Abstract
Free-form printing offers a novel biofabrication approach to generate complex shapes by depositing hydrogel materials within a temporary supportive environment. However, printed hydrogels typically lack the requisite mechanical properties and functionality of the desired tissue, limiting application and, more importantly, safety and efficacy of the implant. The study authors have developed an innovative nanoclay-based bioink to print high shape fidelity functional constructs for potential skeletal application. Laponite® (LAP) nanoclay was combined with gellan gum (GG) to generate a printable hydrogel that was highly stable in vitro, displayed limited swelling ability compared with the silicate-free control and remained stable over time. An agarose fluid gel was found to provide the requisite support for the deposition of the material ink and preservation of the printed structure before crosslinking. Printed C2C12 myoblasts remained viable and displayed extensive proliferation over 21 days in culture. Cell-laden scaffolds demonstrated functionality within 1 day of culture in vitro and that was preserved over 3 weeks. Analysis of absorption and release mechanisms from LAP-GG using model proteins (lysozyme and bovine serum albumin) demonstrated the retention capability of the clay-based materials for compound localisation and absence of burst release. Vascular endothelial growth factor was loaded within the agarose fluid gel and absorbed by the material ink via absorption during deposition. The 3D-printed constructs were implanted on the chorioallantoic membrane of a 10-day-old developing chick. Extensive and preferential vasculature infiltration was observed in LAP-GG–loaded vascular endothelial growth factor constructs compared with controls (p<0.01 and p<0.0001) after only 7 days of incubation. The current studies demonstrate, for the first time, the application of innovative LAP-GG 3D constructs in the generation of growth factor–loaded 3D constructs for potential application in skeletal tissue repair.
Highlights
The printing of cells in three dimensions (3D) for the fabrication of large structures targeted at critical-sized bone tissue defects has proved challenging [1]
Laponite® (LAP) nanoclay was combined with gellan gum (GG) to generate a printable hydrogel that was highly stable in vitro, displayed limited swelling ability compared with the silicate-free control and remained stable over time
Extensive and preferential vasculature infiltration was observed in LAP-GG–loaded vascular endothelial growth factor constructs compared with controls (p
Summary
The printing of cells in three dimensions (3D) for the fabrication of large structures targeted at critical-sized bone tissue defects has proved challenging [1]. The deposition of bioinks within a temporary support material has demonstrated the ability to generate intricate-shaped constructs, at scale, to date impossible to produce with standard 3D printing technology [6,7,8,9]. This novel platform, free-form printing, enables the generation of free-floating fibres in a support gel bath with the capacity for additional processing and removal by crosslinking [6,7] or reverse embedding [8,9]. The use of LAP as a supporting bath allowed the fabrication of complex structures; extrusion of material with pH
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