In Vitro and in Vivo Studies of Selective Laser Melting-Mediated Surface Concave Microwell Treatment to Enhance Osteogenesis and Osseointegration of Nanohydroxyapatite/Polyamide 66 Implants

Yafeng Wen, Yanan Xu, Weikang Zhao, Jun Wu, Qiming Yang, Sinan Chen, Kai Li, B. Qiao
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Abstract

The structural and functional combination of an implant and living bone is greatly influenced by the surface characteristics of the implant. To enhance the implant-bone interface regarding the osteointegration of nanohydroxyapatite/polyamide66 (nHA/PA66) material, concave microwells that were 100 μm, 200 μm and 400 μm in size (denoted by P100, P200, and P400, respectively) were prepared on the nHA/PA66 substrate surface by selective laser melting (SLM) technology. The surface characteristics of these samples were analyzed by scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDS), 3D scanning laser microscopy and static water contact angle measurements. We examined the effects of different concave microwell sizes on the adhesion, proliferation and osteogenesis of C3H10T1/2 cells in vitro by cell counting kit-8 (CCK-8), SEM, alkaline phosphatase assay, alizarin red staining, and western blot, and continued our evaluating through micro-CT, histological analysis and push-out tests to verify the osseointegration ability in vivo. P100, P200 and P400 surface modification led to significant increases in concave microwell diameter, depth and surface roughness, and the contact angle measurements showed that only the hydrophilicity of P100 microwells was improved. In vitro testing revealed that P100 microwells could effectively promote the adhesion, proliferation and osteogenic differentiation of C3H10T1/2 cells; in vivo studies further confirmed that P100 microwells significantly increased new bone volume, enhanced bone remodeling, and improved material-bone interface bonding force and instigated rapid osteointegration of nHA/PA66 with host bone. However, these phenomena were not observed for P200 and P400 microwells. Overall, P100 was associated with improved cell adhesion, proliferation and osteogenic differentiation in vitro and greater implant fixation in vivo. These results suggest that surface concave microwell treatment of nHA/PA66 material through SLM technology provides a fast, simple and effective method for clinical applications involving bone tissue regeneration.
选择性激光熔化介导的表面凹微孔治疗促进纳米羟基磷灰石/聚酰胺66种植体成骨和骨整合的体内外研究
种植体和活骨的结构和功能组合很大程度上受种植体表面特性的影响。为了增强纳米羟基磷灰石/聚酰胺66 (nHA/PA66)材料骨整合的种植-骨界面,采用选择性激光熔化(SLM)技术在nHA/PA66衬底表面制备了尺寸分别为100 μm、200 μm和400 μm的凹微孔(分别用P100、P200和P400表示)。采用扫描电镜(SEM)、能量色散x射线能谱仪(EDS)、三维扫描激光显微镜和静态水接触角测量等方法分析了样品的表面特征。通过细胞计数试剂盒-8 (CCK-8)、扫描电镜(SEM)、碱性磷酸酶试验、茜素红染色、western blot等检测不同凹孔大小对C3H10T1/2细胞体外粘附、增殖和成骨的影响,并通过显微ct、组织学分析和推出试验继续进行评价,验证其体内骨整合能力。P100、P200和P400的表面改性导致凹孔直径、深度和表面粗糙度显著增加,接触角测量结果表明,只有P100微孔的亲水性得到改善。体外实验表明,P100微孔能有效促进C3H10T1/2细胞的粘附、增殖和成骨分化;体内研究进一步证实P100微孔显著增加新骨体积,增强骨重塑,提高材料-骨界面结合力,促进nHA/PA66与宿主骨快速骨整合。而在P200和P400微孔中没有观察到这些现象。总的来说,P100与体外细胞粘附、增殖和成骨分化的改善以及体内更大的植入物固定有关。这些结果表明,采用SLM技术对nHA/PA66材料进行表面凹微孔处理,为骨组织再生的临床应用提供了一种快速、简单、有效的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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