Mineralizing Vesicles: From Biochemical and Biophysical Properties to Their Roles in Physiology and Disease

Mineralizing Vesicles: From Biochemical and Biophysical Properties to Their Roles in Physiology and Disease presents the state-of-the art in the properties of mineralizing EVs and their potential clinical applications.

The first chapter presents foundational biochemical and biological aspects of EVs. Next, the title coves the role of EVs in bone ossification and in cardiovascular and cartilage-related diseases. Considering the unique ability of this class of EVs to form apatite minerals assigned to their special biochemical machinery, three chapters of the book then focus on the enzymes catalysing the inorganic phosphate and calcium turn-over and the dynamic properties of the vesicles’ peripheral proteins. Chapters describe the role of inorganic phosphate and calcium ions and of autophagy on the biogenesis and function of mineralizing EVs. Recent studies show that the lumen of mineralizing EVs is partially filled with miRNA, and a chapter therefore considers research on the possible function of these vesicles as signalosomes. The final five chapters of the book describe practical aspects of working with mineralizing EVs, including their purification, proteomic and biophysical analyses, the use of biomimetic models and mineralizing EVs in regenerative medicine.

This title presents, for the first time, a comprehensive account of mineralizing EVs and their potential clinical applications. It will be invaluable to researchers in the field.

• Covers all aspects of mineralizing EVs, from their composition to their function in physiological   and pathological processes, and their clinical potential
• Presents mineralizing EVs systematically using clear accessible language
• Describes practical aspects of working with mineralizing EVs
• Integrates an account of the biochemistry and biophysics of vesicles with their functions
• Takes a multidisciplinary perspective, offering an exhaustive and cutting-edge account of mineralizing EVs and human health

• Language: English
• Publisher: Academic Press
• Release date: Nov 17, 2023
• ISBN: 9780323991599

Book preview

Mineralizing Vesicles

From Biochemical and Biophysical Properties to Their Roles in Physiology and Disease

Edited by

Massimo Bottini

Department of Experimental Medicine, Faculty, of Medicine and Surgery, University of Rome, Tor Vergata, Rome, Italy

Ana Paula Ramos

Department of Chemistry, Faculty of Philosophy, Science and Letters at Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil

Table of Contents

Cover image

Title page

Copyright

Contributors

Biographies

Preface

Chapter 1. General aspects of extracellular vesicles in bone formation and remodeling

1. Introduction

2. Communications between bone cells

3. Extracellular vesicles

4. Bone cell overview

5. Bone marrow cells interacting with skeletal cells

6. Extracellular vesicle trafficking in the bone microenvironment

7. Extracellular vesicles in fracture healing

8. Extracellular vesicles and bone cancer

9. Medical applications

10. Conclusions

List of abbreviations

Acknowledgments

Chapter 2. Physiological biomineralization. The properties and role of matrix vesicles in skeletal and dental calcifications

1. Introduction

2. Biomineralization in teeth

3. Biomineralization in bones

4. Determinants of mineralization support the functions of matrix vesicles

5. Conclusions

List of abbreviations

Chapter 3. Pathological biomineralization. Part I: Mineralizing extracellular vesicles in cardiovascular diseases

1. Introduction

2. Extracellular vesicles: origin and classification

3. Extracellular vesicles related to cardiovascular diseases

4. Extracellular vesicles: biomarkers of cardiovascular diseases

5. Extracellular vesicles as drug delivery vehicles in cardiovascular disease

6. Conclusions

List of abbreviations

Chapter 4. Pathologic biomineralization: part II: mineralizing extracellular vesicles in osteoarthritis

1. Introduction

2. Historical perspective

3. Characterization of ACVs

4. ACVs from osteoarthritis cartilage

5. Factors affecting ACV mineralization

6. Conclusions and future directions

List of abbreviations

Chapter 5. The biochemistry of mineralizing extracellular vesicles. Part I: The role of phosphatases

1. Inorganic phosphate (Pi) and pyrophosphate (PPi): a physicochemical balance

2. The molecules regulating the Pi/PPi ratio

3. Tissue-nonspecific alkaline phosphatase

4. Ectonucleotide pyrophosphatase/phosphodiesterase 1 and other ENPPs

5. Nucleoside triphosphate diphosphohydrolase 1 (CD39) and ecto-5′-nucleotidase (CD73)

6. PHOSPHO1

7. Can Na,K-ATPase act as a phosphatase?

8. Phosphate transporters

9. Genetic diseases caused by altered Pi/PPi ratio

10. Therapeutic approaches to normalizing the Pi/PPi ratio

11. Conclusions

List of abbreviations

Chapter 6. The biochemistry of mineralizing extracellular vesicles. Part II: Annexins

1. Introduction

2. Annexins

3. Annexin knockout animal models

4. Annexins in mineralization-competent cells

5. Annexins in media vesicles and matrix vesicles

6. In vitro properties of annexins

7. Conclusions

List of abbreviations

Acknowledgments

Chapter 7. Calcium and phosphate and their role in matrix vesicles: A biological view

1. Calcium (Ca2+) and inorganic phosphate (PO43−/Pi) ions are essential for life

2. Ca2+ and phosphate ions in mineralization physiology and pathology

3. Cellular origins of matrix vesicles

4. Formation of mineralization-competent matrix vesicles requires osteo/chondrogenic molecular phenotype of the cells

5. Conclusions

List of abbreviations

Acknowledgments

Chapter 8. Autophagy in bone metabolism and its possible role on the function of mineralizing extracellular vesicles

1. Molecular overview of autophagy

2. The canonical role of autophagy on bone maintenance

3. The noncanonical role of autophagy on bone mineralization

4. The role of autophagy on the release of mineralizing extracellular vesicles

5. Induced autophagy decreases smooth muscles cells calcification

6. Conclusions

List of abbreviations

Chapter 9. The roles of mineralizing extracellular vesicles in cell–cell communication

1. Introduction

2. Proteins in mineralizing EVs

3. miRNAs in mineralizing EVs

4. Conclusion

List of abbreviations

Chapter 10. Working with mineralizing extracellular vesicles. Part I: Purification techniques

1. Discovery of matrix vesicles

2. Purification of matrix vesicles from the growth plates and epiphyseal cartilage of chicken embryos

3. Purification of matrix vesicles from rodent primary chondrocytes

4. Purification of matrix vesicles from osteoblast cell lines and from primary osteoblasts

5. Conclusion

List of abbreviations

Chapter 11. Working with mineralizing extracellular vesicles. Part II: Proteomic profiling

1. Introduction

2. Essential proteins found prior to the proteomic analysis

3. Proteomic analysis of matrix vesicles extracted from growth plate cartilage of chicken embryo

4. Proteomic analysis of matrix vesicles extracted from osteoblast cell lines

5. Comparative analysis of proteomes of matrix vesicles and articular cartilage vesicles

6. Comparative analysis of matrix vesicles and of extracellular vesicles proteomes

7. Conclusions

List of abbreviations

Acknowledgments

Chapter 12. Working with mineralizing extracellular vesicles. Part III: The nucleational core

1. Introduction

2. Physicochemical aspects of biomineralization

3. Discovery of the nucleational core

4. Isolation and characterization of the nucleational core

5. Parameters driving the formation of phosphatidylserine complexes and amorphous calcium phosphate

6. Use of the AFM in the characterization of the nucleational core

7. Conclusions

List of abbreviations

Chapter 13. Working with mineralizing extracellular vesicles. Part IV: Biomimetic models

1. Introduction

2. Proteoliposomes as biomimetic models to assess the role of TNAP in biomineralization

3. Proteoliposomes as biomimetic models to assess the role of Na+,K+-ATPase and NPP1 in biomineralization

4. Proteoliposomes as biomimetic models to assess the role of annexins in biomineralization

5. Langmuir films as biomimetic models of matrix vesicles

6. Conclusions

List of abbreviations

Chapter 14. Working with mineralizing extracellular vesicles. Part V: Use of Mineralizing extracellular vesicles in bone regeneration

1. Introduction

2. Biogenesis of extracellular vesicles

3. Matrix vesicles: A special class of extracellular vesicles

4. Conclusions

List of abbreviations

Index

Copyright

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Contributors

Flavia Amadeu de Oliveira,     Human Genetics Program, Sanford Children's Health Research Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, United States

Luiz Henrique da Silva Andrilli

Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Human Genetics Program, Sanford Children's Health Research Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, United States

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Joanna Bandorowicz-Pikula,     Laboratory of Cellular Metabolism, Nencki Institute of Experimental Biology, Warsaw, Poland

Laurence Bessueille,     Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université de Lyon, Université Lyon 1, UMR CNRS 5246, 69 622 Villeurbanne, Cedex, France

Maytê Bolean,     Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Massimo Bottini

Human Genetics Program, Sanford Children’s Health Research Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, United States

Department of Experimental Medicine, Faculty of Medicine and Surgery, University of Rome Tor Vergata, Rome, Italy

René Buchet,     Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université de Lyon, Université Lyon 1, UMR CNRS 5246, 69 622 Villeurbanne, Cedex, France

Pietro Ciancaglini

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Juçara Gastaldi Cominal,     Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Marcos Antonio Eufrasio Cruz

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Leticia Fernanda Duffles,     Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of Sao Paulo, Ribeirão Preto, SP, Brazil

Colin Farquharson,     Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian, United Kingdom

Bruno Z. Favarin,     Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Sandra Yasuyo Fukada,     Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of Sao Paulo, Ribeirão Preto, SP, Brazil

Thierry Granjon,     Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université de Lyon, Université Lyon 1, UMR CNRS 5246, 69 622 Villeurbanne, Cedex, France

Larwsk Hayann

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Department of Oral and Craniofacial Sciences, Center for Craniofacial Regeneration, University of Pittsburgh School of Dental Medicine, Pittsburgh, PA, United States

Shohei Kohono,     Department of Calcified Tissue Biology, Hiroshima University Graduate School of Biomedical and Health Sciences, Hiroshima, Japan

David Magne,     Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université de Lyon, Université Lyon 1, UMR CNRS 5246, 69 622 Villeurbanne, Cedex, France

Antonio Maurizi,     Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy

Saida Mebarek,     Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université de Lyon, Université Lyon 1, UMR CNRS 5246, 69 622 Villeurbanne, Cedex, France

Maryanne Trafani Melo,     Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

José Luis Millán,     Human Genetics Program, Sanford Children's Health Research Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, United States

Tomoko Minamizaki,     Department of Calcified Tissue Biology, Hiroshima University Graduate School of Biomedical and Health Sciences, Hiroshima, Japan

Dobrawa Napierala,     Department of Oral and Craniofacial Sciences, Center for Craniofacial Regeneration, University of Pittsburgh School of Dental Medicine, Pittsburgh, PA, United States

Sonoko Narisawa,     Human Genetics Program, Sanford Children's Health Research Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, United States

Lucas Fabricio Bahia Nogueira

Department of Experimental Medicine, University of Rome Tor Vergata, Rome, Italy

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Slawomir Pikula,     Laboratory of Biochemistry of Lipids, Nencki Institute of Experimental Biology, Warsaw, Poland

Ana Paula Ramos

Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Ann K. Rosenthal,     Division of Rheumatology, Department of Medicine, Medical College of Wisconsin and the Zablocki VA, Medical Center, Milwaukee, WI, United States

Heitor G. Sebinelli

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Agnieszka Kinga Seliga,     Laboratory of Cellular Metabolism, Nencki Institute of Experimental Biology, Warsaw, Poland

Agnieszka Strzelecka-Kiliszek,     Laboratory of Biochemistry of Lipids, Nencki Institute of Experimental Biology, Warsaw, Poland

Thaise Mayumi Taira,     Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of Sao Paulo, Ribeirão Preto, SP, Brazil

Anna Teti,     Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy

Cintia Kazuko Tokuhara,     Human Genetics Program, Sanford Children's Health Research Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, United States

Ekeveliny A. Veschi,     Department of Chemistry, FFCLRP-University of São Paulo, Ribeirão Preto, SP, Brazil

Lilianna Weremiejczyk,     Laboratory of Biochemistry of Lipids, Nencki Institute of Experimental Biology, Warsaw, Poland

Yuji Yoshiko,     Department of Calcified Tissue Biology, Hiroshima University Graduate School of Biomedical and Health Sciences, Hiroshima, Japan

Biographies

The editors

Massimo Bottini is Associate Professor of Biochemistry at the University of Rome Tor Vergata (Rome, Italy) since 2015. As previous positions, he has been Affiliate Associate Professor at the Sanford Burnham Prebys (La Jolla, USA) from 2015 to 2022, and Visiting Professor at the University of the Chinese Academy of Sciences (Beijing, People's Republic of China) from 2017 to 2019 and at the University of São Paulo (São Paulo, Brazil) in 2020. He has been the recipient of fellowships/grants from the Juvenile Diabetes Research Foundation, the Arthritis National Research Foundation, the European Commission, the University of Rome Tor Vergata, and the Chinese Academy of Sciences. He leads the Laboratory of Biochemical Nanotechnology at the University of Rome Tor Vergata. His research is mostly focused on the characterization of the biochemical and biophysical properties of the extracellular vesicles released during physiologic and ectopic biomineralization processes.

Ana Paula Ramos is an Associate Professor at the University of Paulo-Brazil. She got her first degree in Chemistry in 2004 and a Ph.D. in Physical Chemistry in 2009. She is currently an associate Professor of Physical-Chemistry, at the University of São Paulo-Brazil and a researcher of the Brazilian National Council for Scientific and Technological Development (CNPq). She is the coordinator of the Physical Chemistry of Colloids and Surfaces Laboratory, with expertise in surfaces modification and characterization by microscopic and spectroscopic techniques. Her main research interests are the synthesis of biomaterials for bone replacement and regeneration. In special, her current projects focus on the use of biomimetic matrices inspired by the interaction of phospholipids and osteogenic proteins found in matrix vesicles as modifiers of metals used for dental implants. Moreover, she is currently investigating the role of strontium on physiological and pathological mineralization.

The authors

Flavia Amadeu de Oliveira completed her Ph.D. in 2018 in the field of bone biology at the University of São Paulo, Bauru, Brazil. Currently she is working as a Postdoctoral Associate at Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA. Dr. Amadeu de Oliveira has a long-standing interest in skeletal/dental biology, especially in the molecular mechanisms that regulate bone homeostasis. Her current research focus on the pathophysiological abnormalities in skeletal mineralization of heritable soft bone conditions such as hypophosphatasia (HPP), studying the efficacy of gene/stem cells therapy to validate viral vector/transduced HSCs delivery of mineral-targeted TNAP to correct the soft bones disease in early and late-onset HPP mice while seeking to reduce the risk of ectopic calcification, contributing to devising safe and improved therapies for HPP.

Luiz Henrique Andrilli obtained his first Bachelor of Science degree in Chemistry in 2019 and Master of Science degree in 2022. He is currently a Ph.D. student at the Laboratory of Applied Nanobiotechnology Laboratory: Biomembrane Mimetic Systems, and performing an internship doctorate at the Sanford Burnham Prebys Medical Institute, La Jolla, California, USA. His thesis' studies are focused on membrane mimetic models, with an emphasis on unravelling the mechanisms of bone mineralization and related pathologies. He has papers published in the diverse areas as analytical chemistry, electrochemistry, biochemistry, biophysics, physico-chemistry of membranes, enzymatic kinetics, and formation and characterization of minerals during biological processes, among others.

Joanna Bandorowicz-Pikula, Ph.D. PD, is the Institute professor at the Nencki Institute of Experimental Biology, Laboratory of Cellular Metabolism, Polish Academy of Sciences, Warsaw, Poland. In 1989, Bandorowicz-Pikula received a MSc from Warsaw University and in 1995 completed her Ph.D. at the Nencki Institute of Experimental Biology. Her main research interest is focused on the properties and biological functions of mammalian calcium- and lipid-binding proteins, annexins. In 1996–97, she performed her postdoctoral training at the Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, TX, USA. For many years, she is serving as a member of Doctoral Commission at the Nencki Institute. Since 1992 she is a coauthor of more than 45 experimental papers and 30 review articles.

Laurence Bessueille is currently an associate professor of biochemistry, at the University Claude Bernard Lyon 1; at the Institute of Molecular and Supramolecular Chemistry and Biochemistry, France. Laurence Bessueille completed her Ph.D. at University of Aix-Marseille, France. She did her postdoctoral trainings at Mallinckrodt Medical Inc. Saint Louis, Missouri, USA and at the Enzymatic lipolysis laboratory, CNRS ERS 26, Marseille, France. Her main research projects are centered on the role of alkaline phosphatase in physiological and pathological calcification mechanisms, in particular, in the development of vascular calcifications.

Maytê Bolean got her first degree in Chemistry in 2008, her Master's degree in 2010, and her PhD in 2014, both in Biochemistry and Biophysics fields at the Faculty of Philosophy, Sciences and Letters at Ribeirão Preto, University of São Paulo (FFCLRP-USP), Brazil. She was also a visiting PhD student in Biochemistry at Sanford-Burnham Medical Research Institute (California-USA) in 2013. Her PhD tittle is certificated by International Union for Biophysics Latin American Federation LAFeBS-IUPAB (2013)-POSLATAM. She held a postdoc position at the Department of Chemistry (FFCLRP-USP) funded by FAPESP (2014), and at Katholieke Universiteit Leuven, Leuven, Belgium (2015–16) (BEPE-FAPESP). Currently, she holds a postdoctoral fellowship from CAPES-Brazil at the Applied Nanobiotechnology: Mimetic Systems of Biomembranes Laboratory (FFCLRP-USP). She has experience in Biochemistry, with emphasis on Biophysics and Enzymology, working mainly with structure and function of membrane proteins; expression, purification, and solubilization of membrane proteins and reconstitution in membrane models; and kinetic characterization of membrane enzymes and biophysics characterization of membranes and proteins.

Rene Buchet got a Msc degree in Chemistry in 1980 at University of Geneva, Switzerland, and a Ph D in Chemistry at University of Montreal in 1985. He did his postdoctoral training at the medical school at Univ. Birmingham Alabama, USA, in 1986 and at SUNY Upstate Medical Center at Syracuse, NY, USA, during 1987–90. He was a scientific researcher at ETH, Zurich and at Sandoz, Bern. From 1993 to 2021, he was Professor of Biochemistry at University Claude Bernard Lyon 1; Institute of Molecular and Supramolecular Chemistry and Biochemistry, France. His main research topics were membrane biochemistry and bone biology. He was investigating the mechanisms of mineralization mediated by matrix vesicles. He is now retired.

Pietro Ciancaglini got his first degree in Chemistry in 1985, and a PhD in Biochemistry in 1993. Currently he is a full Professor at the University of São Paulo-Brazil and a researcher of the Brazilian National Council for Scientific and Technological Development (CNPq). He is a coordinator of the Laboratory of Applied Nanobiotechnology: Biomembrane Mimetic Systems. For the last 30 years, he has specialized in obtaining, solubilizing, isolating, purifying, and reconstituting membrane proteins in liposomes (nanostructured vesicles named proteoliposomes). Among the biomolecules of main interest, we can highlight: antimicrobial peptides; antigenic proteins from Leishmania membranes; and alkaline phosphatase (TNAP), ATPase, NPP1, and annexin. He maintains active with international groups from Argentina, United States of America (USA), France, and Italy. In the last 15 years, one important line research has gained focus: construction of matrix vesicles mimic systems. The objective is understanding the function of each protein present in the MVs and their role in the biomineralization process. The final aim is to obtain a mineralizing mimetic vesicle with potential use in nanomedicine. These vesicles could be used in the treatments of bone-related diseases, osteoarthritis, fractures, or deposited on titanium implants.

Juçara Gastaldi Cominal got her first degree in Biomedicine in 2012, followed by a Ph.D. in Biosciences and Biotechnology in 2019. She currently holds a position as a Postdoctoral Research Associate at the Applied Nanobiotechnology Laboratory, which is located in Ribeirão Preto at the University of São Paulo in Brazil. As a member of Dr. Pietro Ciancaglini's team, Juçara brings expertise in native biomembrane systems, extracellular vesicles, mesenchymal stromal cells, primary culture, in vivo models, molecular biology, bone marrow niche, and oncology. Her current project focuses on investigating the role of matrix vesicles during physiological mineralization.

Marcos Cruz obtained a bachelor degree in Chemistry from the University of São Paulo in 2015. He obtained his Master of Science degree from the University of São Paulo in 2018, working in the development of strontium-based biomaterials for bone regeneration. He is currently a PhD candidate at University of São Paulo, under the supervision of Ana Paula Ramos and Pietro Ciancaglini. He is interested in the role of matrix vesicles in bone biomineralization. He has experience on Langmuir monolayers, isolation and characterization of extracellular vesicles, and osteoblast cultures.

Maryanne Trafani de Melo is graduated in Chemistry by the University of São Paulo-Brazil (2012). Master (2015) and PhD in Science (2019) by the University of São Paulo-Brazil, working at the Center for Nanotechnology and Tissue Engineering. Post Doctorate (2022) at the Department of Chemistry at the Faculty of Philosophy, Sciences and Letters of Ribeirão Preto-USP, Laboratory of Surfaces and Colloids. She is currently a postdoctoral fellow at the University Claude Bernard Lyon 1, Department of Chemistry and Biochemistry (CNPq scholarship). She has developed studies mainly on the following topics: production and characterization of scaffolds, isolation of extracellular vesicles, production of biomaterials by 3D printing, and bone regeneration.

Letícia Fernanda Duffles is a bachelor in Dentistry from the Federal University of Minas Gerais—UFMG (2016), has a Master's degree in Pediatric Dentistry from UFMG as well (2018) and a Ph.D. degree in Pediatric Dentistry from the University of São Paulo – USP (2022). Her main area of interest in research is the alveolar bone remodeling, while her current projects focus on the effect of some proteins in the bone remodeling process.

Colin Farquharson completed his Ph.D. at the University of Aberdeen in 1989. His first postdoctoral position led to a career long interest in skeletal biology. He obtained his chair in Skeletal Biology from the University of Edinburgh in 2010 where he is presently a group leader and Director of Postgraduate Research within the Royal (Dick) School of Veterinary Studies. His research program focusses on the cellular mechanisms underpinning bone and cartilage development. In particular, his research aims are to develop a functional understanding of the role of phosphatases in skeletal mineralisation and was the discoverer of PHOSPHO1. He has published over 170 papers/book chapters and supervised 21 Ph.D. students to completion and is presently supervising eight Ph.D. students as principal or cosupervisor. He is past secretary of the Bone Research Society (UK) and member of the Arthritis Research UK Fellowships Implementation Committee. He is at present Co-Editor-in-Chief for the Journal of Endocrinology and Journal of Molecular Endocrinology.

Bruno Favarin holds a BSc degree in Chemistry (2011), as well as a master's degree (2014) and a doctoral degree (2018) in Biochemistry from the University of São Paulo/FFCLRP. He was postdoctoral fellow at the Department of Physics, Faculty of Philosophy, Sciences, and Letters of Ribeirão Preto (2023) and at the Laboratory of Condensed Matter Chemistry pf Paris (Sorbonne University, CNRS, Collège de France) (2021–22). His research experience is primarily in the field of Biochemistry, with a focus on enzymology and biophysics. His main projects have focus on the production of liposomes and proteoliposomes, biochemical and biophysical characterization of biomimetic membranes and extracellular vesicles, kinetic characterization, bone calcification processes, cell culture, and physicochemical analysis (including atomic force microscopy, FTIR, ATR, X-ray, solid-state and liquid-state NMR, cryo-TEM, differential scanning calorimetry—DSC, and dynamic light scattering—DLS analysis).

Sandra Yasuyo Fukada Graduated in Dentistry from the University of São Paulo (1997), PhD in Pharmacology from the University of São Paulo (2004). Postdoctoral fellow at the Laboratory of Inflammation and Pain at Medical School of Ribeirão Preto—USP (2004–07) and at Division of Immunology, Infection, and Inflammation of the University of Glasgow, Scotland (2008–10). Currently is Associate Professor of Pharmacology at the School of Pharmaceutical Sciences of Ribeirão Preto—University of São Paulo. Leads a research group focused in understanding how inflammation affects osteoclasts formation and bone homeostasis leading to bone loss. In particular, her current project investigates the role of matrix vesicles on osteoclastogenesis and its impact on osteolytic diseases.

Thierry Granjon obtained a PhD in biochemistry in 2001. He is currently Associate Professor-HDR in Biochemistry at the University LYON 1-France. He belongs to the Institute of Chemistry and Molecular and Supramolecular Biochemistry, UMR 5246 CNRS in Lyon where he coordinates the Chemistry of Life axe. He leads the Biological Membrane group within the CO2 team, with expertise in fluorescence probes and spectroscopy. One of his main research activities aims to characterize the organization and dynamics of biological membranes and its functional consequences either directly on cells or on mimetic systems with applications in the field of diagnosis, prognosis, and therapeutic targeting, notably in cancer, joint and pulmonary diseases but also in environment. To carry out these studies, the teams work with cells; extracellular vesicles: exosomes, synovial vesicles, matrix vesicles as well as biomimetic systems (liposomes, lipid monolayers, …), and LNPs as nanovectors.

Larwsk Hayann got his bachelor degree in biomedicine in 2012 and started his scientific career studying myelodysplastic syndrome, a pre-leukemia condition. In 2016, he started his Master's at the University of Porto, Portugal, in biochemistry where he developed his thesis on mathematical modeling involved in prebiotic chemistry and primitive molecular signaling. His thesis was carried out at the Center of Astrobiology, Spain, Madrid, in association with NASA. Moving back to Brazil, Larwsk started his Ph.D. in Chemistry in 2022, and currently, he works with strontium-based nanoparticles applied to biomaterials aiming for bone healing, and his major research interest is to understand how ions (especially strontium) affect at the molecular-level mineralizing cell lines (for example, osteoblasts and odontoblasts). Finally, Larwsk was currently enrolled in Napierala Dobrawa's laboratory at the University of Pittsburgh, PA, USA, where he explored molecular signaling pathways activation and matrix vesicle secretion when stimulated with strontium, calcium, and phosphate.

Shohei Kohno received his first degree in nutrition in 2007 and his PhD in nutrition in 2012. He is currently an assistant professor at the Graduate School of Biomedical and Health Sciences, Hiroshima University. His main research interests are the mechanisms of mineralization in hard and soft tissues. His current projects focus on the pathophysiological role of calciprotein particles in abnormal soft tissue mineralization. In addition, he is currently investigating the novel players in osteoblast differentiation using single cell transcriptome analysis.

David Magne has obtained in 2002 a Ph.D. from the University of Nantes (France), focused on the cellular and molecular mechanisms of pathophysiological mineralization. After being a postdoc for two years in the Division of Rheumatology in Geneva (Switzerland), and an assistant professor for five years in the north of France, he obtained in 2010 a position of full professor of Biochemistry in the Claude Bernard University of Lyon. Since 2016, he leads the MEM2 team (Metabolism, Enzymes, and Molecular Mechanisms) at the Institute of Molecular and Supramolecular Chemistry and Biochemistry (ICBMS, CNRS UMR 5246) in Lyon. His group works on the pathophysiological mechanisms of pathological calcification and ossification. He has recently coordinated an international consortium to determine the impact of microcalcification on atherosclerotic plaque development (ERA-NET CVD), and a national collaborative project to determine the role of plaque ossification on plaque stability in the ApoE-/- mouse model.

Antonio Maurizi graduated in Medical Biotechnology from the University of L'Aquila, where he also obtained a Ph.D. in Experimental Medicine in 2019. He has completed scientific training periods at esteemed research institutions in Europe, the United States, and Australia. He currently serves as an Assistant Professor of Histology at the University of L'Aquila. His research primarily focuses on rare genetic diseases that affect the skeletal system, particularly the study of pathogenetic mechanisms and the development of experimental therapies for Cole-Carpenter syndrome. Additionally, Dr. Maurizi has contributed to the study of cellular mechanisms and the development of experimental therapies for Autosomal Dominant Osteopetrosis Type 2. He is currently involved in projects that investigate the mechanisms that mediate breast cancer cell dormancy in the bone microenvironment.

Saida Mebarek has obtained her PhD degree from INSA Lyon in January 2006 in the laboratory of lipids and membrane physiopathology, INSA-INSERM UMR585. From September 2006 to September 2007, she realized one-year postdoctoral internship at Institute of Developmental Biology of Marseilles (UMR CNRS 6126). In September 2007, she joined the University Claude Bernard Lyon 1 as associate professor and obtained an accreditation to supervise research in September 2015. She was appointed Professor at the University Claude Bernard Lyon 1 and the ICBMS in 2022. Saida Mebarek is interested in the vascular disturbances involved in the metabolic syndrome, in particular, those in chronic renal failure and in atherosclerosis. Her research focuses on the role of lipid signaling, in particular, the involvement of phospholipase D in physiological but also pathological mineralization processes, such as vascular calcification and bone metastasis in prostate cancer. She is also interested in the function of extracellular vesicles (exosomes and matrix vesicles). Her work deals with the mechanisms involved in the initiation of mineralization in the so-called matrix vesicles that are released by hypertrophic chondrocytes, osteoblasts, and odontoblasts that are relevant to understanding pathophysiological conditions such as atherosclerosis, chronic kidney disease, and diabetes.

José Luis Millán received his early training in clinical chemistry and biochemistry at the University of Buenos Aires, Argentina, and joined the La Jolla Cancer Research Foundation (LJCRF) in 1977, the predecessor of Sanford Burnham Prebys Medical Discovery Institute (SBP), as a trainee in clinical enzymology. He completed his Ph.D. studies in Medical Biochemistry at the University of Umeå, Sweden, and after postdoctoral stints in Copenhagen and LJCRF, he was appointed to the faculty at SBP in 1986. He served as Professor of Medical Genetics in the Department of Medical Biosciences at his alma mater, Umeå University, Sweden, from 1995 to 2000. He was appointed Sanford Investigator at the Sanford Children's Health Research Center at SBP in 2008. The Millán laboratory works on understanding the mechanisms that control normal skeletal and dental mineralization and elucidating the pathophysiological abnormalities that lead to heritable soft bones conditions such as hypophosphatasia and to soft-tissue calcification.

Tomoko Minamizaki received her DDS (Hiroshima University) and PhD in Dentistry (Hiroshima University) in 2003 and 2007, respectively. She is currently an Assistant Professor of Calcified Tissue Biology at Hiroshima University Graduate School of Biomedical and Health Sciences. Her research interests include osteoblast differentiation, bone metabolism, phosphate metabolism, ectopic mineralization, and bone metastasis. In particular, her current projects focus on the roles of miRNA as a matrix vesicle cargo released from osteoblasts on bone metabolism and bone metastasis.

Dobrawa Napierala is an Associate Professor at the University of Pittsburgh School of Dental Medicine (Pittsburgh, USA). She earned the Ph.D. degree in Biochemistry from the Institute of Bioorganic Chemistry, Polish Academy of Sciences (Poznan, Poland) and completed a postdoctoral training in molecular and human genetics at Baylor College of Medicine (Houston, TX, USA). Currently, she serves as the codirector of the Pittsburgh Center for Interdisciplinary Bone and Mineral Research and the director of Oral and Craniofacial Sciences graduate program. Dr. Napierala's research is focused on molecular interactions and cellular mechanisms regulating the mineralization process in development and homeostasis of skeletal and dental tissues. She is interested in diseases affecting endochondral ossification, bone mineral density, and tooth development. She studies phosphate signaling in physiologic and pathologic mineralization, molecular mechanisms of biogenesis of matrix vesicles, and molecular networks of the Trps1 transcription factor in skeletal and dental tissues.

Sonoko Narisawa obtained her PhD in Medical Science from Tokai University in 1995. She is a Staff Scientist in Sanford Burnham Medical Discovery Institute in La Jolla, California. Sonoko has produced multiple transgenic and knockout mouse lines to study functions of alkaline phosphatase isozymes and their related molecules. Her study administering a recombinant TNAP to TNAP knockout mice (hypophosphatasia model) was an essential step for the establishment of an enzyme replacement therapy to treat human patients, which is currently only effective treatment for hypophosphatasia. She has been involved in studies using those knockout/transgenic mice to investigate in vivo mineralization and is recently exploring possibility of small molecules as a translational approach. She also worked on spermatogenesis as some of alkaline phosphatases are expressed in germ cells. She has also been interested in intestinal alkaline phosphatases which functions include LPS dephosphorylation and lipid absorption.

Lucas Fabrício Bahia Nogueira is graduated (2016) and obtained his MSc in Chemistry (2019) from University of São Paulo (USP, Brazil). He is currently a Ph.D. student enrolled in both the Ph.D. Program in Biochemistry and Molecular Biology at the University of Rome Tor Vergata (Rome,