Regenerative medicine is a branch of translational research in tissue engineering and molecular biology which deals with the process of replacing, engineering or regenerating cells, tissues or organs to restore or establish normal function. At NANOTEC we aim to regenerate damaged tissues by combining cells from the body with highly porous biodegradable scaffolds, which act as templates for tissue regeneration. We also use powerful state of the art techniques to study the bone-tissue regeneration mechanism
Smart Materials for Regenerative Medicine
Current regenerative medicine strategies are mainly focused on the design of improved biomaterials that are non-immunogenic, biocompatible, and biodegradable, and can be functionalized with bioactive proteins and chemicals. Our research is based on alternative approach to produce biomaterials that mimic the extracellular matrix (ECM). For instance artificial biocompatible and biodegradable fibrous extracellular matrices are realized using a variety of natural and synthetic polymers in order to increase cell proliferation and cell adhesion. Another approach involves freeze-dried 3D porous collagen–chitosan scaffolds which are functionalized with protease-responsive capsules loaded with growth factors for guiding and controlling the release of selected bioactive agents that work in concert with the implants.
In association with Maastricht University (Prof. L. Moroni; http://www.moronilab.org/), several biofabrication technologies are also being developed to create smart constructs and direct cell fate. The core activities of the group evolve around acquiring and implementing knowledge for biofabrication technologies based on the following research objectives: (i) develop biofabrication technologies based on additive manufacturing and spinning, and bottom-up methodologies; (ii) integrate neural and vascular cues on current tissue regeneration strategies; (iii) engineer the immune response of biomaterials and biomedical devices.
Another alternative emerging approach involves biomaterials produced by the cells themselves in response to a chemical stimulus. Indeed, we exploited living cells as factories to produce new, functional and intelligent fibrils for tissue engineering, through the aid of synthetic dyes which spontaneously penetrate the membrane of living cells.
Plasma processing of scaffolds for tissue engineering
Plasma processing of scaffolds for tissue engineering can be used mainly to: 1) functionalize the scaffold surface in order to improve, when necessary, its hydrophilic character (i.e. transport of metabolites and waste products through water); 2) produce graded chemical composition from the top to the scaffold’s core to enhance cell colonization of the whole scaffold; 3) “decorate” the scaffold’s surface with biomimetic coatings containing proteins or ceramics (i.e. hydroxyapatite or magnesium) that resembles the natural tissue composition. On the other hand the application of plasma directly on cells or on cell culture media (plasma medicine) should promote a selective improvement of cell proliferation on 3D porous scaffolds stimulating in turn the regeneration of new tissue.
In association with Maastricht University (Prof. L. Moroni), we aim at engineering functional biomaterials to create 3D scaffolds able to control cell fate. This challenge is approached through a biomimetic design inspired by cell niches. Several strategies are being pursued, comprising smart coatings (e.g. layer-by-layer technology), chemical functionalization (e.g. covalent vs dynamic binding), and physical modification of surface properties (e.g. stiffness vs topography).
A deeper comprehension of the biomineralization process is at the basis of tissue engineering and will be instrumental to the further development of regenerative medicine. The achievement of a complete and exhaustive explanation of a process as complex as biomineralization requires the synergy of different advanced experimental techniques. In this framework, we have developed a multi-scale approach–based on different complementary X-ray experimental techniques coupled to new analytical tools, which provide structural and morphological information on the engineered tissues, from the atomic to the micrometric scale. In particular we study the mechanism of mineralized matrix deposition in a tissue engineering approach in which bone tissue is formed when porous ceramic constructs are loaded with bone marrow stromal cells and implanted in vivo.
High resolution X-ray Phase contrast Tomography (XPCT) provides the 3D spatial distribution of the different tissues participating to the biomineralization process; Scanning X-ray micro diffraction (XRmD) exploits the focused sub-micrometer X-ray beam to achieve atomic information with high spatial resolution. Scanning through the beam the organic–inorganic interface, within a porous scaffold of the sample, we are able to distinguish and monitor the evolution of the different ‘players’ of the regeneration process (Collagen, Organic Matrix, Hydroxy Apatite (HA), amorphous calcium phosphate (ACP)). X-ray Fluorescence (XRF) verifies the chemical evolution of the different growing phases and investigates the distribution of Ca in the regenerated bone. This multi-technique approach provides information on the first steps of biomineralization: we investigate the precursor of the biomineralization, the ACP working as a Ca reservoir, the dynamics of the collagen which is anisotropically distributed far from the scaffold interface but strongly packed at the organic–inorganic interface. When Ca ions are sequestered inside the collagen gaps, the mineralization starts to develop and bone nanoparticles appear at the scaffold interface.
To monitor the relation between bone formation and vascularization, it is important to achieve a detailed imaging and a quantitative description of the complete three-dimensional vascular network in such constructs. We imaged the 3D vascularization network inside the scaffold, without any sample sectioning and preparation. This study of angiogenesis in tissue engineering is crucial for the evaluation of the performance of an artificially implanted construct. Indeed the control of the angiogenesis of the micro-vascular network with proper spatial organization is a key step to obtain tissue regeneration and repair.
- F. Spano, A. Quarta, C. Martelli, L. Ottobrini, R. R.M. Rossi, G. Gigli, L. Blasi, Fibrous scaffolds fabricated by emulsion electrospinning: from hosting capacity to in vivo biocompatibility. Nanoscale, 8: 9293-9303, 2016. ISSN: 2040-3364; doi: 10.1039/c6nr00782a
- I. E. Palamà, F. Di Maria, S. D’Amone, G. Barbarella, G. Gigli Biocompatible and biodegradable fluorescent microfibers physiologically secreted by live cells upon spontaneous uptake of thiophene fluorophore. G. Journal of Materials Chemistry B, 3, 151- 158 (2015). ISSN: 2050-750X; doi: 10.1039/c4tb01562b.
- G. Ciasca, M. Papi, L. Businaro, G. Campi, M. Ortolani, V. Palmieri, A. Cedola, A. De Ninno, A. Gerardino, G. Maulucci, M. De Spirito Recent advances in superhydrophobic surfaces and their relevance to biology and medicine. Bioinspiration & biomimetics 11 (1), 011001, 2016. ISSN: 2050-750X; doi:10.1088/1748-3190/11/1/011001.
- F. Intranuovo, R. Gristina, L. Fracassi, L. Lacitignola, A. Crovace, P. Favia; Plasma processing of scaffolds for tissue engineering and regenerative medicine. Plasma Chemistry and Plasma Processing, 36, (2016) 269-280, ISSN: 02724324, doi: 10.1007/s11090-015-9667-0
- M. Fratini, G. Campi, I. Bukreeva, D. Pelliccia, M. Burghammer, G. Tromba, R. Cancedda, M. Mastrogiacomo, A. Cedola. X-ray micro-beam techniques and phase contrast tomography applied to biomaterials. Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms 364, 93-97, 2015. ISSN: 0168-583X; doi: 10.1016/j.nimb.2015.06.023.
- G. Campi, M. Fratini, I. Bukreeva, G. Ciasca, M. Burghammer, F. Brun, G. Tromba, M. Mastrogiacomo, A. Cedola. Imaging collagen packing dynamics during mineralization of engineered bone tissue Acta biomaterialia, 23, 309-316 (2015). ISSN: 1742-7061; doi: 10.1016/j.actbio.2015.05.033.
- I. Bukreeva, M. Fratini, G. Campi, D. Pelliccia, R. Spanò, F. Brun, M. Burghammer, M. Grilli, R. Cancedda, A. Cedola, M. Mastrogiacomo High-resolution X-ray techniques as new tool to investigate the 3D vascularization of engineered-bone tissue. Frontiers in bioengineering and biotechnology, 3, 133 (2015).ISSN: 2296-4185; doi:10.3389/fbioe.2015.00133
- E. Sardella, E. R. Fisher, J. C. Shearer, M. Garzia Trulli, R. Gristina, P. Favia, N2/H2O Plasma Assisted Functionalization of Poly(ε-caprolactone) Porous Scaffolds: Acidic/Basic Character versus Cell Behavior, Plasma Process. Polym. 12-8, 786-798 (2015); ISSN: 16128850, doi: 10.1002/ppap.201400201
- I. Trizio, F. Intranuovo, R. Gristina, G. Dilecce, P. Favia, He/O2 atmospheric pressure plasma jet treatments of PCL scaffolds for tissue engineering and regenerative medicine. Plasma Processes and Polymers 12-12, 1451-1458 (2015) , ISSN: 16128850, doi: 10.1002/ppap.201500104
- I. Trizio, E. Sardella, E. Francioso, G. Dilecce, V. Rizzi, P. Cosma, M. Schmidt, M. Hansch, T. von Woedtke, P. Favia, R. Gristina, Investigation of air-DBD effects on biological liquids for in vitro studies on eukaryotic cells, Clinical Plasma Medicine 3-2, 62-71 (2015) ISSN: 22128166, doi: 10.1016/j.cpme.2015.09.003
- F. Intranuovo, R. Gristina, F. Brun, S. Mohammadi, G. Ceccone, E. Sardella, F.Rossi, G. Tromba, P. Favia, Plasma modification of PCL porous scaffolds fabricated by solvent-casting/particulate-leaching for tissue engineering, Plasma Process. Polym. 11, 184–195 (2014), ISSN: 16128850, doi: 10.1002/ppap.201300149
- R. Spano, I. Bukreeva, G. Campi, G. Tromba, F. Brun, A. Cedola, R. Cancedda, M. Mastrogiacomo Vascular network visualization in bone tissue engineered construct by synchrotron X-ray microtomography. Journal of tissue engineering and regenerative medicine 8, 211-211 (2014). ISSN: 1932-6254;
- Campi, I. Bukreeva, M. Fratini, M. Mastrogiacomo, A. Cedola Imaging tissue regeneration/degeneration by combined X-ray micro-diffraction and phase contrast micro-tomography. In Journal of tissue engineering and regenerative medicine 8, 66-67, 2014. ISSN: 1932-6254;
Other selected publications
- I. E. Palamà, F. Di Maria, I. Viola, E. Fabiano, G. Gigli, C. Bettini, G. Barbarella. Live-cell-permeant thiophene fluorophores and cell-mediated formation of fluorescent fibrils. Journal of the American Chemical Society (JACS), 133, 17777–17785 (2011). ISSN: 0002-7863; doi: 10.1021/la2065522
- I. Viola, I. E. Palamà, A. M. L. Coluccia, M. Biasiucci, B. Dozza, E. Lucarelli, C. Bettini, G. Barbarella, G. Gigli Physiological formation of fluorescent and conductive protein microfibers in live fibroblasts upon spontaneous uptake of biocompatible fluorophores. Integrative Biology, 5, 1057- 1066, 2013. ISSN: 1757-9694; doi: 10.1039/c3ib40064f.
- G. Campi, G. Pezzotti, M. Fratini, A. Ricci, M. Burghammer, R. Cancedda, I. Bukreeva, M. Mastrogiacomo, A. Cedola, Imaging regenerating bone tissue based on neural networks applied to micro-diffraction measurements. Applied Physics Letters,103 (25), 253703 (2013) ISSN: 0003-6951; doi: 10.1063/1.4852056
- G. Campi, A. Ricci, A. Guagliardi, C. Giannini, S. Lagomarsino, R. Cancedda, M. Mastrogiacomo, A. Cedola. Early stage mineralization in tissue engineering mapped by high resolution X-ray microdiffraction. Acta biomaterialia,8(9), 3411-3418. 2012 ISSN: 1742-7061; doi: 10.1016/j.actbio.2012.05.034.
- A. Guagliardi, A. Cedola, C. Giannini, M. Ladisa, A. Cervellino, A. Sorrentino, S. Lagomarsino, R. Cancedda, M. Mastrogiacomo. Debye function analysis and 2D imaging of nanoscaled engineered bone. Biomaterials, 31(32), 8289-8298 (2010). ISSN: 0142-9612; doi: 10.1016/j.biomaterials.2010.07.051
- M. Domingos, F. Intranuovo, A. Gloria, R. Gristina, L. Ambrosio, P.J. Bartolo, P. Favia, Improved osteoblast cell affinity on plasma modified 3-D extruded PCL scaffolds. Acta Biomaterialia 9-4, 5997-6005 (2013) ISSN: 1742-7061; doi: 10.1016/j.actbio.2012.12.031
- R. Cancedda, A. Cedola, A. Giuliani, V. Komlev, S. Lagomarsino, M. Mastrogiacomo, F. Peyrin, F. Rustichelli. Bulk and interface investigations of scaffolds and tissue-engineered bones by X-ray microtomography and X-ray microdiffraction. Biomaterials, 28(15), 2505-2524 (2007). ISSN: 0142-9612; doi: 10.1016/j.biomaterials.2007.01.022
- M. Mastrogiacomo, A. Papadimitropoulos, A. Cedola, F. Peyrin, P. Giannoni, S. G. Pearce, M. Alini, C. Giannini, A. Guagliardi, R. Cancedda Engineering of bone using bone marrow stromal cells and a silicon-stabilized tricalcium phosphate bioceramic: evidence for a coupling between bone formation and scaffold resorption. Biomaterials, 28(7), 1376-1384 (2007). ISSN: 0142-9612; doi: 10.1016/j.biomaterials.2006.10.001
- V. S. Komlev, F. Peyrin, M. Mastrogiacomo, A. Cedola, A. Papadimitropoulos, F. Rustichelli, R. Cancedda, Kinetics of in vivo bone deposition by bone marrow stromal cells into porous calcium phosphate scaffolds: an X-ray computed microtomography study. Tissue engineering, 12(12), 3449-3458, 2006. ISSN: 1076-3279; doi: 10.1089/ten.2006.12.3449
- A. Cedola, M. Mastrogiacomo, S. Lagomarsino, R. Cancedda, C. Giannini, A. Guagliardi, M. Ladisa, M. Burghammer, F. Rustichelli, V. Komlev, Orientation of mineral crystals by collagen fibers during in vivo bone engineering: an X-ray diffraction imaging study Spectrochimica Acta Part B: Atomic Spectroscopy, 62(6), 642-647,2007. ISSN: 0584-8547; doi: 10.1016/j.sab.2007.02.015
1. Special mention to Eloisa Sardella during the competition “Italia Camp-your idea for the country (la tua idea per il paese)” during the BarCamp “ Stati Generali del Mezzogiorno d’Europa” with the idea titled “plasma3D” 2012