CN112479737B - 一种可控多孔生物陶瓷支架及其制备方法、应用 - Google Patents
一种可控多孔生物陶瓷支架及其制备方法、应用 Download PDFInfo
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Abstract
本发明提供了一种可控多孔生物陶瓷支架及其制备方法、应用,制备方法包括如下步骤:绘制具有多孔结构的支架模型;将生物陶瓷和造孔剂混合搅拌得到混合粉末;加入光敏树脂和分散剂得到浆料;将浆料放入料缸槽,将浆料打印制成坯体;将坯体清洗并去除表面液体后对其进行脱脂、烧结,得到支架;将支架放入酸性溶液中进行浸泡,完成后使用清水进行清洗,得到支架;在聚乙烯醇溶液中加入表面涂层粉末并搅拌,得到涂层材料溶液;将支架放入涂层材料溶液中,得到表面附着有涂层材料的支架。本发明的支架中的多孔结构的形状、孔隙率以及形状等参数可控,能够满足植入支架的个性化需求;具有良好的生物活性和生物相容性,适合作为骨修复的材料。
Description
技术领域
本发明涉及生物支架制备技术,具体涉及一种可控多孔生物陶瓷支架及其制备方法、应用。
背景技术
骨组织缺损是一种数量庞大且严重危害健康的一种损伤。创伤、手术、先天畸形、人口老龄化和其他特殊情况造成的骨缺损正影响着世界数百万人的生活质量。在我国骨缺损现存和每年增量都是巨大的,并且我国骨质疏松患者超过一亿人,并且30%的老年骨折与骨质疏松有关,每年新增骨质疏松患者约为300万人。尽管人体骨组织有自我修复能力,但是当骨缺损达到一定的临界值时,骨组织很难实现自我恢复。因此骨组织出现大段缺损时,需要使用适当的骨填充物来填空大段骨缺损的部位来配合治疗。
增材制造,又称为三维打印技术,是一种可以精确制造复杂且指定结构的三维多孔支架的制造方法,弥补了传统方法生产支架时空间结构的不足。利用三维模型的实时数据,通过计算机进行二维切片后,再由计算机控制在平台上逐层制造,最终形成具有一定孔道结构、孔径大小以及孔隙率的三维模型。
生物陶瓷是一种具有特定生物或生理功能的一类陶瓷材料。能够直接用于人体或者参与人体直接相关的生物、医用、生物化学等的陶瓷材料。通常,生物陶瓷具有优异的骨传导性、出色的化学耐腐蚀性和良好的机械特性。生物陶瓷由于具有独特的性质,在骨组织工程中得到了广泛的应用。常用的生物陶瓷材料包括羟基磷灰石、磷酸钙、硅酸钙、磷酸三钙、双相磷酸钙、硫酸钙等一系列材料,在众多的研究中,生物陶瓷采用的多是复合型生物陶瓷,由多种材料组成,弥补不同材料的短板。
医药级聚乙烯醇,不同于化工级别聚乙烯醇,它是一种极安全的高分子有机物,对人体无毒,无副作用,具有良好的生物相容性,尤其在医疗中的如其水性凝胶在眼科、伤口敷料和人工关节方面的有广泛应用,同时在聚乙烯醇薄膜在药用膜,人工肾膜等方面也有使用。其安全性可以从用于伤口皮肤修复,和眼部滴眼液产品可见一斑。其中一些型号也常被用在化妆品中的面膜、洁面膏、化妆水及乳液中,是一种常用的安全性成膜剂。并且聚乙烯醇可以用于造血干细胞的培养液,在此基础上有望大幅降低造血干细胞的培养成本,帮助治疗白血病等疾病。
典型的人类长骨通常由宏观尺度的皮质骨、松质骨、骨膜、骨内膜和关节软骨组成。皮质骨呈现出由微尺度的骨骼,纳米尺度的胶原纤维和亚纳米级的胶原蛋白分子组成的分层结构组织。代谢产物可以通过相互连连通的小管、空隙系统进行输送。松质骨具有多室疏松结构,由大量片状或者针状的骨小梁连接而成的多孔网架结构,位于骨骺内部和骨干的内侧。骨小梁中不存在哈弗斯管。皮质骨中存在的孔径范围为1-100μm,松质骨的孔径范围为200-400μm。现有的研究表明,孔隙的尺寸、连通程度很大程度上影响着物质的运输,以及各种细胞在孔隙间的迁移、粘附和相关成骨基因的表达。
传统的制造多孔结构比较困难,并且缺少对制造出的生物支架进行人体适应性调试的处理方法,使得目前的生物支架的制造难度高、成本高昂以及人体适应性差。
发明内容
为解决上述问题,本发明提供了一种可控多孔生物陶瓷支架及其制备方法、应用。
为实现上述目的,本发明的技术方案为:
一种可控多孔生物陶瓷支架的制备方法,包括如下步骤:
步骤1:建立模型,使用计算机三维绘图软件绘制具有多孔结构的支架模型;
步骤2:配置粉末,将生物陶瓷和造孔剂混合并使用球磨机搅拌得到混合粉末;
步骤3:浆料制备,在由步骤2得到的混合粉末中,加入光敏树脂和分散剂并混合均匀得到浆料;
步骤4:坯体制作,模型导入光固化打印机,将步骤3中得到的浆料放入料缸槽,调整打印层厚和曝光时间,将浆料打印制成具有多孔结构的坯体;
步骤5:坯体成型,将步骤4中得到的坯体清洗并去除表面液体后对其进行脱脂、烧结,得到具有多孔结构的支架;
步骤6:微孔形成,将步骤5中得到的支架放入到酸性溶液中进行浸泡,完成后使用清水进行清洗,得到具有微孔结构的支架;
步骤7:配置涂层材料,在聚乙烯醇溶液中加入表面涂层粉末并搅拌,得到混合均匀的涂层材料溶液;
步骤8:表面附着涂层,将步骤6中得到的支架放入涂层材料溶液中,使用磁力搅拌器搅拌,然后使用离心机离心,去除多余的涂层材料溶液,完成后放入烘干箱进行烘干,得到表面附着有涂层材料的支架;
步骤9:涂层定型,将步骤8中得到的支架进行脱醇处理,之后烧结得到具有多孔生物陶瓷支架。
进一步的,所述步骤2中,所述生物陶瓷的材料包括磷酸钙、硅酸钙、硫酸钙、氧化锆、羟基磷灰石以及氧化铝中的至少一种;所述造孔剂的材料包括氢氧化镁、强氧化锌、碳酸镁、碳酸锌、二合水硫酸钙以及碱式碳酸镁中的至少一种;所述球磨机搅拌的方式依次为正转、停机以及反转;所述陶瓷粉末和造孔剂的粒径均为50~100×103nm,所述陶瓷粉末的含量为混合粉末总质量的70~99.5%,所述造孔剂的含量为混合粉末总质量的0.5~30%。
进一步的,所述步骤3中,光敏树脂由光敏预聚体、活性稀释剂、光引发剂以及光敏剂组成,分散剂为聚丙烯酸钠;所述浆料中各组分的含量分别为光敏树脂15~50%,分散剂0.5~3%,其余组分为混合粉末。
进一步的,所述步骤5中,脱脂的工艺参数为升温至400~500℃并保温1~5h,升温速率为0.5~6℃/min,然后继续升温至700~800℃并保温1~5h,升温速率为0.5~6℃/min;烧结的工艺参数为加热至1000~1700℃并保温0.5~5h,升温速率为0.5~6℃/min。
进一步的,所述步骤6中,酸性溶液包括盐酸、硫酸、硝酸以及乙酸中的至少一种,浸泡的时间为1~24h;所述支架上微孔结构的孔径为0.1~50μm。
进一步的,所述步骤7中,聚乙烯醇溶液中聚乙烯醇的含量为1~20%,其余组分为去离子水;表面涂层溶液中表面涂层粉末的含量为0.5~20%,其余组分为聚乙烯醇溶液,表面涂层粉末包括氧化镁、氧化锌、氧化铜以及TC4钛合金中的至少一种。
进一步的,所述步骤8中,离心机的转速为200~2000r/min,离心时间为2~30min;烘干箱的干燥温度为20~70℃,干燥时间为1~24h。
更进一步的,所述步骤9中,脱醇的工艺参数为升温至200℃~350℃并保温1~6h,继续升温至500~600℃,保温1~12小时,升温速率为0.5~6℃/min;烧结的工艺参数为升温至1000~1500℃,保温1-6小时,升温速率控制在0.5~6℃/min,最后炉冷至常温得到多孔生物陶瓷支架。
一种由上述方法制得的可控多孔生物陶瓷支架。
一种由上述方法制得的可控多孔生物陶瓷支架在骨组织工程中作为骨填充物、置换物或作为体外培养细胞支架的应用。
与现有技术相比,本发明的有益效果在于:
(1)本发明使用光固化3D打印技术制得的多孔结构的孔径为100μm~2000μm,使用酸性溶液浸泡制得的微孔结构的孔径为0.1μm~50μm,孔隙率为20%~90%,并且孔的形状、孔隙率以及形状等参数可控,能够满足植入支架的个性化需求。
(2)本发明的支架中的生物陶瓷本身具有良好的生物活性和生物相容性,适合作为骨修复的材料,作为生物支架,具有良好的生物活性。而表面涂层材料是对细胞增殖和分化有益的元素,有利于细胞的粘附、增殖和分化,同时也能起到抗菌作用。
(3)本发明的支架具有不同孔径的多孔结构,对细胞代谢和营养物质运输具有良好的效果,并且多孔结构扩大了支架的表面积,有利于细胞的粘附。
(4)本发明利用金属氧化物和酸性溶液进行反应,致使支架表面的金属氧化物反应溶解,在支架表面形成微小孔洞,此外,未反应的金属氧化物具有良好的生物活性,能够促进细胞的增殖和分化。
附图说明
图1为本发明实施例所述的可控多孔生物陶瓷支架的制备流程图;
图2为本发明实施例中坯体热重分析示意图。
图3是本发明实施例中表面涂层材料热重分析示意图。
图4为本发明实施例中使用SEM观察多孔结构的示意图。
图5是本发明实施例中使用SEM观察微孔结构的示意图。
图6是本发明实施例中使用MC3T3-E1小鼠前成骨细胞与可控多孔生物陶瓷支架共同培养状态示意图。
图7是本发明实施例中使用MC3T3-E1小鼠前成骨细胞与可控多孔生物陶瓷支架培养14天后使用茜素红S染色细胞分化示意图。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,下面结合具体实施方式对本发明作进一步的详细描述,仅用于解释本发明,但不应将此理解为对本发明的限制。
本发明的实施例提供了一种可控多孔生物陶瓷支架的制备方法,如图1所示,包括如下步骤:
步骤1:使用计算机三维绘图软件Magics绘制具有多孔结构的支架模型;
步骤2:将粒径为40×103nm的硅酸钙粉体和粒径为50×103nm的氢氧化镁粉体以90:10的比例称取并混合,使用行星球磨机进行混合均匀得到混合粉末,其中,行星球磨机搅拌的时间总共为3小时,正转1小时,停机1小时,反转1小时。
步骤3:在由步骤2得到的混合粉末中,加入光敏树脂和分散剂,混合均匀得到浆料,浆料中各组分含量为混合粉末60wt%,光敏树脂38wt%,分散剂2wt%;
步骤4:将支架模型以STL格式导入到光固化打印机中,然后将步骤3中得到的浆料放入料缸槽,调整打印层厚为0.02mm,曝光时间为8s,打印制成具有多孔结构的坯体;
步骤5:将步骤4中得到的坯体清洗并去除表面液体后对其进行脱脂,根据图2所示的热重分析示意图确定脱脂的工艺参数,首先升温至400~500℃保温3小时,升温速率为0.5℃/min,然后继续升温至700~800℃保温2小时,升温速率为0.5℃/min;脱脂完成后对支架进行烧结,烧结工艺参数为加热至1300℃之间,保温1h,升温速率为1℃/min,最终得到具有多孔结构的支架中间体;图中,TG,热重分析,指代的是质量随温度的变化,单位质量损失的百分比。DTG是指对TG质量变化做微分计算,以衡量质量随温度变化的快慢,单位:μV/mW。DSC是指差示扫描量热仪,反映的是样品吸放热的过程,大于0时为吸热,小于0时为放热,单位mW/mg。
步骤6:将步骤5中得到的支架放入到盐酸溶液中进行浸泡,浸泡时间为12小时,完成后使用过量清水进行清洗,得到具有微孔结构的微孔支架;
步骤7:将聚乙烯醇配置成溶液,加入占溶液总质量2%的纳米氧化镁粉末,使用磁力搅拌器对混合溶液进行搅拌,得到混合均匀的涂层材料溶液;
步骤8:将步骤6中得到的支架放入涂层材料溶液中,使用磁力搅拌器搅拌,然后使用离心机离心,去除多余的涂层材料溶液,完成后放入烘干箱进行烘干,得到表面附着有涂层材料的支架;
步骤9:将步骤8中得到的支架进行脱醇处理,脱醇的工艺参数根据图3的热重分析示意图得到,加热温度至320℃保温3h,继续升温至550℃,保温3小时,升温速率控制在0.5℃/min;然后对支架继续进行加热,加热温度至1200℃,保温时间为1小时,升温速率控制在1℃/min,最终炉冷至常温,得到带有涂层的可控多孔生物陶瓷支架。
将由上述步骤得到的可控多孔生物陶瓷支架使用SEM观察表面微观形貌,由图4可知,最终得到的多孔结构的孔径大小约为0.8mm,由图5可知,由氢氧化镁作为造孔剂,得到的微孔结构为不规则的细小微孔,其孔径大小约为10μm。
将可控多孔生物陶瓷支架与MC3T3-E1细胞进行共同培养14天,培养的过程中每隔2-3天更换一次培养基,使用光学显微镜观察细胞和支架的粘附情况,其粘附效果如图6所示,细胞的粘附效果良好。并且在共同培养14天后,使用茜素红S对细胞进行染色,其染色结果如图7所示,由图中可知,细胞分化效果良好,进一步验证了该支架具有较好的生物相容性。
需要说明的是,在本文中,诸如第一和第二等之类的关系术语仅仅用来将一个实体或者操作与另一个实体或操作区分开来,而不一定要求或暗示这些实体或操作之间存在任何这种实际的关系或顺序。而且,术语“包括”、“包含”或者对其任何其他变体在涵盖非排他性的包含,从而使得一系列要素的过程、方法、物品或者设备所固有的要素。在没有更多限制的情况下,由术语“包括一个…”限定的要素,并不排除在包括所述要素的过程、方法、物品或者设备中还存在另外的相同要素。
以上所述仅是本申请的具体实施方式,应当指出,对于本技术的普通技术人员来说,在不脱离本申请原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本保护范围。
Claims (8)
1.一种可控多孔生物陶瓷支架的制备方法,其特征在于,包括如下步骤:
步骤1:建立模型,使用计算机三维绘图软件绘制具有多孔结构的支架模型;
步骤2:配置粉末,将生物陶瓷和造孔剂混合并使用球磨机搅拌得到混合粉末;
所述生物陶瓷的材料包括磷酸钙、硅酸钙、硫酸钙、氧化锆、羟基磷灰石以及氧化铝中的至少一种;所述造孔剂的材料包括氢氧化镁、强氧化锌、碳酸镁、碳酸锌、二合水硫酸钙以及碱式碳酸镁中的至少一种;所述球磨机搅拌的方式依次为正转、停机以及反转;所述陶瓷粉末和造孔剂的粒径均为50~100×103nm,所述陶瓷粉末的含量为混合粉末总质量的70~99.5%,所述造孔剂的含量为混合粉末总质量的0.5~30%;
步骤3:浆料制备,在由步骤2得到的混合粉末中,加入光敏树脂和分散剂并混合均匀得到浆料;
步骤4:坯体制作,模型导入光固化打印机,将步骤3中得到的浆料放入料缸槽,调整打印层厚和曝光时间,将浆料打印制成具有多孔结构的坯体;
步骤5:坯体成型,将步骤4中得到的坯体清洗并去除表面液体后对其进行脱脂、烧结,得到具有多孔结构的支架;
步骤6:微孔形成,将步骤5中得到的支架放入到酸性溶液中进行浸泡,完成后使用清水进行清洗,得到具有微孔结构的支架;
步骤7:配置涂层材料,在聚乙烯醇溶液中加入表面涂层粉末并搅拌,得到混合均匀的涂层材料溶液;
聚乙烯醇溶液中聚乙烯醇的含量为1~20%,其余组分为去离子水;表面涂层溶液中表面涂层粉末的含量为0.5~20%,其余组分为聚乙烯醇溶液,表面涂层粉末包括氧化镁、氧化锌、氧化铜以及TC4钛合金中的至少一种;
步骤8:表面附着涂层,将步骤6中得到的支架放入涂层材料溶液中,使用磁力搅拌器搅拌,然后使用离心机离心,去除多余的涂层材料溶液,完成后放入烘干箱进行烘干,得到表面附着有涂层材料的支架;
步骤9:涂层定型,将步骤8中得到的支架进行脱醇处理,之后烧结得到具有可控多孔生物陶瓷支架。
2.根据权利要求1中的可控多孔生物陶瓷支架的制备方法,其特征在于:所述步骤3中,光敏树脂由光敏预聚体、活性稀释剂、光引发剂以及光敏剂组成,分散剂为聚丙烯酸钠;所述浆料中各组分的含量分别为光敏树脂15~50%,分散剂0.5~3%,其余组分为混合粉末。
3.根据权利要求1中的可控多孔生物陶瓷支架的制备方法,其特征在于:所述步骤5中,脱脂的工艺参数为升温至400~500℃并保温1~5h,升温速率为0.5~6℃/min,然后继续升温至700~800℃并保温1~5h,升温速率为0.5~6℃/min;烧结的工艺参数为加热至1000~1700℃并保温0.5~5h,升温速率为0.5~6℃/min。
4.根据权利要求1中的可控多孔生物陶瓷支架的制备方法,其特征在于:所述步骤6中,酸性溶液包括盐酸、硫酸、硝酸以及乙酸中的至少一种,浸泡的时间为1~24h;所述支架上微孔结构的孔径为0.1~50μm。
5.根据权利要求1中的可控多孔生物陶瓷支架的制备方法,其特征在于:所述步骤8中,离心机的转速为200~2000r/min,离心时间为2~30min;烘干箱的干燥温度为20~70℃,干燥时间为1~24h。
6.根据权利要求1中的可控多孔生物陶瓷支架的制备方法,其特征在于:所述步骤9中,脱醇的工艺参数为升温至200℃~350℃并保温1~6h,继续升温至500~600℃,保温1~12小时,升温速率为0.5~6℃/min;烧结的工艺参数为升温至1000~1500℃,保温1-6小时,升温速率控制在0.5~6℃/min,最后炉冷至常温得到多孔生物陶瓷支架。
7.一种根据权利要求1-6中任一项中制得的可控多孔生物陶瓷支架。
8.一种根据权利要求1-6中任一项中制得的可控多孔生物陶瓷支架在骨组织工程中作为骨填充物、置换物或作为体外培养细胞支架的应用。
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