Gao Chao, Li Chaoming, Xu Yulong, Wang Zhenyong, Li Haojiang, Luo Xujiang, Peng Liqing, Zhang Bin, Shen Shi, Liu Shuyun, Sui Xiang, Guo Quanyi, Yang Jianhua
Department of Orthopedics, the First Affiliated Hospital of Jiamusi University, Jiamusi Heilongjiang, 154007, P.R.China;Institute of Orthopedics, Chinese PLA General Hospital, Beijing Key Lab of Regenerative Medicine in Orthopedics, Key Lab of Musculoskeletal Trauma & War Injuries of Chinese PLA, Beijing, 100853, P.R.China.
College of Mechanical and Electric Engineering, Beijing University of Chemical Technology, Beijing, 100029, P.R.China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2019 May 15;33(5):628-633. doi: 10.7507/1002-1892.201811034.
Electrospinning technique was used to manufacture polycaprolactone (PCL)/collagen typeⅠ nanofibers orientated patches and to study their physical and chemical characterization, discussing their feasibility as synthetic patches for rotator cuff repairing.
PCL patches were prepared by electrospinning with 10% PCL electrospinning solution (control group) and PCL/collagen typeⅠorientated nanofibers patches were prepared by electrospinning with PCL electrospinning solution with 25% collagen type Ⅰ(experimental group). The morphology and microstructure of the two patches were observed by gross and scanning electron microscopy, and the diameter and porosity of the fibers were measured; the mechanical properties of the patches were tested by uniaxial tensile test; the composition of the patches was analyzed by Fourier transform infrared spectroscopy; and the contact angle of the patch surface was measured. Two kinds of patch extracts were co-cultured with the third generation of rabbit tendon stem cells. Cell counting kit 8 (CCK-8) was used to detect the toxicity and cell proliferation of the materials. Normal cultured cells were used as blank control group. Rabbit tendon stem cells were co-cultured with the two patches and stained with dead/living cells after 3 days of culture, and laser confocal scanning microscopy was used to observe the cell adhesion and activity on the patch.
Gross and scanning electron microscopy showed that the two patch fibers were arranged in orientation. The diameter of patch fibers in the experimental group was significantly smaller than that in the control group ( =26.907, =0.000), while the porosity in the experimental group was significantly larger than that in the control group ( =2.506, =0.032). The tensile strength and Young's modulus of the patch in the experimental group were significantly higher than those in the control group ( =3.705, =0.029; =4.064, =0.034). Infrared spectrum analysis showed that PCL and collagen type Ⅰ were successfully mixed in the experimental group. The surface contact angle of the patch in the experimental group was (73.88±4.97)°, which was hydrophilic, while that in the control group was (128.46±5.10) °, which was hydrophobic. There was a significant difference in the surface contact angle between the two groups ( =21.705, =0.002). CCK-8 test showed that with the prolongation of culture time, the cell absorbance ( ) value increased gradually in each group, and there was no significant difference between the experimental group and the control group at each time point ( >0.05). Laser confocal scanning microscopy showed that rabbit tendon stem cells could adhere and grow on the surface of both patches after 3 days of culture. The number of cells adhered to the surface of the patches in the experimental group was more than that in the control group, and the activity was better.
PCL/ collagen type Ⅰ nanofibers orientated patch prepared by electrospinning technology has excellent physical and chemical properties, cell adhesion, and no cytotoxicity. It can be used as an ideal scaffold material in tendon tissue engineering for rotator cuff repair in the future.
采用静电纺丝技术制备聚己内酯（PCL）/Ⅰ型胶原纳米纤维定向贴片，并研究其物理化学特性，探讨其作为肩袖修复合成贴片的可行性。
采用静电纺丝法，用10%的PCL静电纺丝溶液制备PCL贴片（对照组），用含25%Ⅰ型胶原的PCL静电纺丝溶液制备PCL/Ⅰ型胶原定向纳米纤维贴片（实验组）。通过肉眼和扫描电子显微镜观察两种贴片的形态和微观结构，并测量纤维的直径和孔隙率；通过单轴拉伸试验测试贴片的力学性能；用傅里叶变换红外光谱分析贴片的成分；测量贴片表面的接触角。将两种贴片提取物与第三代兔肌腱干细胞共培养。采用细胞计数试剂盒8（CCK-8）检测材料的毒性和细胞增殖情况。以正常培养的细胞作为空白对照组。将兔肌腱干细胞与两种贴片共培养，培养3天后进行活/死细胞染色，用激光共聚焦扫描显微镜观察细胞在贴片上的黏附及活性情况。
肉眼和扫描电子显微镜显示，两种贴片纤维均呈定向排列。实验组贴片纤维直径显著小于对照组（=26.907，=0.000），而实验组孔隙率显著大于对照组（=2.506，=0.032）。实验组贴片的拉伸强度和杨氏模量显著高于对照组（=3.705，=0.029；=4.064，=0.034）。红外光谱分析表明，实验组中PCL与Ⅰ型胶原成功混合。实验组贴片表面接触角为（73.88±4.97）°，呈亲水性，而对照组为（128.46±5.10）°，呈疏水性。两组表面接触角有显著差异（=21.705，=0.002）。CCK-8试验显示，随着培养时间延长，各组细胞吸光度（）值逐渐升高，各时间点实验组与对照组间差异无统计学意义（>0.05）。激光共聚焦扫描显微镜显示，培养3天后兔肌腱干细胞均可在两种贴片表面黏附生长。实验组贴片表面黏附的细胞数量多于对照组，且活性更好。
通过静电纺丝技术制备的PCL/Ⅰ型胶原纳米纤维定向贴片具有优异的物理化学性能、细胞黏附性，且无细胞毒性。未来可作为肌腱组织工程中肩袖修复的理想支架材料。
