The Stratakis Lab

The Stratakis Lab

Ultrafast Laser Micro and Nano Processing Laboratory

Direct Laser biomemetic scafolds and 3D Scaffolds Hosting Neural Stem Cells

3D Scaffolds Hosting Neural Stem Cells

Short Description

The aim is to develop laser-engineered micro/nano scaffolds (3DLS) for hosting 3D cultures of neural stem cells.

Research Topic

RT4. 3D Scaffolds Hosting Neural Stem Cells

Abstract

Neural stem cells (NSCs) are intrinsically capable of differentiating into different neural cell types: neurons, oligodendrocytes and astrocytes, and have emerged as important players in the generation and maintenance of neural tissue as well as in treating neurodegenerative diseases and neurological injuries. However, successful development of NSC-based therapies requires more sophisticated technologies from the ones that are already available and deeper understanding of NSCs’ functions. The NSCs reside in a complex three-dimensional (3D) niche in vivo where they are exposed to a plethora of signals, including physical signals such as tensile, compressive and shear stresses, discontinuities and differences in roughness of the ECM molecules. Topography is capable of inducing different effects on NSCs, such as changes in cell morphology, alignment, adhesion, migration, proliferation, cytoskeleton organization and also differentiation. However, simulating this 3D environment for NSC culture and subsequent development of 3D neuronal networks that maintain functional neuronal properties (synaptogenesis and neurotrophic performance) remains a challenge.

To respond to this challenge we have fabricated 3D laser-engineered micro/nano scaffolds (3DLS) featuring different micro/nano topographies for hosting neurons, glia and NSCs. These are advantageous platforms to study the biology of NSC proliferation, differentiation, neuritogenesis and synaptogenesis. Moreover, we have successfully achieved to replicate these micro/nano topographies on polymeric systems such as poly (lactic-co-glycolic acid) (PLGA) and polydimethylsiloxane (PDMS) in order to investigate the effect of material stiffness on the biology of NSCs. Furthermore, an ongoing study is investigating the effect of functionalized gold nanoparticles on NSCs proliferation and differentiation via their immobilization on these micro/nano patterned scaffolds.

 

 

Publications

  • Laser fabricated discontinuous anisotropic microconical substrates as a new model scaffold to control the directionality of neuronal network outgrowth

Simitzi, C Efstathopoulos P et al., Biomaterials, 67:115-128, 2015

  • Microconical silicon structures influence NGF-induced PC12 cell morphology
  1. Simitzi et.al., J Tissue Eng Regen Med,. PubMed PMID: 24497489, 2014

 

 

Project Members

Dr Kanelina Karali

Dr Paraskevi Kavatzikidou

Dr. Chara Simitzi

Ms. Despina Angelaki

Ms. Eleftheria Babaliari

Ms. Christina Lanara

Dr. Anthi Ranella

Dr. Emmanuel Stratakis

 

 

RT3. Direct Laser Fabrication of Biomimetic Scaffolds

Short Description

The aim is to investigate the cytocompatibility of different cell lines on laser engineered biomimetic 3D scaffolds (e.g. metallic and polymeric) of different micro/nano topographies and/or surface energies.

Research Topic

RT3. Direct Laser Fabrication of Biomimetic Scaffolds

 

Abstract

Micro- and nano- fabrication techniques provide the opportunity to develop novel cell culture platforms, where the effect of various topographical cues on cellular proliferation, orientation, adhesion and differentiation is studied. Biomaterial architecture can drive cellular response via physical and chemical extracellular signals (topographical and chemical cues at micro/nanoscale), mechanical properties of the substrate and adhesion ligands. Such tuning multiple cell instructive cues via micro/nano structuring is significant in Tissue Engineering.

 

Our research focuses on the hierarchical micro/nano texturing on silicon (Si) via ultrashort-pulsed laser irradiation. Ultrashort-pulsed laser-based fabrication enables excellent control over micro- and submicron scales. It is a scalable rapid prototyping technique with its main advantage the control in structure geometry and pattern regularity. Varying laser and irradiation parameters (such as energy fluence, irradiation environment, etc.), changes drastically the Si surface morphology (i.e. geometry, dimensions and density of the structures). Microconical and nanorippled Si morphologies are fabricated and further used as cell culture platforms. Previous studies have shown that controlling surface roughness and wettability of the patterned Si substrates, fibroblast cell adhesion could be tuned. It was also shown that the microconical Si substrates could influence NGF-induced PC12 cell differentiation, sympathetic and sensory neuronal alignment and cortical neural network formation. Furthermore, the functionality of the immune response to Si scaffolds has been studied. The ongoing study of microconical Si morphologies have shown to influence Schwann and Neuro-2a (N2a) cell adhesion and proliferation, while shifting from micro to nano-roughness influences the Schwann and N2a cells outgrowth response.

 

Furthermore ultrafast laser direct writing is used to fabricate polymeric substrates such as PETG at a range of fluences resulting in different roughnesses and geometrical characteristics. Positive replicas on various polymers are successfully reproduced via soft lithography from the laser engineered substrates. Furthermore, dynamic cultures are performed for the study of the cytoskeleton, directionality and proliferation of cells on micro-nano patterns. Finally, a comparison between static and dynamic cultures is performed demonstrating the effect of pattern (geometry and topography) on cell directionality and proliferation.

 

 

SEM images from Si and polymeric laser engineered substrates with Schwann cells

 

Collaborators

  1. Gravanis, I. Charalampopoulos, University Of Crete, Medical School
  2. Athanassakis, University Of Crete, Dept. of Biology
  3. Samara, E. Anastasiadou, S. Pagkakis, Biomedical Research Foundation of the Academy of Athens
  4. G. Kanaras, Physics & Astronomy, University of Southampton.
  5. Hosokawa, Biointerface Research Group, Health Research Institute (HRI) National Institute of Advanced Industrial Science and Technology (AIST)

 

Publications

  • Laser fabricated discontinuous anisotropic microconical substrates as a new model scaffold to control the directionality of neuronal network outgrowth

Simitzi, C Efstathopoulos P et al., Biomaterials, 67:115-128, 2015

  • Microconical silicon structures influence NGF-induced PC12 cell morphology
  1. Simitzi et.al., J Tissue Eng Regen Med,. PubMed PMID: 24497489, 2014
  • Biomimetic micro/nanostructured functional surfaces for microfluidic and tissue engineering applications
    Stratakis, A. Ranella, and C. Fotakis, Biomicrofluidics, 5, 013411, 2011
  • Direct laser writing of 3D scaffolds for neural tissue engineering applications
    Melissinaki, A. A. Gill, I. Ortega, M. Vamvakaki, A. Ranella, J. W. Haycock, C. Fotakis, M. Farsari, and F. Claeyssens, Biofabrication, Vol. 3, 045005, 2011
  • Controlling cell adhesion via replication of laser micro/nano-textured surfaces on polymers
    Koufaki, A. Ranella, K. E Aifantis, M. Barberoglou, S. Psycharakis, C. Fotakis, E. Stratakis , Biofabrication, 3, 045004, 2011
  • Silicon Scaffolds Promoting 3D Neuronal Web of Cytoplasmic Processes
    L. Papadopoulou, A. Samara, M. Barberoglou, A. Manousaki, S.N. Pagakis, E. Anastasiadou, C. Fotakis and E. Stratakis, Tissue Engineering Part C, 16, 497, 2010
  • Tuning cell adhesion by controlling the roughness and wettability of 3D micro/nano silicon structures
    Ranella, M. Barberoglou, S. Bakogianni, C. Fotakis, E. Stratakis, Acta Biomaterialia, 6, 2711, 2010
  • Laser-based micro/nanoengineering for biological applications
    Stratakis, A. Ranella, M. Farsari, and C. Fotakis, Progress in Quantum Electronics, 33, 127, 2009
  • Applications of ultrafast lasers in materials processing: Fabrication on self-cleaning surfaces and scaffolds for tissue engineering
    Fotakis, M. Barberoglou, V. Zorba, E. Stratakis, E. L. Papadopoulou, A. Ranella, K. Terzaki, and M. Farsari, Proceedings of SPIE, Laser Physics and Applications 2008, 2008
  • Novel Aspects of Materials Processing by Ultrafast Lasers: From Electronic to Biological and Cultural Heritage Applications
    Fotakis, V. Zorba, E. Stratakis, P. Tzanetakis, I. Zergioti, D. G. Papagoglou, K. Sambani, G. Filippidis, M. Farsari, P. Pouli, G. Bounos, S. Georgiou, Journal of Physics: Conference Series, 59, 266, 2007

 

Project Members
Dr. Emmanuel Stratakis
Dr. Chara Simitzi
Ms. Xristina Yiannakoy
Mr. Miron Krassas
Dr. Anthi Ranella
Prof. Costas Fotakis

TO BE ADDED

Dr. Paraskevi Kavatzikidou

Dr. Kanelina Karali

Ms. Despina Angelaki

Ms. Eleftheria Babaliari

Ms. Syrago Spanou

Ms. Christina Lanara

Back