A Microfluidic Chip Integrated with Hyaluronic Acid-Functionalized Electrospun Chitosan Nanofibers for Specific Capture and Nondestructive Release of CD44-Overexpressing Circulating Tumor Cells

Abstract

Detection of circulating tumor cells (CTCs) in peripheral blood is of paramount significance for early-stage cancer diagnosis, estimation of cancer development, and individualized cancer therapy. Herein, we report the development of hyaluronic acid (HA)-functionalized electrospun chitosan nanofiber (CNF)-integrated microfludic platform for highly specific capture and nondestructive release of CTCs. First, electrospun CNFs were formed and modified with zwitterion of carboxyl betaine acrylamide (CBAA) via Michael addition reaction and then targeting ligand HA through a disulfide bond. We show that the formed nanofibers still maintain the smooth fibrous morphology after sequential surface modifications, have a good hemocompatibility, and exhibit an excellent antifouling property due to the CBAA modification. After being embedded within a microfluidic chip, the fibrous mat can capture cancer cells (A549, a human lung cancer cell line) with an efficiency of 91% at a flow rate of 1.0 mL/h. Additionally, intact release of cancer cells is able to be achieved after treatment with glutathione for 40 min to have a release efficiency of 90%. Clinical applications show that 9 of 10 nonsmall-cell lung cancer patients and 5 of 5 breast cancer patients are diagnosed to have CTCs (1 to 18 CTCs per mL of blood). Our results suggest that the developed microfluidic system integrated with functionalized CNF mats may be employed for effective CTCs capture for clinical diagnosis of cancer.

Nanostructured Substrates for Detection and Characterization of Circulating Rare Cells: From Materials Research to Clinical Applications

“…A wide spectrum of polymer materials—including PLGA, [129,162–164] chitosan, [136,165,166] PS, [167] and cellulose acetate, [168] polyvinyl alcohol/polyethyleneimine (PVA/PEI), [169–171] nylon-6/poly(sulfobetaine methacrylate)/poly(acrylic acid) (nylon-6/PSBMA/PAA), [172] polystyrene/poly(styrene- co -maleic anhydride), [173] poly(ethylene oxide)/poly(3,4-ethylenedioxyt hiophene):polystyrene sulfonate (PEO/PEDOT:PSS), [174] and poly( N -isopropylacrylamide)/poly(benzophenone) (PNIPAAm/PBP) [175] )—were used in the preparation of polymer nanofiber-embedded substrates for conducting CTC/CFNC capture. Among different polymer nanofiber-embedded substrates, PLGA nanofibers (diameter = 130 nm) were first deposited onto glass substrates for capturing CTCs.…”

“…GSH (50 × 10 −3 m )-mediated disulfide cleavage has been utilized to release CTCs that were captured on anti-EpCAM-modified SS-biotin-Ppy nanowire-embedded substrates, [152] anti-EpCAM-conjugated flowerlike substrates, [207] and HA-functionalized chitosan nanofiber-embedded substrates. [166] The efficiencies were all above 85%, and the viabilities ranged from 80% to 98%. However, the incubation time of 40–60 min for GSH treatment is relatively long.…”

Electrospun Biomaterials in the Treatment and Prevention of Scars in Skin Wound Healing

“…Encouragingly, electrospun nanofibers can be manufactured on an industrial scale, with the production of continuous nanofibers from a variety of polymers already proven (Ramachandran and Gouma, 2008;Zhang et al, 2012;Ma et al, 2015;Wang et al, 2018). Translating nanofiber production from laboratory to commercial scale is readily accommodated through the application of multi-jet nozzle electrospinners, which have been reported to process as much as 6.5 kg/h of polymer to produce fibers (Persano et al, 2013).…”

Beyond Capture: Circulating Tumor Cell Release and Single‐Cell Analysis

“…Redox‐sensitive disulfide bond can be rapidly ruptured with reductants, such as GSH, dithiothreitol, tris(2‐carboxyethyl)phosphine, etc. Disulfide bond‐containing linker can be used to bridge recognition ligands and capture matrix, capacitating to reductant‐triggered cleavage for CTC release . For example, Dou et al developed bioinspired hierarchically structured surfaces modified with anti‐EpCAM Ab via a disulfide bond‐containing linker (Figure c) .…”