Controlling the Spatiotemporal Release of Nerve Growth Factor by Chitosan/Polycaprolactone Conduits for Use in Peripheral Nerve Regeneration

Katarzyna Nawrotek,Radosław Wach,Justyna Gatkowska,Marek Wieczorek,Karolina Rudnicka,Sylwia Michlewska,Monika Kubicka,Adrian Bekier

doi:10.3390/ijms23052852

Abstract

Tubular polymeric structures have been recognized in the treatment of peripheral nerves as comparable to autologous grafting. The best therapeutic outcomes are obtained with conduits releasing therapeutic molecules. In this study, a new approach for the incorporation of biologically active agent-loaded microspheres into the structure of chitosan/polycaprolactone conduits was developed. The support of a polycaprolactone helix formed by 3D melt extrusion was coated with dopamine in order to adsorb nerve growth factor-loaded microspheres. The complex analysis of the influence of process factors on the coverage efficiency of polycaprolactone helix by nerve grow factor-loaded microspheres was analyzed. Thus, the PCL helix characterized with the highest adsorption of microspheres was subjected to nerve growth factor release studies, and finally incorporated into chitosan hydrogel deposit through the process of electrophoretic deposition. It was demonstrated by chemical and physical tests that the chitosan/polycaprolactone conduit meets the requirements imposed on peripheral nerve implants, particularly mimicking mechanical properties of surrounding soft tissue. Moreover, the conduit may support regrowing nerves for a prolonged period, as its structure and integrity persist upon incubation in lysozyme-contained PBS solution up to 28 days at body temperature. In vitro cytocompatibility toward mHippoE-18 embryonic hippocampal cells of the chitosan/polycaprolactone conduit was proven. Most importantly, the developed conduits stimulate axonal growth and support monocyte activation, the latter is advantageous especially at early stages of nerve regeneration. It was demonstrated that, through the described approach for controlling spatiotemporal release of nerve growth factors, these biocompatible structures adjusted to the specific peripheral nerve injury case can be manufactured.

Highlights

A peripheral nerve injury (PNI) relates to damage, crushing, or transection of the peripheral nerve
The chitosan hydrogel tube produced by electrodeposition is perfectly suited for application as a nerve guidance conduit due to high water content and softness, mimicking the surrounding soft tissue
The obtained results indicate that the developed strategy enables a spatiotemporal controlled release of nerve growth factors as the helix dimensions and geometry can be customized

Summary

Introduction

A peripheral nerve injury (PNI) relates to damage, crushing, or transection of the peripheral nerve. PNIs can result from degenerative changes or inflammatory diseases [3,4]. Peripheral nerves are key connections among the brain, spinal cord, and body. This highly complex network provides control over sensation, movement, and motor coordination. Treatment of damaged peripheral nerve tissue may be pharmacologic, by transplantation (i.e., autografting, allografting, or xenografting) or by the implantation of nerve guidance conduits (NGCs) to bridge the nerve stumps, providing room and environment for nerve regeneration [5–8]. Conventional pharmacologic therapy is performed in the early stages of damage caused by inflammation. Innovative solutions allowing for re-generation of damaged tissues with the use of NGCs are being sought

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Results

Conclusion