The Use of Immunohistochemistry in Understanding the Structure and Function of the Extracellular Matrix of Dental Tissues

Abstract

The availability of monoclonal and polyclonal antibodies directed toward the recognition of epitopes in a variety of extracellular matrix components of the dentition represents a powerful tool in the investigation of the structure and biology of dental tissues in health and disease. The immunolocalization of both whole molecule structures and specific regions of molecules has the potential to yield information on tooth development, the effects of aging, changes in tooth structure during the initiation and progression of the caries process, together with the response of the tooth to restorative treatment. This review reports on current research to elucidate the role of extracellular matrices of enamel, dentin, cementum, and bone. Attention is directed at the use of antibodies toward the small leucine-rich proteoglycans such as decorin and biglycan, in addition to their glycosaminoglycan chains. Antibodies are also being developed toward dental tissue-specific macromolecules such as phosphophoryn and amelogenin; the use of these antibodies will increase our understanding of the role of these macromolecules in mineralized tissues.

Immunocytochemical analysis of dentin: A double‐labeling technique

“…CS was localized mostly on the peritubular dentin, as previously shown by other studies on rats 9,16,24,25,32 and humans. 8,13,33,34 CFs and CS were visualized as an intricate network of fibrils (from 80 -90 to 20 -30 nm in diameter) and globules, confirming previous morphologic analyses on the intimate spatial and functional relationships between CFs and proteoglycans.…”

“…12 Recent advances in the purification of reagents and the production of monoclonal antibodies permit the establishment of reproducible and selective protocols that allow a very high level of sensibility. 24,25 The main advantage of these immunocytochemical techniques is the possibility of determining the nature of unknown structures observable under high-resolution microscopes, thus revealing the spatial relationships between the molecules of interest.…”

MMP-2 assay within the hybrid layer created by a two-step etch-and-rinse adhesive: Biochemical and immunohistochemical analysis

“…Indeed the antigenicity of a single protein as revealed by its specific binding to a monoclonal antibody provides definitive evidence of an optimal conservation of the epitope structure. 29,30 As phosphoric acid (1%) has a pH value of 1.0, the acidic conditions could unfold and denature a number of protein/enzymes within the dentine matrix and reduce the antibody binding. 31,32 Although an exact description of the denatured states cannot always be specified, 33 any condition where the physical and chemical properties of the protein are different from the designated native state is called a non-active state.…”

Mechanisms of degradation of the hybrid layer in adhesive dentistry and therapeutic agents to improve bond durability—A literature review

“…Collagen fibrils in the hybrid layer that have been incompletely encapsulated by resin monomer can be identified immune-histochemically [39,42]. Advances in reagent purification and the production of highly specific monoclonal antibodies have permitted the creation of reproducible and selective immune-labeling protocols to analyze collagen or proteoglycans [43].…”