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CN114949343A - 一种可控梯度降解、促组织修复的肌骨系统修复体及其制备方法 - Google Patents

一种可控梯度降解、促组织修复的肌骨系统修复体及其制备方法 Download PDF

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CN114949343A
CN114949343A CN202210098850.4A CN202210098850A CN114949343A CN 114949343 A CN114949343 A CN 114949343A CN 202210098850 A CN202210098850 A CN 202210098850A CN 114949343 A CN114949343 A CN 114949343A
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CN114949343B (zh
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费翔
朱美芳
华英汇
陆腱
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Abstract

本发明涉及一种可控梯度降解、促组织修复的肌骨系统修复体及其制备方法。该修复体包括承力纤维和磁响应加速降解纤维，以及这两种纤维表面的涂层。该方法包括承力纤维制备；磁响应加速降解纤维制备；肌骨系统修复体制备。该方法具有操作简单、连续化制备可行性高的诸多优点；制备得到的肌骨系统修复体具有可控梯度降解以及促组织修复的特点。

Description

一种可控梯度降解、促组织修复的肌骨系统修复体及其制备 方法
技术领域
本发明属于组织修复材料及其制备领域，特别涉及一种可控梯度降解、促组织修复的肌骨系统修复体及其制备方法。
背景技术
随着人们生活水平逐渐提高，体育运动等在生活中逐渐普及，剧烈运动、较高强度活动，以及平时生活中的扭伤、车祸等意外伤，很容易发生关节韧带、肌腱等肌骨组织的损伤。然而，受伤韧带、肌腱位于骨关节之间，难以从血液中获取营养物质，在受损后不易修复；尤其是当损伤程度过大时，需要手术植入人工韧带、人工肌腱等肌骨系统修复体进行修复。目前，临床用肌骨系统修复体产品多以聚酯纤维为基体，虽然其具有足够的力学强度，可满足植入部位日常运动需求，然而其不可降解的特性阻碍了受损组织的生物性愈合，且易引起顽固性滑膜炎，严重影响其服役性能。因此，可降解肌骨系统修复体的开发收到了研究者们的重视。
而目前的可降解肌骨系统修复体主要以聚乳酸、丝素蛋白等可生物吸收的聚合物为基体，侧重于材料的力学性能和生物功能性研究，在替代受损肌骨系统承力功能的同时，促进相应组织生物性修复。然而其缺陷在于，材料的降解速率和肌骨组织再生的速率不匹配，即材料降解导致其力学性能下降过快，而新生组织生长过慢，导致材料和新生组织的总体力学性能在某段时间内无法满足承力需求。因此，通过材料设计，使材料在特定时间加速降解、并促进组织修复，实现材料的这两个速率匹配，是目前肌骨系统修复研究，乃至组织修复研究的热点。
发明内容
本发明所要解决的技术问题是提供一种可控梯度降解、促组织修复的肌骨系统修复体及其制备方法，以提升肌骨系统修复体降解速率和肌骨组织再生速率的匹配程度。
本发明提供一种可控梯度降解、促组织修复的肌骨系统修复体，包括承力纤维和磁响应加速降解纤维，以及这两种纤维表面的涂层；所述承力纤维由100wt％聚合物基体组成；所述磁响应加速降解纤维由80wt％-99wt％聚合物基体和1wt％-20wt％磁响应纳米粒子组成；所述涂层包括无机矿化涂层或有机生物活性涂层。
所述聚合物基体包括合成聚合物和天然聚合物中的一种或几种；所述合成聚合物包括聚乳酸、聚羟基乙酸、聚己内酯及其共聚物中的一种或几种；所述天然聚合物包括丝素蛋白、明胶、壳聚糖中的一种或几种。
所述磁响应纳米粒子包括铁、钴、镍元素的氧化物中的一种或几种。
所述无机矿化涂层为含钙、磷、镁元素无机物中一种或几种的的矿化涂层。
所述有机生物活性涂层为以壳聚糖、明胶、胶原大分子中一种或几种为基体或负载有生长因子小分子的功能涂层。
本发明还提供一种可控梯度降解、促组织修复的肌骨系统修复体的制备方法，包括：
(1)将聚合物通过熔融纺丝或湿法纺丝，得到承力纤维；
(2)将溶液纺聚合物和磁响应纳米粒子均匀分散于溶剂，作为纺丝液，通过湿法纺丝、干法纺丝或静电纺丝，得到磁响应加速降解纤维；
(3)将步骤(1)中承力纤维和步骤(2)中磁响应加速降解纤维进行编织，然后借助静电纺丝、表面矿化或化学键接枝在编织物表面形成无机矿化涂层或有机生物活性涂层，得到肌骨系统修复体。
所述步骤(1)中熔融纺丝的工艺参数为：温度为210-300℃，卷绕速度为500-3000m/min，牵伸倍数为2-5倍。
所述步骤(1)中湿法纺丝的工艺参数为：纺丝液挤出速率为0.5-20mL/h，牵伸倍率为5-10倍。
所述步骤(2)中溶液纺聚合物和磁响应纳米粒子的质量比为80-99：1-20。
所述步骤(2)中溶剂包括NMP、六氟异丙醇、二氯甲烷中的一种或几种。
所述步骤(2)中湿法纺丝的工艺参数为：纺丝液挤出速率为0.5-20mL/h，牵伸倍率为5-10倍。
所述步骤(2)中干法纺丝的工艺参数为：纺丝液挤出速率为0.5-20mL/h，在室温-100℃的温度下使纤维成型，收集速率为10-100m/min。
所述步骤(2)中静电纺丝的工艺参数为：纺丝液挤出速率为0.5-5mL/h，接收距离为10-15cm，施加电压为15-20kV。
所述步骤(3)中编织方法包括加捻、针织、机织中的一种或几种。
本发明还提供一种肌骨系统修复体在制备肌骨组织修复材料中的应用。
本发明采用载磁热纳米粒子纤维与承力纤维组成，可通过体外施加高频交变磁场加速载磁热纳米粒子纤维的降解，使两种纤维材料在降解速率上出现明显梯度，从而通过体外手段可控调节纤维材料构成的拓扑结构，诱导细胞增殖与组织修复；同时，借助负载生物活性物质来促进患处组织修复。因此，本发明可控梯度降解、促组织修复的肌骨系统修复体，具有较高可行性与先进性，有望解决材料植入后降解速率与组织修复速率不匹配的难题。
有益效果
本发明通过磁响应加速降解的载磁热纳米粒子纤维和承力纤维构筑肌骨系统修复材料，在满足修复材料重建目标肌骨组织力学性能的同时，可根据组织修复情况借助体外交变磁场加速载磁热纳米粒子纤维降解，使材料在降解速率上具有可控的降解梯度，以诱导相应细胞增殖及组织重构；同时负载生物活性涂层，以加速组织再生。此外，该修复材料的制备可设计性强、结构性能可控。
本发明肌骨系统修复体品质可控性强、技术独特、先进；同时制备方法具有操作简单、成本低、连续化制备可行性高的诸多优点，可基于现用相关产业设备加以改进实现规模化制备。
具体实施方式
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。此外应理解，在阅读了本发明讲授的内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
实施例1
本实施例提供一种可控梯度降解、促组织修复的肌骨系统修复体，具体用于韧带组织，即人工韧带，包括可降解纤维织物和织物表面涂层。
其中的可降解纤维织物，所用承力纤维为左旋聚乳酸(PLLA，购自济南岱罡生物科技有限公司)的熔纺纤维，所用磁响应加速降解纤维为四氧化三铁(Fe3O4)/再生丝素蛋白(RSF，购自苏州丝美特生物技术有限公司)的静电纺纳米纱线。其制备方法如下：
(1)左旋聚乳酸纤维的熔融纺丝制备
用烘箱将上述左旋聚乳酸切片烘干至含水率为20～30ppm，取1kg加入熔融纺丝设备中，采用36孔喷丝板制得熔纺纤维。熔融纺丝的纺丝工艺如下：
螺杆挤出温度：225℃
箱体温度：230℃
卷绕速度：2500m/min
牵伸温度：60℃
牵伸倍数：2倍
热定型温度：100℃
(2)四氧化三铁/RSF纳米纱线的静电纺丝制备
通过水热法制备四氧化三铁：先配置含5mg/mL氯化铁、10mg/mL柠檬酸钠的乙二醇溶液，待溶液清澈后将其转移至反应釜，在180℃下反应18h，然后用乙醇洗涤，即得所需纳米级Fe3O4。
然后分别配置100mg/mL的纳米Fe3O4/NMP分散液和100mg/mL的RSF/六氟异丙醇溶液，按NMP分散液：六氟异丙醇溶液＝1:9的体积比例复配获得纺丝液，然后进行静电纺丝，纺丝工艺如下：
纺丝液挤出速率：1mL/h
收集距离：15cm
施加电压：19.5kV
通过纱线收集装置将纺得纳米纤维收集成均匀纱线，最后经真空下40℃处理24h去除残余溶剂。
(3)人工韧带基体的编织制备
通过加捻法编织制备人工韧带：将6束PLLA纤维与同等线密度四氧化三铁/RSF纳米纱线借助并捻机复合，再以700tw/m的捻度并捻成1束纱线，再将2束该纱线以700tw/m的捻度并捻成1股纱线，最后将2股该纱线300tw/m的捻度并捻成1条纱线，即为肌骨系统修复体基体。
(4)人工韧带的表面生物功能涂层制备
先用75％(v/v)乙醇浸泡上述肌骨系统修复体基体1h，室温晾干后，对编织物进行表面等离子体处理，在氧气氛围下以200W处理10min；然后将该段浸泡于4mg/mL胶原的0.3％乙酸溶液室温浸泡30min，以获得胶原涂层，用于改善韧带亲水性和细胞相容性。
实验结果表明，在380kHz的交变磁场下，Fe3O4/RSF纳米纱线的降解失重速率(交变磁场处理后的样品，去离子水洗涤3次后，常温真空烘干24h，称重计算样品失重)加快，约为无交变磁场作用下的2倍，因此其可通过外加交变磁场加速RSF纱线的降解，为患处细胞增殖、重构组织提供空间；以小鼠成纤维细胞L929为例，人工韧带表面复合上述生物功能涂层后，细胞活性(样品灭菌处理后置于24孔板，每孔500mL相应完全培养基，以1×105密度种植细胞，72h后将种有细胞的样品转移至空白孔板，PBS洗涤2次后用CCK8试剂盒检测细胞活性)相对提高13.9％。
实施例2
本实施例提供一种可控梯度降解、促组织修复的肌骨系统修复体，具体用于肩袖，即肩袖补片，包括可降解纤维织物和织物表面涂层。
其中的可降解纤维织物，所用承力纤维为RSF纤维(购自苏州丝美特生物技术有限公司)的湿纺纤维，所用磁响应加速降解纤维为四氧化三钴(Co3O4，购自上海麦克林生化科技有限公司)/聚(乙醇酸-乳酸)共聚物(PLGA，购自济南岱罡生物科技有限公司)的干纺纤维。
其制备方法如下：
(1)RSF纤维的湿法纺丝制备
配置20wt％RSF的水溶液作为纺丝液，采用60℃的35wt％硫酸铵水溶液作为凝固浴，进行湿法纺丝(喷丝孔直径150μm，牵伸倍率为7倍)，制得RSF承力纤维。
(2)Co3O4/PLGA干纺纤维的静电纺丝制备
先将PLGA溶于二氯甲烷配置成浓度为150mg/mL的溶液，然后按10mg/mL的浓度加入Co3O4，分散均匀后作为纺丝液进行干法纺丝(纺丝液挤出速率为2mL/h，接收距离为35cm，收集速率为20m/min)，获得Co3O4/PLGA干纺纤维。
(3)肩袖补片基体的编织制备
通过机织法编织制备肩袖补片：先将10束RSF纤维与10束同等线密度Co3O4/PLGA干纺纱线并捻成一股纱线，再以该并捻纱线为经纱，以20束RSF纤维并捻成的纱线为纬纱，进行机织编织，编织结构为平纹结构，所得织物即为肩袖补片基体。
(4)肩袖补片的表面生物功能涂层制备
以bFGF的1wt％水溶液为芯层纺丝液、聚己内酯的10wt％六氟异丙醇溶液为皮层纺丝液，在肩袖补片表面进行同轴静电纺丝(芯层纺丝液流速0.1mL/h，皮层纺丝液流速1mL/h，接收距离14.5cm，施加电压15kV)制备促组织修复涂层，可得肩袖补片。
实验结果表明，在380kHz的交变磁场下，Co3O4/RSF纳米纱线的降解失重速率加快，约为无交变磁场作用下的1.2倍，因此其可通过外加交变磁场加速RSF纱线的降解，为患处细胞增殖、重构组织提供空间；以小鼠成纤维细胞L929为例，人工韧带表面复合上述生物功能涂层后，细胞活性相对提高14.1％。

Claims (9)

1.一种可控梯度降解、促组织修复的肌骨系统修复体，其特征在于，包括承力纤维和磁响应加速降解纤维，以及这两种纤维表面的涂层；所述承力纤维由100wt％聚合物基体组成；所述磁响应加速降解纤维由80wt％-99wt％聚合物基体和1wt％-20wt％磁响应纳米粒子组成；所述涂层为无机矿化涂层或有机生物活性涂层。
2.根据权利要求1所述的修复体，其特征在于，所述聚合物基体包括合成聚合物和天然聚合物中的一种或几种；所述合成聚合物包括聚乳酸、聚羟基乙酸、聚己内酯及其共聚物中的一种或几种；所述天然聚合物包括丝素蛋白、明胶、壳聚糖中的一种或几种。
3.根据权利要求1所述的修复体，其特征在于，所述磁响应纳米粒子包括铁、钴、镍元素的氧化物中的一种或几种。
4.根据权利要求1所述的修复体，其特征在于，所述无机矿化涂层为含钙、磷、镁元素无机物中一种或几种的矿化涂层；有机生物活性涂层为以壳聚糖、明胶、胶原大分子中一种或几种为基体或负载有生长因子小分子的功能涂层。
5.一种可控梯度降解、促组织修复的肌骨系统修复体的制备方法，包括：
(1)将聚合物通过熔融纺丝或湿法纺丝，得到承力纤维；
(2)将溶液纺聚合物和磁响应纳米粒子均匀分散于溶剂，作为纺丝液，通过湿法纺丝、干法纺丝或静电纺丝，得到磁响应加速降解纤维；
(3)将步骤(1)中承力纤维和步骤(2)中磁响应加速降解纤维进行编织组合，然后借助静电纺丝、表面矿化或化学键接枝在编织物表面形成无机矿化涂层或有机生物活性涂层，得到肌骨系统修复体。
6.根据权利要求5所述的制备方法，其特征在于，所述步骤(1)中熔融纺丝的工艺参数为：温度为210-300℃，卷绕速度为500-3000m/min，牵伸倍数为2-5倍；湿法纺丝的工艺参数为：纺丝液挤出速率为0.5-20mL/h，牵伸倍率为5-10倍。
7.根据权利要求5所述的制备方法，其特征在于，所述步骤(2)中溶液纺聚合物和磁响应纳米粒子的质量比为80-99：1-20；溶剂包括NMP、六氟异丙醇、二氯甲烷中的一种或几种。
8.根据权利要求5所述的制备方法，其特征在于，所述步骤(3)中编织方法包括加捻、针织、机织中的一种或几种。
9.一种如权利要求1所述的肌骨系统修复体在制备肌骨组织修复材料中的应用。
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