CN107185040A - 纳米磷酸三钙‑水凝胶‑ⅰ型胶原生物材料的制备方法及其用途和用法 - Google Patents

纳米磷酸三钙‑水凝胶‑ⅰ型胶原生物材料的制备方法及其用途和用法 Download PDF

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CN107185040A
CN107185040A CN201710373395.3A CN201710373395A CN107185040A CN 107185040 A CN107185040 A CN 107185040A CN 201710373395 A CN201710373395 A CN 201710373395A CN 107185040 A CN107185040 A CN 107185040A
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tricalcium phosphate
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程丽佳
时政
雨田
鄢硕
朱江
蔡佩玲
王婷
曾怡乐
廖羽
杨明
谢欣蓉
唐椿鑫
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Beijing Haokun Kangyuan Medical Science And Technology Development Co ltd
Dragon Totem Technology Hefei Co ltd
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Abstract

本发明属于医用生物材料技术领域,具体为纳米磷酸三钙‑水凝胶‑Ⅰ型胶原复合材料的制备方法及其用途和用法。将水凝胶在低温下溶解,按25胶原:15水凝胶:60磷酸三钙的比例依次分别加入纳米磷酸三钙粉体和Ⅰ型胶原,最后用藻酸盐翻模法进行塑形。该材料用于制备人工骨。本发明提供的纳米磷酸三钙‑水凝胶‑Ⅰ型胶原复合材料的制备方法及其用途和用法,改进材料的弹性、硬度和孔隙率,有利于血管的生成,提供营养,更加有利于骨组织的形成,提高成骨效率。

Description

纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料的制备方法及其用 途和用法
技术领域
本发明属于医用生物材料技术领域,具体为纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料的制备方法及其用途和用法。
背景技术
在创伤骨科以及外科临床治疗邻域,治疗骨缺损、骨不连等修复问题中,需要在缺损部位以及不连部位植入或填充自体骨、异体骨以及人工骨来达到治愈的目的。目前,自体骨移植由于无排斥和免疫反应成为临床上骨移植的金标准,但是,有限的自体骨由于大小和形状与缺损区域并不匹配,会引起患者自身疼痛、感染和血肿等并发症,因此其未能在临床上广泛应用;同时,同种异体骨移植又会使受者发生严重的免疫反应甚至染上传染性疾病,再加上供者骨来源有限,使其应用也受到了限制。在骨移植物供不应求时,人工合成生物材料作为骨替代材料已经被广泛应用来代替自体或同种异体骨移植。磷酸三钙(Tricalcium Phosphate,TCP)作为具有和骨组织相似的物理性质,分子式为Ca3(PO4)2,是一种生物相容性、生物降解性较好的陶瓷材料,其在生物医学领域具有重要的应用价值;而纳米磷酸三钙(n-TCP)较TCP粒径更均一,成骨效果更好,可有效替代TCP;水凝胶成份是PLGA-PEG-PLGA聚合物,是由一定比例的DL-丙交酯、乙交酯和聚乙二醇合成,其特点是在温度低于相转变温度(35±2)℃时,聚合物可溶于水形成自由流动液体,温度升高至相转变温度以上,聚合物的水溶液发生相变,形成非化学交联的凝胶,并且形成凝胶的过程是可逆的,在温度降低时又可发生凝胶到溶液的转变,水凝胶作为一种具有网状结构三维材料,具有和细胞外基质相似的微结构,被广泛用于组织工程支架。Ⅰ型胶原是人体骨组织分泌的主要矿物质,其具有高张力、低延展性、生物相容性好、可刺激细胞增殖、分化和生物可降解性的特性。利用水凝胶的水溶性可作为固化液,胶原的促新生骨形成能力,以及纳米磷酸三钙可缓慢释放Ca、P离子的特性,将三种材料采用不同比例,不同方式混合处理,在不需要高温烧结的条件下,制备一种可快速成形,成骨效果好的生物复合材料,即磷酸三钙-水凝胶-Ⅰ型胶原复合材料。三种材料单独使用时具有不同程度的缺陷,不能满足生物力学的性能和较好的成骨性能。
发明内容
本发明的目的在于提供一种纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料的制备方法,旨在改进以磷酸钙和高分子材料为基底的人工骨的弹性、硬度和孔隙率,从而有利于血管的生成,提高成骨效率。
具体的技术方案为:
纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料,包括纳米磷酸三钙、温敏性的水凝胶和Ⅰ型胶原;所述的水凝胶成份是PLGA-PEG-PLGA聚合物,由DL-丙交酯、乙交酯和聚乙二醇合成。
将水凝胶与蒸馏水进行混合,称量温敏水凝胶1g后加入蒸馏水4ml,在2-10℃下搅拌混合,搅拌时间30min/d,顺时针搅拌,连续搅拌3d,得到水凝胶溶液,呈透明胶体悬浊液;
称取纳米磷酸三钙粉体,在20℃条件下,将水凝胶溶液与纳米磷酸三钙粉体按20:80的比例进行混合,混合时将纳米磷酸三钙缓慢加入水凝胶溶液,边加边搅拌,搅拌时温度不低于20℃,搅拌时间5-10min,得到纳米磷酸三钙-水凝胶混合材料;
将Ⅰ型胶原溶于纳米磷酸三钙-水凝胶混合液中,按胶原与纳米磷酸三钙-水凝胶混合液按1:3的比例进行混合,混合时将胶原加入纳米磷酸三钙-水凝胶混合液中,在温度不低于20℃的条件搅拌混合,在搅拌过程中滴中稀盐酸,使混合液最后pH=1,搅拌时间5-10min,得到纳米磷酸三钙-水凝胶-Ⅰ型胶原混合液,各成份最终比例为25:15:60,此时材料混合液仍为混合浆液;
将得到的混合浆液用藻酸盐翻模法进行塑形,藻酸盐是由海带中提取的天然多糖碳水化合物,具体生物安全性。首先,制备藻酸盐类模具,将藻酸盐与水混合,将糊状的藻酸盐类材料迅速覆盖于任何形状的模型上,待其干燥固化后取下藻酸盐类模具。然后,将纳米磷酸三钙-水凝胶-Ⅰ型胶原混合浆液注射入藻酸盐模具中,进行倒模,将温度升到37℃以上(此温度可模拟人体温度),得到纳米磷酸三钙-水凝胶-Ⅰ型胶原复合材料,最后将藻酸盐溶解掉。
最后成形的材料放入2.5%的戊二醛溶液中,戊二醛可作为一种有机交联剂,使纳米磷酸三钙-水凝胶-Ⅰ型胶原材料交联形成具有一定孔隙的网状结构,有利于新生血管的长入,为新骨组织提供营养。交联12h后,取出材料,用95%的乙醇冲洗,最后将材料放入-20℃下冷冻2h,然后冻干24h得到纳米磷酸三钙-水凝胶-Ⅰ型胶原复合材料。
本发明的制备方法解决了磷酸三钙粉体需要烧结成型的缺点,做到了随意塑形,且生物力学性能强,空隙结构均匀,孔径在200-400μm,孔隙率为50%左右,有利于新生血管长入及新生骨组织形成。
本发明提供的纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料,采用温敏性的水凝胶作为一种具有网状结构三维材料,又是一种好的固化液,在体内可起到缓释磷酸三钙的作用,将Ca、P离子缓慢释放以形成新生骨组织,解决磷酸三钙粉体需要烧结成型的缺点,且有利于血管形成,从而有利于骨组织的再生;Ⅰ型胶原作为骨细胞的生物支架,可帮助磷酸三钙加速骨细胞的形成,从而加速诱导机体内新生骨组织形成,利用水凝胶的温度敏感性,在35℃上下2℃可进行溶液和凝胶的转变,可塑形成任何形状,制备方法简单,无缺任何烧结过程。
具体实施方式
结合实施例说明本发明的具体实施方式。
将水凝胶与蒸馏水按1:4的比例进行混合,称量温敏水凝胶1g后加入蒸馏水4ml,在2-10℃下搅拌混合,搅拌时间30min/d,顺时针搅拌,连续搅拌3d,得到水凝胶溶液,呈透明胶体悬浊液;
称取纳米磷酸三钙粉体,在20℃条件下,将水凝胶溶液与纳米磷酸三钙粉体按20:80的比例进行混合,混合时将纳米磷酸三钙缓慢加入水凝胶溶液,边加边搅拌,搅拌时温度不低于20℃,搅拌时间5-10min,得到纳米磷酸三钙-水凝胶混合材料;
将Ⅰ型胶原溶于纳米磷酸三钙-水凝胶混合液中,按胶原与纳米磷酸三钙-水凝胶混合液按1:3的比例进行混合,混合时将胶原加入纳米磷酸三钙-水凝胶混合液中,在温度不低于20℃的条件搅拌混合,在搅拌过程中滴中稀盐酸,使混合液最后pH=1,搅拌时间5-10min,得到纳米磷酸三钙-水凝胶-Ⅰ型胶原混合液,各成份最终比例为25:15:60,此时材料混合液仍为混合浆液;
将得到的混合浆液用藻酸盐翻模法进行塑形,藻酸盐是由海带中提取的天然多糖碳水化合物,具体生物安全性。首先,制备藻酸盐类模具,将藻酸盐与水混合,将糊状的藻酸盐类材料迅速覆盖于任何形状的模型上,待其干燥固化后取下藻酸盐类模具。然后,将纳米磷酸三钙-水凝胶-Ⅰ型胶原混合浆液注射入藻酸盐模具中,进行倒模,将温度升到37℃以上(此温度可模拟人体温度),得到纳米磷酸三钙-水凝胶-Ⅰ型胶原复合材料,最后将藻酸盐溶解掉。
最后成形的材料放入2.5%的戊二醛溶液中,戊二醛可作为一种有机交联剂,使纳米磷酸三钙-水凝胶-Ⅰ型胶原材料交联形成具有一定孔隙的网状结构,有利于新生血管的长入,为新骨组织提供营养。交联12h后,取出材料,用95%的乙醇冲洗,最后将材料放入-20℃下冷冻2h,然后冻干24h得到纳米磷酸三钙-水凝胶-Ⅰ型胶原复合材料。

Claims (4)

1.纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料,其特征在于,包括纳米磷酸三钙、温敏性的水凝胶和Ⅰ型胶原;所述的水凝胶成份是PLGA-PEG-PLGA聚合物,由DL-丙交酯、乙交酯和聚乙二醇合成。
2.根据权利要求1所述的纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料的制备方法,其特征在于,包括以下过程:
水凝胶1g加入蒸馏水4ml,在2-10℃下搅拌混合,搅拌时间30min/d,顺时针搅拌,连续搅拌3d,得到水凝胶溶液,呈透明胶体悬浊液;
称取纳米磷酸三钙粉体,在20℃条件下,将水凝胶溶液与纳米磷酸三钙粉体按20:80的比例进行混合,混合时将纳米磷酸三钙缓慢加入水凝胶溶液,边加边搅拌,搅拌时温度不低于20℃,搅拌时间5-10min,得到纳米磷酸三钙-水凝胶混合材料;
将Ⅰ型胶原溶于纳米磷酸三钙-水凝胶混合液中,Ⅰ型胶原与纳米磷酸三钙-水凝胶混合液按体积1:3的比例进行混合,混合时将胶原加入纳米磷酸三钙-水凝胶混合液中,在温度不低于20℃的条件搅拌混合,在搅拌过程中滴中稀盐酸,使混合液pH=1,搅拌时间5-10min,得到纳米磷酸三钙-水凝胶-Ⅰ型胶原混合液;
将得到的混合浆液用藻酸盐翻模法进行塑形;首先,制备藻酸盐类模具,将藻酸盐与水混合,将糊状的藻酸盐类材料迅速覆盖于任何形状的模型上,待其干燥固化后取下藻酸盐类模具;然后,将纳米磷酸三钙-水凝胶-Ⅰ型胶原混合浆液注射入藻酸盐模具中,进行倒模,将温度升到模拟的人体温度,得到纳米磷酸三钙-水凝胶-Ⅰ型胶原复合材料,最后将藻酸盐溶解掉;
最后成形的材料放入2.5%的戊二醛溶液中,使纳米磷酸三钙-水凝胶-Ⅰ型胶原材料交联形成具有一定孔隙的网状结构,交联12h后,取出材料,用95%的乙醇冲洗,最后将材料放入-20℃下冷冻2h,然后冻干24h得到纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料。
3.根据权利要求1所述的纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料的用途,其特征在于,作为人工骨材料。
4.根据权利要求1所述的纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料的用法,其特征在于,根据骨缺损的形状,先对患者进行骨缺损处进行CT扫描,然后采用3D打印的方式把缺损模型打印出来,再根据此模型制备相应形状的纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料,将纳米磷酸三钙-水凝胶-Ⅰ型胶原生物材料安装到骨缺损处。
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