Schweitzer Luiz, Schoon Janosch, Bläß Niklas, Huesker Katrin, Neufend Janine V, Siemens Nikolai, Bekeschus Sander, Schlüter Rabea, Schneider Peter, Uhlmann Eckart, Wassilew Georgi, Schulze Frank
Fraunhofer Institute for Production Systems and Design Technology, Berlin, Germany.
Center for Orthopaedics, Trauma Surgery and Rehabilitation Medicine, University Medicine Greifswald, Greifswald, Germany.
Front Bioeng Biotechnol. 2024 Oct 28;12:1462232. doi: 10.3389/fbioe.2024.1462232. eCollection 2024.
BACKGROUND/OBJECTIVE: Endoprostheses might fail due to complications such as implant loosening or periprosthetic infections. The surface topography of implant materials is known to influence osseointegration and attachment of pathogenic bacteria. Laser-Induced Periodic Surface Structures (LIPSS) can improve the surface topography of orthopedic implant materials. In this preclinical study, laser pulses with a wavelength in the ultraviolet (UV) spectrum were applied for the generation of LIPSS to positively influence formation of extracellular matrix by primary human Osteoblasts (hOBs) and to reduce microbial biofilm formation .
Laser machining was employed for generating UV-LIPSS on sample disks made of Ti6Al4V and Ti6Al7Nb alloys. Sample disks with polished surfaces were used as controls. Scanning electron microscopy was used for visualization of surface topography and adherent cells. Metal ion release and cellular metal levels were investigated by inductively coupled plasma mass spectrometry. Cell culture of hOBs on sample disks with and without UV-LIPSS surface treatments was performed. Cells were investigated for their viability, proliferation, osteogenic function and cytokine release. Biofilm formation was facilitated by seeding on sample disks and quantified by wheat germ agglutinin (WGA) staining.
UV-LIPSS modification results in topographies with a periodicity of 223 nm ≤ λ ≤ 278 nm. The release of metal ions was found increased for UV-LIPSS on Ti6Al4V and decreased for UV-LIPSS on Ti6Al7Nb, while cellular metal levels remain unaffected. Cellular adherence was decreased for hOBs on UV-LIPSS Ti6Al4V when compared to controls while proliferation rate was unaffected. Metabolic activity was lower on UV-LIPSS Ti6Al7Nb when compared to the control. Alkaline phosphatase activity was upregulated for hOBs grown on UV-LIPSS on both alloys. Less pro-inflammatory cytokines were released for cells grown on UV-LIPSS Ti6Al7Nb when compared to polished surfaces. WGA signals were significantly lower on UV-LIPSS Ti6Al7Nb indicating reduced formation of a biofilm.
Our results suggest that UV-LIPSS texturing of Ti6Al7Nb positively influence bone forming function and cytokine secretion profile of hOBs . In addition, our results indicate diminished biofilm formation on UV-LIPSS treated Ti6Al7Nb surfaces. These effects might prove beneficial in the context of long-term arthroplasty outcomes.
背景/目的：由于植入物松动或假体周围感染等并发症，内置假体可能会失效。已知植入材料的表面形貌会影响骨整合和病原菌的附着。激光诱导周期性表面结构（LIPSS）可以改善骨科植入材料的表面形貌。在这项临床前研究中，应用紫外线（UV）光谱中的激光脉冲来生成LIPSS，以积极影响原代人成骨细胞（hOBs）的细胞外基质形成，并减少微生物生物膜的形成。
采用激光加工在由Ti6Al4V和Ti6Al7Nb合金制成的样品盘上生成UV-LIPSS。将具有抛光表面的样品盘用作对照。使用扫描电子显微镜观察表面形貌和贴壁细胞。通过电感耦合等离子体质谱法研究金属离子释放和细胞内金属水平。在有和没有UV-LIPSS表面处理的样品盘上进行hOBs的细胞培养。研究细胞的活力、增殖、成骨功能和细胞因子释放。通过接种在样品盘上促进生物膜形成，并通过麦胚凝集素（WGA）染色进行定量。
UV-LIPSS修饰导致形成周期性为223nm≤λ≤278nm的形貌。发现Ti6Al4V上的UV-LIPSS使金属离子释放增加，而Ti6Al7Nb上的UV-LIPSS使金属离子释放减少，而细胞内金属水平不受影响。与对照相比，UV-LIPSS Ti6Al4V上的hOBs细胞黏附减少，而增殖率不受影响。与对照相比，UV-LIPSS Ti6Al7Nb上的代谢活性较低。两种合金上在UV-LIPSS上生长的hOBs的碱性磷酸酶活性均上调。与抛光表面相比，在UV-LIPSS Ti6Al7Nb上生长的细胞释放的促炎细胞因子较少。UV-LIPSS Ti6Al7Nb上的WGA信号明显较低，表明生物膜形成减少。
我们的结果表明，Ti6Al7Nb的UV-LIPSS纹理化对hOBs的骨形成功能和细胞因子分泌谱有积极影响。此外，我们的结果表明，在UV-LIPSS处理的Ti6Al7Nb表面上生物膜形成减少。这些效应在长期关节置换结果方面可能是有益的。
