所述伤科黄水由黄连、黄柏、栀子、紫草、薄荷、白矾共6味中药组成的洗剂。在一个实施例中,所述伤科黄水的质量浓度为1~5%。在一个实施例中,所述伤科黄水的质量浓度为1.5~3%。
在一个实施例中,本发明还以水为溶剂。在一个实施例中,伤科黄水占溶剂的10%~100%。在一个实施例中,伤科黄水占溶剂的25%~100%。在一个实施例中,伤科黄水占溶剂的50%~100%。
在一个实施例中,聚乙烯醇在聚乙烯醇和溶剂的混合液中的质量分数为4%~20%。在一个实施例中,聚乙烯醇在混合液中的质量分数为6%~15%。在一个实施例中,聚乙烯醇在混合液中的质量分数为8%~12%。
本发明还提供了一种担载伤科黄水的水凝胶的制备方法,包括:
将聚乙烯醇溶解于溶剂中,得到混合液;所述溶剂包括10~100wt%的伤科黄水和余量的水;
将所述混合液进行冷冻-溶解,得到担载伤科黄水的水凝胶。
本发明首先将聚乙烯醇溶解于溶剂中,所述溶剂包括伤科黄水和水。具体而言,可以将聚乙烯醇在加热的条件下溶解于溶剂中,静置脱泡后得到混合液。
得到混合液后,将所述混合液进行冷冻-溶解,得到担载伤科黄水的水凝胶。具体而言,本发明将混合液放入模具中,水平置于冷冻装置中进行冷冻,冷冻完毕后在室温溶解,即可得到担载伤科黄水的水凝胶。在一个实施例中,冷冻-溶解可以循环进行。在一个实施例中,所述循环的次数为1~5次。在一个实施例中,所述循环的次数1~3次。在一个实施例中,所述循环的次数为1~2次。
在一个实施例中,所述单次冷冻的温度为-40~-20℃,所述单次冷冻的时间为0.5~12h。在一个实施例中,单次冷冻的时间为1~6小时。在一个实施例中,单次冷冻的时间为2~4小时。
本发明以聚乙烯醇和伤科黄水为主要原料,经过冷冻交联法制备得到担载有伤科黄水有效成分的PVA水凝胶,具有缓释效果。本发明提供的PVA水凝胶可以用作伤口敷料,能够提供湿性的愈合环境,吸收渗出液,防止积液形成;能够提供有效的微生物屏障和持续的、持久的抗菌抗炎活性;能够持续释放药物,提供持续治疗效果;使用时具有舒适性,贴附性,使用方便和可避免去除时创面损坏的优点。实验结果表明,本发明提供的水凝胶具有良好的缓释性。
附图说明
图1为纯黄水稀释50倍后的HPLC在340nm荧光检测曲线;
图2为实施例15制备的伤科黄水/PVA水凝胶释放后的HPLC在340nm荧光检测曲线;
图3为10%PVA含量的伤科黄水/PVA水凝胶的释放结果;
图4为12%PVA含量的伤科黄水/PVA水凝胶的释放结果。
图5为发明提供的水凝胶释放6h后的细胞存活率;
图6为发明提供的水凝胶释放12h后的细胞存活率;
图7为发明提供的水凝胶释放24h的细胞存活率。
具体实施方式
以下各实施例中,伤科黄水由佛山市中医院提供,其质量浓度为2%。
实施例1~3:醇解度与冷冻成胶情况验证
分别选取三种相同分子量,不同醇解度的PVA,与水配制成质量分数为15%的PVA溶液,并在-20℃的冰箱中进行不同冷冻时间、不同循环次数的成胶实验,其成胶情况如表1所示,表1为本发明实施例1~3制备水凝胶的成胶结果。
制备过程如下:
将PVA与水混合,加热溶解后,静置除泡;
将PVA水溶液定量的加入塑料模具中,水平放置于冷冻装置中,冷冻一定时间,取出置于室温溶解一段时间。冷冻一次溶解一次为一个循环。将PVA溶液经过一定循环次数后,制备成PVA水凝胶。
表1本发明实施例1~3制备水凝胶的成胶结果
表1中,能够成水凝胶划√,不能成水凝胶划×。
表1中,1次,2h表示单次冷冻时间为2h,冷冻-解冻循环1次。
实施例1~3的实验结果显示,醇解度为99%以上的PVA能够通过冷冻法制备得到PVA水凝胶,因而上述实施例优化出最优成胶醇解度为99%-100%
实施例4~6:分子量与冷冻成胶情况验证:
分别选取三种相同醇解度,不同分子量的PVA,配制成质量分数为15%的PVA溶液,并在-20℃的冰箱中进行不同冷冻时间、不同循环次数的成胶实验,其成胶情况如表2所示,表2为本发明实施例4~6制备水凝胶的成胶结果。
制备过程如下:
将PVA与水混合,加热溶解后,静置除泡;
将PVA水溶液定量的加入塑料模具中,水平放置于冷冻装置中,冷冻一定时间,取出置于室温溶解一段时间。冷冻一次溶解一次为一个循环。将PVA溶液经过一定循环次数后,制备成PVA水凝胶。
表2本发明实施例4~6制备水凝胶的成胶结果
实施例4-6的实验结果显示,所选的三种分子量的PVA,除实施例4中1次2h组不能成胶外,其余各组均能够成胶。实际使用中可根据需求(粘度,强度等)选择分子量。本申请后续试验选取1799型进行含“伤科黄水”的水凝胶制备及优化实验。
实施例7~10不同浓度的PVA含量对成胶情况的影响
选取1799型PVA,并以佛山市中医院的伤科黄水为溶剂,分别配制成5%,10%,15%,20%的溶液,在-20℃的冰箱中进行不同冷冻时间、不同循环次数的成胶实验,检验各组成胶情况及成胶后水凝胶的粘附能力,结果参见表3,表3为本发明实施例7~10制备水凝胶的成胶结果及性能测试结果。
制备过程如下:
将PVA与伤科黄水混合,加热溶解后,静置除泡;
将伤科黄水/PVA溶液定量的加入塑料模具中,水平放置于冷冻装置中,冷冻一定时间,取出置于室温溶解一段时间。冷冻一次溶解一次为一个循环。将伤科黄水/PVA溶液经过一定循环次数后,制备成伤科黄水/PVA水凝胶。
表3本发明实施例7~10制备水凝胶的成胶结果及性能测试结果
表3中,/表示未能形成成型的水凝胶;
表3中,以+表示形成水凝胶的粘性,其中+++表示很粘,++表示粘度适中,+表示粘度较低,-表示基本无粘性。
实施例7~10结果显示,较高PVA含量时PVA溶解不完全,不能用来检测其成胶情况,而过低的PVA含量(实施例7)则很难获得水凝胶。实施例8中成胶状况较好。
实施例11~14:不同浓度PVA含量单次冷冻成胶形成的伤科黄水/PVA水凝胶拉伸强度测试:
选取1799型PVA,并以佛山市中医院的伤科黄水为溶剂,分别配制成8%,10%,12%,14%的PVA-伤科黄水溶液,在-20℃的冰箱中单次冷冻不同时间,成胶后,以哑铃型裁刀制成样条,测试其拉伸性能,其结果如表4所示,表4为本发明实施例11~14制备水凝胶的成胶结果及拉伸性能测试结果。
表4为本发明实施例11~14制备水凝胶的成胶结果及拉伸性能测试结果
实施例 | PVA含量(%) | 2h拉伸强度(KPa) | 4h拉伸强度(KPa) |
11 | 8 | 未测得 | 21.79±3.18 |
12 | 10 | 31.68±3.66 | 42.34±1.73 |
13 | 12 | 42.95±0.22 | 19.49±3.71 |
14 | 14 | 52.77±3.16 | 75.96±13.50 |
实施例15:伤科黄水/PVA水凝胶的释放实验
选取1799型PVA,并以佛山市中医院的伤科黄水为溶剂,配制成PVA的质量分数为12%的伤科黄水/PVA溶液,在-20℃的冰箱中单次冷冻2h制成伤科黄水/PVA水凝胶。
以圆形裁刀裁取直径为8mm的水凝胶4片,称重。置于100mL的蓝盖瓶中,加入50mL的PBS溶液,旋紧后放置到恒温震荡箱中,37℃,60rpm下震荡,每隔一段时间取样1mL,取样后补加1mL的PBS,样品以HPLC,检测各取样点中盐酸小檗碱、栀子苷、绿原酸、表小檗碱、黄柏碱、黄连碱、药根碱、巴马汀的含量,并计算各点中各组分的释放百分比,结果参见图1和图2,图1为纯黄水稀释50倍后的HPLC在340nm荧光检测曲线,图2为实施例15制备的伤科黄水/PVA水凝胶释放后的HPLC在340nm荧光检测曲线。图2中,峰上标记X为盐酸小檗碱;B为表小檗碱,Y为药根碱,L为绿原酸。
实施例16
选取1799型PVA,并以佛山市中医院的伤科黄水和水为溶剂,配制成PVA的质量分数为10%、伤科黄水的质量分数分别为90%、45%、22.5%的伤科黄水/PVA溶液,在-20℃的冰箱中单次冷冻2h或4h制成伤科黄水/PVA水凝胶,分别计为:10PVA90HS2H、10PVA90HS4H、10PVA45HS2H、10PVA45HS4H、10PVA22.5HS2H和10PVA22.5HS4H。
选取1799型PVA,并以佛山市中医院的伤科黄水和水为溶剂,配制成PVA的质量分数为12%、伤科黄水的质量分数分别为88%、44%、22%的伤科黄水/PVA溶液,在-20℃的冰箱中单次冷冻2h或4h制成伤科黄水/PVA水凝胶,分别计为:12PVA88HS2H、12PVA88HS4H、12PVA44HS2H、12PVA44HS4H、12PVA22HS2H和12PVA22HS4H。
水凝胶的释放实验:将上述制备的伤科黄水/PVA水凝胶,37℃下在PBS中释放不同时间(0.25h,0.5h,1h,1.5h,2h,4h,6h,12h,24h,36h)并通过HPLC检测各点盐酸小檗碱的累计释放量,评估伤科黄水/PVA水凝胶的释放行为(模拟体液释放),具体方法如下:
1.将不同条件制备的伤科黄水/PVA水凝胶裁取3-4块,称重;
2. 100mL的蓝盖瓶中,加入50mL的PBS溶液,旋紧后放置到恒温震荡箱中,37℃,60rpm下震荡,每隔一段时间取样1mL,取样后补加1mL的PBS;
3.样品以HPLC检测,检测各取样点中盐酸小檗碱、栀子苷、绿原酸、表小檗碱、药根碱(黄水中部分主要成分),并计算各点中各组分的释放百分比。
结果参见图3和图4,图3为10%PVA含量的伤科黄水/PVA水凝胶的释放结果,图4为12%PVA含量的伤科黄水/PVA水凝胶的释放结果。由图3和图4可知,本发明提供的伤科黄水/PVA水凝胶可实现有效成分的缓释效果。
实施例17伤科黄水/PVA水凝胶的细胞毒性实验
实验目的:验证黄水/PVA水凝胶对皮肤相关细胞是否具有毒性;
细胞选择:NIH-3T3;
组别:(1)空白组:无处理;
(2)对照组:以0.4%DMEM培养基处理;
(3)10%PVA组:选取1799型PVA,并以水为溶剂,配制成10%的溶液,单次冷冻4h,得到10%PVA水凝胶;按照每毫升0.4%DMEM培养基中加入0.1g水凝胶,在37℃、60rad/m条件下释放,得到10%PVA组;
(4)1.125%黄水组:以0.4%DMEM培养基与伤科黄水配置,伤科黄水的质量浓度为1.125%;
(5)2.25%黄水组:以0.4%DMEM培养基与伤科黄水配置,伤科黄水的质量浓度为2.25%;
(6)4.5%黄水组:以0.4%DMEM培养基与伤科黄水配置,伤科黄水的质量浓度为4.5%;
(7)9%黄水组:以0.4%DMEM培养基与伤科黄水配置,伤科黄水的质量浓度为9%;
(8)12.5%释放液组:选取1799型PVA,并以伤科黄水为溶剂,配制成10%的溶液,单次冷冻4h,得到90%黄水/10%PVA水凝胶;按照每毫升0.4%DMEM培养基中加入0.1g水凝胶,在37℃、60rad/m条件下释放6h,释放液以0.4%DMEM培养基稀释8倍,得到12.5%释放液组;
(9)25%释放液组:选取1799型PVA,并以伤科黄水为溶剂,配制成10%的溶液,单次冷冻4h,得到90%黄水/10%PVA水凝胶;按照每毫升0.4%DMEM培养基中加入0.1g水凝胶,在37℃、60rad/m条件下释放6h,释放液以0.4%DMEM培养基稀释4倍,得到25%释放液组;
(10)50%释放液组:选取1799型PVA,并以伤科黄水为溶剂,配制成10%的溶液,单次冷冻4h,得到90%黄水/10%PVA水凝胶;按照每毫升0.4%DMEM培养基中加入0.1g水凝胶,在37℃、60rad/m条件下释放6h,释放液以0.4%DMEM培养基稀释2倍,得到50%释放液组;
(11)100%释放液组:选取1799型PVA,并以伤科黄水为溶剂,配制成10%的溶液,单次冷冻4h,得到90%黄水/10%PVA水凝胶;按照每毫升0.4%DMEM培养基中加入0.1g水凝胶,在37℃、60rad/m条件下释放6h,得到100%释放液组;
实验步骤:
1、96孔板中以5*10^3cell/孔种细胞,加入10%的DMEM培养基培养24h;
2、各组吸取200uL加入1中96孔板中培养24h。
3、吸弃各孔培养基,加入200μL的无菌PBS洗去孔板底部沉降物2次,吸弃PBS,加入100μL20%MTT(浓度5mg/mL)的0.4%DMEM培养基,孵育4h。吸弃培养基,加入150μL的DMSO,振荡溶解15min,转入新的96孔板中,酶标仪492nm读数。结果参见图5,图5为发明提供的水凝胶释放6h后的细胞存活率。
与上述实验过程相同,区别在于,释放液组由90%黄水/10%PVA水凝胶释放12h得到,结果参见图6,图6为发明提供的水凝胶释放12h后的细胞存活率。
与上述实验过程相同,区别在于,释放液组由90%黄水/10%PVA水凝胶释放24h得到,结果参见图7,图7为发明提供的水凝胶释放24h的细胞存活率。由图5、图6和图7可知,本发明提供的水凝胶对皮肤相关细胞不具备毒性。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (6)
1.一种担载伤科黄水的水凝胶,由醇解度为99~100%的聚乙烯醇和伤科黄水经过冷冻交联法制备,所述伤科黄水的质量浓度为2%;所述聚乙烯醇占聚乙烯醇和伤科黄水总量的10~15wt%。
2.根据权利要求1所述的水凝胶,其特征在于,所述聚乙烯醇的分子量为2×104~3×105。
3.一种权利要求1所述的担载伤科黄水的水凝胶的制备方法,包括:
将聚乙烯醇溶解于伤科黄水中,得到混合液;将所述混合液进行冷冻-溶解,得到担载伤科黄水的水凝胶。
4.根据权利要求3所述的制备方法,其特征在于,所述聚乙烯醇的分子量为2×104~3×105。
5.根据权利要求3所述的制备方法,其特征在于,所述冷冻的时间为0.5~12h。
6.根据权利要求3所述的制备方法,其特征在于,将所述混合液进行冷冻-溶解,循环1~5次。
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