In vitro effects of beta-titanium alloys on biological behavior of macrophages and their cross-talk with osteoblasts
Assoc. Prof. Anisoara Cimpean
Phone number: 021.318.15.75/int 106
Fax : 021.318.15.75/int 102
University of Bucharest
Faculty of Biology, Department of Biochemistry and Molecular Biology
Contractual authority: UEFISCDI
Project code: PN-II-ID-PCE-2011-3-0737
Project type and number: PCE 188/2011
Project title: In vitro effects of beta-titanium alloys on biological behavior of macrophages and their cross-talk with osteoblasts
Duration: 36 months
Budget: 1.400.000 lei
The project aims to elucidate the modulation of macrophage function and of the cross-talk between macrophages and osteoblasts by new beta-titanium alloys which are based on titanium and nontoxic alloying elements, such as Ta, Nb/Zr. For comparative purposes, a commercial Ti-6Al-4V alloy, will be also investigated. In the first stage, we will investigate if the macrophages cultured in contact with the tested substrates display differences regarding cell viability, proliferation, apoptosis, cytoskeleton and adhesive structures (focal contact, podosomes) that may be important for the interpretation of macrophage function in chronic inflammatory responses. Next, we will evaluate macrophage cytokine, chemokine and matrixin expression at transcriptional and translational levels. The proposed project is an innovative approach which will bring new knowledge on signaling pathways (such as NFκB and MAP-Kinase) that could be activated by various Ti alloy compositions and on materials’ effects upon the cross-talk between macrophages and osteoblasts using a Transwell co-culture model. This cross-talk will be studied in terms of modulation of: i) soluble factors involved in bone turnover (RANKL, OPG); ii) osteogenesis process (alkaline phosphatase activity, mineralization process); iii) macrophage and osteoblast cytokine production. These studies can conduct to the development of cellular models for in vitro assessment of the inflammatory response to osteoprothetic materials.
Since chemistry of implant materials has the capacity to influence cell behavior, it is important to understand the responses of macrophages to the various compositions. Thus, our in vitro studies have as a general objective to elucidate the modulation of macrophage function and of the cross-talk between macrophages and osteoblasts by new beta-titanium alloys of different chemical composition.
The specific objectives of this project are: i) Scientific fundamentation of the necessity to in vitro evaluate the cellular inflamatory response to implant materials in biocompatibility studies and of the celullar interactions and signaling pathways that could be activated by surfaces with different characteristics; ii) Determination of the TiTaZr/Nb based materials effects on macrophage viability, proliferation and apoptosis; iii) Characterization of macrophage cytoskeleton and adhesion structures, which may be important for the interpretation of macrophage function in chronic inflammatory responses; iv) Assessment of the material induced gene expression regulation by evaluation of mRNA and protein expression of cytokines and chemokines secreted by the macrophages; v) Investigation of beta-Ti alloys’ implication in induction of matrixins such as matrix metalloproteinases (MMPs) and ADAM-9; vi) Determination of signaling pathways that could be activated by the different TiTaZr/Nb compositions; vii) Evaluation of beta-Ti alloys’ effects on the cross-talk between macrophages and osteoblasts in a Transwell co-culture model.
|Name and surname||Position in the project|
|Cimpean Anisoara||Project director/Senior researcher II|
|Costache Marieta||Senior researcher I|
|Ion Raluca||Young researcher III|
|Mitran Valentina||Research assistent|
|Galateanu Bianca||Research assistent|
|Neacsu Patricia||Research assistent (phD student)|
|Dinescu Sorina||Research assistent (phD student)|
|Serban Mirela||Research assistent|
1. Mindroiu M, Ion R, Pirvu C, Cimpean A, Surfactant-dependent macrophage response to polypyrrole-based coatings electrodeposited on Ti6Al7Nb alloy, Mater Sci Eng C, 33, 3353-3361 (2013).
2. Ion R, Gordin DM, Mitran V, Osiceanu P, Dinescu S, Gloriant T, Cimpean A, In vitro bio-functional performances of the novel superelastic beta-type Ti-23Nb-0.7Ta-2Zr-0.5N alloy, Mater Sci Eng C Mater Biol Appl, 35, 411-419 (2014).
3. Neacsu P, Mazare A, Cimpean A, Park J, Costache M, Schmuki P, Demetrescu I, Reduced inflammatory activity of RAW 264.6 macrophages on titania nanotube modified Ti surface, Int J Biochem Cell Biol, 55, 187-195, (2014).
4. Neacsu P, Gordin DM, Mitran V, Gloriant T, Costache M, Cimpean A. In vitro performance assessment of new beta Ti-Mo-Nb alloy compositions. Mater Sci Eng C Mater Biol Applic, 47, 105-113 (2015).
5. Neacsu P, Mazare A, Schmuki P, Cimpean A. Attenuation of the macrophage inflammatory activity by TiO2 nanotubes via inhibition of MAPK and NF-kB pathways. Int J Nanomedicine, 10, 6455-6476 (2015).
6. Ion R, Stoian AB, Dumitriu C, Grigorescu S, Mazare A, Cimpean A, Demetrescu I, Schmuki P. Nanochannels formed on TiZr alloy improve biological response. Acta Biomater, 24, 270-377 (2015).
BOOK CHAPTERS PUBLISHED IN COLLECTIVE VOLUMES AT PRESTIGIOUS INTERNATIONAL PRINTING HOUSES
1. A. Cimpean, R. Ion, D.M. Gordin, P. Neacsu, V. Mitran, T. Gloriant, Biomaterials as modulators of macrophage inflammatory response, Biomaterials (vol. 11) in: N.K. Navani, S. Sinha (Eds.), J.N. Govil (Executive Ed.), Nanotechnology series, Studium Press LLC Houston (USA), 273-310, 2013, ISBN-1-62699-011-5.
COMMUNICATIONS TO NATIONAL SCIENTIFIC REUNIONS
1. Neacsu P, Sebestyen I, Mitran V, Ion R, Cimpean A, Modulation of macrophage biological behavior by TiO2 nanotubes, Scientific communication session of the students from Faculty of Biology, 25th May 2012, Bucharest, Romania (oral presentation).
2. Neacsu P, Cimpean A, Costache M, Reduced inflammatory response to TiO2 nanotubes. Scientific communication session of the students from Faculty of Biology, 31st May 2013, Bucharest, Romania (oral presentation).
3. Neacsu P, Ion R, Cimpean A, Costache M, Macrophage response to TiO2 nanochannels, Scientific communication session of the students from Faculty of Biology, 30th May 2014, Bucharest, Romania (oral presentation).
4. Neacsu P, Ion RN, Mitran V, Cimpean A. MAPK and NF-kB signaling pathways modulation by TiO2 nanotubes. Scientific communication session of the students from Faculty of Biology, 29th May 2015, Bucharest, Romania (oral presentation) – 2ndprize.
COMMUNICATIONS TO INTERNATIONAL SCIENTIFIC REUNIONS
1. Ion R, Neacsu P, Gordin DM, Gloriant T, Cimpean A, RAW 264.7 macrophage behavior on the new beta-type Ti-23Nb-0.7Ta-2Zr- 0.5N alloy, 5th International Conference “Biomaterials, Tissue Engineering & Medical Devices” – BioMedD 2012, Constanta, 29th Aug. – 1st Sept., p. 191 (poster presentation).
2. Neacsu P, Ion R, Mitran V, Raducanu D, Dan I, Cimpean A, Osteoblast and macrophage responses to Ti-25Ta-5Zr alloy, 5th International Conference “Biomaterials, Tissue Engineering & Medical Devices” – BioMedD 2012, Constanta, 29th Aug. – 1st Sept., p. 215 (poster presentation).
3. Ion R, Mitran V, Neacsu P, Gordin D, Dinescu S, Costache M, Gloriant T, Cimpean A, Modulation of macrophage inflammatory response by titanium alloy chemistry, 5th International Conference on Drug Discovery and Therapy, Dubai, 18th-21st Feb. 2013 (poster presentation).
4. Neacsu P, Mitran V, Gordin D, Ion R, Costache M, Gloriant T, Cimpean A, In vitro bioactivity of Ti-Mo-Nb alloys, 1stInternational Symposium on Modern Biotechnological Advances for Human Health – BAHH, Bucharest, 20th-22nd May 2014 (poster presentation).
5. Neacsu P, Mazare A, Park J, Costache M, Schmuki P, Demetrescu I, Cimpean A, Mitigating the inflammatory response by TiO2 nanotubes, 50th International Conference FEBS EMBO, Paris, 30th-4th September 2014, p. 143 (poster presentation).
6. Neacsu P, Mazare A, Ion RN, Mitran V, Costache M, Schmuki P, Cimpean A. Modulation of MAPK and NF-kB signaling pathways by TiO2 nanotubes Ti-modified surface, 40th FEBS Congress, Berlin, Germany, 4-9 iuly 2015, FEBS Journal, 282 (Suppl. 1), p. 197 (poster presentation).
PhD/Master/Bachelor Theses on project’s topics
Patricia Neacsu: In vitro evaluation of macrophage response to titanium-based biomaterials (2012-2015)
Patricia Neacsu: Modulation of macrophages behavior by TiO2 nanotubes (2012)
Andreea Iulia Sebestyen: Macrophage response to titanium surfaces modified with TiO2 nanotubes (2013)
Jalba Nicoleta: Assesment of the inflammatory response induced by beta Ti-Mo-Nb alloys (2014)
Irina Antoanela Boaru: In vitro macrophages response to Ti23Nb0.7Ta2Zr0.5N (2014)
Bachelor thesis: Raluca Mudrag: Macrophages implications in the inflammatory response to implantable materials (2015)