@article{chansoria_asif_gupta_piedrahita_shirwaiker_2022, title={Multiscale Anisotropic Tissue Biofabrication via Bulk Acoustic Patterning of Cells and Functional Additives in Hybrid Bioinks}, volume={1}, ISSN={["2192-2659"]}, DOI={10.1002/adhm.202102351}, abstractNote={Recapitulation of the microstructural organization of cellular and extracellular components found in natural tissues is an important but challenging feat for tissue engineering, which demands innovation across both process and material fronts. In this work, a highly versatile ultrasound‐assisted biofabrication (UAB) approach is demonstrated that utilizes radiation forces generated by superimposing ultrasonic bulk acoustic waves to rapidly organize arrays of cells and other biomaterial additives within single and multilayered hydrogel constructs. UAB is used in conjunction with a novel hybrid bioink system, comprising of cartilage‐forming cells (human adipose‐derived stem cells or chondrocytes) and additives to promote cell adhesion (collagen microaggregates or polycaprolactone microfibers) encapsulated within gelatin methacryloyl (GelMA) hydrogels, to fabricate cartilaginous tissue constructs featuring bulk anisotropy. The hybrid matrices fabricated under the appropriate synergistic thermo‐reversible and photocrosslinking conditions demonstrate enhanced mechanical stiffness, stretchability, strength, construct shape fidelity and aligned encapsulated cell morphology and collagen II secretion in long‐term culture. Hybridization of UAB is also shown with extrusion and stereolithography printing to fabricate constructs featuring 3D perfusable channels for vasculature combined with a crisscross or circumferential organization of cells and adhesive bioadditives, which is relevant for further translation of UAB toward complex physiological‐scale biomimetic tissue fabrication.}, journal={ADVANCED HEALTHCARE MATERIALS}, author={Chansoria, Parth and Asif, Suleman and Gupta, Nithin and Piedrahita, Jorge and Shirwaiker, Rohan A.}, year={2022}, month={Jan} }
 @article{maturavongsadit_narayanan_chansoria_shirwaiker_benhabbour_2021, title={Cell-Laden Nanocellulose/Chitosan-Based Bioinks for 3D Bioprinting and Enhanced Osteogenic Cell Differentiation}, volume={4}, ISSN={["2576-6422"]}, DOI={10.1021/acsabm.0c01108}, abstractNote={3D bioprinting has recently emerged as a very useful tool in tissue engineering and regenerative medicine. However, developing suitable bioinks to fabricate specific tissue constructs remains a challenging task. Herein, we report on a nanocellulose/chitosan-based bioink, which is compatible with a 3D extrusion-based bioprinting technology, to design and engineer constructs for bone tissue engineering and regeneration applications. Bioinks were prepared using thermogelling chitosan, glycerophosphate, hydroxyethyl cellulose, and cellulose nanocrystals (CNCs). Formulations were optimized by varying the concentrations of glycerophosphate (80-300 mM), hydroxyethyl cellulose (0-0.5 mg/mL), and CNCs (0-2% w/v) to promote fast gelation kinetics (<7 s) at 37 °C and retain the shape integrity of constructs post 3D bioprinting. We investigated the effect of CNCs and pre-osteoblast cells (MC3T3-E1) on the rheological properties of bioinks, bioink printability, and mechanical properties of bioprinted scaffolds. We demonstrate that the addition of CNCs and cells (5 million cells/mL) significantly improved the viscosity of bioinks and the mechanical properties of chitosan scaffolds post-fabrication. The bioinks were biocompatible and printable at an optimized range of printing pressures (12-20 kPa) that did not compromise cell viability. The presence of CNCs promoted greater osteogenesis of MC3T3-E1 cells in chitosan scaffolds as shown by the upregulation of alkaline phosphatase activity, higher calcium mineralization, and extracellular matrix formation. The versatility of this CNCs-incorporated chitosan hydrogel makes it attractive as a bioink for 3D bioprinting to engineer scaffolds for bone tissue engineering and other therapeutic applications.}, number={3}, journal={ACS APPLIED BIO MATERIALS}, author={Maturavongsadit, Panita and Narayanan, Lokesh Karthik and Chansoria, Parth and Shirwaiker, Rohan and Benhabbour, S. Rahima}, year={2021}, month={Mar}, pages={2342–2353} }
 @article{chansoria_asif_polkoff_chung_piedrahita_shirwaiker_2021, title={Characterizing the Effects of Synergistic Thermal and Photo-Cross-Linking during Biofabrication on the Structural and Functional Properties of Gelatin Methacryloyl (GeIMA) Hydrogels}, volume={7}, ISSN={["2373-9878"]}, DOI={10.1021/acsbiomaterials.1c00635}, abstractNote={Gelatin methacryloyl (GelMA) hydrogels have emerged as promising and versatile biomaterial matrices with applications spanning drug delivery, disease modeling, and tissue engineering and regenerative medicine. GelMA exhibits reversible thermal cross-linking at temperatures below 37 °C due to the entanglement of constitutive polymeric chains, and subsequent ultraviolet (UV) photo-cross-linking can covalently bind neighboring chains to create irreversibly cross-linked hydrogels. However, how these cross-linking modalities interact and can be modulated during biofabrication to control the structural and functional characteristics of this versatile biomaterial is not well explored yet. Accordingly, this work characterizes the effects of synergistic thermal and photo-cross-linking as a function of GelMA solution temperature and UV photo-cross-linking duration during biofabrication on the hydrogels' stiffness, microstructure, proteolytic degradation, and responses of NIH 3T3 and human adipose-derived stem cells (hASC). Smaller pore size, lower degradation rate, and increased stiffness are reported in hydrogels processed at lower temperature or prolonged UV exposure. In hydrogels with low stiffness, the cells were found to shear the matrix and cluster into microspheroids, while poor cell attachment was noted in high stiffness hydrogels. In hydrogels with moderate stiffness, ones processed at lower temperature demonstrated better shape fidelity and cell proliferation over time. Analysis of gene expression of hASC encapsulated within the hydrogels showed that, while the GelMA matrix assisted in maintenance of stem cell phenotype (CD44), a higher matrix stiffness resulted in higher pro-inflammatory marker (ICAM1) and markers for cell-matrix interaction (ITGA1 and ITGA10). Analysis of constructs with ultrasonically patterned hASC showed that hydrogels processed at higher temperature possessed lower structural fidelity but resulted in more cell elongation and greater anisotropy over time. These findings demonstrate the significant impact of GelMA material formulation and processing conditions on the structural and functional properties of the hydrogels. The understanding of these material-process-structure-function interactions is critical toward optimizing the functional properties of GelMA hydrogels for different targeted applications.}, number={11}, journal={ACS BIOMATERIALS SCIENCE & ENGINEERING}, author={Chansoria, Parth and Asif, Suleman and Polkoff, Kathryn and Chung, Jaewook and Piedrahita, Jorge A. and Shirwaiker, Rohan A.}, year={2021}, month={Nov}, pages={5175–5188} }
 @misc{chansoria_schuchard_shirwaiker_2021, title={Process hybridization schemes for multiscale engineered tissue biofabrication}, volume={13}, ISSN={["1939-0041"]}, url={http://dx.doi.org/10.1002/wnan.1673}, DOI={10.1002/wnan.1673}, abstractNote={Recapitulation of multiscale structure-function properties of cells, cell-secreted extracellular matrix, and 3D architecture of natural tissues is central to engineering biomimetic tissue substitutes. Toward achieving biomimicry, a variety of biofabrication processes have been developed, which can be broadly classified into five categories-fiber and fabric formation, additive manufacturing, surface modification, remote fields, and other notable processes-each with specific advantages and limitations. The majority of biofabrication literature has focused on using a single process at a time, which often limits the range of tissues that could be created with relevant features that span nano to macro scales. With multiscale biomimicry as the goal, development of hybrid biofabrication strategies that synergistically unite two or more processes to complement each other's strengths and limitations has been steadily increasing. This work discusses recent literature in this domain and attempts to equip the reader with the understanding of selecting appropriate processes that can harmonize toward creating engineered tissues with appropriate multiscale structure-function properties. Opportunities related to various hybridization schemes and a future outlook on scale-up biofabrication have also been discussed. This article is categorized under: Nanotechnology Approaches to Biology > Nanoscale Systems in Biology Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement.}, number={2}, journal={WILEY INTERDISCIPLINARY REVIEWS-NANOMEDICINE AND NANOBIOTECHNOLOGY}, publisher={Wiley}, author={Chansoria, Parth and Schuchard, Karl and Shirwaiker, Rohan A.}, year={2021}, month={Mar} }
 @article{chansoria_shirwaiker_2020, title={3D bioprinting of anisotropic engineered tissue constructs with ultrasonically induced cell patterning}, volume={32}, ISSN={["2214-7810"]}, DOI={10.1016/j.addma.2020.101042}, abstractNote={As 3D bioprinting continues to evolve as a promising alternative to engineer complex human tissues in-vitro, there is a need to augment bioprinting processes to achieve the requisite cellular and extracellular organizational characteristics found in the original tissues. While the cell distribution within bioinks is typically homogeneous, incorporating appropriate cellular patterning within the bioprinted constructs is an essential first step towards the eventual formation of anisotropically organized tissue matrix essential to its biomechanical form and function. This study describes a new bioprinting technique that uses ultrasonic standing bulk acoustic waves (SBAW) to organize cells into controllable anisotropic patterns within viscous bioinks while maintaining high cell viability. First, we develop a 3D computational model to discern the SBAW pressure pattern in response to multiple ultrasonic frequencies (0.71–2 MHz). We then experimentally analyze the patterns and viabilities of human adipose-derived stem cells (hASC) and human osteosarcoma cells (MG63) in alginate as a function of the SBAW frequency. Computational results indicate the formation of parallel pressure strands with higher pressure amplitudes near the bottom of the deposited layer, which is corroborated by experimental images of cell patterning. The inter-strand spacing is found to be affected by the frequency (p < 0.0001), while an interaction effect between the cell type and frequency governs the width of the strands (p = 0.02). Further, we demonstrate the synergistic bioprinting and SBAW-induced patterning of hASC within alginate and gelatin methacrylate (GelMA) constructs in tandem with chemical and photo-crosslinking, respectively. Pertinent cellular patterning and viability of at least 80 % were noted in the alginate and GelMA constructs across the experimental design space. Finally, we demonstrate the vat photo-polymerization-based bioprinting of a 3-layered GelMA construct with hASC strand lay pattern of 0-45-90° across the layers. This work represents a step forward in advancing bioprinting capabilities to achieve biomimetic tissue constructs.}, journal={ADDITIVE MANUFACTURING}, author={Chansoria, Parth and Shirwaiker, Rohan}, year={2020}, month={Mar} }
 @article{chansoria_narayanan_wood_alvarado_lin_shirwaiker_2020, title={Effects of Autoclaving, EtOH, and UV Sterilization on the Chemical, Mechanical, Printability, and Biocompatibility Characteristics of Alginate}, volume={6}, ISSN={["2373-9878"]}, DOI={10.1021/acsbiomaterials.0c00806}, abstractNote={Sterilization is a necessary step during the processing of biomaterials, but it can affect the materials' functional characteristics. This study characterizes the effects of three commonly used sterilization processes-autoclaving (heat-based), ethanol (EtOH; chemical-based), and ultraviolet (UV; radiation-based)-on the chemical, mechanical, printability, and biocompatibility properties of alginate, a widely used biopolymer for drug delivery, tissue engineering, and other biomedical applications. Sterility assessment tests showed that autoclaving was effective against Gram-positive and Gram-negative bacteria at loads up to 108 CFU/mL, while EtOH was the least effective. Nuclear magnetic-resonance spectroscopy showed that the sterilization processes did not affect the monomeric content in the alginate solutions. The differences in compressive stiffness of the three sterilized hydrogels were also not significant. However, autoclaving significantly reduced the molecular weight and polydispersity index, as determined via gel permeation chromatography, as well as the dynamic viscosity of alginate. Printability analyses showed that the sterilization process as well as the extrusion pressure and speed affected the number of discontinuities and spreading ratio in printed and cross-linked strands. Finally, human adipose-derived stem cells demonstrated over 90% viability in all sterilized hydrogels over 7 days, but the differences in cellular metabolic activity in the three groups were significant. Taken together, the autoclaving process, while demonstrating broad spectrum sterility effectiveness, also resulted in most notable changes in alginate's key properties. In addition to the specific results with the three sterilization processes and alginate, this study serves as a roadmap to characterize the interrelationships between sterilization processes, fundamental chemical properties, and resulting functional characteristics and processability of hydrogels.}, number={9}, journal={ACS BIOMATERIALS SCIENCE & ENGINEERING}, author={Chansoria, Parth and Narayanan, Lokesh Karthik and Wood, Madison and Alvarado, Claudia and Lin, Annie and Shirwaiker, Rohan A.}, year={2020}, month={Sep}, pages={5191–5201} }
 @article{asif_chansoria_shirwaiker_2020, title={Ultrasound-assisted vat photopolymerization 3D printing of preferentially organized carbon fiber reinforced polymer composites}, volume={56}, ISSN={["2212-4616"]}, DOI={10.1016/j.jmapro.2020.04.029}, abstractNote={In this study, we present a new vat photopolymerization 3D printing process that uses acoustic radiation forces from ultrasonic standing waves to organize carbon short fibers within a photocurable resin. A chamber was developed to generate the standing bulk acoustic wave in the resin to align the carbon fibers along the nodes of the standing wave. The resin was then selectively cured to create constructs in the shape of a dog bone specimen by exposing to UV. The effect of fiber concentration (0.5 %, 1 %, 2 %, and 4 % w/v) and direction of alignment (parallel, perpendicular) on tensile strength of the carbon fiber reinforced polymer composites was determined. The constructs with 1 % w/v showed the highest gain in tensile strength due to the fiber alignment. For two-layered constructs with 1 % fiber concentration, 0°–0° constructs (fibers aligned along the uniaxial testing direction) demonstrated significantly higher tensile strength followed by 0°–90°constructs compared to constructs with randomly distributed fibers and without fibers.}, journal={JOURNAL OF MANUFACTURING PROCESSES}, author={Asif, Suleman and Chansoria, Parth and Shirwaiker, Rohan}, year={2020}, month={Aug}, pages={1340–1343} }