Controlled and tuneable drug release from electrospun fibers and a non-invasive approach for cytotoxicity testing

G Piccirillo,B Bochicchio,A Laurita,A Pepe,D A Carvajal Berrio,K Schenke-Layland,S Hinderer

doi:10.1038/s41598-019-40079-7

Abstract

Electrospinning is an attractive method to generate drug releasing systems. In this work, we encapsulated the cell death-inducing drug Diclofenac (DCF) in an electrospun poly-L-lactide (PLA) scaffold. The scaffold offers a system for a sustained and controlled delivery of the cytotoxic DCF over time making it clinically favourable by achieving a prolonged therapeutic effect. We exposed human dermal fibroblasts (HDFs) to the drug-eluting scaffold and employed multiphoton microscopy and fluorescence lifetime imaging microscopy. These methods were suitable for non-invasive and marker-independent assessment of the cytotoxic effects. Released DCF induced changes in cell morphology and glycolytic activity. Furthermore, we showed that drug release can be influenced by adding dimethyl sulfoxide as a co-solvent for electrospinning. Interestingly, without affecting the drug diffusion mechanism, the resulting PLA scaffolds showed altered fibre morphology and enhanced initial DCF burst release. The here described model could represent an interesting way to control the diffusion of encapsulated bio-active molecules and test them using a marker-independent, non-invasive approach.

Highlights

Diclofenac (2-2-(2,6-dichloroanilino)phenylacetic acid, DCF) is one of the most sold and used non-steroidal anti-inflammatory drugs prescribed to millions of people worldwide[1,2] for the treatment of osteoarthritis, rheumatoid arthritis[3,4], and muscle pain[5], as well as other applications[6]
In this work we aimed to describe DCF induced cell death in human dermal fibroblasts (HDFs) using a new effective, non-destructive in vitro model
Absorbance was constant after 24 hours and close to 100% (DCF released: 93 ± 4% leading to a DCFONa concentration of 1.5 ± 0.1 mg/mL in the extracts)

Summary

Introduction

Diclofenac (2-2-(2,6-dichloroanilino)phenylacetic acid, DCF) is one of the most sold and used non-steroidal anti-inflammatory drugs prescribed to millions of people worldwide[1,2] for the treatment of osteoarthritis, rheumatoid arthritis[3,4], and muscle pain[5], as well as other applications[6]. Studies with cultured hepatocytes from various species demonstrated that high DCF concentrations are able to induce acute cell injury[22,23,24,25,26,27,28,29]. While DCF hepato-26–29,32–35 and nephro-toxicity[17,18,21,36] has been widely investigated, not that much is known about its activity as an anti-cancer drug[6,7,8,9,10,37]. In this work we aimed to describe DCF induced cell death in human dermal fibroblasts (HDFs) using a new effective, non-destructive in vitro model. In the present study we aimed to generate a model scaffold with electrospinning that allows a controlled and tuneable diffusion of encapsulated bio-active molecules and test them using a marker-independent, non-invasive approach

Objectives

Methods

Results

Conclusion