Non-mineralized Tissues Research Articles

Phosphotungstic acid-enhanced micro-computed tomography and RNA sequencing provide a new perspective on temporomandibular joint arthritis induced by complete Freund's adjuvant and collagen-induced arthritis in rat models

Background/PurposeTemporomandibular joint (TMJ) arthritis causes inflammation and degradation of the mandibular condylar cartilage and subchondral bone. Complete Freund's adjuvant (CFA) and collagen-induced arthritis (CIA) are models for studying TMJ arthritis. While micro-computed tomography (micro-CT) is crucial for three-dimensional (3D) bone analysis, it has limitations in imaging nonmineralized tissues. Phosphotungstic acid (PTA) enhances soft tissue contrast. However, research on the 3D imaging of mandibular condylar cartilage and the molecular mechanisms of CFA- and CIA-induced arthritis remains unclear. This study aimed to investigate the bone and PTA-stained cartilage in the mandibular condyle using 3D reconstruction and explore the characteristics of enriched gene ontology terms underlying CFA- and CIA-induced TMJ arthritis in rat models. Materials and methodsRat mandibular condyles were collected from control, CFA, and CIA groups. Live micro-CT created 3D bone structures, and PTA-enhanced micro-CT constructed 3D mandibular condylar cartilage. Gene ontology enrichment analysis identified enriched gene ontology terms from differentially expressed genes through RNA sequencing. ResultsMajor deformities in cartilage volume and bone morphology were observed in the arthritis-induced groups. The CIA group exhibited significant correlations between cartilage volume and bone parameters changes. Gene ontology enrichment analysis indicated fewer terms with upregulated differentially expressed genes related to inflammation and immune response in the CIA group than in the CFA group. ConclusionThis study reveals distinct responses between CFA- and CIA-induced TMJ arthritis models. The CIA group exhibited strong correlations between cartilage volume and bone parameter changes and had less pronounced inflammation and immune response than the CFA group.

Hyaluronic acid in Dentoalveolar regeneration: Biological rationale and clinical applications

BackgroundHyaluronic acid (HA) is found in different locations in the periodontium, including mineralized tissues (i.e., cementum and alveolar bone) and non-mineralized tissues (i.e., gingiva and periodontal ligament). In addition, it seems to play an essential part in regulating the underlying mechanisms involved in tissue inflammatory reactions and wound healing. HA has the potential to regulate periodontal tissue regeneration and treat periodontal disease. AimThe current review of the literature was conducted to assess how HA plays its part in periodontal therapy and examine the contemporary literature's viewpoint on its use in periodontal regeneration. ConclusionHA has a multifunctional character in periodontal regeneration, and healing and appears to provide promising outcomes in different periodontal regenerative applications.

Comparison of Histological, Clinical, and Radiographic Outcomes of Postextraction Ridge Preservation by Allogenic Bone Grafting With and Without Injectable Platelet-Rich Fibrin: A Double-Blinded Randomized Controlled Clinical Trial.

Objectives: This randomized controlled clinical trial compared the histological, clinical, and radiographic outcomes of postextraction ridge preservation by allogenic bone grafting with and without injectable platelet-rich fibrin (I-PRF). Materials and Methods: Twenty single-rooted maxillary and mandibular teeth to be extracted and replaced by dental implants were randomly divided into two groups (n = 10). Cone-beam computed tomography (CBCT) scans were obtained preoperatively to assess bone dimensions and ridge width. The teeth were then extracted, and tooth socket preservation was performed with allograft and collagen type 1 in the control group and allograft, collagen type 1, and I-PRF in the intervention group. CBCT scans were obtained again 3 months after the first stage of surgery, and the second stage of surgery was performed for implant placement, ridge width measurement, and obtaining a biopsy sample. Radiographic bone width, clinical bone width, and radiographic bone height were measured. A histomorphometric method was applied to quantify residual graft material, new bone formation, and nonmineralized tissues. The data were analyzed with Student's t-test and Mann-Whitney U test (α = 0.05). Results: The intervention group showed a significantly smaller reduction in radiographic bone width (P=0.038) and clinical bone width (P=0.033), reduction in radiographic bone height (P=0.213) was not significant. A significantly lower percentage of residual graft particles (P=0.021) and a significantly higher mean percentage of newly formed bone (P=0.038) than the control group. However, the difference in the percentage of nonmineralized tissue (P=0.208) was not significant. Conclusion: Despite the optimal outcome of ridge preservation in both groups, the application of allograft plus I-PRF yielded superior histological, clinical, and radiographic results compared with allograft alone, and this difference was significant in most variables.

Geochemical extraction of ceratopsian remains from ironstone

ABSTRACT Ironstone surrounds many fossils and has a hardness that provides a significant challenge to fossil conservators globally. There are various forms of ironstone, with the carbonate forms of siderite and silicates most often containing vertebrate fossils and potentially preserved soft tissues. The ironstone itself is much harder than the fossils preserved within, leading to the mechanical preparation of the fossils––typically using pin vises or airscribes––being time-consuming and presenting the risk of damage. Existing chemical methods for softening ironstone to prepare the fossils have varied success and also pose a risk of damaging the fossil itself. Here we show that carbonic acid can soften ironstone without damaging permineralized bone or potentially preserved soft tissues. Carbonic acid treatments reduced the hardness of ironstone without causing any significant change in hardness, color, chemical composition, or weight to permineralized ceratopsian bones or a modern cervid bone that retained nonmineralized internal tissues. In addition, we found that solutions of sodium hydroxide and hydrogen peroxide were viable for softening ironstone to remove from the permineralized bone when preserved soft tissue recovery is not a priority. The treatments proposed in this study are important as they are applicable to a range of preparation scenarios, are cost-efficient, are relatively safe to handle, and cause no damage to permineralized fossil bones. Putting these methods into practice can lead to more efficient and safe preparation of fossils in ironstone.

Characterization of the developing axolotl nasal cavity supports multiple evolution of the vertebrate choana.

All tetrapods (mammals, birds, reptiles, and amphibians) share the ability to breathe with their mouths closed due to the formation of choanae, which are openings that allow communication between the nasal and oral cavities. In most fishes, the nasal cavities serve a strictly olfactory function, possessing incurrent and excurrent nares that lie outside of the mouth and therefore, never communicate with the respiratory system. It is not until the evolution of tetrapods, in which the nasal cavities consistently open into the mouth, that they are used both for olfaction and for respiration. However, this developmental transition is poorly understood, with no consensus on the evolutionary origin of the choana in various groups despite decades of debate. Here, we use high-contrast 3D imaging in conjunction with histology and apoptotic cell analysis in non-mineralized embryonic tissues to study the formation of the choana in the axolotl (Ambystoma mexicanum), an aquatic salamander species. We show that the axolotl choana forms from an extension of the embryonic nasal sac, which pushes through intervening mesenchyme and connects with the palate epithelium of the oral cavity, eventually breaking through. This mechanism differs from caecilians, mammals and reptiles, where fusion across a bucconasal groove plays an active role in choana formation. Nevertheless, caecilians, mammals and axolotls converge on the development of a transient epithelial tissue that has to break down in order to develop a patent choana, adding another twist to the intriguing arguments on the evolutionary history of the choana.

A brief overview on the applications of hyaluronic acid in periodontal therapy

Hyaluronic acid (HA) is a component of the extracellular matrix of the non-mineralized and mineralized tissues, including the periodontium. HA have important physiological and biological properties. HA maintains the tissue volume, presents bacteriostatic, anti-inflammatory, osteoinductive, and angiogenetic properties, as well as stimulates many cell functions having a role in wound healing. HA has been used as an adjunctive in the treatment of gingivitis, periodontitis, gingival recessions, or in the reconstruction of papilla deficiencies. In periodontitis patients, the association of HA with subgingival mechanical instrumentation reduces local inflammation and its use with access flap results in larger attachment level gain of infrabony defects compared to control groups non treated with HA. HA application in conjunction with advanced flaps may improve gingival recession levels. HA injection for papillary deficiency seems to be a viable treatment option that improves clinical volume, esthetic outcomes and is associated with minimal pain.

Characterization of pulp calcifications and changes in their composition after treatments with citric acid and ethylenediaminetetraacetic acid solutions.

The ectopic calcifications of non-mineralized tissues can occur in several forms throughout life, such as pulpal calcification. The presence of pulp stones is a challenge in endodontic treatment because they partially or fully obliterate the pulp chamber hindering access to root canals and their subsequent shaping. This study aimed to determine their crystallographic properties and evaluate the capacity of citric acid (CA) and ethylenediaminetetraacetic acid (EDTA) to promote the demineralization of pulp calcifications. The samples were obtained from patients with indications of endodontic treatment, and the radiographic examination was suggestive of pulp stone in at least one permanent tooth. The samples were isolated and analyzed by scanning electron microscopy/energy-dispersive x-ray spectroscopy (SEM/EDX). The Fourier Transform by high resolution-transmission electron microscopy, Raman microscopy, and X-ray diffraction (XRD) were used to identify the mineral phase and crystallographic characteristics. To evaluate the effect of CA and EDTA on the crystallinity of calcifications, they were submerged into these two individual solutions and the changes were assessed in situ by Raman spectroscopy. The SEM images obtained from calcifications demonstrated irregular morphologies. EDX of sample surfaces shows a high presence of oxygen, carbon, calcium, and phosphorous, however, other elements such as sodium, magnesium, nitrogen, chlorine, potassium, sulfur, and zinc were identified in less quantity. According to Raman, XRD, and high-resolution transmission electron microscopy, the predominant mineral phase identified in the pulpal calcification was a poor crystallinity apatite. According to in situ analyses, the effect of CA and EDTA was observed on the signals of PO4 3- and CH2 groups corresponding to inorganic and organic components. The changes with CA were evident at 7min while the effect of EDTA was observed until 15 min of treatment. All results indicate that pulp stones have a heterogeneous composition principally composed of apatite with low crystallinity. The solubility of these pathological minerals is adequate using solutions such as EDTA or CA; however, the effectivity depends on the mineralization grade of calcifications, time, and concentration of exposition to this chemical.

Role of hyaluronic acid in periodontal therapy (Review).

Hyaluronic acid (HA) is essential for the function of extracellular matrices in both hard and soft periodontal components. HA plays an important role in the mechanisms underlying inflammation and wound healing. HA is located in periodontal tissues in differing amounts, including non-mineralized tissues, such as gingiva and periodontal ligament, and lower levels located in mineralized tissues, such as cementum and alveolar bone. According to preliminary findings, HA exhibits potential in the regulation of periodontal tissue regeneration and in the treatment of periodontal disease. HA promotes symptomatic relief in both marginal gingiva and deeper periodontal tissues. The present review aimed to examine the role of HA in periodontal therapy, and investigate the current literature supporting its use in periodontal regeneration.

Characterization of SIBLING Proteins in the Mineralized Tissues.

The SIBLING proteins are a family of non-collagenous proteins (NCPs) previously thought to be expressed only in dentin but have been demonstrated in other mineralized and non-mineralized tissues. They are believed to play vital roles in both osteogenesis and dentinogenesis. Since they are tightly regulated lifelong processes and involve a peak of mineralization, three different age groups were investigated. Fifteen wild-type (WT) mice were euthanized at ages 1, 3, and 6 months. Hematoxylin and eosin staining (H&E) was performed to localize various microscopic structures in the mice mandibles and tibias. The immunostaining pattern was compared using antibodies for dentin sialoprotein (DSP), dentin matrix protein 1 (DMP1), bone sialoprotein (BSP), and osteopontin (OPN). Immunostaining of DSP in tibia showed its most noticeable staining in the 3-month age group. DSP was expressed in alveolar bone, cellular cementum, and PDL. A similar expression of DMP1 was seen in the tibia and dentin. BSP was most noticeably detected in the tibia and acellular cementum. OPN was mainly expressed in the bone. A lower level of OPN was observed at all age groups in the teeth. The immunostaining intensity was the least detected for all proteins in the 6-month tibia sample. The expression patterns of the four SIBLING proteins showed variations in their staining intensity and temporospatial patterning concordant with skeletal and dental maturity. These findings suggest some role in this tightly regulated mineralization process.

Diverse stem cells for periodontal tissue formation and regeneration.

The periodontium is comprised of multiple units of mineralized and nonmineralized tissues including the cementum on the root surface, the alveolar bone, periodontal ligament (PDL), and the gingiva. PDL contains a variety of cell populations including mesenchymal stem/progenitor cells (MSCs) termed PDLSCs, which contribute to periodontal regeneration. Recent studies utilizing mouse genetic models shed light on the identities of these mesenchymal progenitors in their native environment, particularly regarding how they contribute to homeostasis and repair of the periodontium. The current concept is that mesenchymal progenitors in the PDL are localized to the perivascular niche. Single-cell RNA sequencing (scRNA-seq) analyses reveal heterogeneity and cell-type specific markers of cells in the periodontium, as well as their developmental relationship with precursor cells in the dental follicle. The characteristics of PDLSCs and their diversity in vivo are now beginning to be unraveled thanks to insights from mouse genetic models and scRNA-seq analyses, which aid to uncover the fundamental properties of stem cells in the human PDL. The new knowledge will be highly important for developing more effective stem cell-based regenerative therapies to repair periodontal tissues in the future.

Between a rock and a hard place: Regulation of mineralization in the periodontium.

The periodontium supports and attaches teeth via mineralized and nonmineralized tissues. It consists of two, unique mineralized tissues, cementum and alveolar bone. In between these tissues, lies an unmineralized, fibrous periodontal ligament (PDL), which distributes occlusal forces, nourishes and invests teeth, and harbors progenitor cells for dentoalveolar repair. Many unanswered questions remain regarding periodontal biology. This review will focus on recent research providing insights into one enduring mystery: the precise regulation of the hard-soft tissue borders in the periodontium which define the interfaces of the cementum-PDL-alveolar bone structure. We will focus on advances in understanding the molecular mechanisms that maintain the unmineralized PDL "between a rock and a hard place" by regulating the mineralization of cementum and alveolar bone.

Cortical Versus Cancellous Solvent-Dehydrated Bone Allograft for Alveolar Ridge Preservation: A Histologic Study.

Allografts have been routinely used for immediate grafting of extraction sites as modalities of alveolar ridge preservation (ARP). Solvent-dehydrated bone allograft (SDBA), which is commonly utilized for socket grafting, exists in the form of cortical and cancellous particles. This study aims to provide a histologic comparison of cortical and cancellous SDBA for ARP. A total of 35 extraction sockets were allocated to receive either a cortical (17 sites) or cancellous (18 sites) SDBA, followed by application of a resorbable collagen wound dressing in both groups. At approximately 4 months, a bone core biopsy sample was obtained during implant placement. Histomorphometric assessment was then conducted to compare the differences between both forms of SDBA. Within its limitations, a higher percentage of vital bone was observed in the cortical bone group compared to the cancellous bone group (28.6% vs 20.1%, respectively, P = .042), while there was a lack of statistically significant differences among other fractions of the bone biopsy sample (residual graft particles and nonmineralized tissues such as connective tissue or other components).

Single-Cell Transcriptomic Analysis Reveals Developmental Relationships and Specific Markers of Mouse Periodontium Cellular Subsets.

The periodontium is essential for supporting the functionality of the tooth, composed of diversity of mineralized and non-mineralized tissues such as the cementum, the periodontal ligament (PDL) and the alveolar bone. The periodontium is developmentally derived from the dental follicle (DF), a fibrous tissue surrounding the developing tooth bud. We previously showed through in vivo lineage-tracing experiments that DF contains mesenchymal progenitor cells expressing parathyroid hormone-related protein (PTHrP), which give rise to cells forming the periodontal attachment apparatus in a manner regulated by autocrine signaling through the PTH/PTHrP receptor. However, the developmental relationships between PTHrP+ DF cells and diverse cell populations constituting the periodontium remain undefined. Here, we performed single-cell RNA-sequencing (scRNA-seq) analyses of cells in the periodontium by integrating the two datasets, i.e. PTHrP-mCherry+ DF cells at P6 and 2.3kb Col1a1 promoter-driven GFP+ periodontal cells at P25 that include descendants of PTHrP+ DF cells, cementoblasts, osteoblasts and periodontal ligament cells. This integrative scRNA-seq analysis revealed heterogeneity of cells of the periodontium and their cell type-specific markers, as well as their relationships with DF cells. Most importantly, our analysis identified a cementoblast-specific metagene that discriminate cementoblasts from alveolar bone osteoblasts, including Pthlh (encoding PTHrP) and Tubb3. RNA velocity analysis indicated that cementoblasts were directly derived from PTHrP+ DF cells in the early developmental stage and did not interconvert with other cell types. Further, CellPhoneDB cell-cell communication analysis indicated that PTHrP derived from cementoblasts acts on diversity of cells in the periodontium in an autocrine and paracrine manner. Collectively, our findings provide insights into the lineage hierarchy and intercellular interactions of cells in the periodontium at a single-cell level, aiding to understand cellular and molecular basis of periodontal tissue formation.

Iodine staining outperforms phosphotungstic acid in high-resolution micro-CT scanning of post-natal mice cardiac structures.

Purpose: Micro-computed tomography (micro-CT) scan provides high-resolution three-dimensional images of mineralized tissues in small animal models. Contrast enhancement is essential to visualize non-mineralized tissues with micro-CT scan. We attempted to compare the two most common contrast agents to stain and image mouse cardiac structures. Approach: Ex-vivo micro-CT scan images of the mouse hearts were obtained following staining by potassium iodide or phosphotungstic acid (PTA). PTA-stained samples were imaged after various durations following staining (14days, 25days, 187days, and 780days), whereas iodine-stained samples were imaged after 72hours. We compared median staining intensity between PTA and iodine at 0.1-mm intervals from the edge using the Mann Whitney test with correction for multiple comparisons. Results: Sixty post-natal mice hearts were stained with either PTA or iodine and imaged using micro-CT scan. Iodine proved to be faster and more uniform in complete enhancement of cardiac tissue in as short as 72h, whereas PTA required a significantly longer time period to penetrate mouse cardiac structure ( ). Median staining intensity with iodine was strongly higher than that with PTA from 0.1- to 1.5-mm distance from the epicardial edge (2-tailed value or lower throughout). Conclusions: Iodine-stained soft tissue imaging by micro-CT scan provides a non-destructive, efficient, and accurate visualization tool for anatomical analysis of animal heart models of human cardiovascular conditions. Iodine is more efficient compared to PTA to achieve complete murine myocardial staining in a significantly shorter time period.

Guidelines for Micro-Computed Tomography Analysis of Rodent Dentoalveolar Tissues.

ABSTRACTMicro–computed tomography (μCT) has become essential for analysis of mineralized as well as nonmineralized tissues and is therefore widely applicable in the life sciences. However, lack of standardized approaches and protocols for scanning, analyzing, and reporting data often makes it difficult to understand exactly how analyses were performed, how to interpret results, and if findings can be broadly compared with other models and studies. This problem is compounded in analysis of the dentoalveolar complex by the presence of four distinct mineralized tissues: enamel, dentin, cementum, and alveolar bone. Furthermore, these hard tissues interface with adjacent soft tissues, the dental pulp and periodontal ligament (PDL), making for a complex organ. Drawing on others' and our own experience analyzing rodent dentoalveolar tissues by μCT, we introduce techniques to successfully analyze dentoalveolar tissues with similar or disparate compositions, densities, and morphological characteristics. Our goal is to provide practical guidelines for μCT analysis of rodent dentoalveolar tissues, including approaches to optimize scan parameters (filters, voltage, voxel size, and integration time), reproducibly orient samples, define regions and volumes of interest, segment and subdivide tissues, interpret findings, and report methods and results. We include illustrative examples of analyses performed on genetically engineered mouse models with phenotypes in enamel, dentin, cementum, and alveolar bone. The recommendations are designed to increase transparency and reproducibility, promote best practices, and provide a basic framework to apply μCT analysis to the dentoalveolar complex that can also be extrapolated to a variety of other tissues of the body. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC. on behalf of American Society for Bone and Mineral Research.

Spatial survey of non-collagenous proteins in mineralizing and non-mineralizing vertebrate tissues ex vivo

Bone biomineralization is a complex process in which type I collagen and associated non-collagenous proteins (NCPs), including glycoproteins and proteoglycans, interact closely with inorganic calcium and phosphate ions to control the precipitation of nanosized, non-stoichiometric hydroxyapatite (HAP, idealized stoichiometry Ca10(PO4)6(OH)2) within the organic matrix of a tissue. The ability of certain vertebrate tissues to mineralize is critically related to several aspects of their function. The goal of this study was to identify specific NCPs in mineralizing and non-mineralizing tissues of two animal models, rat and turkey, and to determine whether some NCPs are unique to each type of tissue. The tissues investigated were rat femur (mineralizing) and tail tendon (non-mineralizing) and turkey leg tendon (having both mineralizing and non-mineralizing regions in the same individual specimen). An experimental approach ex vivo was designed for this investigation by combining sequential protein extraction with comprehensive protein mapping using proteomics and Western blotting. The extraction method enabled separation of various NCPs based on their association with either the extracellular organic collagenous matrix phases or the inorganic mineral phases of the tissues. The proteomics work generated a complete picture of NCPs in different tissues and animal species. Subsequently, Western blotting provided validation for some of the proteomics findings. The survey then yielded generalized results relevant to various protein families, rather than only individual NCPs. This study focused primarily on the NCPs belonging to the small leucine-rich proteoglycan (SLRP) family and the small integrin-binding ligand N-linked glycoproteins (SIBLINGs). SLRPs were found to be associated only with the collagenous matrix, a result suggesting that they are mainly involved in structural matrix organization and not in mineralization. SIBLINGs as well as matrix Gla (γ-carboxyglutamate) protein were strictly localized within the inorganic mineral phase of mineralizing tissues, a finding suggesting that their roles are limited to mineralization. The results from this study indicated that osteocalcin was closely involved in mineralization but did not preclude possible additional roles as a hormone. This report provides for the first time a spatial survey and comparison of NCPs from mineralizing and non-mineralizing tissues ex vivo and defines the proteome of turkey leg tendons as a model for vertebrate mineralization.

Jawbone remodeling: a conceptual study based on Synchrotron High-resolution Tomography

One of the most important aspects of bone remodeling is the constant turnover mainly driven by the mechanical loading stimulus. The remodeling process produces changes not only in the bone microarchitecture but also in the density distribution of the mineralized matrix - i.e. in calcium concentrations- and in the osteocyte lacunar network. Synchrotron radiation-based X-ray microtomography (microCT) has proven to be an efficient technique, capable to achieve the analysis of 3D bone architecture and of local mineralization at different hierarchical length scales, including the imaging of the lacuno-canalicular network. In the present study, we used microCT within a conceptual study of jawbone remodeling, demonstratively focusing the investigation in two critical contexts, namely in the peri-dental and the peri-implant tissues. The microCT analysis showed that a relevant inhomogeneity was clearly present in both peri-dental and peri-implant biopsies, not only in terms of microarchitecture and mineralization degree, but also considering the lacunar network, i.e. size and numerical density of the osteocyte lacunae. The correlated histological results obtained on the same samples confirmed these observations, also adding information related to non-mineralized tissues. Despite its demonstrative nature, it was concluded that the proposed method was powerful in studying jawbone remodeling because it revealed a direct correlation of its rate with the lacunar density, as achieved by the analysis of the osteocyte lacunar network, and an inverse correlation with the local bone mineral density, as revealed with the Roschger approach.

Bone formation following sinus grafting with an alloplastic biphasic calcium phosphate in Lanyu Taiwanese mini-pigs.

To evaluate the new bone formation after grafting with a synthetic biphasic calcium phosphate in sinuses with minimal bone height, the alloplastic and xenograft materials were compared after grafting into Lanyu Taiwanese mini-pig sinuses via split-mouth design. In six mini-pigs, synthetic hydroxyapatite/tricalcium phosphate (HA/TCP) particles were inserted into one of the sinus cavities using the extra-oral approach, where deproteinized bovine bone mineral (DBBM) particles were placed contralaterally. Fluorescent bony labels of Alizarin and Calcein green were delivered at weeks 4 and 8, respectively. Animals were sacrificed at week12 and the augmented tissues were evaluated by cone-beam computed tomography, microcomputed tomography, and histology. By radiographic examination, the mean thicknesses of sinus cortexes for DBBM and HA/TCP groups were similar (0.35versus 0.38cm) and the mean volumes augmented were also indifferent (1.29versus 1.64 cm3 ). The distributions of bones, residual particles, and non-mineralized tissues in augmented masses between groups were undistinguishable. Under microscopy, however, macroporosities of osteons were filled with HA/TCP residual particles, whereas the newly formed bones lay on top of DBBM particle surfaces. Although the mineral deposition rates between groups were indifferent, the mean labeled surface in the HA/TCP group was significantly greater than those in the DBBM group at week4 (35.16%versus 14.00% for HA/TCP and DBBM, respectively) but less than that at week8 (19.33%versus 39.16%, respectively). Sinus augmentation with synthetic HA/TCP and DBBM exhibited similar effectiveness in new bone formation.

Linkage of Periodontitis and Rheumatoid Arthritis: Current Evidence and Potential Biological Interactions.

The association between rheumatoid arthritis (RA) and periodontal disease (PD) has been the focus of numerous investigations driven by their common pathological features. RA is an autoimmune disease characterized by chronic inflammation, the production of anti-citrullinated proteins antibodies (ACPA) leading to synovial joint inflammation and destruction. PD is a chronic inflammatory condition associated with a dysbiotic microbial biofilm affecting the supporting tissues around the teeth leading to the destruction of mineralized and non-mineralized connective tissues. Chronic inflammation associated with both RA and PD is similar in the predominant adaptive immune phenotype, in the imbalance between pro- and anti-inflammatory cytokines and in the role of smoking and genetic background as risk factors. Structural damage that occurs in consequence of chronic inflammation is the ultimate cause of loss of function and disability observed with the progression of RA and PD. Interestingly, the periodontal pathogen Porphyromonas gingivalis has been implicated in the generation of ACPA in RA patients, suggesting a direct biological intersection between PD and RA. However, more studies are warranted to confirm this link, elucidate potential mechanisms involved, and ascertain temporal associations between RA and PD. This review is mainly focused on recent clinical and translational research intends to discuss and provide an overview of the relationship between RA and PD, exploring the similarities in the immune-pathological aspects and the possible mechanisms linking the development and progression of both diseases. In addition, the current available treatments targeting both RA and PD were revised.

Tuftelin's involvement in embryonic development.

Little is known about tuftelin expression in the developing embryo, previously it was thought to play a role in tooth enamel mineralization. In this study we show tuftelin's spatio-temporal expression in mineralizing and nonmineralizing tissues of the craniofacial complex in the developing mouse embryo. Embryos aged E10.5-E18.5 and newborns aged P3 were used in this study. Polymerase chain reaction (PCR), Real-time PCR, sequencing, and in-situ hybridization were used to detect and quantify messenger RNA (mRNA) expression in different developmental stages. We applied indirect immunohistochemistry and western-blot analyses to investigate protein expression.Two tuftelin mRNA transcripts and a single 64KDa protein were detected throughout embryonic development. Tuftelin was detected in tissues which develop from different embryonic origins; ectoderm, ectomesenchyme, and mesoderm.Tuftelin mRNA and protein were expressed already at E10.5, before the initiation of tooth formation and earlier than previously described. The expression pattern of tuftelin mRNA and protein exhibits dynamic spatio-temporal changes in various tissues.Tuftelin is expressed in neuronal tissues, thus fitting with its described correlation to nerve growth factor. A shift between cytoplasmatic and perinuclear/nuclear expression implies a possible role in regulation of transcription.Recent studies showed tuftelin is induced under hypoxic conditions in-vitro and in-vivo, through the hypoxia-inducible factor 1-α pathway. These results led to the hypothesis that tuftelin is involved in adaptation to hypoxic conditions. The fact that much of mammalian embryogenesis occurs at O 2 concentrations of 1-5%, raises the possibility that tuftelin expression throughout development is due to its role in the adaptivemechanisms in response to hypoxia.