Hyaluronic acid-functionalized electrospun PLGA nanofibers embedded in a microfluidic chip for cancer cell capture and culture

Abstract

Circulating tumor cells (CTCs) are important markers of metastatic cancer. The isolation and detection of CTCs from peripheral blood provides valuable information for cancer diagnosis and precision medicine. However, cost-efficient targeted separation of CTCs of different origins with clinically significant specificity and efficiency remains a major challenge. In this study, a facile approach was developed to fabricate a thin sheet of hyaluronic acid (HA)-functionalized PLGA nanofibrous membrane and integrate it into a microfluidic chamber. The HA was covalently conjugated onto polyethyleneimine (PEI)-modified electrospun poly(lactic-co-glycolic acid) (PLGA) nanofibers. Different techniques were employed to characterize the resulted nanofibers. The results show that the CD44+ carcinoma of various origins (HeLa, KB, A549, and MCF-7 cells) could be selectively captured by the PLGA-PEI-HA nanofibers in the microfluidic platform. Importantly, the PLGA-PEI-HA nanofibrous membrane was more efficient to capture HeLa cancer cells under flowing conditions than in static dishes, and at a really low density (20 cells per mL). Furthermore, with constant media perfusion, the captured HeLa cells could grow on the nanofibrous membrane in the microchip for days without compromised cell viability. This is the first trial of using HA-functionalized electrospun nanofibers in a lab-chip device for cancer cell capture and culture. Compared to conventional CTC capture methods, the integration of inexpensive functional electrospun nanofibers and microfluidic technologies may expand the frontiers of using advanced nanomaterials in portable diagnostic applications.

Hyaluronan, Cancer-Associated Fibroblasts and the Tumor Microenvironment in Malignant Progression

“…The role of HA and its receptors in circulating CAFs and tumor cells is currently understudied. However, several studies have reported that circulating tumor cells from cancer patients express the HA receptor CD44 (Grillet et al, 2017 ) and can be captured from circulation by adhering to HA, a process that is mediated by HA receptors (Xu et al, 2017 ). Interestingly, circulating cells with this dual phenotype are EpCAM- and are therefore distinct form the more commonly studied EpCAM+ circulating tumor cells (Mirza et al, 2017 ).…”

Smart Carriers and Nanohealers: A Nanomedical Insight on Natural Polymers

“…HA nanofibers provide the required air permeation at the wound site, better than native solid HA, which aids in the cellular migration and proliferation to promote the growth of tissue and faster wound healing [ 158 ]. With regard to the significance of HA in wound healing, the material was explored in combination with PLGA, PCL, PEO, CS [ 159 , 160 , 161 ]. HA nanofibers have also been explored for anti-infective properties.…”

Nanomaterial-based Microfluidic Chips for the Capture and Detection of Circulating Tumor Cells

“…The electrospun nanofibers are easy to fabricate, have a large surface-to-volume ratio and high porosity. Recently, electrospun nanofibers have been used as substrates to capture CTCs 90 - 94 . For example, Shi's group used hyaluronic acid-functionalized electrospun PLGA nanofibers embedded in a microfluidic chip for the capture and detection of CTCs 94 .…”

“…Recently, electrospun nanofibers have been used as substrates to capture CTCs 90 - 94 . For example, Shi's group used hyaluronic acid-functionalized electrospun PLGA nanofibers embedded in a microfluidic chip for the capture and detection of CTCs 94 . The hyaluronic acid was covalently conjugated onto polyethyleneimine-modified electrospun PLGA nanofibers.…”