In Vitro 3D Cell-Cultured Arterial Models for Studying Vascular Drug Targeting Under Flow
March 14th, 2021
2.9K views
Here, we present a new protocol to study and map the targeted deposition of drug carriers to endothelial cells in fabricated, real-sized, three-dimensional human artery models under physiological flow. The presented method may serve as a new platform for targeting drug carriers within the vascular system.
Overview
This article presents a detailed protocol for fabricating real-sized, three-dimensional (3D) human arterial models using 3D printing, lining them with human endothelial cells, and studying particle targeting under physiological flow conditions. The approach enables the investigation of drug carrier accumulation and vascular targeting in anatomically accurate, patient-specific artery models, providing a valuable platform for optimizing targeted therapies for cardiovascular diseases.
Key Study Components
Area of Science
- Cardiovascular biology
- Biomedical engineering
- Drug delivery research
Background
- 3D models of human arteries allow for realistic simulation of cardiovascular processes.
- Existing 3D models have not been widely used for vascular targeting studies.
- Understanding drug carrier behavior in physiologically relevant models is crucial for improving targeted therapies.
- Low-cost, anatomically accurate models can facilitate translational research in vascular drug delivery.
Purpose of Study
- To develop a method for fabricating 3D-printed, real-sized human arterial models lined with endothelial cells.
- To study the targeting and accumulation of particles (as drug carrier surrogates) under physiological flow conditions.
- To provide a platform for optimizing drug carriers for vascular disease treatment and diagnosis.
Methods Used
- Selection of patient-derived or standard carotid artery geometries for model design.
- 3D printing of artery molds, followed by post-processing (sanding, rinsing, lacquering).
- Casting silicone rubber into the mold and dissolving the plastic to create the vessel lumen.
- Sterilization and coating of the model with fibronectin, followed by seeding with human endothelial cells.
- Incubation on a rotator to ensure homogeneous cell distribution and subsequent fixation.
- Establishment of closed and open circuit flow systems using a peristaltic pump to simulate physiological flow.
- Introduction of fluorescent particles and real-time imaging to assess particle deposition and adhesion.
- Image analysis using custom software to quantify particle targeting.
Main Results
- Successful fabrication of 3D arterial models lined with viable endothelial cells.
- Fluorescent particles exhibited recirculation patterns, mimicking physiological flow.
- Higher particle adhesion was observed in regions of high wall shear stress, outside recirculation zones.
- The platform allows for the study of functionalized particles and different artery models to optimize targeting strategies.
Conclusions
- The described technique enables realistic modeling of vascular targeting under physiological conditions.
- This platform can be used to optimize drug carriers for targeted delivery in cardiovascular diseases.
- It provides a valuable tool for studying the interaction of drug carriers with endothelial cells in anatomically accurate human artery models.
Frequently Asked Questions
What is the main advantage of using 3D-printed arterial models in this protocol?
The main advantage is the ability to study drug carrier accumulation and targeting in anatomically accurate, physiologically relevant human artery models under controlled flow conditions.
How are the 3D artery models fabricated?
Models are designed from patient images or standard geometries, 3D printed, post-processed, cast with silicone rubber, and the plastic mold is dissolved to create the vessel lumen.
How are endothelial cells incorporated into the model?
The inner surface of the model is coated with fibronectin, then seeded with human endothelial cells, which are incubated on a rotator to ensure even distribution and attachment.
How is physiological flow simulated in the model?
A peristaltic pump and tubing system are used to circulate fluid through the model, replicating physiological flow rates and shear stresses found in human arteries.
What types of particles are used to study targeting?
Fluorescent carboxylated polystyrene particles and glass beads are used as surrogates for drug carriers to assess deposition and adhesion under flow.
How is particle targeting quantified?
Images of the model are captured during and after flow experiments, and custom software is used to analyze and count the number of particles adhering to the endothelial surface.
Can this platform be adapted for different vascular diseases?
Yes, the protocol allows for the use of different artery geometries and functionalized particles, making it suitable for studying targeted delivery in various cardiovascular disease models.
Executive Industry Relevance
Physiologically accurate 3D cell-cultured arterial models enable predictive evaluation of vascular drug targeting strategies under controlled flow, directly addressing early-stage de-risking for cardiovascular therapeutics. This platform supports mechanistic interrogation of drug carrier behavior in disease-relevant geometries, improving translational confidence and portfolio triage for vascular-targeted therapies. Integration of patient-specific flow and anatomical fidelity enhances the relevance of preclinical data for downstream development decisions.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables direct assessment of drug carrier accumulation and adhesion in human-replicated arterial environments.
- Supports mechanistic de-risking by modeling physiological flow and wall shear stress effects on targeting.
- Facilitates functional validation of vascular targets in anatomically accurate, disease-relevant systems.
- Improves predictive confidence for candidate selection and early portfolio triage.
Screening & Assay Development
- Provides a standardized, reproducible platform for quantitative measurement of particle deposition and targeting efficiency.
- Supports assay development for screening functionalized drug carriers under physiologically relevant flow conditions.
- Enables scalable evaluation of multiple formulations or targeting strategies in a controlled 3D system.
- Delivers quantitative outputs suitable for cross-comparison and downstream analytics.
Translational & Preclinical Research
- Aligns preclinical modeling with human vascular anatomy and flow, enhancing translational continuity.
- Permits adaptation to patient-specific geometries for disease-relevant targeting studies.
- Supports risk-adjusted advancement decisions by providing mechanistic insight into delivery and adhesion profiles.
- Enables exploration of targeted delivery strategies for atherosclerotic and other vascular disease models.
Pipeline & Workflow Integration
This method bridges early discovery, assay development, and preclinical validation by providing a reusable, physiologically relevant platform for vascular drug targeting studies.
- Discovery Biology: Supports hypothesis testing and pathway clarification for vascular targeting mechanisms.
- Screening: Delivers reproducible, quantitative readouts of particle adhesion and deposition under flow.
- Analytics: Enables image-based quantification and statistical comparison of targeting efficiency across conditions.
- Translational Research: Facilitates continuity from in vitro modeling to preclinical validation in disease-relevant systems.
- Enterprise Reuse: Offers a modular platform adaptable to various artery models and targeting agents.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in vascular targeting.
- Operational Value: Standardizes experimental conditions and supports reproducibility across teams.
- Strategic Value: Informs go/no-go decisions and enhances capital efficiency by de-risking early candidates.
- Portfolio Impact: Enables risk-adjusted prioritization and advancement of vascular-targeted assets.
Implementation Considerations
- Requires expertise in 3D printing, cell culture, and vascular biology.
- Needs access to imaging systems and custom image analysis software for quantitative outputs.
- Demands cross-team standardization of model preparation and flow conditions for reproducibility.
- Adaptable to different artery geometries and disease models with appropriate design modifications.
- Dependent on accurate simulation of physiological flow and wall shear stress for translational relevance.
Biopharma FAQs
Why does null hypothesis testing matter for particle adhesion quantification?
Null hypothesis testing enables objective evaluation of whether observed particle adhesion differs significantly between experimental conditions, supporting robust target validation and mechanistic de-risking in vascular targeting studies.
How does independent variable isolation fit the flow-based targeting workflow?
Isolating variables such as flow rate or particle functionalization allows teams to attribute changes in deposition or adhesion directly to specific factors, strengthening mechanistic insight and predictive confidence for candidate selection.
What do quantitative dependent variable measurements enable in this 3D model?
Quantitative measurements of particle deposition and adhesion provide actionable data for comparing targeting strategies, optimizing formulations, and informing go/no-go decisions in early discovery and screening pipelines.
Why are replication requirements critical for cross-functional collaboration in vascular targeting studies?
Replication ensures that observed targeting effects are reproducible and not artifacts of model preparation or flow conditions, enabling reliable data sharing and decision-making across discovery, screening, and translational teams.
What statistical analysis capabilities are required before implementing image-based targeting quantification?
Robust statistical analysis, including threshold setting and particle counting validation, is essential to ensure that image-derived targeting metrics are accurate, comparable, and suitable for portfolio-level decision support.
Chapters in this video
0:04
Introduction
0:44
Design and Fabrication of a 3D Bifurcation of the Human Carotid Artery Model
2:18
Human Umbilical Vein Endothelial Cell (HUVEC) Seeding
3:26
Closed-Circuit Configuration: Perfusion Experiment and Imaging
4:42
Open-Circuit Configuration: Model Washing, Data Acquisition, and Analysis
5:36
Results: Microscopic Analysis of 3D Carotid Artery Model-Seeded Endothelial Cells
6:17
Conclusion
