3D打印聚醚醚酮/羟基磷灰石复合物的生物相容性评价

Biocompatibility of 3D printed polyetheretherketone/hydroxyapatite composites

背景:聚醚醚酮材料具有稳定的物理、化学、力学性质,辐射透明性及与人体骨骼相似的弹性模量等优点,但存在着生物惰性的问题,限制了其在临床中的应用。目的:评价熔融沉积型3D打印技术制备聚醚醚酮/羟基磷灰石复合材料的生物相容性。方法:以聚醚醚酮、聚醚醚酮/羟基磷灰石(羟基磷灰石质量分数为10%)丝材为原料,应用熔融沉积型3D打印技术分别制备聚醚醚酮材料(对照组)、聚醚醚酮/羟基磷灰石复合材料(实验组)。将第3代小鼠前成骨细胞MC3T3-E1分别与两组材料浸提液共培养,以单独培养的细胞为阳性对照,通过Live/Dead荧光染色和CCK-8实验检测细胞的存活与增殖情况;将第3代小鼠前成骨细胞MC3T3-E1分别接种于两种材料表面,利用扫描电镜观察MC3T3-E1细胞在材料表面的黏附生长、增殖情况。结果与结论:①Live/Dead荧光染色显示,MC3T3-E1细胞在两组材料浸提液中生长良好,细胞活力良好,细胞存活率在90%以上,说明两种材料均无明显细胞毒性,随着培养时间的延长,细胞增殖良好;②CCK-8实验结果显示,两种材料无明显的细胞毒性,其中实验组培养3,5,7 d的细胞增殖吸光度值大于阳性对照组、对照组(P<0.05);③扫描电镜下可见,接种第1天时,MC3T3-E1细胞在两组材料表面黏附生长,细胞铺展良好,其中以实验组细胞分泌的基质较多;第3天时,黏附于两组材料表面的细胞增多,同时分泌更多的基质,其中以实验组材料表面黏附细胞数量及基质分泌更多;第7天时,两组材料表面出现细胞重叠生长现象,实验组材料上细胞铺展生长、分泌基质的效率更高;④结果表明,3D打印制备聚醚醚酮/羟基磷灰石复合材料具有良好的生物相容性。

BACKGROUND:Polyetheretherketone materials have the advantages of stable physical,chemical and mechanical properties,radiation transparency and elastic modulus similar to human bones,but there is a biological inertia problem,which limits its clinical application.OBJECTIVE:To evaluate the biocompatibility of polyetheretherketone/hydroxyapatite composites prepared by fused deposition molding 3D printing.METHODS:Polyetheretherketone and polyetheretherketone/hydroxyapatite composite materials(hydroxyapatite mass fraction 10%)were prepared by fused deposition molding 3D printing technology,which were denoted as polyetheretherketone group(control group)and polyetheretherketone/hydroxyapatite group(experimental group).Passage 3 mouse preosteoblasts(MC3T3-E1)were co-cultured with the extracts of the two groups of materials,separately.Cells cultured alone were used as positive controls.The survival and proliferation of MC3T3-E1 were detected by Live/Dead fluorescence staining and CCK-8 assay.The adhesion growth and proliferation of MC3T3-E1 on the surface of the materials were observed by scanning electron microscopy.RESULTS AND CONCLUSION:(1)Live/Dead fluorescence staining exhibited that the MC3T3-E1 cells in two kinds of extracts grew well and presented good viability.The cell survival rate was higher than 90%.The two materials had no obvious cytotoxicity.The cells proliferated well with the extension of culture time.(2)The results of CCK-8 assay showed that the cytotoxicity of the two kinds of materials was qualified.The proliferation absorbance value of cells in the experimental group was higher than that of positive control group and control group at 3,5 and 7 days of culture(P<0.05).(3)Scanning electron microscope observation showed the good adhesion and cell spreading.Matrix secretion of MC3T3-E1 cells was numerous in the experimental group at 1 day of inoculation.The number of cells adhering to the surface of the two groups of materials increased,and more matrixes were secreted at the same time,among which the number of cells adhering to the surface of the experimental group and the secretion of matrix were more at 3 days of inoculation.The overlapping growth of cells appeared on the surface of the two groups of materials,and the cells in the experimental group were more efficient in spreading growth and secreting matrix at 7 days of inoculation.(4)These findings verify that the polyetheretherketone/hydroxyapatite composite prepared by 3D printing has good biocompatibility.