ELITISS

Project name: Electrically stimulated scaffolds for tissue engineering

Contract no. 6 / 2012

Project code: PN-II-PT-PCCA-2011-3.1-1187

PN II Program Name: Partnerships

Subprogram: PCCA1

Contracting authority: Executive Agency for Higher Education, Research, Development and Innovation Funding

Project director: Maria DINESCU

Contract value: 2,000,000 lei

Consortium:
CO - INFLPR
P1 - IFIN-HH
P2 - ICMPP
P3 - UPB

Project summary:

The project relies on an innovative, sound and unique combination of clean, environmental friendly and spatiotemporally accurate laser techniques for producing biofunctionalized conducting tridimensional scaffolds for tissue engineering. The project addresses the synergistic combination of major pillars of science and technology (Micro-Nano-Bio-Info), in the framework of Convergent Technologies for Improving Human Performance: (a) nano-micro science and technology-by using laser processing techniques for scaffolds fabrication; (b) biotechnology and biomedicine-by scaffolds biofunctionalization with biologically active agents and by analyzing the cells cultured within the scaffolds. (c) information technology-by developing advanced computing and application codes for tridimensional scaffolds fabrication and for monitoring the cells cultured within them. The laser techniques will be integrated in a three-step protocol, comprising scaffolds fabrication, biofunctionalization and testing. Rigid (titanium) and flexible organic-inorganic hybrid polymeric scaffolds will be obtained. Each of them will be biofunctionalized with polypyrrole-based conducting polymers combined with biologically active agents e.g. growth factors, drugs. Electrical stimulation of the scaffolds will trigger a spatiotemporally localized control of cell growth and the delivery of biologically active agents. The process occurs outside the body i.e. non invasive, ex vivo tissue engineering. This will lead to a new generation of biofunctionalized conducting tridimensional scaffolds with potential for bone/cartilage replacements, skin grafts and nerve regeneration. In all, the integrated platform proposed by the project will emerge as a new tissue engineering approach for modulating cell growth, proliferation and organization into functional tissues. The proposed methods based on clean, environmental friendly laser techniques will improve the therapeutic effects of tissue engineering technologies.

Project objectives:

The general aim of the project is the development of cutting-edge technologies based on clean, environmental friendly, accurate, easy-to-handle laser techniques for the fabrication, biofunctionalization and testing of PPy-based conducting scaffolds for tissue engineering.

The objectives of the project are in agreement with the specific objectives of the program: ▲ To speed up the development in the research field of laser-based techniques for material processing and imaging existing in Romania. ▲ To align this field at the modern European standards, in order to participate to European Research Area (ERA) and to increase the chances of success in European projects competitions. ▲To develop and consolidate the Romanian expertise in this field. ▲To improve the communications between research institutes and universities which led to effects on human resources and on knowledge transfer. ▲To develop interregional connections by including partners from other parts of the country. The scientific objective is to achieve new knowledge in scientific areas under the framework of Convergent Technologies for Improving Human Performance, emerging from synergistic combination of major provinces of science and technology (Micro-Nano-Bio-Info): (a) nano-micro science and technology- laser processing at micro- and nanoscale of various classes of materials (metals, polymers) for fabricating scaffolds for tissue engineering; (b) biotechnology and biomedicine- temporally and spatially localized control of cell behavior via electrical stimulation of the scaffolds, procedures to analyze cellular processes; (c) information technology- advanced computing, elaboration of application codes in MATLAB, for non-invasive, marker free imaging of cells cultured within the scaffolds (by digital holographic microscopy). Technological objectives: · Setting high technology experimental setups; · Developping laser processing and imaging techniques; · Producing a new generation of 3D conducting scaffolds (rigid and flexible), with applications for the engineering of hard and soft tissues · Testing the scaffolds in cell cultures. The following operational objectives will be addressed: I. Producing 3D conducting scaffolds from various substrate materials (titanium, organic-inorganic hybrid polymers) with well-defined, porous architectures coated with PPy-based conducting polymers suitable for engineering hard (i.e. bone, cartilage) and soft (i.e. epidermal, neuronal) tissues. II. Biofunctionalization of the scaffolds with coatings of PPy-based polymers combined with biologically active agents (growth factors, proteins, components of extracellular matrix, antibiotics, anti-inflammatory drugs), in the form of blends or multilayers. The biofunctionalized coatings will promote positive interactions with the cells cultured within the scaffolds as well as with the surrounding tissues (following implantation). III. Controlling the behavior of the cells cultured within the scaffolds by electrical stimulation of the biofunctionalized scaffolds. Electrical stimuli will be used for: i) promoting the organization of cells into functional tissues (via electrically controlled release of growth factors, proteins and components of extracellular matrix); ii) enhancing scaffolds biointegration by fighting bacterial infections and inflammation (via electrically controlled release of antibiotics and anti-inflammatory drugs).

Publications (in ISI ranked journals):

1. MAPLE deposition of 3D micropatterned polymeric substrates for cell cultures
Irina Alexandra Paun, Mona Mihailescu, BogdanCalenic, Catalin Romeo Luculescu, Maria Greabu and Maria Dinescu, Applied Surface Science, 278, 166–172 (2013)

2. Laser micro-patterning of biodegradable polymer blends for tissue engineering
Irina Alexandra Paun, Marian Zamfirescu, Mona Mihailescu, Catalin Romeo Luculescu, Cosmin Catalin Mustaciosu, Ion Dorobantu, Bogdan Calenic, Maria Dinescu, Journal of Material Science, Journal of Materials Science 2015 50 (2) 923-936 (accepted for publication on October 21, 2014)

3. Investigation of osteoblast cells behavior in polymeric 3D micropatterned scaffolds using digital holographic microscopy
Mihailescu M, Popescu RC, Matei A, Acasandrei A, Paun IA, Dinescu M, Appl Opt. 2014 Aug 1;53(22):4850-8

4. I.A. Paun, A.M. Acasandrei, C.R. Luculescu, CC. Mustaciosu, V. Ion, M. MIhailescu, E. Vasile, M. Dinescu
MAPLE deposition of polypyrrole-based composite layers for bone regeneration, Appl. Surf. Sci. 357A 975-984 (2015)

5. I.A. Paun, F. Stokker-Cheregi, C.R. Luculescu, A.M. Acasandrei, V. Ion, M. Zamfirescu, C.C. Mustaciosu, M. Mihailescu, M. Dinescu
Electrically stimulated osteogenesis on Ti-PPy/PLGA constructs prepared by laser-assisted processes, Mater Sci Eng C Mater Biol Appl. 2015 ;55:61-9. doi: 10.1016/j.msec.2015.05.059 (2015)

6. M. Mihailescu, I.A. Paun, M. Zamfirescu, C. R. Luculescu, A. M. Acasandrei, M. Dinescu
Laser-assisted fabrication and noninvasive imaging of 3D cell-seeding constructs for bone tissue engineering - to be published in Journal of Materials Science.

Proceedings papers:

1. MG63 cells behavior on rough polypyrrole scaffolds investigated by digital holographic microscopy
M. Mihailescu ; A. Matei ; A. Acasandrei ; R. C. Popescu ; I. A. Paun ; Maria Dinescu, SPIE 9204, Interferometry XVII: Advanced Applications, 92040N (18 August 2014)

Conference presentations:
1. COLA Conference, 6-11 October 2013, Ischia, Italia (poster)
Electrically stimulated scaffolds for tissue engineering applications
I. A.Paun, A Matei, M.D. Dumitru, E.C. Buruiana, M.A. Acasandrei, M. Mihailescu, M. Dinescu

2. International Vacuum Congress IVC-19/ICSS-15 and ICN+T, 9-13 September 2013, Paris, France (oral presentation)
Functional polymeric coatings for tissue engineering applications
Irina Alexandra Paun, Andreea Matei, Adriana Acasandrei, Flavian Stokker-Cheregi, Marian Zamfirescu, Antoniu Moldovan and Maria Dinescu

3. The 2nd Biophotonics Conference, 17-19 July, Taipei, Taiwan (poster)
Cells features on polymeric blends substrate analyzed using digital holographic microscopy
M. Mihailescu, R.C. Popescu, I.A. Paun, M.A. Acasandrei, A. Matei

4. The 10th Conference on Lasers and Electro-Optics Pacific Rim(CLEO-PR 2013), 30 June-4 July 2013 Kyoto, Japan (poster)
Deep Walls Microscaffold Characterization Using Digital Holographic Microscopy
M. Mihailescu, I. A. Paun, R. C. Popescu, A. Matei, A. Acasandrei, M. Dinescu, E. I. Scarlat

5. E-MRS Conference, Symposium R, 26-31 May 2013, Strasbourg, France (poster)
Laser based techniques for electrically stimulated scaffolds preparation
A Matei, M.D. Dumitru, I. A.Paun, E.C. Buruiana, M.A. Acasandrei, M. Mihailescu, M. Dinescu

6. Frontiers in Polymer Science 2013, 21-23 Mai 2013, Sitges- Barcelona, Spania
Photobehavior of some urethane methacrylates of Ormosil type used in formation of hybrid composites by UV/laser irradiation
E. C. Buruiana, V. Melinte, F. Jitaru, A. Matei, M. Dinescu, T. Buruiana

7. Advanced Topics in Optoelectronics, Microelectronics and Nanotechnologies, Constanta, Romania, 21-24 August 2014
Focusing criterion in digital holographic microscopy image reconstruction
M. Mihailescu, N. Mihale, R. C. Popescu, A. Acasandrei, I. A. Paun, M. Dinescu, E. Scarlat

8. Conferinta Facultatii de Chimie, Universitatea Al. I. Cuza din Iasi, 31 oct.-1 nov., Iasi
Hybrid composites obtained by photopolymerization of poly(propylene glycol) dimethacrylate in monomer mixtures
V. Podasca, T. Buruiana, E.C. Buruiana

9. 12th IUVSTA School on Lasers in Materials Science, 13-20.07.2014, Venice, Italy
Functional oxide thin films obtained by Pulsed Laser Deposition
M. Dinescu

10. High Power Laser Ablation, 18-28.04.2014, Santa Fe, USA
MAPLE for Functional Coatings with Applications in Tissue Engineering
F. Stokker, M. Dinescu, I. Paun, A. Matei

11. E-MRS 2014 SPRING MEETING,27-29.05.2014, Lille, France
Laser processing of soft materials
M. Dinescu, I. Paun, V. Dinca, A. Palla-Papavlu

12. I.A Paun, M. Zamfirescu, C.R. Luculescu, M. Mihailescu, A. M. Acasandrei, C.C. Mustaciosu and M. Dinescu
Laser-assisted nano/micro-fabrication of electrically responsive, biomimetic scaffolds for bone tissue regeneration
POSTER presented at EMRS 2015

13. M. Mihailescu, I.A. Paun , M. Zamfirescu, C. R. Luculescu, A. Acasandrei , M. Dinescu
Nanoscale laser-based fabrication and holographic imaging of 3D engineered constructs for bone regeneration
POSTER presented at EMRS 2015

14. M. Mihailescu, E I. Scarlat, I.A. Paun, I. Grigorescu, A. Acasandrei
Fractal Properties of Osteoblast-Like Cells From Holographic Phase Images
S5 P19, 15 th Int. Balkan Workshop on Applied Physics IBWAP, Constanta 2-4 July 2015

First stage - 31.12.2012 (click for the scientific abstract)

The objective of this stage is to produce and to test electrically conducting scaffolds consisting of 3D titanium, respectively of organic-inorganic hybrid polymeric constructs with well-defined, porous architectures coated with PPy-based conducting polymers.

Value of first stage: 789,000 lei

Second stage - 31.12.2013

The objective of this stage is to control the behavior of cells cultured within the scaffolds (adhesion, growth, proliferation, organization into tissues) by electrical stimulation.

Value of second stage: 692,750 lei

Third stage - 31.12.2014

The objectives of this stage are:

to produce PPy-based conducting scaffolds with biofunctionalized surfaces
to use electrical stimuli for : a) accelerating the organization of cells in functional tissues (via electrically controlled release of growth factors, proteins and components of extracellular matrix); b) enhancing scaffolds biointegration i.e. fighting bacterial infections and inflammation via electrically controlled release of antibiotics and anti-inflammatory drugs.

Value of third stage: 518,250 lei

Fourth stage - 31.12.2015 (click for the scientific abstract)

Value of fourth stage: 150,401 lei

Fifth stage - 31.12.2016 (click for the scientific abstract)

Value of fifth stage: 282,174 lei