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Concept and objectives

The primary motivation of NEUROSCAFFOLDS is to address the most challenging tissue to be repaired, i.e. the nervous system, where neurons cannot be assembled randomly, as in other tissues such as bones and liver. NEUROSCAFFOLDS is based on three requirements dictated by the specific properties of the nervous system.

When neurons or neuronal precursor stem cells are cultivated on dishes with specific nanopatterns, such as lines, grooves or pillars (see Fig.1) it is possible to guide neurite growth, but also the height where the neurons establish synaptic contact and form a neuronal network, as proved by members of the present consortium. These networks are primarily 2D but the nervous system is formed by the assembly of neurons in a genuine 3D architecture, therefore


In order to proceed to the repair of injuries in the nervous system it is necessary to control the growth and functional connectivity of 3D neuronal networks

This investigation will be carried out, initially, in vitro, where we will control the mechanical, geometrical and chemical properties of scaffolds. The present state of the art of RP is ready to include in their fabrication structures specific for driving the formation of 3D neuronal networks, based on carbon nanotubes (CNTs), specific 3D nanopatterns and the decoration with chemical guidance molecules and growth factors. Therefore

Fig.1: SEM images of neuronal filopodia which grew on nanopillars with different periods and height( from (21)): a-c: 1 µm period and respectively 50, 220 and 400 nm height; d-f 500nm period and respectively 35, 180 and 360 nm height. Scale bar 2 µm. Pillars with a height below 220 nm can guide neurite directions, while neurons cultivated over higher pillars grow over then 300-400 nm above the floor of the dish. 



3D scaffolds for neuronal network must have a hierarchical level of complexity in which: i- the macroscopic structure is determined by imaging at a milli (and micro) meter resolution; ii - the scaffold substrate is patterned and decorated with guidance molecules at a nanometer resolution.

These 3D neuronal networks will be integrated with glial cells providing a physical and a biological support. By combining expertise from Neuroscience, Nanotechnology and Tissue Engineering we will attempt the repair of lesions of the Nervous System. Therefore


  Neuroscaffolds for the repair of lesions must be initially implanted in relatively simple but meaningful parts of the peripheral nervous system (PNS) such as the sciatic nerve and


And, for the China coordinated project:

of the central nervous system (CNS) such as the spinal cord.

These scaffolds will be implanted in laboratory animals. As we envisage also implants in humans, these scaffolds will be aimed at a Marketing Authorisation Application, respecting the standards of Good Clinical and Laboratory Practice (GCP & GLP) provided by the International Conference on Harmonisation (ICH) and other industrial standards currently in force for possible use in humans.



Objectives and biological applications

In order to satisfy the 3 requirements illustrated in the previous section, NEUROSCAFFOLDS will have 2 main complementary and interconnected objectives. 


RP Fabrication of scaffolds for the growth and network formation of neurons.

Objective 1 has the following specific aims:

»       Development of RP procedures for the construction of 3D scaffolds

»       Investigation of different chemical composition of these 3D scaffolds

»       Development of 3D scaffolds with multichannel conduits

»       Investigation of biodegradable 3D scaffolds

»       Decoration of these scaffolds with nanopatterns of different chemical composition

»       Decoration of these scaffolds with guidance molecules and/or growth factors

»       Formation of 3D neuronal networks embedded in these scaffolds

»       Testing biocompatibility and long term survival of these neuronal networks



Fabrication and implant of   scaffolds for the repair of sciatic nerve and spinal cord injuries

Objective 2 has the following specific aims:

»       Implantation of these scaffolds for the repair of the sciatic nerve

»       For the China coordinated project:

Implantation of these scaffolds for the repair of the spinal cord

»       Development of suitable multichannel electrodes for recording the electrical activity in the dorsal root ganglia/spinal cord after sciatic nerve injury

»       Good Laboratory and Clinical Practice of these implants for perspective human implants


A success of these two objectives will be very relevant for basic research and for regenerative medicine with major health implications. NEUROSCAFFOLDS has several biological applications, namely:

»       Construction of in vitro 3D neuronal networks with specific properties

»       Analysis of the histochemical, electrical, mechanical properties of neurons growing in these 3D scaffolds

»       Analysis of the survival, proliferation and differentiation of neurons grown on these 3D scaffolds


In order to achieve these objectives we have established a strict coordination and cooperation of two clusters in China and in Europe. These two projects are separate and funded by two different entities.

 The cluster in China is composed of the Institute of Genetics and Developmental Biology of the Chinese Academy of Science in Bejing (Jianwu Dai and Zhifeng Xiao) able to modify and functionalize the scaffolds with appropriate molecules, the Institute of NanoTech and NanoBionics of the Chinese Academy of Science in Suzhou (Guosheng Cheng and Zhijun Zhang) experts in manufacturing and imaging 3D Neuroscaffolds and the Department of Neurosurgery, Drum Tower Hospital of the Nanjing University (Weibang Liang) expert in implants in the sciatic nerve and spinal cord.

The cluster in Europe is formed by Nanotechologists and Neuroscientists at the IIT in Genova (Fabio Benfenati, Enzo DiFabrizio and Fernando Brandi) and in Trieste – both at SISSA (Vincent Torre) and at Trieste University (Laura Ballerini) – and at the Ecole Normale Supérieure (ENS, Yong Chen) in Paris. The European team will also be formed by University of Lund (Jens Shoenbourg and Martin Kanje), who will study the repair of the sciatic nerve and the spinal cord. These two clusters have all the expertise to tackle and reach our objectives.

Relevance to the Call

The Call NMP.2013.2.2-2 Biomaterials: Imaging and rapid prototyping technology for custom made scaffolds – coordinated call with China requires the development of novel scaffolds based on SFF fabrication for tissue repair. The Call requires a balanced cooperation from the European and the Chinese centers. NEUROSCAFFOLDS  matches these requirements perfectly, as it will develop new scaffolds for neurons, neuronal networks and for the repair of spinal cord and sciatic nerve injuries within a well-balanced EU-China partnership in a highly integrated and cooperative way. NEUROSCAFFOLDS  also fulfills all expected impacts (see section 3), namely:

(i)                  More robust European - Chinese research cooperation

(ii)                More intensive exchange and training of researchers between EU and China

(iii)               Development of technologies for the production of scaffolds specific for neurons and neuronal networks

(iv)              Improved manufacturing of custom-made scaffolds

For the sciatic nerve repair in Europe.

And, in the China coordinated project

For the spinal cord repair and for the sciatic nerve repair.



The Project Approach: five interconnected activities

To reach the identified objectives we plan to carry out the following five interconnected activities

»       RP fabrication of  3D scaffolds for the Nervous system

»       Fabrication of multichannel conduits

»       Decoration of 3D Scaffolds and Multichannel conduits

»       In vitro testing

»       Implants


ProposedScientific and Technological Activities

The following sections describe in detail scientific and technological activities of our NEUROSCAFFOLDS project. A summary of the present state of art, relevant to NEUROSCAFFOLDS is presented in section 1.2.1. References can be found at the end of the project description.