Impact of 3-D printed PLA- and chitosan-based scaffolds on human monocyte/macrophage responses: unraveling the effect of 3-D structures on inflammation.

Recent studies have pointed towards a decisive role of inflammation in triggering tissue repair and regeneration, while at the same time it is accepted that an exacerbated inflammatory response may lead to rejection of an implant. Within this context, understanding and having the capacity to regulate the inflammatory response elicited by 3-D scaffolds aimed for tissue regeneration is crucial. This work reports on the analysis of the cytokine profile of human monocytes/macrophages in contact with biodegradable 3-D scaffolds with different surface properties, architecture and controlled pore geometry, fabricated by 3-D printing technology. Fabrication processes were optimized to create four different 3-D platforms based on polylactic acid (PLA), PLA/calcium phosphate glass or chitosan. Cytokine secretion and cell morphology of human peripheral blood monocytes allowed to differentiate on the different matrices were analyzed. While all scaffolds supported monocyte/macrophage adhesion and stimulated cytokine production, striking differences between PLA-based and chitosan scaffolds were found, with chitosan eliciting increased secretion of tumor necrosis factor (TNF)-α, while PLA-based scaffolds induced higher production of interleukin (IL)-6, IL-12/23 and IL-10. Even though the material itself induced the biggest differences, the scaffold geometry also impacted on TNF-α and IL-12/23 production, with chitosan scaffolds having larger pores and wider angles leading to a higher secretion of these pro-inflammatory cytokines. These findings strengthen the appropriateness of these 3-D platforms to study modulation of macrophage responses by specific parameters (chemistry, topography, scaffold architecture).

[1]  M. Lamghari,et al.  Adsorbed fibrinogen leads to improved bone regeneration and correlates with differences in the systemic immune response. , 2013, Acta biomaterialia.

[2]  Melba Navarro,et al.  Development of a Biodegradable Composite Scaffold for Bone Tissue Engineering: Physicochemical, Topographical, Mechanical, Degradation, and Biological Properties , 2006 .

[3]  M. Neil,et al.  Structurally Distinct Membrane Nanotubes between Human Macrophages Support Long-Distance Vesicular Traffic or Surfing of Bacteria1 , 2006, The Journal of Immunology.

[4]  V. Kuchroo,et al.  IL-12 family cytokines: immunological playmakers , 2012, Nature Immunology.

[5]  S. Hollister Porous scaffold design for tissue engineering , 2005, Nature materials.

[6]  Alberto Mantovani,et al.  Orchestration of metabolism by macrophages. , 2012, Cell metabolism.

[7]  B. Brown,et al.  Macrophage polarization: an opportunity for improved outcomes in biomaterials and regenerative medicine. , 2012, Biomaterials.

[8]  J. Simon,et al.  Immune responses to implants - a review of the implications for the design of immunomodulatory biomaterials. , 2011, Biomaterials.

[9]  Han Tong Loh,et al.  Fabrication of 3D chitosan–hydroxyapatite scaffolds using a robotic dispensing system , 2002 .

[10]  M. Bissell Cellular Plasticity of Inflammatory Myeloid Cells in the Peritoneal Foreign Body Response , 2011 .

[11]  D. Brunette,et al.  The effect of surface topography on early NFκB signaling in macrophages. , 2010, Journal of biomedical materials research. Part A.

[12]  James M. Anderson,et al.  Foreign body reaction to biomaterials. , 2008, Seminars in immunology.

[13]  Antonios G Mikos,et al.  Harnessing and modulating inflammation in strategies for bone regeneration. , 2011, Tissue engineering. Part B, Reviews.

[14]  D. Dormont,et al.  Macrophage activation switching: an asset for the resolution of inflammation , 2005, Clinical and experimental immunology.

[15]  R. Soares,et al.  Immobilization of human mesenchymal stem cells within RGD-grafted alginate microspheres and assessment of their angiogenic potential. , 2010, Biomacromolecules.

[16]  Jan Feijen,et al.  A poly(D,L-lactide) resin for the preparation of tissue engineering scaffolds by stereolithography. , 2009, Biomaterials.

[17]  T. Turvey,et al.  Biodegradable fixation for craniomaxillofacial surgery: a 10-year experience involving 761 operations and 745 patients. , 2011, International journal of oral and maxillofacial surgery.

[18]  Melba Navarro,et al.  Physicochemical Degradation of Titania‐Stabilized Soluble Phosphate Glasses for Medical Applications , 2003 .

[19]  K. Heffels,et al.  Inducing healing-like human primary macrophage phenotypes by 3D hydrogel coated nanofibres. , 2012, Biomaterials.

[20]  Buddy D. Ratner,et al.  Biomaterial topography alters healing in vivo and monocyte/macrophage activation in vitro. , 2010, Journal of biomedical materials research. Part A.

[21]  D. Kaplan,et al.  Porosity of 3D biomaterial scaffolds and osteogenesis. , 2005, Biomaterials.

[22]  Marcello Imbriani,et al.  Effect of electrospun fiber diameter and alignment on macrophage activation and secretion of proinflammatory cytokines and chemokines. , 2011, Biomacromolecules.

[23]  Melba Navarro,et al.  Cellular response to calcium phosphate glasses with controlled solubility. , 2003, Journal of biomedical materials research. Part A.

[24]  Jacqueline A. Jones,et al.  Characterization of topographical effects on macrophage behavior in a foreign body response model. , 2010, Biomaterials.

[25]  M. Barbosa,et al.  Enhanced mesenchymal stromal cell recruitment via natural killer cells by incorporation of inflammatory signals in biomaterials , 2012, Journal of the Royal Society Interface.

[26]  Y. Wong,et al.  Direct writing of chitosan scaffolds using a robotic system , 2005 .

[27]  Yongnian Yan,et al.  Multinozzle low-temperature deposition system for construction of gradient tissue engineering scaffolds. , 2009, Journal of biomedical materials research. Part B, Applied biomaterials.

[28]  I. Hammami,et al.  Biodegradable chitosan particles induce chemokine release and negligible arginase-1 activity compared to IL-4 in murine bone marrow-derived macrophages. , 2011, Biochemical and biophysical research communications.

[29]  Mayte Suárez-Fariñas,et al.  Tumor-associated macrophages in the cutaneous SCC microenvironment are heterogeneously activated. , 2011, The Journal of investigative dermatology.

[30]  L. Ambrosio,et al.  Layer-by-layer self-assembly of chitosan and poly(γ-glutamic acid) into polyelectrolyte complexes. , 2011, Biomacromolecules.

[31]  C. Hughes,et al.  Of Mice and Not Men: Differences between Mouse and Human Immunology , 2004, The Journal of Immunology.

[32]  M. Barbosa,et al.  The effect of adsorbed fibronectin and osteopontin on macrophage adhesion and morphology on hydrophilic and hydrophobic model surfaces. , 2012, Acta biomaterialia.

[33]  B. Saramago,et al.  Functionalization of chitosan membranes through phosphorylation: Atomic force microscopy, wettability, and cytotoxicity studies , 2006 .

[34]  Weiliam Chen,et al.  A fibroblast/macrophage co-culture model to evaluate the biocompatibility of an electrospun Dextran/PLGA scaffold and its potential to induce inflammatory responses , 2011, Biomedical materials.

[35]  R. Hernández-Pando,et al.  Inflammatory cytokine production by immunological and foreign body multinucleated giant cells , 2000, Immunology.

[36]  Silvano Sozzani,et al.  The chemokine system in diverse forms of macrophage activation and polarization. , 2004, Trends in immunology.

[37]  M. Barbosa,et al.  Evaluation of the effect of the degree of acetylation on the inflammatory response to 3D porous chitosan scaffolds. , 2009, Journal of biomedical materials research. Part A.

[38]  V. Pascual,et al.  From IL-2 to IL-37: the expanding spectrum of anti-inflammatory cytokines , 2012, Nature Immunology.

[39]  Y. Nakayama,et al.  Spatial regulation and surface chemistry control of monocyte/macrophage adhesion and foreign body giant cell formation by photochemically micropatterned surfaces. , 1999, Journal of biomedical materials research.

[40]  Philip Kollmannsberger,et al.  Geometry as a Factor for Tissue Growth: Towards Shape Optimization of Tissue Engineering Scaffolds , 2013, Advanced healthcare materials.

[41]  J. Planell,et al.  High-resolution PLA-based composite scaffolds via 3-D printing technology. , 2013, Acta biomaterialia.

[42]  Stanley J Stachelek,et al.  Correlating macrophage morphology and cytokine production resulting from biomaterial contact. , 2013, Journal of biomedical materials research. Part A.

[43]  Yongnian Yan,et al.  Fabrication of porous scaffolds for bone tissue engineering via low-temperature deposition , 2002 .

[44]  P H Krebsbach,et al.  Indirect solid free form fabrication of local and global porous, biomimetic and composite 3D polymer-ceramic scaffolds. , 2003, Biomaterials.

[45]  M. Oliveira,et al.  Chitosan drives anti-inflammatory macrophage polarisation and pro-inflammatory dendritic cell stimulation. , 2012, European cells & materials.

[46]  W Kenneth Ward,et al.  The effect of microgeometry, implant thickness and polyurethane chemistry on the foreign body response to subcutaneous implants. , 2002, Biomaterials.

[47]  S F Hulbert,et al.  Tissue reaction to three ceramics of porous and non-porous structures. , 1972, Journal of biomedical materials research.

[48]  Milan Mrksich,et al.  Geometric cues for directing the differentiation of mesenchymal stem cells , 2010, Proceedings of the National Academy of Sciences.

[49]  J. Hamuro,et al.  The polarization of T(h)1/T(h)2 balance is dependent on the intracellular thiol redox status of macrophages due to the distinctive cytokine production. , 2002, International immunology.