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Binding of Plasmodium falciparum-Infected Red Blood Cells to Engineered 3D Microvessels
Livia Piatti1, Caitlin C Howard2, Ying Zheng2
1European Molecular Biology Laboratory (EMBL) Barcelona, Barcelona, Spain.
Abstract:
P. falciparum-infected red blood cell (iRBC) sequestration in the microvasculature is a pivotal event in severe malaria pathogenesis. In vitro binding assays using endothelial cell monolayers under static and flow conditions have revealed key ligand-receptor interactions for iRBC sequestration. However, mechanisms remain elusive for iRBC sequestration in specific vascular locations, which prevents further development of effective therapies. New models are needed to better recapitulate the complex geometry of blood flow in human blood vessels and organ-specific vascular signatures. Recent advances in engineering 3D microvessels in vitro have emerged as promising technologies to not only model complex human vascular structures but also allow for precise and step-wise control of individual biological and biomechanical parameters. By designing networks with different branching structures and change of vessel diameter along the flow path, these models recapitulate pressure and flow changes occurring in vivo. Here, we describe the methodology employed to build 3D microvessels using soft lithography and injection molding techniques, as well as the protocol to fabricate capillary-size vessels through collagen photoablation. Furthermore, we describe the methodology of using these models to study malaria and narrate necessary steps for perfusion of P. falciparum through 3D microvessels and different options to quantify P. falciparum-iRBC binding.
Insights
Developing advanced 3D microvessel models allows researchers to study Plasmodium falciparum-infected red blood cell sequestration in malaria. These models better mimic human vasculature for improved therapeutic development.
Area of Science:
- Biomedical Engineering
- Parasitology
- Vascular Biology
Background:
- Sequestration of Plasmodium falciparum-infected red blood cells (iRBCs) in microvasculature is key to severe malaria.
- Current in vitro models lack the complexity of human blood vessels, hindering understanding of iRBC sequestration mechanisms.
- Novel models are required to replicate in vivo vascular geometry and organ-specific signatures.
Purpose of the Study:
- To present a methodology for engineering 3D microvessels that recapitulate human vascular complexity.
- To demonstrate the application of these 3D microvessel models for studying malaria pathogenesis.
- To enable precise control over biological and biomechanical parameters in malaria research.
Main Methods:
- Fabrication of 3D microvessels using soft lithography and injection molding.
- Collagen photoablation technique for creating capillary-size vessels.
- Perfusion of P. falciparum through 3D microvessels and quantification of iRBC binding.
Main Results:
- Engineered 3D microvessels successfully mimic complex vascular structures and flow dynamics.
- The models allow for controlled study of P. falciparum-iRBC interactions within a vascular context.
- Methodologies for perfusion and quantification of binding in 3D microvessels are established.
Conclusions:
- Engineered 3D microvessels offer a powerful platform for investigating malaria pathogenesis.
- These advanced models overcome limitations of traditional in vitro assays.
- This technology facilitates the development of targeted therapies for severe malaria.
