Investigation of surface topography and stiffness on adhesion and neurites extension of PC12 cells on crosslinked silica aerogel substrates.

Kyle J Lynch,Omar Skalli,Firouzeh Sabri,Nic D Leipzig

doi:10.1371/journal.pone.0185978

Abstract

Fundamental understanding and characterization of neural response to substrate topography is essential in the development of next generation biomaterials for nerve repair. Aerogels are a new class of materials with great potential as a biomaterial. In this work, we examine the extension of neurites by PC12 cells plated on matrigel-coated and collagen-coated mesoporous aerogel surfaces. We have successfully established the methodology for adhesion and growth of PC12 cells on polyurea crosslinked silica aerogels. Additionally, we have quantified neurite behaviors and compared their response on aerogel substrates with their behavior on tissue culture (TC) plastic, and polydimethylsiloxane (PDMS). We found that, on average, PC12 cells extend longer neurites on crosslinked silica aerogels than on tissue culture plastic, and, that the average number of neurites per cluster is lower on aerogels than on tissue culture plastic. Aerogels are an attractive candidate for future development of smart neural implants and the work presented here creates a platform for future work with this class of materials as a substrate for bioelectronic interfacing.

Highlights

One of the first steps towards the development of modern biomaterials to engineer neuronal scaffolds is to characterize the biophysical interactions between neuronal cell and the surface of the material
Recent studies have shown that substrates with micro- [1, 2] and nanostructured [3, 4] surfaces provide topographical cues that can positively influence cellular response in tissue culture systems
We were able to successfully culture and image PC12-C41 and PC12-ATCC cells plated on PDMS, aerogel, and tissue culture (TC) plastic Matrigel-coated substrates

Summary

Introduction

One of the first steps towards the development of modern biomaterials to engineer neuronal scaffolds is to characterize the biophysical interactions between neuronal cell and the surface of the material. Mechanical properties, such as stiffness [1,2,3,4,5,6,7,8,9,10,11,12], and topographical features of the substrate onto which cells attach influence parameters including cell survival, proliferation, adhesion, differentiation and metabolism [1,2,3,4,5]. Topography and mechanical properties of the substrate onto which cells are attached can be engineered to control and regulate specific cellular functions and activities [13]. The ability to precisely control the adhesion, proliferation, and growth rate of cells and neurons, to a substrate is an important stage of creating and utilizing novel materials for tissue engineering applications [17]. The design and successful implementation of smart electrically active implants is currently limited by the availability of biostable and biocompatible substrate materials that can tolerate all the required processing steps involved in fabricating suitable bioelectronic interfaces [17]

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Results