T35 - Microphysiological System for Investigating Wound Healing Responses

Friday, October 10, 2025

10:00 AM - 11:00 AM PDT

Location: Exhibit Hall F, G & H

Presenting Author(s)

EB

Elizabeth Byrnes

Graduate Student
North Carolina State University, United States

Co-Author(s)

Halston Deal

Post-Doc
NC State University
Knightdale, North Carolina, United States
HC

Halle J. Cantor

PhD Candidate
North Carolina State University, United States
MP

Megan G. Palmisano

Post Doctoral Research Fellow
North Carolina State University, United States
AO

Aubrey M. O'Quinn

Student
North Carolina State University, United States
Preethi Coulter

Undergraduate Research Assistant
Lampe Joint Department of Biomedical Engineering at North Carolina State University and University of North Carolina at Chapel Hill
Chapel Hill, North Carolina, United States
JF

Joshua Furr

Undergraduate Researcher
North Carolina State University, United States

Primary Investigator(s)

Ashley C. Brown

Lampe Distinguished Professor
North Carolina State University and University of North Carolina at Chapel Hill
Raleigh, North Carolina, United States

Last Author(s)

MD

Michael Daniele

Faculty Researcher
University of North Carolina at Chapel Hill & North Carolina State University, United States

Introduction: : Fibrin is the end product of the coagulation cascade and plays a critical role in hemostasis, tissue repair, and thrombosis. The molecular and mechanical properties of fibrin differ significantly during physiologic versus pathophysiological states. These differences impact clot stability, polymerization kinetics, and interactions with molecular components, such as extracellular matrix (ECM) factors that support vascular repair. Despite its importance, our ability to monitor clot formation and degradation in real time remains limited to bulky apparatuses that require large sample volumes, requiring up to 6mL. Preclinical animal models of hemostasis and thrombosis have provided valuable clinical insights into the role of fibrin in hemostasis; however, species-specific differences in coagulation cascade dynamics, platelet biology, and immune responses contribute to a poor correlation between animal studies and human clinical outcomes. Microphysiological systems (MPS) have been quickly adopted as tools to study cells and tissues by providing chemically-defined platforms to investigate fundamental biomaterial processes, such as coagulation and fibrinolysis. MPS overcomes the limitations inherent to traditional animal models by isolating target effects. Herein, we present the development and application of a versatile H-channel MPS for the characterization of fibrin, fibrin-derived biomaterials, and cellular responses. Our device captures high-density data on real-time biochemical interactions and polymerization kinetics under simulated physiological conditions. This device will ultimately lead to improvements in our understanding of the physico-chemical mechanisms underlying hemostasis and thrombosis to improve clinical outcomes.

Materials and

Methods: : This study details the development and application of a versatile H-channel microphysiological system (MPS). The device was designed with Fusion 360, followed by mold fabrication using a CADworks 3D printer, and subsequent polydimethylsiloxane (PDMS) casting. The PDMS was then plasma-bonded to glass to form the microfluidic channels seen in a multiplexed version in Figure 1A. The top channel was loaded with a fibrin clot volume ranging from 20uL to 50uL, and the flow channel was either left for the perfusion of various agents, including fibrinolytic solutions for degradation studies, dyes for permeability assessments, and treatments for studying responses to various coagulopathies through controlled flow (Figure 1B). To monitor cell migration within fibrin clots, cells were cultured in the flow channel. This device was then used to study fibrin permeability, clot degradation, and cellular migration by confocal imaging the clot interface (Figure 1C). Continuing work shows that multiplexing enables a higher rate of data output.

Results, Conclusions, and Discussions:: The interfacial clot monitoring device offers a highly controlled platform for dissecting fundamental biomaterial processes involved in hemostasis and thrombosis. Its design enables precise modulation of flow conditions such as shear stresses and rates, targeted delivery of molecular components, incorporation of cellular elements, and real-time monitoring of dynamic events, facilitating detailed characterization of fibrin formation, structure, and interactions. For instance, we have used this MPS to study cellular migration, permeability, and fibrinolysis. The versatility of this device has been demonstrated by its ability to elucidate results for equine studies comparing antithrombotics by quantifying their impact on clot density and leukocyte infiltration, human sex-dependent studies by revealing differences in clot response, particle delivery, and cellular responses to various species’ fibrin. In studies comparing the permeability of increasing fibrin densities in clots, the MPS demonstrated a decrease in permeability as fibrin density increases. This shows a simple, yet descriptive, aspect of fibrin that is applicable to multiple other physiological processes. This MPS has the potential for providing a foundation to radically improve clinical outcomes in hemostasis and thrombosis. By providing a platform to dissect mechanisms underlying clot formation and degradation, the system can accelerate the development and optimization of antithrombotic and procoagulant therapies. Moreover, the capacity for culturing cells within the device permits analysis of the interplay between fibrin and cellular components involved in vascular repair, enabling investigation into new strategies for wound healing and regenerative medicine. While the current focus is on fibrin, the versatile nature of the H-channel design suggests its adaptability for investigating other biomaterials and biological processes, positioning it as a foundational tool for future microphysiological research.

Acknowledgements and/or References (Optional): :