Hyaluronic acid-based hydrogels as 3D matrices for in vitro evaluation of chemotherapeutic drugs using poorly adherent prostate cancer cells

Abstract

The current investigation aimed to develop a biomimetic, three-dimensional (3D) culture system for poorly adherent bone metastatic prostate cancer cells (C4-2B) for use as an in vitro platform for anti-cancer drug screening. To this end, hyaluronic acid (HA) derivatives carrying complementary aldehyde (HAALD) and hydrazide (HAADH) groups were synthesized and characterized. In situ encapsulation of C4-2B cells was achieved by simple mixing of HAALD and HAADH in the presence of the cells. Unlike two-dimensional (2D) monolayer culture in which cells adopt an atypical spread morphology, cells residing in the HA matrix formed distinct clustered structures which grew and merged, reminiscent of real tumors. Anti-cancer drugs added to the media surrounding the cell/gel construct diffused into the gel and killed the embedded cells. The HA hydrogel system was used successfully to test the efficacy of anti-cancer drugs including camptothecin, docetaxel, and rapamycin, alone and in combination, including specificity, dose and time responses. Responses of cells to anti-neoplastics differed between the 3D HA hydrogel and 2D monolayer systems. We suggest that the data obtained from 3D HA systems is superior to that from conventional 2D monolayers as the 3D system better reflects the bone metastatic microenvironment of the cancer cells.

In Vitro Modeling of Tissue-Specific 3D Microenvironments and Possibile Application to Pediatric Cancer Research

“…Poly(lactidecoglycolide), poly(lactic acid), poly(vinyl alcohol), poly( -caprolactone) or polyethylene glycol) (e.g. Poly(lactide-coglycolide), poly(lactic acid), poly(vinyl alcohol), poly( -caprolactone) or polyethylene glycol) [77,78,99,[115][116][117][118][119][120][121]. One advantage of scaffold-based strategies is that it is possible to better manage matrix stiffness/compliance, porosity, and biochemical composition.…”

Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

“…17 Most studies have shown that the cell viability in 3D cultures during the first few days (1-5 days) was not significantly different when compared to that in 2D culture. 30,34 In some cases, 3D cultures showed slightly reduced cell viability 29 when the culture time prolonged, and this was related to the structure of spheroids, which may result in the lack of oxygen and nutrients, and the accumulation of waste at the core of the spheroid as they grow larger. 25 The structures of 3D spheroids.…”

Spheroids Formation on Non‐Adhesive Surfaces by Liquid Overlay Technique: Considerations and Practical Approaches

“…Also, some therapeutics can become adsorbed to the scaffolds, which may lead to a reduction of the amount of therapeutics that reach cancer cells and, consequently, disturb the therapeutic outcome. [35] Furthermore, some of the animal-sourced natural ECM extracts used for the production of matrices may contain residual growth factors and other undefined constituents that may affect spheroids formation, as well as cancer cells responsiveness to drugs. [31] Lastly, the analysis of spheroids that grew on artificial matrices can be difficult, as the imaging of the spheroids by optical and fluorescence techniques depends on the scaffold size and transparency.…”