Abstract
Efficient separation of sub-micrometer synthetic or biological components is imperative in particle-based drug delivery systems and purification of extracellular vesicles for point-of-care diagnostics. Herein, we report a novel phenomenon in spiral inertial microfluidics, in which the particle transient innermost distance (Dinner) varies with size during Dean vortices-induced migration and can be utilized for small microparticle (MP) separation; aptly termed as high-resolution Dean flow fractionation (HiDFF). The developed technology was optimized using binary bead mixtures (1–3 μm) to achieve ~100- to 1000-fold enrichment of smaller particles. We demonstrated tunable size fractionation of polydispersed drug-loaded poly(lactic-co-glycolic acid) particles for enhanced drug release and anti-tumor effects. As a proof-of-concept for microvesicles studies, circulating extracellular vesicles/MPs were isolated directly from whole blood using HiDFF. Purified MPs exhibited well-preserved surface morphology with efficient isolation within minutes as compared with multi-step centrifugation. In a cohort of type 2 diabetes mellitus subjects, we observed strong associations of immune cell-derived MPs with cardiovascular risk factors including body mass index, carotid intima-media thickness and triglyceride levels (P<0.05). Overall, HiDFF represents a key technological progress toward high-throughput, single-step purification of engineered or cell-derived MPs with the potential for quantitative MP-based health profiling.
Highlights
Enabling technologies for continuous, size-based separation of submicrometer engineered or biological components are highly desirable in clinical applications, such as particle-based drug delivery systems[1] and the purification of extracellular vesicles in clinical diagnostics.[2]
We clinically validated the developed technology in a cohort of patients with type 2 diabetes mellitus (T2DM) and observed that immune cell-derived MPs were strongly correlated with established cardiovascular risk factors, including body mass index, carotid intimamedia thickness and triglyceride levels (Po0.05). These results clearly demonstrate the capabilities of high-resolution Dean flow fractionation (HiDFF) for highthroughput separation of small engineered or biological targets and can be further developed into a clinical tool for rapid quantitative assessment of immune and vascular health using MPs
Both immune MPs (LMPs and neutrophil-derived MPs (NMPs)) were strongly correlated to established cardiovascular risk factors, including body mass index, carotid intima-media thickness and triglyceride levels (Po0.05, Po0.01 and Po0.001, respectively) in T2DM patients (Figure 5d), suggesting a link between inflammation and lipid metabolism in diabetes patients. This link was further evident when we observed a negative association of the endothelial-derived MPs (EMPs) with the high-density lipoprotein cholesterol (Po0.01) (Supplementary Figure S8). These results demonstrated the efficacy of HiDFF technology for rapid MPs sorting and phenotyping, and further studies are warranted to study the role of leukocyte-derived MPs (LMPs) and NMPs in relation to lipid patterns and endothelial dysfunction in T2DM, as well as micro- and macrovascular disorders in general
Summary
Enabling technologies for continuous, size-based separation of submicrometer engineered or biological components are highly desirable in clinical applications, such as particle-based drug delivery systems[1] and the purification of extracellular vesicles in clinical diagnostics.[2] In microparticle fabrication, conventional ‘bottom-up’ self-assembly emulsification techniques yield a broad particle size distribution, which can affect the drug release kinetics and biotransport in blood.[3,4] well-controlled and monodisperse particles can be produced by ‘top–down’ approaches using specific lithographic techniques[5,6] and microfluidics,[7,8] microfabricated particles are prone to damage during mechanical harvesting, a problem further aggravated at the smaller/nanoscale level. Developing novel tools to achieve tunable size fractionation of polydispersed synthetic particles would enable optimal biodistribution and controlled drug release. Such technologies facilitate physical isolation of smaller biological targets (o2 μm) including platelets, microbes and extracellular vesicles in a label-free manner for unbiased downstream analysis
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