The Experts below are selected from a list of 288 Experts worldwide ranked by ideXlab platform
Ron Shahar - One of the best experts on this subject based on the ideXlab platform.
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High resolution 3D structures of Mineralized Tissues in health and disease
Nature Reviews Endocrinology, 2021Co-Authors: Steve Weiner, Emeline Raguin, Ron ShaharAbstract:A thorough knowledge of the structures of healthy Mineralized Tissues, such as bone or cartilage, is key to understanding the pathological changes occurring during disease. Such knowledge enables the underlying mechanisms that are responsible for pathology to be pinpointed. One high-resolution 3D method in particular — focused ion beam-scanning electron microscopy (FIB-SEM) — has fundamentally changed our understanding of healthy vertebrate Mineralized Tissues. FIB-SEM can be used to study deMineralized matrix, the hydrated components of tissue (including cells) using cryo-fixation and even untreated Mineralized tissue. The latter requires minimal sample preparation, making it possible to study enough samples to carry out studies capable of detecting statistically significant differences — a pre-requisite for the study of pathological Tissues. Here, we present an imaging and characterization strategy for tissue structures at different length scales, describe new insights obtained on healthy Mineralized Tissues using FIB-SEM, and suggest future research directions for both healthy and diseased Mineralized Tissues. Understanding Mineralized Tissues, such as bone and cartilage, in health and disease requires a thorough knowledge of their structures. This Perspective proposes a new imaging and characterization strategy for Mineralized tissue research that utilizes focused ion beam-scanning electron microscopy.
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high resolution 3d structures of Mineralized Tissues in health and disease
Nature Reviews Endocrinology, 2021Co-Authors: Steve Weiner, Emeline Raguin, Ron ShaharAbstract:A thorough knowledge of the structures of healthy Mineralized Tissues, such as bone or cartilage, is key to understanding the pathological changes occurring during disease. Such knowledge enables the underlying mechanisms that are responsible for pathology to be pinpointed. One high-resolution 3D method in particular — focused ion beam-scanning electron microscopy (FIB-SEM) — has fundamentally changed our understanding of healthy vertebrate Mineralized Tissues. FIB-SEM can be used to study deMineralized matrix, the hydrated components of tissue (including cells) using cryo-fixation and even untreated Mineralized tissue. The latter requires minimal sample preparation, making it possible to study enough samples to carry out studies capable of detecting statistically significant differences — a pre-requisite for the study of pathological Tissues. Here, we present an imaging and characterization strategy for tissue structures at different length scales, describe new insights obtained on healthy Mineralized Tissues using FIB-SEM, and suggest future research directions for both healthy and diseased Mineralized Tissues. Understanding Mineralized Tissues, such as bone and cartilage, in health and disease requires a thorough knowledge of their structures. This Perspective proposes a new imaging and characterization strategy for Mineralized tissue research that utilizes focused ion beam-scanning electron microscopy.
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High resolution 3D structures of Mineralized Tissues in health and disease.
Nature reviews. Endocrinology, 2021Co-Authors: Steve Weiner, Emeline Raguin, Ron ShaharAbstract:A thorough knowledge of the structures of healthy Mineralized Tissues, such as bone or cartilage, is key to understanding the pathological changes occurring during disease. Such knowledge enables the underlying mechanisms that are responsible for pathology to be pinpointed. One high-resolution 3D method in particular - focused ion beam-scanning electron microscopy (FIB-SEM) - has fundamentally changed our understanding of healthy vertebrate Mineralized Tissues. FIB-SEM can be used to study deMineralized matrix, the hydrated components of tissue (including cells) using cryo-fixation and even untreated Mineralized tissue. The latter requires minimal sample preparation, making it possible to study enough samples to carry out studies capable of detecting statistically significant differences - a pre-requisite for the study of pathological Tissues. Here, we present an imaging and characterization strategy for tissue structures at different length scales, describe new insights obtained on healthy Mineralized Tissues using FIB-SEM, and suggest future research directions for both healthy and diseased Mineralized Tissues.
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Open questions on the 3D structures of collagen containing vertebrate Mineralized Tissues: A perspective.
Journal of structural biology, 2017Co-Authors: Ron Shahar, Steve WeinerAbstract:Our current understanding of the structures of vertebrate Mineralized Tissues is largely based on light microscopy/histology and projections of 3D structures onto 2D planes using electron microscopy. We know little about the fine details of these structures in 3D at the length scales of their basic building blocks, the inherent variations of structure within a tissue and the cell-extracellular tissue interfaces. This limits progress in understanding tissue formation, relating structure to mechanical and metabolic functions, and obtaining deeper insights into pathologies and the evolution of these Tissues. In this perspective we identify and discuss a series of open questions pertaining to collagen containing vertebrate Mineralized Tissues that can be addressed using appropriate 3D structural determination methods. By so doing we hope to encourage more research into the 3D structures of Mineralized vertebrate Tissues.
Steve Weiner - One of the best experts on this subject based on the ideXlab platform.
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High resolution 3D structures of Mineralized Tissues in health and disease
Nature Reviews Endocrinology, 2021Co-Authors: Steve Weiner, Emeline Raguin, Ron ShaharAbstract:A thorough knowledge of the structures of healthy Mineralized Tissues, such as bone or cartilage, is key to understanding the pathological changes occurring during disease. Such knowledge enables the underlying mechanisms that are responsible for pathology to be pinpointed. One high-resolution 3D method in particular — focused ion beam-scanning electron microscopy (FIB-SEM) — has fundamentally changed our understanding of healthy vertebrate Mineralized Tissues. FIB-SEM can be used to study deMineralized matrix, the hydrated components of tissue (including cells) using cryo-fixation and even untreated Mineralized tissue. The latter requires minimal sample preparation, making it possible to study enough samples to carry out studies capable of detecting statistically significant differences — a pre-requisite for the study of pathological Tissues. Here, we present an imaging and characterization strategy for tissue structures at different length scales, describe new insights obtained on healthy Mineralized Tissues using FIB-SEM, and suggest future research directions for both healthy and diseased Mineralized Tissues. Understanding Mineralized Tissues, such as bone and cartilage, in health and disease requires a thorough knowledge of their structures. This Perspective proposes a new imaging and characterization strategy for Mineralized tissue research that utilizes focused ion beam-scanning electron microscopy.
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high resolution 3d structures of Mineralized Tissues in health and disease
Nature Reviews Endocrinology, 2021Co-Authors: Steve Weiner, Emeline Raguin, Ron ShaharAbstract:A thorough knowledge of the structures of healthy Mineralized Tissues, such as bone or cartilage, is key to understanding the pathological changes occurring during disease. Such knowledge enables the underlying mechanisms that are responsible for pathology to be pinpointed. One high-resolution 3D method in particular — focused ion beam-scanning electron microscopy (FIB-SEM) — has fundamentally changed our understanding of healthy vertebrate Mineralized Tissues. FIB-SEM can be used to study deMineralized matrix, the hydrated components of tissue (including cells) using cryo-fixation and even untreated Mineralized tissue. The latter requires minimal sample preparation, making it possible to study enough samples to carry out studies capable of detecting statistically significant differences — a pre-requisite for the study of pathological Tissues. Here, we present an imaging and characterization strategy for tissue structures at different length scales, describe new insights obtained on healthy Mineralized Tissues using FIB-SEM, and suggest future research directions for both healthy and diseased Mineralized Tissues. Understanding Mineralized Tissues, such as bone and cartilage, in health and disease requires a thorough knowledge of their structures. This Perspective proposes a new imaging and characterization strategy for Mineralized tissue research that utilizes focused ion beam-scanning electron microscopy.
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High resolution 3D structures of Mineralized Tissues in health and disease.
Nature reviews. Endocrinology, 2021Co-Authors: Steve Weiner, Emeline Raguin, Ron ShaharAbstract:A thorough knowledge of the structures of healthy Mineralized Tissues, such as bone or cartilage, is key to understanding the pathological changes occurring during disease. Such knowledge enables the underlying mechanisms that are responsible for pathology to be pinpointed. One high-resolution 3D method in particular - focused ion beam-scanning electron microscopy (FIB-SEM) - has fundamentally changed our understanding of healthy vertebrate Mineralized Tissues. FIB-SEM can be used to study deMineralized matrix, the hydrated components of tissue (including cells) using cryo-fixation and even untreated Mineralized tissue. The latter requires minimal sample preparation, making it possible to study enough samples to carry out studies capable of detecting statistically significant differences - a pre-requisite for the study of pathological Tissues. Here, we present an imaging and characterization strategy for tissue structures at different length scales, describe new insights obtained on healthy Mineralized Tissues using FIB-SEM, and suggest future research directions for both healthy and diseased Mineralized Tissues.
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Open questions on the 3D structures of collagen containing vertebrate Mineralized Tissues: A perspective.
Journal of structural biology, 2017Co-Authors: Ron Shahar, Steve WeinerAbstract:Our current understanding of the structures of vertebrate Mineralized Tissues is largely based on light microscopy/histology and projections of 3D structures onto 2D planes using electron microscopy. We know little about the fine details of these structures in 3D at the length scales of their basic building blocks, the inherent variations of structure within a tissue and the cell-extracellular tissue interfaces. This limits progress in understanding tissue formation, relating structure to mechanical and metabolic functions, and obtaining deeper insights into pathologies and the evolution of these Tissues. In this perspective we identify and discuss a series of open questions pertaining to collagen containing vertebrate Mineralized Tissues that can be addressed using appropriate 3D structural determination methods. By so doing we hope to encourage more research into the 3D structures of Mineralized vertebrate Tissues.
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interactions of matrix proteins from Mineralized Tissues with octacalcium phosphate
Connective Tissue Research, 1994Co-Authors: H Furedimilhofer, Steve Weiner, Janet Moradianoldak, Arthur Veis, K P Mintz, Lia AddadiAbstract:Acidic matrix macromolecules, present in many Mineralized Tissues, including those of vertebrates, are thought to be involved in controlling crystal formation. Little, however, is known about their in vivo functions, particularly in relation to calcium-phosphate-containing crystals. The manner in which a variety of synthetic and natural acidic macromolecules interact in vitro with crystals of octacalcium phosphate (OCP) has been studied. Interactions were assessed by examining changes in morphology of the crystals resulting from preferential interaction of the additive with some crystal faces and not others. Macromolecules rich in acidic amino acids, with or without polysaccharides, such as polyaspartate and mollusk shell proteins respectively, were shown to interact preferentially with rows of Ca ions exposed on the hydrated plate surface of OCP crystals. In contrast, the phosphorylated proteins, phosphophoryn and phosvitin, interacted specifically with the apatite-like motifs on the OCP side faces. BSP ...
Antonio Nanci - One of the best experts on this subject based on the ideXlab platform.
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Calbindin-D9k and calbindin-D28k expression in rat Mineralized Tissues in vivo.
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 2009Co-Authors: Ariane Berdal, D. Hotton, J.l. Saffar, Monique Thomasset, Antonio NanciAbstract:Following their terminal differentiation, highly specialized cells, ameloblasts, odontoblasts, and osteoblasts sequentially elaborate Mineralized Tissues. While the developmental expression pattern of matrix proteins has been studied extensively, less attention has been paid to the molecules involved in calcium handling, such as calcium-binding proteins. This shortcoming, as well as previous conflicting data, led us to conduct studies on calbindin-D 9k and calbindin-D 28k in rat mandibular bone and incisor based on several methods established on rat ameloblasts in vivo. Radioimmunoassays showed that calbindin-D 28k accounts for approximately 0.1% of cytosolic proteins in the ectomesenchymal fraction and 1% in the epithelial fraction of the rat incisor and is 100-fold more concentrated than calbindin-D 9k in both tissue types. Western blot analysis confirmed that the anticalbindin-D 28k reactive species corresponded to the well characterized renal calbindin-D 28k in the ectomesenchyme. In this tissue, calbindin-D 28k was ultrastructurally immunolocalized in the odontoblasts. Quantitative immunocytochemistry showed that labeling was distributed throughout their nucleus and cytoplasm. The similar cytoplasmic distribution of both calbindin-D proteins and mRNAs suggests that their expression is regulated at the subcellular level. In particular, immunoreactive calbindin-D,,, appeared to be associated with rough endoplasmic reticulum. Calbindin-D 9k antisense probe showed negligible labeling in odontoblasts, in parallel with the protein quantities measured (∼10 ng/mg of total protein). Finally, in situ hybridization showed transcripts for both calbindins-D in ameloblasts and also in osteoblasts. In summary, the present results support the concept that an elevated expression of these vitamin D-dependent calcium-binding proteins may characterize the phenotype of cells directly involved in the elaboration of Mineralized Tissues, enamel, dentine, and bone.
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osteopontin an interfacial extracellular matrix protein in Mineralized Tissues
Connective Tissue Research, 1996Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Among the noncollagenous matrix proteins found in Mineralized Tissues (MTs), colloidal-gold immunocytochemistry has demonstrated that the ultrastructural distribution of osteopontin (OPN) is unique in that this protein preferentially accumulates at MT interfaces. In bone, OPN is present as a major component of cell- and matrix-matrix interfacial structures termed laminae limitantes and cement lines, respectively. Here, we review the implications of this distinct, interfacial tissue distribution as it relates to the properties and functional motifs of OPN (e.g. RGD, polyAsp, phosphorylation) in different MTs, and more specifically, how it pertains to current theory on the cellular and extracellular matrix (ECM) events associated with bone remodeling. The production of OPN as one of the earliest, and latest, secretory activities of the osteoblast lineage is discussed, together with a consideration of the role of OPN in cement lines and laminae limitantes in bone and in other normal, pathological and healing MTs such as teeth, kidney stones, bone wound healing and implant osseointegration. Further to its ability to influence cell dynamics, calcification and possibly tissue cohesion in MTs, it is proposed that OPN in cement lines may also promote adhesion between apposing substrata. With regard to this latter function, it is suggested that the molecular interactions within, and biomechanical properties of, such an OPN-rich interfacial zone may be important in minimizing strain-induced fatigue damage and microcrack propagation in bone and across other MT interfaces.
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Secretion of Osteopontin by macrophages and its accumulation at tissue surfaces during wound healing in Mineralized Tissues: a potential requirement for macrophage adhesion and phagocytosis.
The Anatomical record, 1996Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Osteopontin (OPN), a noncollagenous, extracellular matrix sialoprotein found at relatively high levels in both normal and pathological Mineralized Tissues, is expressed by tissue-specific cells in bone, calcified cartilage, and teeth. On the other hand, a hallmark of OPN expression in pathologically mineralizing tissue, and in other soft Tissues experiencing a more generalized type of necrotic injury, is the production of OPN by macrophages at the lesion site. In the present study, we have localized OPN and other noncollagenous proteins by ultrastructural colloidal-gold immunocytochemistry using a rat model in which Mineralized tissue defects are surgically created in mandibular bone and teeth. The healing response was examined by immunocytochemistry and transmission electron microscopy at 10 min, 3 days and 7 days post-surgery using antibodies against OPN, bone sialoprotein, osteocalcin, bone acidic glycoprotein-75, fibronectin, and amelogenin. Whereas most of these proteins were characteristically distributed within their respective extracellular matrices as described previously, OPN was additionally observed to accumulate as a lamina limitans at surgically exposed bone and tooth surfaces, as well as at the surface of particulate, Mineralized tissue debris. Intracellular labeling of the Golgi apparatus and secretory granules of macrophages at the lesion site demonstrated that OPN production by macrophages was a prominent secretory event of the inflammatory response during wound healing in Mineralized Tissues. Pseudopodal and lamellipodal cytoplasmic extensions of macrophages were observed in direct contact with the OPN-containing lamina limitans at these surfaces. Particulate, calcified debris internalized by macrophages also displayed a prominent surface "coating" of OPN. In conclusion, our interpretation of the present data is that OPN secreted by macrophages may serve as a macrophage adhesion protein, and where concentrated at the surface of small particulate, Mineralized tissue debris, may act as an opsonin, thereby facilitating cell adhesion and phagocytosis by macrophages, a process likely mediated by integrin-binding, signal transduction, and cytoskeletal restructuring.
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Postembedding colloidal‐gold immunocytochemistry of noncollagenous extracellular matrix proteins in Mineralized Tissues
Microscopy research and technique, 1995Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Immunocytochemistry is a powerful tool for investigating protein secretion, extracellular matrix assembly, and cell-matrix and matrix-matrix/mineral relationships. When applied to the Tissues of bones (bone and calcified cartilage) and teeth (dentin, cementum, and enamel), where calcium phosphate–containing extracellular matrices are the predominant structural component related to their weight-bearing and masticatory roles, respectively, data from immunocytochemical studies have been prominent in advancing our understanding of Mineralized tissue modeling and remodeling. The present review on the application of postembedding, colloidal-gold immunocytochemistry to Mineralized Tissues focuses on the advantages of this approach and relates them to conceptual, theoretical, and experimental data currently available discussing matrix-mineral interactions and extracellular matrix formation and turnover in these Tissues. More specifically, data are summarized regarding the distribution and role of noncollagenous proteins in different Mineralized Tissues, particularly in the context of how they interface with mineral, and how this relationship might be affected by the various tissue-processing steps and immunocytochemical strategies commonly implemented to examine the distribution and function of tissue proteins. Furthermore, a technical discussion is presented that outlines several different possibilities for epitope exposure in Mineralized Tissues during preparation of thin sections for transmission electron microscopy. Cell biological concepts of protein secretion by cells of the Mineralized Tissues, and subsequent extracellular matrix assembly and organization, are illustrated by examples of high-resolution, colloidal-gold immunolabeling for osteopontin, bone sialoprotein, and osteocalcin in the collagen-based Mineralized Tissues and for enamel protein (amelogenin) in enamel. © 1995 Wiley-Liss, Inc.
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postembedding colloidal gold immunocytochemistry of noncollagenous extracellular matrix proteins in Mineralized Tissues
Microscopy Research and Technique, 1995Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Immunocytochemistry is a powerful tool for investigating protein secretion, extracellular matrix assembly, and cell-matrix and matrix-matrix/mineral relationships. When applied to the Tissues of bones (bone and calcified cartilage) and teeth (dentin, cementum, and enamel), where calcium phosphate–containing extracellular matrices are the predominant structural component related to their weight-bearing and masticatory roles, respectively, data from immunocytochemical studies have been prominent in advancing our understanding of Mineralized tissue modeling and remodeling. The present review on the application of postembedding, colloidal-gold immunocytochemistry to Mineralized Tissues focuses on the advantages of this approach and relates them to conceptual, theoretical, and experimental data currently available discussing matrix-mineral interactions and extracellular matrix formation and turnover in these Tissues. More specifically, data are summarized regarding the distribution and role of noncollagenous proteins in different Mineralized Tissues, particularly in the context of how they interface with mineral, and how this relationship might be affected by the various tissue-processing steps and immunocytochemical strategies commonly implemented to examine the distribution and function of tissue proteins. Furthermore, a technical discussion is presented that outlines several different possibilities for epitope exposure in Mineralized Tissues during preparation of thin sections for transmission electron microscopy. Cell biological concepts of protein secretion by cells of the Mineralized Tissues, and subsequent extracellular matrix assembly and organization, are illustrated by examples of high-resolution, colloidal-gold immunolabeling for osteopontin, bone sialoprotein, and osteocalcin in the collagen-based Mineralized Tissues and for enamel protein (amelogenin) in enamel. © 1995 Wiley-Liss, Inc.
Marc D. Mckee - One of the best experts on this subject based on the ideXlab platform.
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osteopontin an interfacial extracellular matrix protein in Mineralized Tissues
Connective Tissue Research, 1996Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Among the noncollagenous matrix proteins found in Mineralized Tissues (MTs), colloidal-gold immunocytochemistry has demonstrated that the ultrastructural distribution of osteopontin (OPN) is unique in that this protein preferentially accumulates at MT interfaces. In bone, OPN is present as a major component of cell- and matrix-matrix interfacial structures termed laminae limitantes and cement lines, respectively. Here, we review the implications of this distinct, interfacial tissue distribution as it relates to the properties and functional motifs of OPN (e.g. RGD, polyAsp, phosphorylation) in different MTs, and more specifically, how it pertains to current theory on the cellular and extracellular matrix (ECM) events associated with bone remodeling. The production of OPN as one of the earliest, and latest, secretory activities of the osteoblast lineage is discussed, together with a consideration of the role of OPN in cement lines and laminae limitantes in bone and in other normal, pathological and healing MTs such as teeth, kidney stones, bone wound healing and implant osseointegration. Further to its ability to influence cell dynamics, calcification and possibly tissue cohesion in MTs, it is proposed that OPN in cement lines may also promote adhesion between apposing substrata. With regard to this latter function, it is suggested that the molecular interactions within, and biomechanical properties of, such an OPN-rich interfacial zone may be important in minimizing strain-induced fatigue damage and microcrack propagation in bone and across other MT interfaces.
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Secretion of Osteopontin by macrophages and its accumulation at tissue surfaces during wound healing in Mineralized Tissues: a potential requirement for macrophage adhesion and phagocytosis.
The Anatomical record, 1996Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Osteopontin (OPN), a noncollagenous, extracellular matrix sialoprotein found at relatively high levels in both normal and pathological Mineralized Tissues, is expressed by tissue-specific cells in bone, calcified cartilage, and teeth. On the other hand, a hallmark of OPN expression in pathologically mineralizing tissue, and in other soft Tissues experiencing a more generalized type of necrotic injury, is the production of OPN by macrophages at the lesion site. In the present study, we have localized OPN and other noncollagenous proteins by ultrastructural colloidal-gold immunocytochemistry using a rat model in which Mineralized tissue defects are surgically created in mandibular bone and teeth. The healing response was examined by immunocytochemistry and transmission electron microscopy at 10 min, 3 days and 7 days post-surgery using antibodies against OPN, bone sialoprotein, osteocalcin, bone acidic glycoprotein-75, fibronectin, and amelogenin. Whereas most of these proteins were characteristically distributed within their respective extracellular matrices as described previously, OPN was additionally observed to accumulate as a lamina limitans at surgically exposed bone and tooth surfaces, as well as at the surface of particulate, Mineralized tissue debris. Intracellular labeling of the Golgi apparatus and secretory granules of macrophages at the lesion site demonstrated that OPN production by macrophages was a prominent secretory event of the inflammatory response during wound healing in Mineralized Tissues. Pseudopodal and lamellipodal cytoplasmic extensions of macrophages were observed in direct contact with the OPN-containing lamina limitans at these surfaces. Particulate, calcified debris internalized by macrophages also displayed a prominent surface "coating" of OPN. In conclusion, our interpretation of the present data is that OPN secreted by macrophages may serve as a macrophage adhesion protein, and where concentrated at the surface of small particulate, Mineralized tissue debris, may act as an opsonin, thereby facilitating cell adhesion and phagocytosis by macrophages, a process likely mediated by integrin-binding, signal transduction, and cytoskeletal restructuring.
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Postembedding colloidal‐gold immunocytochemistry of noncollagenous extracellular matrix proteins in Mineralized Tissues
Microscopy research and technique, 1995Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Immunocytochemistry is a powerful tool for investigating protein secretion, extracellular matrix assembly, and cell-matrix and matrix-matrix/mineral relationships. When applied to the Tissues of bones (bone and calcified cartilage) and teeth (dentin, cementum, and enamel), where calcium phosphate–containing extracellular matrices are the predominant structural component related to their weight-bearing and masticatory roles, respectively, data from immunocytochemical studies have been prominent in advancing our understanding of Mineralized tissue modeling and remodeling. The present review on the application of postembedding, colloidal-gold immunocytochemistry to Mineralized Tissues focuses on the advantages of this approach and relates them to conceptual, theoretical, and experimental data currently available discussing matrix-mineral interactions and extracellular matrix formation and turnover in these Tissues. More specifically, data are summarized regarding the distribution and role of noncollagenous proteins in different Mineralized Tissues, particularly in the context of how they interface with mineral, and how this relationship might be affected by the various tissue-processing steps and immunocytochemical strategies commonly implemented to examine the distribution and function of tissue proteins. Furthermore, a technical discussion is presented that outlines several different possibilities for epitope exposure in Mineralized Tissues during preparation of thin sections for transmission electron microscopy. Cell biological concepts of protein secretion by cells of the Mineralized Tissues, and subsequent extracellular matrix assembly and organization, are illustrated by examples of high-resolution, colloidal-gold immunolabeling for osteopontin, bone sialoprotein, and osteocalcin in the collagen-based Mineralized Tissues and for enamel protein (amelogenin) in enamel. © 1995 Wiley-Liss, Inc.
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postembedding colloidal gold immunocytochemistry of noncollagenous extracellular matrix proteins in Mineralized Tissues
Microscopy Research and Technique, 1995Co-Authors: Marc D. Mckee, Antonio NanciAbstract:Immunocytochemistry is a powerful tool for investigating protein secretion, extracellular matrix assembly, and cell-matrix and matrix-matrix/mineral relationships. When applied to the Tissues of bones (bone and calcified cartilage) and teeth (dentin, cementum, and enamel), where calcium phosphate–containing extracellular matrices are the predominant structural component related to their weight-bearing and masticatory roles, respectively, data from immunocytochemical studies have been prominent in advancing our understanding of Mineralized tissue modeling and remodeling. The present review on the application of postembedding, colloidal-gold immunocytochemistry to Mineralized Tissues focuses on the advantages of this approach and relates them to conceptual, theoretical, and experimental data currently available discussing matrix-mineral interactions and extracellular matrix formation and turnover in these Tissues. More specifically, data are summarized regarding the distribution and role of noncollagenous proteins in different Mineralized Tissues, particularly in the context of how they interface with mineral, and how this relationship might be affected by the various tissue-processing steps and immunocytochemical strategies commonly implemented to examine the distribution and function of tissue proteins. Furthermore, a technical discussion is presented that outlines several different possibilities for epitope exposure in Mineralized Tissues during preparation of thin sections for transmission electron microscopy. Cell biological concepts of protein secretion by cells of the Mineralized Tissues, and subsequent extracellular matrix assembly and organization, are illustrated by examples of high-resolution, colloidal-gold immunolabeling for osteopontin, bone sialoprotein, and osteocalcin in the collagen-based Mineralized Tissues and for enamel protein (amelogenin) in enamel. © 1995 Wiley-Liss, Inc.
Ariane Berdal - One of the best experts on this subject based on the ideXlab platform.
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Methods for In Situ Protein Visualization in Dental Mineralized Tissues.
Methods in molecular biology (Clifton N.J.), 2019Co-Authors: D. Hotton, Ariane Berdal, A. BolañosAbstract:Immunohistochemistry (IHC) is a technique based on the specificity of antibody-antigen principle used commonly to detect antigens in tissue sections. The immune labeling can be performed in paraffin sections, cryostat sections, and ultrathin sections and can be observed in light confocal and transmission electron microscopy. However, the use of immunohistochemical techniques for the study of Mineralized Tissues has been a challenge for decades (Berdal et al., Arch Oral Biol 36:715-725, 1991; Nanci et al., Eur J Histochem 52:201-214, 2008). Specific procedures are necessary when compared with soft tissue immunohistochemistry. This chapter describes methods for IHC on Tissue-Tek O.C.T. compound and paraffin-embedded sections to detect antigens in the dental Mineralized Tissues.
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in situ hybridization in Mineralized Tissues the added value of lna probes for rna detection
Methods of Molecular Biology, 2019Co-Authors: G Lignon, D. Hotton, Ariane Berdal, A. BolañosAbstract:In situ hybridization (ISH) is one of the fundamental methods in developmental biology and neurobiology. Their first ISH protocols were reported in 1969 (Gall and Pardue, Proc Natl AcadSci USA 63:378-83, 1969). Since several decades, ISH based on the specific hybridization of 100-2000 nucleotides long probes enabled the localization of DNA/RNA sequences in Tissues and cells with high cellular resolution. But sometimes a limited sensitivity notably in Mineralized Tissues (Obernosterer et al., Nature Protocols 2:1508-14, 2007).Here we describe a recent improvement of in situ hybridization efficiency by applying nucleotide locked nucleic acid (LNA)-incorporated oligodeoxynucleotide probes (20 LNA/DNA nucleotide probes) essentially used for noncoding miRNA and messenger RNAs.
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Calbindin-D9k and calbindin-D28k expression in rat Mineralized Tissues in vivo.
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 2009Co-Authors: Ariane Berdal, D. Hotton, J.l. Saffar, Monique Thomasset, Antonio NanciAbstract:Following their terminal differentiation, highly specialized cells, ameloblasts, odontoblasts, and osteoblasts sequentially elaborate Mineralized Tissues. While the developmental expression pattern of matrix proteins has been studied extensively, less attention has been paid to the molecules involved in calcium handling, such as calcium-binding proteins. This shortcoming, as well as previous conflicting data, led us to conduct studies on calbindin-D 9k and calbindin-D 28k in rat mandibular bone and incisor based on several methods established on rat ameloblasts in vivo. Radioimmunoassays showed that calbindin-D 28k accounts for approximately 0.1% of cytosolic proteins in the ectomesenchymal fraction and 1% in the epithelial fraction of the rat incisor and is 100-fold more concentrated than calbindin-D 9k in both tissue types. Western blot analysis confirmed that the anticalbindin-D 28k reactive species corresponded to the well characterized renal calbindin-D 28k in the ectomesenchyme. In this tissue, calbindin-D 28k was ultrastructurally immunolocalized in the odontoblasts. Quantitative immunocytochemistry showed that labeling was distributed throughout their nucleus and cytoplasm. The similar cytoplasmic distribution of both calbindin-D proteins and mRNAs suggests that their expression is regulated at the subcellular level. In particular, immunoreactive calbindin-D,,, appeared to be associated with rough endoplasmic reticulum. Calbindin-D 9k antisense probe showed negligible labeling in odontoblasts, in parallel with the protein quantities measured (∼10 ng/mg of total protein). Finally, in situ hybridization showed transcripts for both calbindins-D in ameloblasts and also in osteoblasts. In summary, the present results support the concept that an elevated expression of these vitamin D-dependent calcium-binding proteins may characterize the phenotype of cells directly involved in the elaboration of Mineralized Tissues, enamel, dentine, and bone.
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expression pattern of dlx3 during cell differentiation in Mineralized Tissues
Bone, 2005Co-Authors: Sonia Ghoulmazgar, D. Hotton, Frederic Lezot, Claudine Blinwakkach, Audrey Asselin, J M Sautier, Ariane BerdalAbstract:The present study was designed to compare the expression pattern of Dlx3 in four different Mineralized Tissues because of: 1-its role in skeleton patterning, 2-its expression in dental epithelium and mesenchyme during morphogenesis, 3-the membranous and endochondral bone and tooth phenotype of tricho-dento-osseous syndrome related to Dlx3 gene mutation and 4-recently emerging knowledge on Dlx family members in the bone field. Ameloblasts, odontoblasts, osteoblasts and chondrocytes were analyzed in vitro and in vivo. Dlx3 transcripts were detected by RT-PCR in established model systems (microdissected dental epithelium and mesenchyme; primary cultures of rat chondrocytes), as recently performed in osteoblasts in vitro. A human 414-bp Dlx3 probe was generated. A 4.5-kb human Dlx3 sense RNA was identified in maxillo-facial samples by Northern blotting. Immunolabeling and in situ hybridization were performed in mice from Theiler stage E 14.5 until birth. In teeth, although Dlx3 was still expressed in differentiated ameloblasts, it was down regulated during odontoblast polarization. During endochondral bone formation, Dlx3 protein was detected in chondrocytes and was most strongly expressed in the prehypertrophic cartilage zone and in differentiating and differentiated osteoblasts of metaphyseal periosteum. In vitro, real-time PCR studies supported this upregulation in prehypertrophic chondrocytes, closely correlated with Ihh variations. In membranous bone, Dlx3 was present in preosteoblasts, osteoblasts and osteoid-osteocytes. The present data on Dlx3 and recently published functional studies show that this transcription factor may be instrumental during growth in the control of matrix deposition and biomineralization in the entire skeleton.
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putative membrane receptor for 1 25 oh 2 vitamin d3 in human Mineralized Tissues during prenatal development
Connective Tissue Research, 2003Co-Authors: Ariane Berdal, M Mesbah, Petros Papagerakis, Ilka NemereAbstract:The calciotropic hormone, 1,25(OH) 2 vitamin D 3 [1,25(OH) 2 D 3 ], controls the formation of dental and bone Mineralized Tissues. The role of nuclear 1,25(OH) 2 D 3 receptor has been extensively studied in the diverse secretory cells, i.e., osteoblasts, chondrocytes, ameloblasts, and odontoblasts. A nongenomic pathway also has been characterized and related to the interactions of 1,25(OH) 2 D 3 ligand with a putative cell membrane receptor. This recognition moiety called 1,25(OH) 2 vitamin D 3 membrane-associated, rapid-response steroid-binding [1,25D 3 -MARRS] protein is investigated here in the craniofacial skeleton of human embryos and fetuses. Immunolocalization using a specific Ab099 against chick intestinal basolateral 1,25D 3 -MARRS protein was performed. The data show a complementary expression pattern of the membrane receptor when compared with published data on the nuclear receptor, notably during amelogenesis. In mandible, membrane receptors for 1,25(OH) 2 D 3 were identified in the heterogeno...