The Effects of Titanium Dioxide Nanoparticles on Osteoblasts Mineralization: A Comparison between 2D and 3D Cell Culture Models

Although several studies assess the biological effects of micro and titanium dioxide nanoparticles (TiO2 NPs), the literature shows controversial results regarding their effect on bone cell behavior. Studies on the effects of nanoparticles on mammalian cells on two-dimensional (2D) cell cultures display several disadvantages, such as changes in cell morphology, function, and metabolism and fewer cell–cell contacts. This highlights the need to explore the effects of TiO2 NPs in more complex 3D environments, to better mimic the bone microenvironment. This study aims to compare the differentiation and mineralized matrix production of human osteoblasts SAOS-2 in a monolayer or 3D models after exposure to different concentrations of TiO2 NPs. Nanoparticles were characterized, and their internalization and effects on the SAOS-2 monolayer and 3D spheroid cells were evaluated with morphological analysis. The mineralization of human osteoblasts upon exposure to TiO2 NPs was evaluated by alizarin red staining, demonstrating a dose-dependent increase in mineralized matrix in human primary osteoblasts and SAOS-2 both in the monolayer and 3D models. Furthermore, our results reveal that, after high exposure to TiO2 NPs, the dose-dependent increase in the bone mineralized matrix in the 3D cells model is higher than in the 2D culture, showing a promising model to test the effect on bone osteointegration.

[1]  Hiroaki Nakamura,et al.  3D-cultured small size adipose-derived stem cell spheroids promote bone regeneration in the critical-sized bone defect rat model. , 2022, Biochemical and biophysical research communications.

[2]  M. Álvarez-Pérez,et al.  3D Spheroid Cell Cultures and Their Role in Bone Regeneration: a Systematic Review , 2021, Odovtos - International Journal of Dental Sciences.

[3]  J. Ohno,et al.  Three-dimensional spheroids of dedifferentiated fat cells enhance bone regeneration , 2021, Regenerative therapy.

[4]  Yoon-Hee Park,et al.  Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells , 2021, Medicina.

[5]  K. Khalaf,et al.  Impact of tribocorrosion and titanium particles release on dental implant complications — A narrative review , 2021, The Japanese dental science review.

[6]  V. Quinelato,et al.  Epidermal Growth Factor Is Associated with Loss of Mucosae Sealing and Peri-Implant Mucositis: A Pilot Study , 2021, Healthcare.

[7]  M. Álvarez-Pérez,et al.  Case Report: Formation of 3D Osteoblast Spheroid Under Magnetic Levitation for Bone Tissue Engineering , 2021, Frontiers in Molecular Biosciences.

[8]  D. Serrano,et al.  Mimicking bone microenvironment: 2D and 3D in vitro models of human osteoblasts. , 2021, Pharmacological research.

[9]  Panyu Zhou,et al.  Investigation of Mg–xLi–Zn alloys for potential application of biodegradable bone implant materials , 2021, Journal of Materials Science: Materials in Medicine.

[10]  A. Awasthi,et al.  Nanomaterials in Biology , 2021, Environmental Science and Pollution Research.

[11]  Yuan Zhang,et al.  The unfavorable role of titanium particles released from dental implants , 2021, Nanotheranostics.

[12]  Luis Amengual-Peñafiel,et al.  Osteoimmunology drives dental implant osseointegration: A new paradigm for implant dentistry , 2021, The Japanese dental science review.

[13]  G. Romanos,et al.  Titanium Wear of Dental Implants from Placement, under Loading and Maintenance Protocols , 2021, International journal of molecular sciences.

[14]  J. Burdick,et al.  3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels , 2020, Nature Communications.

[15]  R. Thiré,et al.  Biomimetic Mineralization on 3D Printed PLA Scaffolds: On the Response of Human Primary Osteoblasts Spheroids and In Vivo Implantation , 2020, Polymers.

[16]  Philippe Bédard,et al.  Innovative Human Three-Dimensional Tissue-Engineered Models as an Alternative to Animal Testing , 2020, Bioengineering.

[17]  K. Miłowska,et al.  Spheroids as a Type of Three-Dimensional Cell Cultures—Examples of Methods of Preparation and the Most Important Application , 2020, International journal of molecular sciences.

[18]  F. Erogbogbo,et al.  Assessing Advantages and Drawbacks of Rapidly Generated Ultra-Large 3D Breast Cancer Spheroids: Studies with Chemotherapeutics and Nanoparticles , 2020, International journal of molecular sciences.

[19]  Yong Teng,et al.  Is It Time to Start Transitioning From 2D to 3D Cell Culture? , 2020, Frontiers in Molecular Biosciences.

[20]  Tiziano Tuccinardi,et al.  The History of Nanoscience and Nanotechnology: From Chemical–Physical Applications to Nanomedicine , 2019, Molecules.

[21]  T. Silva,et al.  Inhibitory effects of dabigatran etexilate, a direct thrombin inhibitor, on osteoclasts and osteoblasts. , 2019, Thrombosis research.

[22]  J. Yamashita,et al.  Alternating Differentiation and Dedifferentiation between Mature Osteoblasts and Osteocytes , 2019, Scientific Reports.

[23]  M. H. Fernandes,et al.  The two faces of titanium dioxide nanoparticles bio-camouflage in 3D bone spheroids , 2019, Scientific Reports.

[24]  Truc Thi Hoang Nguyen,et al.  General review of titanium toxicity , 2019, International journal of implant dentistry.

[25]  G. Romanos,et al.  Potential Causes of Titanium Particle and Ion Release in Implant Dentistry: A Systematic Review , 2018, International journal of molecular sciences.

[26]  R. Ghodsi,et al.  Bio-camouflage of anatase nanoparticles explored by in situ high-resolution electron microscopy. , 2017, Nanoscale.

[27]  Li-Hsin Han,et al.  Modeling Physiological Events in 2D vs. 3D Cell Culture. , 2017, Physiology.

[28]  A. Janorkar,et al.  Spheroid model for functional osteogenic evaluation of human adipose derived stem cells. , 2017, Journal of biomedical materials research. Part A.

[29]  N. Benkirane-Jessel,et al.  Nanoengineered implant as a new platform for regenerative nanomedicine using 3D well-organized human cell spheroids , 2017, International journal of nanomedicine.

[30]  M. Chatzinikolaidou.  Cell spheroids: the new frontiers in in vitro models for cancer drug validation. , 2016, Drug discovery today.

[31]  J. K. Leach,et al.  Increased Survival and Function of Mesenchymal Stem Cell Spheroids Entrapped in Instructive Alginate Hydrogels , 2016, Stem cells translational medicine.

[32]  J. Werckmann,et al.  Trojan-Like Internalization of Anatase Titanium Dioxide Nanoparticles by Human Osteoblast Cells , 2016, Scientific Reports.

[33]  Alexander Tsouknidas,et al.  New Ti-Alloys and Surface Modifications to Improve the Mechanical Properties and the Biological Response to Orthopedic and Dental Implants: A Review , 2016, BioMed research international.

[34]  C. Gabriel,et al.  Morphological and Immunohistochemical Characterization of Canine Osteosarcoma Spheroid Cell Cultures , 2015, Anatomia, histologia, embryologia.

[35]  Francesco Salvatore,et al.  The impact of nanoparticle protein corona on cytotoxicity, immunotoxicity and target drug delivery. , 2016, Nanomedicine.

[36]  H. Kim,et al.  Nanotechnology in dentistry: prevention, diagnosis, and therapy , 2015, International journal of nanomedicine.

[37]  G. Romanos.  Current concepts in the use of lasers in periodontal and implant dentistry , 2015, Journal of Indian Society of Periodontology.

[38]  S. Kates,et al.  Surface Damage on Dental Implants with Release of Loose Particles after Insertion into Bone. , 2015, Clinical implant dentistry and related research.

[39]  Krasimir Vasilev,et al.  Innate Immunity and Biomaterials at the Nexus: Friends or Foes , 2015, BioMed research international.

[40]  Junji Fukuda,et al.  An oxygen-permeable spheroid culture system for the prevention of central hypoxia and necrosis of spheroids. , 2012, Biomaterials.

[41]  Sumit Arora,et al.  Nanotoxicology and in vitro studies: the need of the hour. , 2012, Toxicology and applied pharmacology.

[42]  Rocky S. Tuan,et al.  What are the local and systemic biologic reactions and mediators to wear debris, and what host factors determine or modulate the biologic response to wear particles? , 2008, The Journal of the American Academy of Orthopaedic Surgeons.

[43]  K. Robbie,et al.  Nanomaterials and nanoparticles: Sources and toxicity , 2007, Biointerphases.

[44]  JONG BIN Kim,et al.  Three-dimensional tissue culture models in cancer biology. , 2005, Seminars in cancer biology.

[45]  G. Truskey,et al.  Effects of titanium particle size on osteoblast functions in vitro and in vivo. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[46]  T. Webster,et al.  Increased viable osteoblast density in the presence of nanophase compared to conventional alumina and titania particles. , 2004, Biomaterials.

[47]  E. Schwarz,et al.  PGE2 and IL‐6 production by fibroblasts in response to titanium wear debris particles is mediated through a Cox‐2 dependent pathway , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[48]  S. Goodman,et al.  Histomorphological reaction of bone to different concentrations of phagocytosable particles of high-density polyethylene and Ti-6Al-4V alloy in vivo. , 1996, Biomaterials.