三维打印双相磷酸钙陶瓷支架在骨组织工程中的应用

发布时间：2018-03-24 05:25

本文选题：三维打印技术　切入点：纳米羟基磷灰石　出处：《吉林大学》2017年硕士论文

【摘要】：目的:因意外伤害、炎症、肿瘤及先天畸形所致骨缺损在临床上十分常见,这种缺损常常会影响患者的正常生理形态和生理功能,从而严重影响患者的生活[1]。目前临床上用于治疗这类骨缺损的方式有自体骨移植,异体骨移植以及人工骨移植等传统治疗方法。但是这些传统的治疗方法都存在着各自的局限性[2]。随着材料科学、生命科学和工程技术的发展,组织工程技术在应用于骨缺损的修复方面越来越受到人们的重视。组织工程包括三个基本要素,分别为种子细胞、支架、生物因子。其中支架作为联系细胞与周围环境的的桥梁,对于组织工程的成败有重要的影响。因而,支架的选择十分重要。理想的骨组织工程支架材料的研究范围十分广泛,它应具有良好的生物相容性、骨诱导性、骨传导性,与人骨相适宜的生物力学性能,合理的生物降解性以及足够的孔隙率和孔隙尺寸结构。同时在临床应用方面,支架材料又应具备较高的可塑性,来源充足,加工简单,价格低廉等优点。在本实验中,采用三维打印技术制备的羟基磷灰石(HA)/β-磷酸三钙(β-TCP)复合双相磷酸钙陶瓷(BCP)支架,探究其在骨组织工程中的应用潜力。方法:通过三维打印技术制备HA/β-TCP复合纳米双相磷酸钙陶瓷支架,其中HA与β-TCP以质量比为HA/β-TCP:3/7的比例混合。并以相同材料制备的压模片。使用扫描电镜(SEM)观察支架形态,X线衍射(XRD)检测支架物相。分离培养兔骨髓间充质干细胞(BMSC),将支架与兔BMSC共培养;通过CCK-8实验、碱性磷酸酶实验检测支架的生物相容性以及种子细胞在支架上的增殖分化。结果:三维打印技术制备的BCP支架高3mm、直径分别3mm和8mm,圆柱形。压模片的形状与BCP支架相同。三维打印支架的孔隙结构大小均匀,互相连通,孔隙的孔径大小在350-450μm之间。孔隙壁表面可见许多微孔结构。CCK-8结果显示:在体外培养1d、3d、5d后,空白组的细胞数量明显高于实验组及对照组(P0.05)。培养1d后,对照组细胞数量高于实验组(P0.05),而3d、5d后,对照组与实验组细胞数量已无明显差异(P0.05)。培养7d后,对照组的细胞数量低于实验组及空白组(P0.05),实验组与空白组的细胞数量无明显差异。ALP活性检测试验显示:在体外培养4、7、14d后,实验组的ALP活性均明显高于对照组(P0.05)。培养4d后对照组的ALP活性则高于空白组(P0.05)。而培养7、14d后,对照组与空白组的ALP活性无明显差异。结论:通过上述实验,结果表明:通过三维打印技术制备的HA/β-TCP复合双相磷酸钙陶瓷支架,具有良好的三维孔隙结构且孔隙间互相交通。支架的生物相容性良好,BMSC能够在支架上很好的粘附、增殖及分化。相较于平面结构的支架,BCP支架的多孔结构以及微孔形成了更大的表面积,有利于细胞粘附增殖分化。通过本实验,证明了三维打印技术制备的双相磷酸钙陶瓷支架在骨组织工程中的应用潜力,并为后续的实验及临床应用奠定了前期基础。
[Abstract]:Objective: bone defects caused by accidental injury, inflammation, tumor and congenital malformation are very common in clinical practice. These defects often affect the normal physiological morphology and function of patients. This seriously affected the life of the patients [1] .At present, autogenous bone transplantation is used to treat this kind of bone defect in clinic. Traditional treatments such as allogeneic bone transplantation and artificial bone transplantation have their own limitations. [2] with the development of material science, life science and engineering technology, Tissue engineering technology has attracted more and more attention in the application of bone defect repair. Tissue engineering includes three basic elements: seed cells and scaffolds. Biological factors. Scaffolds, as a bridge between cells and surrounding environment, play an important role in the success or failure of tissue engineering. Therefore, the selection of scaffolds is very important. It should have good biocompatibility, bone inductivity, bone conductivity, biomechanical properties suitable for human bone, reasonable biodegradability and adequate porosity and pore size structure. In this experiment, hydroxyapatite / 尾 -tricalcium phosphate (尾 -TCP) composite biphasic calcium phosphate ceramic scaffold was prepared by three-dimensional printing technique. Methods: HA/ 尾 -TCP composite nano-biphasic calcium phosphate ceramic scaffolds were prepared by three-dimensional printing technique. The HA and 尾 -TCP were mixed in the ratio of HA/ 尾 -TCPW / 3 / 7 and the compression tablets were prepared from the same material. The morphology of the scaffold was observed by scanning electron microscopy (SEM) and X-ray diffraction (XRD) was used to detect the phase of the scaffold. The rabbit bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured. The scaffold was co-cultured with rabbit BMSC. Through the CCK-8 experiment, Alkaline phosphatase assay was used to detect the biocompatibility of the scaffold and the proliferation and differentiation of seed cells on the scaffold. Results: the BCP scaffold prepared by 3D printing technique was 3 mm high, with diameter of 8 mm and 3mm diameter of 8 mm. The same rack. The pore structure of the three-dimensional printing bracket is uniform, The pore size ranged from 350 渭 m to 450 渭 m. The results of CCK-8 showed that the number of cells in the blank group was significantly higher than that in the experimental group and the control group after 1 d of culture for 3 d or 5 d. After 1 day of culture, the number of cells in the blank group was significantly higher than that in the experimental group and the control group. The number of cells in the control group was higher than that in the experimental group, but there was no significant difference in the number of cells between the control group and the experimental group after 3 days. The number of cells in the control group was lower than that in the experimental group and the blank group, and there was no significant difference in the number of cells between the experimental group and the blank group. The activity of ALP in the experimental group was significantly higher than that in the control group (P 0.05). The activity of ALP in the control group was higher than that in the blank group after 4 days of culture. However, there was no significant difference in the activity of ALP between the control group and the blank group after 7d of culture. The results show that the HA/ 尾 -TCP composite biphasic calcium phosphate ceramic scaffold prepared by 3D printing technique has good three-dimensional pore structure and good biocompatibility. Proliferation and differentiation. Compared with planar scaffold BCP scaffolds, the porous structure and micropores form a larger surface area, which is conducive to cell adhesion, proliferation and differentiation. The application potential of the biphasic calcium phosphate ceramic scaffold prepared by 3D printing technology in bone tissue engineering is proved, and the preliminary foundation for subsequent experiment and clinical application is established.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R318.08;TP391.73

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