Backes Eduardo H, Fernandes Emanuel M, Diogo Gabriela S, Marques Catarina F, Silva Tiago H, Costa Lidiane C, Passador Fabio R, Reis Rui L, Pessan Luiz A
Graduate Program in Materials Science and Engineering, Federal University of São Carlos, via Washington Luiz, Km 235, 13565-905 São Carlos, SP, Brazil; 3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal.
3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal; ICVS/3B's - PT Government Associate Laboratory, Braga, Guimarães, Portugal.
Mater Sci Eng C Mater Biol Appl. 2021 Mar;122:111928. doi: 10.1016/j.msec.2021.111928. Epub 2021 Feb 3.
In this study, polylactic acid (PLA) filled with hydroxyapatite (HA) or beta-tricalcium phosphate (TCP) in 5 wt% and 10 wt% of concentration were produced employing twin-screw extrusion followed by fused filament fabrication in two different architectures, varying the orientation of fibers of adjacent layers. The extruded 3D filaments presented suitable rheological and thermal properties to manufacture of 3D scaffolds envisaging bone tissue engineering. The produced scaffolds exhibited a high level of printing accuracy related to the 3D model; confirmed by micro-CT and electron microscopy analysis. The developed architectures presented mechanical properties compatible with human bone replacement. The addition of HA and TCP made the filaments bioactive, and the deposition of new calcium phosphates was observed upon 7 days of incubation in simulated body fluid, exemplifying a microenvironment suitable for cell attachment and proliferation. After 7 days of cell culture, the constructs with a higher percentage of HA and TCP demonstrated a significantly superior amount of DNA when compared to neat PLA, indicating that higher concentrations of HA and TCP could guide a good cellular response and increasing cell cytocompatibility. Differentiation tests were performed, and the biocomposites of PLA/HA and PLA/TCP exhibited earlier markers of cell differentiation as confirmed by alkaline phosphatase and alizarin red assays. The 3D printed composite scaffolds, manufactured with bioactive materials and adequate porous size, supported cell attachment, proliferation, and differentiation, which together with their scalability, promise a high potential for bone tissue engineering applications.
在本研究中,采用双螺杆挤出法制备了分别填充有5 wt%和10 wt%浓度的羟基磷灰石(HA)或β-磷酸三钙(TCP)的聚乳酸(PLA),随后通过熔丝制造法以两种不同结构制造,改变相邻层纤维的取向。挤出的3D长丝呈现出适合制造用于骨组织工程的3D支架的流变学和热性能。所制备的支架显示出与3D模型相关的高打印精度;这通过显微CT和电子显微镜分析得到证实。所开发的结构呈现出与人体骨替代物相容的力学性能。HA和TCP的添加使长丝具有生物活性,并且在模拟体液中孵育7天后观察到新的磷酸钙沉积,这例证了适合细胞附着和增殖的微环境。细胞培养7天后,与纯PLA相比,含有较高百分比HA和TCP的构建体显示出显著更高量的DNA,表明较高浓度的HA和TCP可以引导良好的细胞反应并提高细胞的细胞相容性。进行了分化测试,并且通过碱性磷酸酶和茜素红测定证实,PLA/HA和PLA/TCP的生物复合材料表现出更早的细胞分化标志物。用生物活性材料和合适的孔径制造的3D打印复合支架支持细胞附着、增殖和分化,并且它们的可扩展性使其在骨组织工程应用中具有很高的潜力。