基于3D打印与静电纺织技术的血管化组织工程骨构建
发布时间：2019-03-05 08:33
【摘要】：目的:利用3D打印技术及高压静电纺织技术联合构建组织工程支架-细胞复合物,并通过体内外实验验证其在构建血管化组织工程骨中的优势。方法:利用医学图像三维重建软件(Mimics)及3D打印机将断层扫描数据(CT)进行处理、编辑制作成个性化三维立体模型,在模型打印的同时利用同轴高压静电纺织技术制备芯层为P3HB4HB、壳层为PVA与人骨髓间充质干细胞的(PVA-细胞)/P3HB4HB纤维。将一次成型的支架-细胞复合物向成骨方向诱导14日,植入前复合内皮细胞。体外实验中通过力学测试观察经3D打印技术制作的P3HB4HB外部支架的力学性能;通过光镜、透射电镜、亲水性实验及扫描电镜观察经静电纺织技术制备的PVA/P3HB4HB支架的微观结构及生物相容性;通过扫描电镜、DAPI免疫荧光染色、吖啶橙免疫荧光染色以及CCK-8实验观察细胞在支架结构中的黏附、增殖情况;体内实验中将支架-细胞复合物作为实验组,不含细胞的支架材料作为对照组,标本分别于12周与24周后取出,行HE、VonKossa、天狼星红、马松三色、CD31免疫组织化学及Ⅰ型胶原免疫组织化学染色,观察支架-细胞复合物成骨及血管的能力。结果:利用3D打印技术能够制造外观个性化很强的模型,模型能够为组织工程支架提供一定的力学支撑;通过扫描电镜及亲水性实验可观察到通过静电纺织技术制备的PVA/P3HB4HB支架具有较高的孔隙率、良好的生物相容性及仿生细胞外基质的三维立体结构;利用吖啶橙免疫荧光染色、DAPI免疫荧光染色、扫描电镜及CCK-8可显示细胞与支架复合后仍具有增值、分化的能力,实现了材料与细胞的一次成型与精确种植;在体内实验中,分别对实验组12周和24周的标本行HE染色、Von Kossa染色、天狼星红染色、马松三色染色、CD31免疫组织化学染色、Ⅰ型胶原免疫组织化学染色,结果均呈阳性,并随着时间的延长骨组织及血管结构的数量及质量明显增加,对照组阴性。结论:利用3D打印技术及高压静电纺织技术联合构建的支架-细胞复合物具有一定的力学性能及仿生细胞外基质结构的结构与功能,通过复合内皮细胞后,能够在体内构建出含血管的组织工程骨。
[Abstract]:Aim: to construct tissue engineering scaffold-cell complex using 3D printing technique and high voltage electrostatic textile technology, and to verify its advantages in the construction of vascularized tissue engineering bone in vitro and in vivo. Methods: the medical image 3D reconstruction software (Mimics) and 3D printer were used to process the scanning data (CT), and the 3D 3D model was edited and made into personalized 3D model. The PVA- cells / P3HB4HB fibers of PVA and human bone marrow mesenchymal stem cells (PVA- cells) were prepared by co-axial high-voltage electrostatic spinning technique and the core layer was P3HB4HB.The core layer was P3HB4HB. the core layer was P3HB4HB. The scaffold-cell complex was induced to osteogenic direction for 14 days, and the endothelial cells were mixed before implantation. The mechanical properties of P3HB4HB external scaffolds fabricated by 3D printing technology were observed by mechanical test in vitro. The microstructure and biocompatibility of PVA/P3HB4HB scaffolds were observed by light microscopy, transmission electron microscopy, hydrophilicity test and scanning electron microscopy. Scanning electron microscopy, DAPI immunofluorescence staining, acridine orange immunofluorescence staining and CCK-8 test were used to observe the adhesion and proliferation of the cells in the scaffolds. In vivo, the scaffold-cell complex was used as the experimental group, and the cell-free scaffold material was used as the control group. After 12 weeks and 24 weeks respectively, the specimens were taken out, HE,VonKossa, Sirius Red and Ma Song trichromatic, respectively. CD31 immunohistochemical staining and type 鈪，
本文编号：2434715
[Abstract]:Aim: to construct tissue engineering scaffold-cell complex using 3D printing technique and high voltage electrostatic textile technology, and to verify its advantages in the construction of vascularized tissue engineering bone in vitro and in vivo. Methods: the medical image 3D reconstruction software (Mimics) and 3D printer were used to process the scanning data (CT), and the 3D 3D model was edited and made into personalized 3D model. The PVA- cells / P3HB4HB fibers of PVA and human bone marrow mesenchymal stem cells (PVA- cells) were prepared by co-axial high-voltage electrostatic spinning technique and the core layer was P3HB4HB.The core layer was P3HB4HB. the core layer was P3HB4HB. The scaffold-cell complex was induced to osteogenic direction for 14 days, and the endothelial cells were mixed before implantation. The mechanical properties of P3HB4HB external scaffolds fabricated by 3D printing technology were observed by mechanical test in vitro. The microstructure and biocompatibility of PVA/P3HB4HB scaffolds were observed by light microscopy, transmission electron microscopy, hydrophilicity test and scanning electron microscopy. Scanning electron microscopy, DAPI immunofluorescence staining, acridine orange immunofluorescence staining and CCK-8 test were used to observe the adhesion and proliferation of the cells in the scaffolds. In vivo, the scaffold-cell complex was used as the experimental group, and the cell-free scaffold material was used as the control group. After 12 weeks and 24 weeks respectively, the specimens were taken out, HE,VonKossa, Sirius Red and Ma Song trichromatic, respectively. CD31 immunohistochemical staining and type 鈪，
本文编号：2434715
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