Synthetic microfluidic systems for wound healing
Inventors
Prabhakarpandian, Balabhaskar • Pant, Kapil
Assignees
Publication Number
US-9784727-B2
Publication Date
2017-10-10
Expiration Date
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Abstract
A method of assaying wound healing can include: growing cells on the matrix in the first flow channel; introducing an agent that removes the matrix from the junction; introducing a matrix material into the second flow channel so as to form the second matrix in the second flow channel and junction; and detecting cellular migration into the junction onto the second matrix. The agent that removes the matrix can include a biomolecule or chemical agent. The method can include removing cells in the matrix in the junction before introducing the matrix material into the second flow channel. A bioactive agent can be introduced into the junction to determine if it modulates cellular migration and/or clot formation into the intersection openings of tissue and vascular channels.
Core Innovation
The invention provides a device and methodology to study and characterize wound healing or hemostasis in idealized and physiologically realistic microenvironments. The device comprises flow channels that intersect at junctions where internal surfaces can be coated with a matrix material, cells can be grown on the matrix in a first flow channel, cells and/or matrix in the junction can be injured and/or removed, a second matrix material can be introduced into the second flow channel and junction, and cellular migration from the first flow channel into the junction onto the second matrix can be detected. The method includes introducing an agent that removes the matrix and/or cells from the junction, introducing a bioactive agent into the junction to determine modulation of cellular migration, and comparing different matrix materials for cellular migration into the junction.
The device can be an idealized microfluidic network (IMN) or a synthetic microvascular network (SMN) and can include multiple multi-channel constructs with central chambers surrounded by capillary channels, porous walls with gaps, and inlet and outlet ports. The device can reproduce morphological size, physiological blood flow, and cellular make-up of circulatory vessels, and can be used with primary cells or cell lines. The device and methodologies allow for study and visualization in real time of wound healing or hemostasis and provide an environment to facilitate studying the interplay between cellular activities.
The background identifies a need for a better system and methodology for studying wound healing and hemostasis because current in vitro models rely primarily on static experiments and fail to reproduce injury characteristics and the variety of physiological (e.g., blood flow, platelet adhesion) and biological (e.g., endothelial response, coagulation pathway) mechanisms involved in wound healing and stopping hemorrhage. The invention addresses this need by providing an in vitro device and assay that can create reproducible wounds, model bleeding wounds of varying sizes, and simulate physiologically realistic fluid dynamic environments to study restoration of hemostasis and wound closure. The device can be used for conventional imaging-based end-point determinations, real time tracking, and medium to high-throughput screening.
Claims Coverage
The independent claim recites a method for assaying cell migration with several inventive features.
Device with intersecting flow channels and matrix coating
Providing a device having a first flow channel that intersects a second flow channel at a junction, the first flow channel having an internal surface that is coated with a matrix material.
Growth of cells on matrix in first flow channel
Growing cells on the matrix material in the first flow channel.
Creation of a cell-free junction by injuring or removing cells
Injuring and/or removing cells in the junction if cells are in the junction.
Detection of cellular migration into junction
Detecting cellular migration of the cells from the first flow channel into the junction.
The independent claim is directed to a microfluidic assay method combining a matrix-coated intersecting-channel device, cultivation and removal of cells/matrix at a junction, and detection of cell migration into the junction.
Stated Advantages
Allows study and characterization of wound healing and hemostasis in idealized and physiologically realistic microenvironments.
Enables study and visualization in real time and quantitative real-time visualization of wound healing or hemostasis.
Provides a platform for developing and testing therapeutics, including screening hemostatic and anti-thrombotic drugs and dressings and testing drugs that promote wound healing.
Can reproduce physiological and biological mechanisms involved in wound healing and hemostasis, providing data predictive of in vivo efficacy and reducing the need for animal experiments.
Supports creation of uniform and repeatable wounds and varied wound models, enables medium to high-throughput screening, and reduces reagent/cell use with disposable chips.
Documented Applications
Studying wound healing phenomena, including creating reproducible wounds, studying cell migration into wounded areas, and comparing cellular migration on different 3D matrix materials.
Modeling hemostasis and thrombosis, including creating bleeding wounds of varying sizes, monitoring thrombus formation and restoration of hemostasis, and modeling clot formation and occlusion after induced injury.
Screening and testing hemostatic agents, anti-thrombotic drugs, hemostatic dressings, and other therapeutics in physiologically realistic fluid dynamic environments.
Drug delivery vehicle screening assays with a system comprising pumps, automated stages, and optical visualization for real-time monitoring.
Studying effects of altered physiological conditions relevant to hemorrhagic shock, including dilution of coagulation factors, pH and temperature changes, and metabolic changes.
High to medium-throughput screening and real-time visualization for quantitative assessment of clot size, rate of clot formation, and flow/pressure changes in microfluidic assays.
Studying migration of cells such as leukocytes and cases where a cancer cell can leave one chamber and travel through fluid channels to a different chamber.
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