CN113521381A - 促进细胞增殖的医用膜材料及其制备方法和应用 - Google Patents
促进细胞增殖的医用膜材料及其制备方法和应用 Download PDFInfo
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Abstract
本发明属于医药新材料技术领域,本发明提供的促进细胞增殖的医用膜材料的制备方法,通过将量子点的氨基和海藻酸钠的羧基结合,在海藻酸钠的表面形成一层致密的膜,工艺简洁,原料容易获取。本发明提供的促进细胞增殖的医用膜材料,避免直接滴入量子点溶液进入细胞,因为量子点具有细胞毒性会导致细胞的死亡,同时弥补了海藻酸盐敷料的缺点,并且有促进细胞增殖的功能,可缝合,有利于上皮细胞在材料上生长以促进伤口的愈合,减少瘢痕形成。此外,本发明提供的促进细胞增殖的医用膜材料应用于伤口覆盖,其少量的量子点产生的ROS少对细胞影响不大,结合NIR照射下量子点产生的ROS以促进细胞增殖,又不具有细胞毒性。
Description
技术领域
本发明属于医药新材料技术领域,具体涉及促进细胞增殖的医用膜材料及其制备方法。
背景技术
近年来,科学家们正在大力研究治疗伤口损伤,加速伤口愈合的方法。伤口即上皮细胞损伤,包括外科手术后的伤口、烧伤科的伤口,由于炎症的反复刺激,容易形成瘢痕,因此需要敷料对伤口进行临时覆盖保护,以促进伤口修复。对于伤口的修复所采用的敷料最好具有能够良好地促进细胞生长的功能的材料,由于组织工程材料相当于人工细胞外基质,具有诱导组织再生、调节细胞生长和功能分化,无疑是伤口覆盖保护膜的理想材料。
现有技术中会选择海藻酸盐材料作为敷料,海藻酸盐是从褐藻类的海带或马尾藻中提取的一种多聚糖,是一种天然的三维网状高分子材料。具有无毒、无刺激性、高吸湿性、生物相容性等优良性能,但是依然不具备良好地促进细胞增殖的功效,从而无法更高效率的促进伤口的愈合。
发明内容
针对上述现有技术的不足,本发明提供了促进细胞增殖的医用膜材料及其制备方法和应用,目的是为了解决现有作为伤口敷料的海藻酸盐材料,不具备良好地促进细胞增殖的功效,从而无法更高效率的促进伤口的愈合的技术问题。
本发明提供的促进细胞增殖的医用膜材料的制备方法,具体技术方案如下:
促进细胞增殖的医用膜材料的制备方法,包括如下步骤:
S1,将3-氨丙基三甲氧基硅烷、1,8-萘酰亚胺、超纯水混合并搅拌,获得硅量子点前体溶液;
S2,将步骤S1的硅量子点前体溶液经过紫外光照射,获得硅量子点样品溶液;
S3,将步骤S2中的硅量子点样品溶液进行低速离心,收集上清液,并对上清液进行透析,收集截留液,将所述截留液用超纯水稀释后干燥,获得高荧光SiNPs;
S4,将步骤S3中的高荧光SiNPs用超纯水配置为量子点溶液,将海藻酸钠用超纯水配置为海藻酸钠溶液;
S5,将步骤S4中的量子点溶液滴洒入在步骤S5中的海藻酸钠溶液中,获得促进细胞增殖的医用膜材料。
在某些实施方式中,步骤S1中,3-氨丙基三甲氧基硅烷与超纯水的体积比为1:9,1,8-萘酰亚胺与超纯水的质量比为1:45,所述搅拌的时间为10min。
在某些实施方式中,步骤S2中,所述硅量子点样品溶液是由硅量子点前体溶液经过紫外光照射30min获得,所述硅量子点样品溶液为绿色荧光。
在某些实施方式中,步骤S2中,经过紫外光照射后的所述硅量子点样品溶液需要冷却至30℃以下。
在某些实施方式中,步骤S3中,所述低速离心的速度为3000-6000rpm,所述低速离心的时间为10-20min;所述透析膜的截留分子量为1-5kDa。
在某些实施方式中,步骤S4中,量子点溶液的浓度为5-30%,海藻酸钠溶液的浓度为2-10%。
在某些实施方式中,步骤S5中,所述促进细胞增殖的医用膜材料的厚度为0.1mm。
本发明还提供了促进细胞增殖的医用膜材料,利用上述的方法制备的医用膜材料。
本发明还提供了促进细胞增殖的医用膜材料的应用,基于上述的促进细胞增殖的医用膜材料,将促进细胞增殖的医用膜材料用于制作伤口的敷料。
本发明具有以下有益效果:本发明提供的促进细胞增殖的医用膜材料的制备方法,通过将量子点的氨基和海藻酸钠的羧基结合,在海藻酸钠的表面形成一层致密的膜,工艺简洁,原料容易获取。本发明提供的促进细胞增殖的医用膜材料,避免直接滴入量子点溶液进入细胞,因为量子点具有细胞毒性会导致细胞的死亡,同时弥补了海藻酸盐敷料的缺点,并且有促进细胞增殖的功能,可缝合,有利于上皮细胞在材料上生长以促进伤口的愈合,减少瘢痕形成。此外,本发明提供的促进细胞增殖的医用膜材料应用于伤口覆盖,其少量的量子点产生的ROS少对细胞影响不大,结合NIR照射下量子点产生的ROS以促进细胞增殖,又不具有细胞毒性。
附图说明
图1是本发明提供的促进细胞增殖的医用膜材料的制备方法的流程图;
图2是本发明实施例2中FTIR检测的光谱图;
图3是本发明实施例2中硅量子点与海藻酸钠连接示意图;
图4是本发明实施例2中促进细胞增殖的医用膜材料与海藻酸钠膜材料实物图;
图5是本发明实施例2中细胞染色荧光显微镜图;
图6是本发明实施例3中不同培养时间后的细胞的增殖情况。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚明白,以下结合具体实施例,并参照附图1-6,对本发明进一步详细说明。
实施例1
本实施例提供的促进细胞增殖的医用膜材料的制备方法,具体技术方案如下:
促进细胞增殖的医用膜材料的制备方法,包括如下步骤:
S1,将100ml的3-氨丙基三甲氧基硅烷(APS)加入至溶解了20g的1,8-萘酰亚胺的900ml的超纯水中,搅拌10min,获得硅量子点前体溶液;
S2,将步骤S1的硅量子点前体溶液经过紫外光照射30min获得绿色荧光的硅量子点样品溶液,后冷却至30℃以下。
S3,将步骤S2中的硅量子点样品溶液进行6000rpm,15min的低速离心,低速离心的目的在于使1,8-萘酰亚胺沉淀下来,硅量子点在这种低离心率下不会沉淀,因此所得硅量子点和APS保留在上清液中,收集上清液,并对上清液进行透析(MWCO,1000,Spectra/Pro),APS由于分子量较小(<1kda)而被充分过滤,收集截留液,收集的SiNPs的分子量远大于1kda,将截留液用超纯水稀释后干燥,称重获得10g高荧光SiNPs。
S4,将步骤S3中的高荧光SiNPs用超纯水配置为10%量子点溶液,将海藻酸钠(SA)用超纯水配置为2%海藻酸钠溶液;
S5,将步骤S4中的量子点溶液滴洒入在步骤S5中的海藻酸钠溶液中,获得促进细胞增殖的医用膜材料。获得促进细胞增殖的医用膜材料,其厚度为0.1mm。以上操作均在超净台进行,全程保持无菌。
实施例2
本实施例提供了促进细胞增殖的医用膜材料,利用实施例1的方法制备的医用膜材料。
通过傅里叶转换红外光谱(FTIR)对本实施例提供的促进细胞增殖的医用膜材料进行检测,如图2所示,量子点接在SA上,其中QD表示硅量子点,QDP表示促进细胞增殖的医用膜材料。本实施例的促进细胞增殖的医用膜材料连接结构图如图3所示,量子点的氨基和海藻酸钠的羧基结合。实物图如图4所示,QDP相较于SA更加的透明和轻薄。
对本实施例中的促进细胞增殖的医用膜材料(QDP)、海藻酸钠(SA)通过Calcein-AM、PI和Merge染料进行安全性检测,如图5所示,在QDP膜上的细胞存活量明显高于SA膜的,说明本实施例中的促进细胞增殖的医用膜材料更具安全性。
实施例3
本实施例提供的促进细胞增殖的医用膜材料的应用,将促进细胞增殖的医用膜材料用于制作伤口的敷料。
从狗腹股沟脂肪中提取脂肪干细胞原代培养,增殖、传代后,用P3代时传至96孔板中从狗腹股沟脂肪中提取脂肪干细胞原代培养,增殖、传代后,用P3代时传至96孔板,培养基:低糖DMEM+10%胎牛血清+1%双抗。细胞在37℃、5%CO2的恒温细胞培养箱中培养。将实施例1制备好的膜剪成0.3*0.3mm大小,以便放入96孔板中,以上操作均在超净台进行,全程保持无菌。利用上述的促进细胞增殖的医用膜材料和NIR(Near Infrared)处理过的细胞分别在第1、3、7天用CCK-8检测试剂进行处理4h后测吸光度以判断细胞量来说明细胞增殖程度。如图6所示,其中,control代表空白对照既没有促进细胞增殖的医用膜材料也没有NIR,NIR代表仅有红外光照射,DQP代表覆盖了促进细胞增殖的医用膜材料,DQP+NIR代表覆盖了促进细胞增殖的医用膜材料后并进行红外光照射,单独的近红外光对细胞增殖没有影响,近红外光(NIR)照射量子点可以产生活性氧自由基(ROS)一定量的ROS可以促进细胞增殖。
实施例4
本实施例提供的促进细胞增殖的医用膜材料的制备方法,具体技术方案如下:
促进细胞增殖的医用膜材料的制备方法,包括如下步骤:
S1,将100ml的3-氨丙基三甲氧基硅烷(APS)加入至溶解了20g的1,8-萘酰亚胺的900ml的超纯水中,搅拌10min,获得硅量子点前体溶液;
S2,将步骤S1的硅量子点前体溶液经过紫外光照射30min获得绿色荧光的硅量子点样品溶液,后冷却至30℃以下。
S3,将步骤S2中的硅量子点样品溶液进行2000rpm,20min的低速离心,低速离心的目的在于使1,8-萘酰亚胺沉淀下来,硅量子点在这种低离心率下不会沉淀,因此所得硅量子点和APS保留在上清液中,收集上清液,并对上清液进行透析(MWCO,5000,Spectra/Pro),APS由于分子量较小(<5kda)而被充分过滤,收集截留液,收集的SiNPs的分子量大于5kda,将截留液用超纯水稀释后干燥,称重获得10g高荧光SiNPs。
S4,将步骤S3中的高荧光SiNPs用超纯水配置为30%量子点溶液,将海藻酸钠(SA)用超纯水配置为10%海藻酸钠溶液;
S5,将步骤S4中的量子点溶液滴洒入在步骤S5中的海藻酸钠溶液中,获得促进细胞增殖的医用膜材料。获得促进细胞增殖的医用膜材料,其厚度为0.1mm。以上操作均在超净台进行,全程保持无菌。
实施例5
本实施例提供的促进细胞增殖的医用膜材料的制备方法,具体技术方案如下:
促进细胞增殖的医用膜材料的制备方法,包括如下步骤:
S1,将100ml的3-氨丙基三甲氧基硅烷(APS)加入至溶解了20g的1,8-萘酰亚胺的900ml的超纯水中,搅拌10min,获得硅量子点前体溶液;
S2,将步骤S1的硅量子点前体溶液经过紫外光照射30min获得绿色荧光的硅量子点样品溶液,后冷却至30℃以下。
S3,将步骤S2中的硅量子点样品溶液进行5000rpm,10min的低速离心,低速离心的目的在于使1,8-萘酰亚胺沉淀下来,硅量子点在这种低离心率下不会沉淀,因此所得硅量子点和APS保留在上清液中,收集上清液,并对上清液进行透析(MWCO,2000,Spectra/Pro),APS由于分子量较小(<2kda)而被充分过滤,收集截留液,收集的SiNPs的分子量大于2kda,将截留液用超纯水稀释后干燥,称重获得10g高荧光SiNPs。
S4,将步骤S3中的高荧光SiNPs用超纯水配置为5%量子点溶液,将海藻酸钠(SA)用超纯水配置为5%海藻酸钠溶液;
S5,将步骤S4中的量子点溶液滴洒入在步骤S5中的海藻酸钠溶液中,获得促进细胞增殖的医用膜材料。获得促进细胞增殖的医用膜材料,其厚度为0.1mm。以上操作均在超净台进行,全程保持无菌。
上述仅本发明较佳可行实施例,并非是对本发明的限制,本发明也并不限于上述举例,本技术领域的技术人员,在本发明的实质范围内,所作出的变化、改型、添加或替换,也应属于本发明的保护范围。
Claims (9)
1.促进细胞增殖的医用膜材料的制备方法,其特征在于,包括如下步骤:
S1,将3-氨丙基三甲氧基硅烷、1,8-萘酰亚胺、超纯水混合并搅拌,获得硅量子点前体溶液;
S2,将步骤S1的硅量子点前体溶液经过紫外光照射,获得硅量子点样品溶液;
S3,将步骤S2中的硅量子点样品溶液进行低速离心,收集上清液,并对上清液进行透析,收集截留液,将所述截留液用超纯水稀释后干燥,获得高荧光SiNPs;
S4,将步骤S3中的高荧光SiNPs用超纯水配置为量子点溶液,将海藻酸钠用超纯水配置为海藻酸钠溶液;
S5,将步骤S4中的量子点溶液滴洒入在步骤S5中的海藻酸钠溶液中,获得促进细胞增殖的医用膜材料。
2.根据权利要求1所述的促进细胞增殖的医用膜材料的制备方法,其特征在于,步骤S1中,3-氨丙基三甲氧基硅烷与超纯水的体积比为1:9,1,8-萘酰亚胺与超纯水的质量比为1:45,所述搅拌的时间为10min。
3.根据权利要求1所述的促进细胞增殖的医用膜材料的制备方法,其特征在于,步骤S2中,所述硅量子点样品溶液是由硅量子点前体溶液经过紫外光照射30min获得,所述硅量子点样品溶液为绿色荧光。
4.根据权利要求1所述的促进细胞增殖的医用膜材料的制备方法,其特征在于,步骤S2中,经过紫外光照射后的所述硅量子点样品溶液需要冷却至30℃以下。
5.根据权利要求1所述的促进细胞增殖的医用膜材料的制备方法,其特征在于,步骤S3中,所述低速离心的速度为3000-6000rpm,所述低速离心的时间为10-20min;所述透析膜的截留分子量为1-5kDa。
6.根据权利要求1所述的促进细胞增殖的医用膜材料的制备方法,其特征在于,步骤S4中,量子点溶液的浓度为5-30%,海藻酸钠溶液的浓度为2-10%。
7.根据权利要求1所述的促进细胞增殖的医用膜材料的制备方法,其特征在于,步骤S5中,所述促进细胞增殖的医用膜材料的厚度为0.1mm。
8.促进细胞增殖的医用膜材料,其特征在于,利用权利要求1-7任一项所述的方法制备的医用膜材料。
9.促进细胞增殖的医用膜材料的应用,基于权利要求8所述的促进细胞增殖的医用膜材料,其特征在于,将促进细胞增殖的医用膜材料用于制作伤口的敷料。
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