Enhanced chemoresistance of squamous carcinoma cells grown in 3D cryogenic electrospun scaffolds

Abstract

It is critically important to study head and neck squamous cell carcinoma tumorigenic mechanisms in order to gain a better understanding of tumor development, progression, and treatment. Unfortunately, a representative three-dimensional (3D) model for these evaluations has yet to be developed. The purpose of this study was to replicate tumor extracellular matrix (ECM) morphology utilizing electrospinning technology. First, the tumor ECM was evaluated by decellularizing tumor samples and analyzing the fibrous structure of the ECM by scanning electron microscopy. Cryogenic electrospun silk scaffolds were then fabricated to mimic the tumor ECM, and were found to be similar in fiber orientation and fiber dimensions to the native tumor ECM. Tumor cells were cultured on these ECM mimicking scaffolds and compared to an in vivo model of the same derivative human tumor in terms of proliferation and differentiation. The tumor cells in the 3D model show similar phenotypes to those found in vivo, contrasting to the same cells grown in two-dimensional (2D) culture. The sensitivity of the tumor cells to paclitaxel was compared between 2D culture and 3D culture. The results indicate that increased drug concentrations, orders of magnitude higher than the IC90 for 2D culture, had minimal effects on HN12 cell viability in the 3D model. In conclusion, an in vitro tumor model has been developed that will allow for a better understanding of tumor biology and aid chemotherapeutic drug development and accurate evaluation of drug efficacy.

The effect of electrospun polycaprolactone scaffold morphology on human kidney epithelial cells

“…The diameter of the fibre determines the pore size of the scaffold, and this determines how well cells will penetrate into a scaffold (Balguid et al 2009;Baker et al 2008;Ju et al 2010). Techniques such as cryogenic electrospinning have shown to create structures with a much greater porosity, enabling cell infiltration throughout the scaffold (Leong et al 2013;Bulysheva et al 2013;Leong et al 2009); this can be controlled by modifying the humidity of the spinning environment, changing the amount of ice crystal formation on the cooled mandrel (Leong et al 2013). It is essential in the culture of a complex 3D system for a distribution of cells throughout the scaffold, increasing the porosity within electrospun scaffolds by cryogenic electrospinning allows for this greater cell distribution.…”

“…Fibre diameter has shown to affect the gene expression of cells (Hodgkinson et al 2014;Wang et al 2014;Bean & Tuan 2015), and cryogenic scaffolds have shown to effect the behaviour of scaffolds and give a better representation on an in vivo environment for drug development (Bulysheva et al 2013). We found that small diameter fibres at 7 days had a significant reduction in KIM-1 expression compared to tissue culture plastic in both aligned and cryogenic scaffolds, possibly indicating healthier cells (Waanders et al 2010).…”

“…Other methods have been proposed to increase the porosity of scaffolds to allow for greater cell integration; dual-spinning using a water soluble sacrificial material allows for a scaffold with greater porosity (Lowery et al 2010;Phipps et al 2012) and techniques such as cell electrospinning enable cell to be directly integrated within the scaffold (Jayasinghe 2013;Townsend-Nicholson & Jayasinghe 2006). Techniques such as cryogenic electrospinning have also been proposed, this utilises the deposition of ice crystals on a cooled surface as a template for electrospun fibres, giving greater porosity (Leong et al 2013;Bulysheva et al 2013). …”

Fibronectin-matrix sandwich-like microenvironments to manipulate cell fate

“…5A, cell proliferation was similar for both 2D and sandwich microenvironments after 6 h of culture, but significantly lower within sandwich conditions after 24 h, as reported to occur in vivo. 32 Smaller but significant differences can be also observed after 24 hours depending on whether the sandwich was assembled immediately (SW t0 ) or after 3 h of culture (SW).…”

Recreating complex pathophysiologies in vitro with extracellular matrix surrogates for anticancer therapeutics screening

“…Collagen-based electrospun scaffolds were used with C4-2B cells (prostate cancer) to recapitulate tumour progression and were Page 16 of 44 A c c e p t e d M a n u s c r i p t shown to be resistant to docetaxel and camptothecin [167]. Electrospun silk fibres loaded with HN12 cells (metastatic squamous cell carcinoma) presented comparable cell proliferation, differentiation, infiltration and chemoresistance to that of in vivo HN12 mouse xenografts [168].…”