静电纺纳米纤维PLCL/纤维蛋白原人工韧带的生物相容性评价

Biocompatibility evaluation of electrospun PLCL/fibrinogen nanofibers in anterior cruciate ligament reconstruction

本研究旨在评价聚乳酸-己内酯(PLCL)/纤维蛋白原纳米纤维膜(P/F-Ns)的安全性和功能性,为临床应用提供理论支撑。通过扫描电镜、材料测试机、接触角测量仪和酶标仪分别研究P/F-Ns的表面形态、力学性能、亲水性和纤维蛋白原含量。通过CCK-8法和实时荧光定量PCR评价Hig-82细胞在P/F-Ns上的黏附、增殖及韧带修复相关基因表达。结果显示,随着纤维蛋白原含量增加,P/F-Ns表面纤维孔隙增大,力学性能下降,亲水性增强,其中P/F-N-2最有利于细胞黏附和增殖。以不含纤维蛋白原的P/F-N-0为对照,含纤维蛋白原的P/F-N-1、P/F-N-2和P/F-N-3上细胞韧带修复相关基因在第3天和第7天表达均上调。与P/F-N-0相比,含纤维蛋白原的P/F-Ns具有更好的生物相容性,能够有效应用于前交叉韧带重建。

The study aimed to evaluate the safety and function of poly(lactic-acid-co-ε-caprolactone)(PLCL)/fibrinogen nanofibers(P/F-Ns), and provide theoretical basis for the clinical application. The surface morphology,mechanical properties, the hydrophilicity and the fibrinogen content of P/F-Ns were tested by scanning electron microscope, the material testing machine, the contact angle meter and the microplate reader, respectively. The cell adhesion, proliferation and ligament remodeling genes expression of Hig-82 cells on P/F-Ns were conducted through cell counting kit-8(CCK-8) and real-time quantitative PCR analyses, respectively. The results showed that with the increase of the fibrinogen content, the pore sizes and hydrophilicity of three P/F-Ns increased, but the mechanical properties decreased. Cell adhesion and proliferation tests showed that P/F-N-2 held the best ability to promote cell adhesion and proliferation. The ligament remodeling genes expressions of Hig-82 cells on P/F-N-1, P/F-N-2 and P/F-N-3 were all upregulated compared to P/F-N-0 on days 3 and 7. All the three P/F-Ns containing fibrinogen(P/F-N-1, P/F-N-2 and P/F-N-3) had better biocompatibility compared to P/F-N-0, and could be efficiently applied to the reconstruction of anterior cruciate ligament.