中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (30): 4780-4786.doi: 10.3969/j.issn.2095-4344.1421

• 复合支架材料 composite scaffold materials • 上一篇    下一篇

基于3D打印的双相磷酸钙胶原改性支架

张海鸽1，索海瑞1，2，王  玲1，2，徐铭恩1，2
  

  1. 1杭州电子科技大学浙江省医学信息与生物三维打印重点实验室，浙江省杭州市  310018；2杭州电子科技大学自动化学院，浙江省杭州市  310018
  • 收稿日期:2019-05-13 出版日期:2019-10-28 发布日期:2019-10-28
  • 通讯作者: 徐铭恩，博士后，教授，杭州电子科技大学自动化学院；杭州电子科技大学浙江省医学信息与生物三维打印重点实验室，浙江省杭州市 310018
  • 作者简介:张海鸽，女，1991年生，河南省三门峡市人，汉族，杭州电子科技大学在读硕士，主要从事生物3D打印研究。
  • 基金资助:

    国家自然科学基金(61675059)，项目负责人：王玲；国家重点研发计划(2017YFC1103400)，项目负责人：徐铭恩

Biphasic calcium phosphate scaffolds modified with collagen based on three-dimensional printing technology

Zhang Haige1, Suo Hairui1,2, Wang Ling1,2, Xu Mingen1,2
  

  1. 1Zhejiang Provincial Key Lab of Medical Information and Three-Dimensional Bio-Printing, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China; 2College of Automation, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China
  • Received:2019-05-13 Online:2019-10-28 Published:2019-10-28
  • Contact: Xu Mingen, MD, Professor, Zhejiang Provincial Key Lab of Medical Information and Three-Dimensional Bio-Printing, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China; College of Automation, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China
  • About author:Zhang Haige, Master candidate, Zhejiang Provincial Key Lab of Medical Information and Three-Dimensional Bio-Printing, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 61675059 (to WL); the National Key R & D Program of China, No. 2017YFC1103400 (to XME)

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摘要：

文章快速阅读：

 

文题释义：
改性：使用物理或化学等方法改变原材料物质形态或性质的方法。对于生物材料，为保证更好地与植入体融合，材料的表面具有重要的作用。表面改性即在保证原材料原有性能的条件下，更好的改善其表面性能，如亲水性、生物活性等，为细胞提供良好的生长环境。
双相磷酸钙：是由羟基磷灰石和磷酸三钙混合而成，它的钙磷比、溶解性和降解速率都在羟基磷灰石和磷酸三钙之间，可通过调节两种物质的比例来控制支架的溶解性、再吸收性及降解速率等，确保在促进骨骼向内生长的同时保持良好的稳定性。
 
 
背景：制作出类似天然骨成分和机械强度、表面活性良好且具有多孔结构的三维胶原/双相磷酸钙支架是骨组织工程研究的一个难点。
目的：制备兼具高孔隙率、高机械强度和高表面生物活性的三维多孔骨组织支架。
方法：以质量比为60∶40的羟基磷灰石与β-磷酸三钙为浆料，利用3D打印技术制备双相磷酸钙支架，高温烧结后，分别以0.5，1.0，1.5 g/L的Ⅰ型胶原溶液对支架进行涂覆处理，制备胶原/双相磷酸钙支架，通过表观形貌、孔隙率、吸水率、机械性能测试筛选最佳Ⅰ型胶原溶液涂覆质量浓度，进行细胞实验。将大鼠骨髓间充质干细胞分别接种于双相磷酸钙支架与胶原/双相磷酸钙支架上，培养1，7 d，Calcein-AM染色观察细胞活性；培养1，3，7 d，Alamar Blue法检测细胞增殖。
结果与结论：①当Ⅰ型胶原溶液涂覆质量浓度为0.5 g/L时，胶原/双相磷酸钙支架具有良好的孔隙率与吸水率，抗压强度为(4.99±0.15) MPa，压缩模量为(95.24±0.57) MPa，符合天然松质骨的机械强度要求，细胞实验选择此涂覆质量浓度；②两种支架都支持骨髓间充质干细胞的生长，培养至7 d时，细胞在胶原/双相磷酸钙支架表面已基本长满，大多呈梭形或星形，细胞伸展良好，排列紧密，在支架上黏附形成网状结构，但双相磷酸钙支架上还有部分区域无细胞黏附；③两组支架中培养1，3 d的细胞增殖比较差异无显著性意义(P > 0.05)，胶原/双相磷酸钙支架上培养7 d的细胞增殖快于双相磷酸钙支架(P < 0.05)；④结果表明在保证孔隙率、高机械强度的情况下，Ⅰ型胶原溶液涂覆可提高双相磷酸钙支架的生物活性。

关键词: 3D打印, 双相磷酸钙, 胶原, 涂覆, 细胞活性, 生物材料, 骨组织, 胶原/双相磷酸钙支架, 细胞增殖

Abstract:

BACKGROUND: Three-dimensional collagen/biphasic calcium phosphate scaffolds with natural bone composition and mechanical strength, good surface activity and porous structure are a difficult point in bone tissue engineering research.
OBJECTIVE: To manufacture a three-dimensional porous bone tissue scaffold with high porosity, high mechanical strength and high surface biological activity.
METHODS: Biphasic calcium phosphate scaffold was manufactured by three-dimensional printing technology using hydroxyapatite and β-tricalcium phosphate with a mass ratio of 60∶40 as slurry. After high temperature sintering, the biphasic calcium phosphate scaffolds were coated with type I collagen at the concentrations of 0.5, 1.0 and 1.5 g/L. The optimal concentration of type I collagen solution was screened by apparent morphology, porosity, water absorption and mechanical properties to conduct cell experiments. Rat bone marrow mesenchymal stem cells were seeded onto biphasic calcium phosphate scaffolds and collagen/biphasic calcium phosphate scaffolds, separately. Cell viability was observed by Calcein-AM staining at 1 and 7 days; cell proliferation was detected by Alamar Blue method at 1, 3 and 7 days.
RESULTS AND CONCLUSION: When the mass concentration of type I collagen solution was 0.5 g/L, the collagen/biphasic calcium phosphate scaffold had good porosity and water absorption, the compressive strength was (4.99±0.15) MPa and the compressive modulus was (95.24±0.57) MPa, which met the mechanical strength requirements of natural cancellous bone. Therefore, the coating mass concentration 0.5 g/L was selected for cell experiments. Both kinds of scaffolds supported the growth of mesenchymal stem cells. After 7 days of culture, cells almost covered the surface of the collagen/biphasic calcium phosphate scaffold. Most of them were fusiform or star-shaped, and the cells were well stretched, arranged closely and adhered to form network structure. However, there was still no cell adhesion in some areas of the biphasic calcium phosphate scaffold. There was no significant difference in cell proliferation between two kinds of cells at 1 and 3 days (P > 0.05). The proliferation of cells cultured on the collagen/biphasic calcium phosphate scaffold for 7 days was faster than that cultured on the biphasic calcium phosphate scaffold for the same duration (P < 0.05). The results show that the coating of type I collagen solution can improve the biological activity of the biphasic calcium phosphate scaffold under the condition of ensuring porosity and high mechanical strength.

Key words: 3D printing, biphasic calcium phosphate, collagen, coating, cell viability, biological material, bone tissue, collagen/biphasic calcium phosphate scaffold, cell proliferation

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