聚多巴胺涂层3D打印双相磷酸钙组织工程支架的制备及其评价

doi:10.13481/j.1671-587x.20190438

摘要/Abstract

摘要： 目的：在3D打印制备的双相磷酸钙（BCP）支架的表面进行聚多巴胺（PDA）涂层，构建成骨活性较高的新型骨组织工程支架，并对其应用潜力进行初步评价。方法：通过3D打印技术制备BCP生物陶瓷支架，将制备完成的组织工程支架在多巴胺溶液中浸泡，以使支架表面形成纳米级的PDA覆膜结构，无PDA涂层的支架为3DBCP组，有PDA涂层的支架为PDA-3DBCP组。利用扫描电子显微镜观察各组支架表面微观形貌，通过测定支架表面的水接触角表征材料的亲水性，采用比重法测定支架的孔隙度，万能试验机检测支架的机械强度，CCK-8方法检测支架上细胞的增殖活性。结果：与3DBCP组比较，PDA-3DBCP组支架表面水接触角明显缩小（t=5.06，P<0.05），支架孔隙度和机械强度差异无统计学意义（t=0.103，P>0.05；t=0.002，P>0.05）。将细胞接种至支架并进行1、3和5 d的复合培养，CCK-8法检测，随着培养时间的延长，2组细胞增殖活性逐渐升高，第5天时与3DBCP组比较，PDA-3DBCP组支架细胞增殖活性明显升高（t=39.3，P<0.05）。结论：PDA涂层的3D打印BCP多孔支架符合骨组织工程支架材料的表征，并且可以提高细胞的增殖能力，具备作为骨组织工程支架的潜力。

关键词: 3D打印, 聚多巴胺, 骨组织工程, 生物陶瓷, 双相磷酸钙

Abstract: Objective:To coat polydopamine (PDA) on the surface of biphasic calcium phosphate (BCP) scaffold prepared by 3D printing and construct a novel bone tissue engineering scaffold with high osteogenic activity,and to conduct a preliminary evaluation on its application potential. Methods:BCP bioceramic scaffolds were prepared by 3D printing technology, and then they were immersed in dopamine solution for a certain period to form a nanoscale PDA film structure on the surface.The PDA-free scaffolds were named 3DBCP group, and the PDA-coated scaffolds were named PDA-3DBCP group. Scanning electron microscope (SEM) was used to observe the surface topography of the scaffolds in each group; the hydrophilicity of the material was characterized by measuring the water contact angle of the scaffold surface; the scaffold porosity was determined by the specific gravity method, and the mechanical strength was tested by the universal testing machine; CCK-8 assay was adopted to detect the cell proliferation activity on the scaffold. Results:Compared with 3DBCP group, the surface water contact angle of the the scaffolds in PDA-3DBCP group was significantly reduced (t=5.06, P<0.05);there were no significant differences in the porosity and the mechanical strength between two groups(t=0.103, P>0.05; t=0.002, P>0.05).The cells were inoculated into the scaffolds and cultured for 1, 3 and 5 d;the CCK-8 assay results showed that the proliferation activities of the cells in two groups were gradually increased with the prolongation of the culture time. Furthermore, compared with 3DBCP group, the cell proliferation activity of the cells in PDA-3DBCP group on the 5th day was significantly increased (t=39.3, P<0.05). Conclusion:The PDA-coated 3D printed BCP porous scaffold has the advantages in characterization,it can improve the cell proliferation ability, which shows the potential as bone tissue engineering scaffolds.

Key words: 3D printing, polydopamine, bone tissue engineering, bioceramics, biphasic calcium phosphate

中图分类号:

R318

王宁宁, 徐志民, 郑叶, 张赢心, 韩冰. 聚多巴胺涂层3D打印双相磷酸钙组织工程支架的制备及其评价[J]. 吉林大学学报(医学版), 2019, 45(04): 955-959.

WANG Ningning, XU Zhimin, ZHENG Ye, ZHANG Yingxin, HAN Bing. Preparation of polydopamine coated 3D printed biphasic calcium phosphate tissue engineering scaffold and its evaluation[J]. Journal of Jilin University(Medicine Edition), 2019, 45(04): 955-959.

参考文献

[1] SHAO H,KE X,LIU A. Bone regeneration in 3D printing bioactive ceramic scaffolds with improved tissue/material interface pore architecture in thin-wall bone defect[J]. Biofabrication, 2017, 9(2):1-23.
[2] CHEN C L, ZENGA J, Roland L T. Complications of double free flap and free flap combined with locoregional flap in head and neck reconstruction:A systematic review[J]. J Sci Special Head Neck, 2018, 40(3):632-646.
[3] SATTESON E S, SATTESON A C, WALTONEN J D,et al. Donor-site outcomes for the osteocutaneous radial forearm free flap[J]. J Reconstruct Microsurg, 2017, 33(8):544-548.
[4] ZHONG J, SHAO Z. Growth and crystallization of calcium phosohate mediated by bombyx mori silk fibrion[J]. Acta Polymer Sinica, 2014, (10):1428-1434.
[5] 贺超良,汤朝晖,田华雨,等. 3D打印技术制备生物医用高分子材料的研究进展[J]. 高分子学报,2013(6):722-723.
[6] TANG Q, HU Z C, JIN H M,et al. Microporous polysaccharide multilayer coated BCP composite scaffoldds with immobilised calcitriol promote osteoporotic bone regeneration both in vitro and in vivo[J]. Theranostics,2019, 9(4):1125-1143.
[7] JIANG J, HAO W, LI Y Z, et al. Hydroxyapatite/regenerated silk fibroin scaffold-enhanced osteoinductivity and osteoconductivity of bone marrow-derived mesenchymal stromal cells[J]. Biotechnol Lett,2013, 35(4):657-661.
[8] ZAN X J, SITASUWAN P, FENG S, et al. Effect of roughness on in situ biomineralized CaP-collagen coating on the osteogenesis of mesenchymal stem cells[J]. Langmuir,2016, 32(7):1808-1817.
[9] KIM J W, SHIN Y C, LEE J J, et al. The effect of reduced graphene oxide-coated biphasic calcium phosphate bone graft material on osteogenesis[J]. Int J Mol Sci, 2017, 18(8):1-17.
[10] HUANG L X, XIAO L, POUDEL A J, et al. Porous chitosan microspheres as microcarriers for 3D cell culture[J]. Carbohydrate Polymers, 2018, 202:611-620.
[11] WANG H, HU M. Recent progress on 3D printed tissue engineering scaffolds of the jaws[J]. J Oral Maxillofac Surg,2017, 27(3):218-222.
[12] PINA S, CANADAS R F, JIMÉNEZ G, et al. Biofunctional ionic-doped calcium phosphates:silk fibroin composites for bone tissue engineering scaffolding[J]. Cells Tissues Organs(Print),2017, 204(3/4):150-163.
[13] DADHICH P,DAS B,PAL P,et al. A simple approach for an eggshell-based 3D-printed osteoinductive multiphasic calcium phosphate scaffold[J]. ACS Appl Mater Interfaces, 2016, 8(19):11910-11924.
[14] HU J X, CAI X, MO S B, et al. Fabrication and characterization of chitosan-silk fibroin/hydroxyapatite composites via in situ precipitation for bone tissue engineering[J]. Chin J Polymer Sci,2015, 33(12):1661-1671.
[15] 尤琦. 三维打印技术构建双相磷酸钙陶瓷支架及其性能研究[D]. 长春:吉林大学,2016.
[16] 夏轶超,澈力格尔,李宝印,等. 壳聚糖膜覆盖3D打印双相磷酸钙骨组织工程支架的制备和性能[J]. 吉林大学学报:医学版, 2018,44(4):770-775.
[17] LI C, ZHANG Z, HE CL. Biodegradable thermo-sensitive hydrogels for controlled delivery of parathyroid hormone related peptide[J]. Acta Polymer Sinica,2012, 12(7):778-783.
[18] ZHANG X, KONG M, TIAN MP. The temperature-responsive hydroxybutyl chitosan hydrogels with poly-dopamine coating for cell sheet transplantation[J]. IntJ Biol Macromol,2018, 120:152-158.
[19] KAOCT, CHEN Y J, NG H Y, et al. Surface modification of calcium silicate via mussel-inspired polydopamine and effective adsorption of extracellular matrix to promote osteogenesis differentiation for bone tissue engineering[J]. Materials(Basel), 2018, 11(9):E1664.
[20] MA H S, LUO J, SUN Z, et al. 3D printing of biomaterials with mussel-inspired nanostructures for tumor therapy and tissue regeneration[J]. Biomaterials, 2016, 111:138-148.
[21] PAN Z, QU Z H, ZHANG Z, et al. Particle-collision and porogen-leaching technique to fabricate polymeric porous scaffolds with microscale roughness of interior surfaces[J]. Chin J Polymer Sci, 2013, 31(5):737-747.
[22] 龚华俊,杨小平,陈国强,等. 电纺丝法制备聚乳酸/多壁碳纳米管/羟基磷灰石杂化纳米纤维的研究[J]. 高分子学报,2005(2):297-300.
[23] 李保强,胡巧玲,钱秀珍,等. 原位沉析法制备可吸收壳聚糖/羟基磷灰石棒材[J]. 高分子学报,2002(6):828-833.
[24] ZHU J, LUO J J, ZHAO X Y, et al. Electrospun homogeneous silk fibroin/poly(-caprolactone) nanofibrous scaffolds by addition of acetic acid for tissue engineering[J]. J Biomater Appl,2016, 31(3):421-437.
[25] KAO C T,LIN C C, CHEN Y W, et al. Poly (dopamine) coating of 3D printed poly(lactic acid) scaffolds for bone tissue engineering[J]. Mater Sci Engineer C,2015, 56:165-173.
[26] 殷俊飞扬,钟静,陈莉智,等. 3D打印技术在颌面整形外科的应用进展[J].中国医用物理学杂志, 2018, 35(12):1479-1482.
[27] GE M, XUE L,NIE T T,et al. The precision structural regulation of PLLA porous scaffold and its influence on the proliferation and differentiation of MC3T3-E1 cells[J]. J Biomater Sci Polym Ed,2016, 27(17):1685-1697.
[28] JO A R,NONG M W, CHO Y S, et al. Assessment of cell proliferation in knitting scaffolds with respect to pore-size heterogeneity, surface wettability, and surface roughness[J]. J Appl Polymer Sci, 2015, 32(38):1-13.
[29] GOLIZADEH M,KARIMA A, GANDOMI-RAVANDI S,et al. Evaluation of cellular attachment and proliferation on different surface charged functional cellulose electrospun nanofibers[J]. Carbohydrate Polymers,2019, 207:796-805.
[30] ZHU Y L, ZHU R Q, MA J. In vitro cell proliferation evaluation of porous nano-zirconia scaffolds with different porosity for bone tissue engineering[J]. Biomed Mater, 2015, 10(5):055009.
[31] DANILEVICIUS P,GEORGIADI L,PATEMAN C J,et al. The effect of porosity on cell ingrowth into accurately defined, laser-made, polylactide-based 3D scaffolds[J]. Appl Surface Sci, 2015,336:2-10.
[32] MICHAEL S, LAURA K, ANKE B. 3D powder printed bioglass and beta-tricalcium phosphate bone scaffolds[J]. Materials, 2018, 11(2):1-21.
[33] GETHIN R O, MICHEL D, HANNU L. Hydoxyapatite/beta-tricalcium phosphate biphasic ceramics as regenerative material for the repair of complex bone defects[J]. J Biomed Mater Res Part B-Appl Biomater, 2018, 106(6):2493-2512.