73 research outputs found
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Direct Freeform Fabrication of Spatially Heterogeneous Living Cell-Impregnated Implants
The objectives of this work are the development of the processes, materials, and tooling to
directly “3-D print” living, pre-seeded, patient-specific implants of spatially heterogeneous
compositions. The research presented herein attempts to overcome some of the challenges to
scaffolding, such as the difficulty of producing spatially heterogeneous implants that require
varied seeding densities and/or cell-type distributions. In the proposed approach, living implants
are fabricated by the layer-wise deposition of pre-cell-seeded alginate hydrogel. Although
alginate hydrogels have been previously used to mold living implants, the properties of the
alginate formulations used for molding were not suitable for 3-D printing. In addition to changing
the formulation to make the alginate hydrogels “printable,” we developed a robotic hydrogel
deposition system and supporting CAD software to deposit the gel in arbitrary geometries. We
demonstrated this technology’s capabilities by printing alginate gel implants of multiple materials
with various spatial heterogeneities, including, implants with completely embedded material
clusters. The process was determined to be both viable (94±5% n=15) and sterile (less than one
bacterium per 0.9 µL after 8 days of incubation). Additionally, we demonstrated the printing of a
meniscus cartilage-shaped gel generated directly from a CT Scan. The proposed approach may
hold advantages over other tissue printing efforts [5,9]. This technology has the potential to
overcome challenges to scaffolding and could enable the efficient fabrication of spatially
heterogeneous, patient-specific, living implants.Mechanical Engineerin
Characterization of mesenchymal stem cells and fibrochondrocytes in three-dimensional co-culture: analysis of cell shape, matrix production, and mechanical performance
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A Study of Variable Stiffness Alginate Printing for Medical Applications
Technologies for multi-material 3D-printing of anatomical shapes are useful
both for fabrication of heterogeneous cell-seeded implants as well as for
fabrication of synthetic models for surgical planning and training. For both these
applications, it would be desirable to print directly with biological materials to
best emulate the target’s properties. Using a novel material platform, we
describe a series of experiments attempting to print variable-stiffness hydrogels.
We vary compliances by alternating 2% alginate hydrogel and a Dextran-infused
calcium chloride post-crosslinker. Stiffness throughout the construct ranged
from 4 kPa to 20 kPa as a function of post-crosslinker concentration, which was
spatially specified by the user.Mechanical Engineerin
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Improved Quality of 3D-Printed Tissue Constructs Through Enhanced Mixing of Alginate Hydrogels
While alginate hydrogel is a desirable material platform for Solid Freeform Fabrication (SFF) of
cell-seeded tissue engineering scaffolds, achieving consistently high-quality results can be
challenging. Local variations in the material properties cause inconsistent material deposition
behavior and consequently decrease the resultant geometric fidelity of the construct. The effects
of gel mixing on material property consistency, geometric fidelity, and cell viability were
characterized in an attempt to improve the formulation’s compatibility with SFF processing.
Material homogeneity was quantified through a novel experimental setup composed of an
EnduraTEC mechanical test-frame and custom syringe-extrusion jig. Cell viability and
geometric fidelity were assessed using standard protocol. The baseline mechanical stiffness of
the printed samples was 16±3 kPa (n=6). We found that increasing mixing reduced material
inconsistency and improved geometric fidelity, without adversely affecting cell viability: the
printed construct quality was drastically improved by increasing mixing well beyond previously
established limits.Mechanical Engineerin
Cell and protein compatible 3D bioprinting of mechanically strong constructs for bone repair
Rapid prototyping of bone tissue engineering constructs often utilizes elevated temperatures, organic solvents and/or UV light for materials processing. These harsh conditions may prevent the incorporation of cells and therapeutic proteins in the fabrication processes. Here we developed a method for using bioprinting to produce constructs from a thermoresponsive microparticulate material based on poly(lactic-co-glycolic acid) at ambient conditions. These constructs could be engineered with yield stresses of up to 1.22 MPa and Young's moduli of up to 57.3 MPa which are within the range of properties of human cancellous bone. Further study showed that protein-releasing microspheres could be incorporated into the bioprinted constructs. The release of the model protein lysozyme from bioprinted constructs was sustainted for a period of 15 days and a high degree of protein activity could be measured up to day 9. This work suggests that bioprinting is a viable route to the production of mechanically strong constructs for bone repair under mild conditions which allow the inclusion of viable cells and active proteins
Effect of dynamic compressive loading and its combination with a growth factor on the chondrocytic phenotype of 3-dimensional scaffold-embedded chondrocytes
Background and purpose Three-dimensionally (3D-) embedded chondrocytes have been suggested to maintain the chondrocytic phenotype. Furthermore, mechanical stress and growth factors have been found to be capable of enhancing cell proliferation and ECM synthesis. We investigated the effect of mechanical loading and growth factors on reactivation of the 3D-embedded chondrocytes
Articular cartilage and changes in Arthritis: Cell biology of osteoarthritis
The reaction patterns of chondrocytes in osteoarthritis can be summarized in five categories: (1) proliferation and cell death (apoptosis); changes in (2) synthetic activity and (3) degradation; (4) phenotypic modulation of the articular chondrocytes; and (5) formation of osteophytes. In osteoarthritis, the primary responses are reinitiation of synthesis of cartilage macromolecules, the initiation of synthesis of types IIA and III procollagens as markers of a more primitive phenotype, and synthesis of active proteolytic enzymes. Reversion to a fibroblast-like phenotype, known as 'dedifferentiation', does not appear to be an important component. Proliferation plays a role in forming characteristic chondrocyte clusters near the surface, while apoptosis probably occurs primarily in the calcified cartilage
Effects of Matrix Metalloproteinases on Cartilage Biophysical Properties in Vitro and in Vivo
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