Syndecan from embryonic tooth mesenchyme binds tenascin

A stem cell aging framework, from mechanisms to interventions.

Stem cells maintain a subtle balance between self-renew and differentiate under the regulatory network supported by both intracellular and extracellular components. Proteoglycans are large glycoproteins presented abundantly on the cell surface and in the extracellular matrix where they play pivotal roles in facilitating signaling transduction and maintaining stem cell homeostasis. In this review, we outlined the distinct proteoglycans profiles and their functions in the regulation of stem cell homeostasis, as well as recent progresses and prospects of utilizing proteoglycans /glycosaminoglycans as a novel glycomics carrier or bio-active molecules in bone regeneration.

Proteoglycans and Glycosaminoglycans in Stem Cell Homeostasis and Bone Tissue Regeneration

Human in vitro bone models can create the possibility for investigation of physiological bone remodeling while addressing the principle of replacement, reduction and refinement of animal experiments (3R). Current in vitro models lack cell-matrix interactions and their spatiotemporal complexity. To facilitate these analyses, a bone-mimetic template was developed in this study, inspired by bone’s extracellular matrix composition and organization. Silk fibroin (SF) was used as an organic matrix, poly-aspartic acid (pAsp) was used to mimic the functionality of non-collagenous proteins, and 10x simulated body fluid served as mineralization solution. By using pAsp in the mineralization solution, minerals were guided towards the SF material resulting in mineralization inside and as a coating on top of the SF. After cytocompatibility testing, remodeling experiments were performed in which mineralized scaffold remodeling by osteoclasts and osteoblasts was tracked with non-destructive micro-computed tomography and medium analyses over a period of 42 days. The mineralized scaffolds supported osteoclastic resorption and osteoblastic mineralization, in the physiological bone remodeling specific sequence. This model could therefore facilitate the investigation of cell-matrix interactions and may thus reduce animal experiments and advance in vitro drug testing for bone remodeling pathologies like osteoporosis, where cell-matrix interactions need to be targeted.

/pdf/bioinspired-silk-fibroin-mineralization-for-advanced-in-2f8ud292.pdf

Bioinspired Silk Fibroin Mineralization for Advanced In Vitro Bone Remodeling Models

Tooth number abnormality is one of the most common dental developmental diseases, which includes both tooth agenesis and supernumerary teeth. Tooth development is regulated by numerous developmental signals, such as the well-known Wnt, BMP, FGF, Shh and Eda pathways, which mediate the ongoing complex interactions between epithelium and mesenchyme. Abnormal expression of these crutial signalling during this process may eventually lead to the development of anomalies in tooth number; however, the underlying mechanisms remain elusive. In this review, we summarized the major process of tooth development, the latest progress of mechanism studies and newly reported clinical investigations of tooth number abnormality. In addition, potential treatment approaches for tooth number abnormality based on developmental biology are also discussed. This review not only provides a reference for the diagnosis and treatment of tooth number abnormality in clinical practice but also facilitates the translation of basic research to the clinical application. 

/pdf/tooth-number-abnormality-from-bench-to-bedside-24dblnee.pdf

Tooth number abnormality: from bench to bedside

Whole-mount in situ hybridization (WMISH) is a commonly used technique for visualizing the expression profile of mRNAs in embryos. Unlike traditional in situ hybridization techniques, which require thin tissue sections, the WMISH technique allows gene expression patterns to be assessed over the entire embryo and structure. Here, we describe the detailed procedural steps of WMISH, including probe production, embryo fixation and staining, and post-hybridization signal detection. Using this protocol, we visualized highly specific expression patterns of Sonic hedgehog and Bmp4 mRNAs in E12.5 mouse embryos.

Whole-Mount In Situ Hybridization of Mouse Embryos Using DIG-Labeled RNA Probes.

In situ hybridization is a commonly used technique using an antisense RNA probe to localize a specific RNA sequence on histological sections. This approach can visualize the expression pattern of a gene of interest in a portion of tissues. Here, we detail an optimized method for performing in situ hybridization on mouse paraffin sections using digoxigenin (DIG)-labeled RNA probes.

In Situ Hybridization on Mouse Paraffin Sections Using DIG-Labeled RNA Probes.

The extracellular matrix of the bone and dentin contains several non-collagenous proteins (NCPs). One category of NCPs is termed the SIBLING (small integrin-binding ligand, N-linked glycoprotein) family, which includes osteopontin (OPN), bone sialoprotein (BSP), dentin matrix protein 1 (DMP1), dentin sialophosphoprotein (DSPP), etc. These proteins have abundant phosphoserines, aspartic acids, and glutamic acids. In this protocol, we describe the extraction of NCPs from the bone and dentin matrices using guanidine-HCl/EDTA and the isolation of polyanionic SIBLINGs from NCPs using ion-exchange fast protein liquid chromatography (FPLC) to separate the differentially charged proteins into different fractions through a gradient elution by NaCl.

Isolation of SIBLING Proteins from Bone and Dentin Matrices.

The spatiotemporal expression pattern of Syndecans in murine embryonic teeth

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