A novel approach for efficient manufacturing of three-dimensional (3D) microstructured scaffolds designed for cell studies and tissue engineering applications is presented. A thermal extrusion (fused filament fabrication) 3D printer is employed as a simple and low-cost tabletop device enabling rapid materialization of CAD models out of biocompatible and biodegradable polylactic acid (PLA). Here it was used to produce cm- scale microporous (pore size varying from 100 to 400 µm) scaffolds. The fabricated objects were further laser processed in a direct laser writing (DLW) subtractive (ablation) and additive (lithography) manners. The first approach enables precise surface modification by creating micro-craters, holes and grooves thus increasing the surface roughness. An alternative way is to immerse the 3D PLA scaffold in a monomer solution and use the same DLW setup to refine its inner structure by fabricating dots, lines or a fine mesh on top as well as inside the pores of previously produced scaffolds. The DLW technique is empowered by ultrafast lasers - it allows 3D structuring with high spatial resolution in a great variety of photosensitive materials. Structure geometry on macro- to micro- scales could be finely tuned by combining these two fabrication techniques. Such artificial 3D substrates could be used for cell growth or as biocompatible-biodegradable implants. This combination of distinct material processing techniques enables rapid fabrication of diverse functional micro- featured and integrated devices. Hopefully, the proposed approach will find numerous applications in the field of ms, microfluidics, microoptics and many others.
We present a novel approach to manufacturing 3D microstructured composite scaffolds for tissue engineering applications. A thermal extrusion 3D printer – a simple, low-cost tabletop device enabling rapid materialization of CAD models in plastics – was used to produce cm-scale microporous scaffolds out of polylactic acid (PLA). The fabricated objects were subsequently immersed in a photosensitive monomer solution and direct laser writing technique (DLW) was used to refine its inner structure by fabricating a fine mesh inside the previously produced scaffold. In addition, a composite material structure out of four different materials fabricated via DLW is presented. This technique, empowered by ultrafast lasers allows 3D structuring with high spatial resolution in a great variety of photosensitive materials. A composite scaffold made of distinct materials and periodicities is acquired after the development process used to wash out non-linked monomers. Another way to modify the 3D printed PLA surfaces was also demonstrated - ablation with femtosecond laser beam. Structure geometry on macro- to micro- scales could be finely tuned by combining these fabrication techniques. Such artificial 3D substrates could be used for cell growth or as biocompatible-biodegradable implants. To our best knowledge, this is the first experimental demonstration showing the creation of composite 3D scaffolds using convenient 3D printing combined with DLW. This combination of distinct material processing techniques enables rapid fabrication of diverse functional micro-featured and integrated devices. Hopefully, the proposed approach will find numerous applications in the field of tissue engineering, as well as in microelectromechanical systems, microfluidics, microoptics and others.
This work presents the latest results on direct laser writing of polymeric materials for tissue engineering applications. A femtosecond Yb:KGW laser (300 fs, 200 kHz, 515 nm) was used as a light source for non-linear lithography. Fabrication was implemented in various photosensitive polymeric materials, such as: hybrid organic-inorganic sol-gel based on silicon-zirconium oxides, commercial ORMOCER® class photoresins. These materials were structured via multi-photon polymerization technique with submicron resolution. Porous three-dimensional scaffolds for artificial tissue engineering were fabricated with constructed system and were up to several millimeters in overall size with 10 to 100 μm internal pores. Biocompatibility of the used materials was tested in primary rabbit muscle-derived stem cell culture in vitro and using laboratory rats in vivo. This interdisciplinary study suggests that proposed technique and materials are suitable for tissue engineering applications.
Fabrication of scaffolds for cell growth with appropriate mechanical characteristics is top-most important for successful
creation of tissue. Due to ability of fast fabrication of periodic structures with a different period, the holographic
lithography technique is a suitable tool for scaffolds fabrication. The scaffolds fabricated by holographic lithography can
be used in various biomedical investigations such as the cellular adhesion, proliferation and viability. These
investigations allow selection of the suitable material and geometry of scaffolds which can be used in creation of tissue.
Scaffolds fabricated from di-acrylated poly(ethylene glycol) (PEG-DA-258) over a large area by holographic lithography
technique are presented in this paper. The PEG-DA scaffolds fabricated by holographic lithography showed good
cytocompatibility for rabbit myogenic stem cells. It was observed that adult rabbit muscle-derived myogenic stem cells
grew onto PEG-DA scaffolds. They were attached to the pillars and formed cell-cell interactions. It demonstrates that the
fabricated structures have potential to be an interconnection channel network for cell-to-cell interactions, flow transport
of nutrients and metabolic waste as well as vascular capillary ingrowth. These results are encouraging for further
development of holographic lithography by improving its efficiency for microstructuring three-dimensional scaffolds out
of biodegradable hydrogels
In this work, a combination of Direct Laser Writing (DLW), PoliDiMethylSiloxane (PDMS) soft lithography and
UV lithography was used to create cm- scale microstructured polymer scaolds for cell culture experiments out of
dierent biocompatible materials: novel hybrid organic-inorganic SZ2080, PDMS elastomer, biodegradable PEG-
DA-258 and SU-8. Rabbit muscle-derived stem cells were seeded on the fabricated dierent periodicity scaolds
to evaluate if the relief surface had any eect on cell proliferation. An array of microlenses was fabricated using
DLW out of SZ2080 and replicated in PDMS and PEG-DA-258, showing good potential applicability of the used
techniques in many other elds like micro- and nano-
uidics, photonics, and MicroElectroMechanical Systems
(MEMS). The synergetic employment of three dierent fabrication techniques allowed to produce desired objects
with low cost, high throughput and precision as well as use materials that are dicult to process by other means
(PDMS and PEG-DA-258). DLW is a relatively slow fabrication method, since the object has to be written
point-by-point. By applying PDMS soft lithography, we were enabled to replicate laser-fabricated scaolds for
stem cell growth and micro-optical elements for lab-on-a-chip applications with high speed, low cost and good
reproducible quality.
In this work we present the latest results in the application of multi-photon polymerization for tissue engineering,
by fabricating microstructured artificial 3D scaffolds for stem cell growth. Microstructuring of large scale 3D
scaffolds is investigated and the direct laser writing technique is supplemented by fabrication by multi-beam
interference and micromolding of large scale structures. Within the limitation of our study, we conclude that the
proposed nonlinear direct laser writing technique offers rapid and flexible fabrication of biomedical components
with required shape, pore size and general porosity. The applications could target biostable and biodegradable
implants applied for bone or tissue replacement as well as drug delivery or release agents.
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