Browsing by Subject "tissue engineering"
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Item Fluorescent microparticles for sensing cell microenvironment oxygen levels within 3D scaffolds(Elsevier B.V, 2009-03-14) Acosta, Miguel A.; Ymele-Leki, Patrick; Kostov, Yordan V.; Leach, Jennie B.We present the development and characterization of fluorescent oxygen-sensing microparticles designed for measuring oxygen concentration in microenvironments existing within standard cell culture and transparent three-dimensional (3D) cell scaffolds. The microparticle synthesis employs poly(dimethylsiloxane) to encapsulate silica gel particles bound with an oxygen-sensitive luminophore as well as a reference or normalization fluorophore that is insensitive to oxygen. We developed a rapid, automated and non-invasive sensor analysis method based on fluorescence microscopy to measure oxygen concentration in a hydrogel scaffold. We demonstrate that the microparticles are non-cytotoxic and that their response is comparable to that of a traditional dissolved oxygen meter. Microparticle size (5–40 μm) was selected for microscale-mapping of oxygen concentration to allow measurements local to individual cells. Two methods of calibration were evaluated and revealed that the sensor system enables characterization of a range of hypoxic to hyperoxic conditions relevant to cell and tissue biology (i.e., pO₂ 10–160 mm Hg). The calibration analysis also revealed that the microparticles have a high fraction of quenched luminophore (0.90 ± 0.02), indicating that the reported approach provides significant advantages for sensor performance. This study thus reports a versatile oxygen-sensing technology that enables future correlations of local oxygen concentration with individual cell response in cultured engineered tissues.Item Hydrolytically degradable poly(ethylene glycol) hydrogel scaffolds with tunable degradation and mechanical properties(American Chemical Society, 2010-05-10) Zustiak, Silviya P.; Leach, Jennie B.The objective of this work was to create three-dimensional (3D) hydrogel matrices with defined mechanical properties, as well as tunable degradability for use in applications involving protein delivery and cell encapsulation. Thus, we report the synthesis and characterization of a novel hydrolytically degradable poly(ethylene glycol) (PEG) hydrogel composed of PEG vinyl sulfone (PEG-VS) cross-linked with PEG-diester-dithiol. Unlike previously reported degradable PEGbased hydrogels, these materials are homogeneous in structure, fully hydrophilic and have highly specific cross-linking chemistry. We characterized hydrogel degradation and associated trends in mechanical properties, i.e., storage modulus (G′), swelling ratio (QM), and mesh size (ξ). Degradation time and the monitored mechanical properties of the hydrogel correlated with crosslinker molecular weight, cross-linker functionality, and total polymer density; these properties changed predictably as degradation proceeded (G′ decreased, whereas QM and ξ increased) until the gels reached complete degradation. Balb/3T3 fibroblast adhesion and proliferation within the 3D hydrogel matrices were also verified. In sum, these unique properties indicate that the reported degradable PEG hydrogels are well poised for specific applications in protein and cell delivery to repair soft tissue.Item Protein–Hydrogel Interactions in Tissue Engineering: Mechanisms and Applications(Mary Ann Liebert, Inc, 2012-11-13) Zustiak, Silviya P.; Wei, Yunqian; Leach, Jennie B.Recent advances in our understanding of the sophistication of the cellular microenvironment and the dynamics of tissue remodeling during development, disease, and regeneration have increased our appreciation of the current challenges facing tissue engineering. As this appreciation advances, we are better equipped to approach problems in the biology and therapeutics of even more complex fields, such as stem cells and cancer. To aid in these studies, as well as the established areas of tissue engineering, including cardiovascular, musculoskeletal, and neural applications, biomaterials scientists have developed an extensive array of materials with specifically designed chemical, mechanical, and biological properties. Herein, we highlight an important topic within this area of biomaterials research, protein–hydrogel interactions. Due to inherent advantages of hydrated scaffolds for soft tissue engineering as well as specialized bioactivity of proteins and peptides, this field is well-posed to tackle major needs within emerging areas of tissue engineering. We provide an overview of the major modes of interactions between hydrogels and proteins (e.g., weak forces, covalent binding, affinity binding), examples of applications within growth factor delivery and three-dimensional scaffolds, and finally future directions within the area of hydrogel–protein interactions that will advance our ability to control the cell–biomaterial interface.Item Reporter Scaffolds for Clinically Relevant Cell Transplantation Studies(Springer US, 2019-11-04) Bolger, Morgan; Groynom, Rebecca; Bogie, Kath; Lavik, ErinThere are a number of cell therapies that are either in clinical trials or moving toward clinical trials, particularly for diseases of the retina. One of the challenges with cell therapies is tracking the status of cells over time. Genetic manipulation can facilitate this, but it can limit the clinical application of the cells. There are a host of fluorophores that have been developed to assess the status of cells, but these molecules tend to be cleared rapidly from cells. There are preclinical strategies that use degradable scaffolds, and we hypothesized that these scaffolds could be used to track the state of cells during preclinical studies. In this work, we explored whether fluorophores could be delivered from simple scaffolds fabricated under extremely harsh conditions, be active upon release, and report on the cells growing on the scaffolds over time. We encapsulated CellROX® Green Reagent, and pHrodo™ Red AM in poly(lactic-co-glycolic acid) (PLGA) scaffolds, showed that they could be delivered over weeks and were still active upon release and taken up by cells. These experiments provide the foundation for using scaffolds to deliver molecules to report on cells.Item Surface Modification of Nerve Guide Conduits with ECM Coatings and Investigating Their Impact on Schwann Cell Response(Wiley, 2024-02-23) Pinzon-Herrera, Luis; Magness, John; Apodaca-Reyes, Hector; Sanchez, Jesus; Almodovar, JorgeIn this study, layer-by-layer coatings composed of heparin and collagen are proposed as an extracellular mimetic environment on nerve guide conduits (NGC) to modulate the behavior of Schwann cells (hSCs). The authors evaluated the stability, degradation over time, and bioactivity of six bilayers of heparin/collagen layer-by-layer coatings, denoted as (HEP/COL)₆. The stability study reveals that (HEP/COL)₆ is stable after incubating the coatings in cell media for up to 21 days. The impact of (HEP/COL)₆ on hSCs viability, protein expression, and migration is evaluated. These assays show that hSCs cultured in (HEP/COL)₆ have enhanced protein expression and migration. This condition increases the expression of neurotrophic and immunomodulatory factors up to 1.5-fold compared to controls, and hSCs migrated 1.34 times faster than in the uncoated surfaces. Finally, (HEP/COL)₆ is also applied to a commercial collagen-based NGC, NeuraGen, and hSC viability and adhesion are studied after 6 days of culture. The morphology of NeuraGen is not altered by the presence of (HEP/COL)₆ and a nearly 170% increase of the cell viability is observed in the condition where NeuraGen is used with (HEP/COL)₆. Additionally, cell adhesion on the coated samples is successfully demonstrated. This work demonstrates the reparative enhancing potential of extracellular mimetic coatings.