Influence of Hyaluronic Acid in Periodontal Tissue Regeneration

Hyaluronic acid is a high molecular weight polysaccharide - glycosaminoglycan, which plays a vital role in the functioning of extracellular matrices, including those of mineralized and non-mineralized periodontal tissues. Hyaluronic acid is also important because of its numerous actions in the mechanisms associated with inflammation and the wound healing process. Hyaluronic acid has been identified in all periodontal tissues in varying quantities, being more prominent in the non-mineralized tissues, such as gingiva and periodontal ligament, compared to mineralized tissues, such as the cement and alveolar bone. Preliminary evidence suggests that hyaluronic acid is a very promising candidate as a mediator of periodontal tissue regeneration and periodontal disease treatment, by promoting a rapid remission of symptoms, not only to the marginal gingiva, but also to the deeper seated periodontal tissues. However, further researches for the therapeutic effects of hyaluronic acid in periodontal disease are essential for realization of true benefits of hyaluronic administration in periodontal tissue regeneration. Keywords: Hyaluronic Acid; Gingival Inflammation; Periodontal Disease; Periodontal Reparatio

Can a continuous mineral foam explain the stiffening of aged bone tissue? A micromechanical approach to mineral fusion in musculoskeletal tissues

Recent experimental data revealed a stiffening of aged cortical bone tissue, which could not be explained by common multiscale elastic material models. We explain this data by incorporating the role of mineral fusion via a new hierarchical modeling approach exploiting the asymptotic (periodic) homogenization (AH) technique for three-dimensional linear elastic composites. We quantify for the first time the stiffening that is obtained by considering a fused mineral structure in a softer matrix in comparison with a composite having non-fused cubic mineral inclusions. We integrate the AH approach in the Eshelby-based hierarchical mineralized turkey leg tendon model (Tiburtius et al 2014 Biomech. Model. Mechanobiol. 13 1003–23), which can be considered as a base for musculoskeletal mineralized tissue modeling. We model the finest scale compartments, i.e. the extrafibrillar space and the mineralized collagen fibril, by replacing the self-consistent scheme with our AH approach. This way, we perform a parametric analysis at increasing mineral volume fraction, by varying the amount of mineral that is fusing in the axial and transverse tissue directions in both compartments. Our effective stiffness results are in good agreement with those reported for aged human radius and support the argument that the axial stiffening in aged bone tissue is caused by the formation of a continuous mineral foam. Moreover, the proposed theoretical and computational approach supports the design of biomimetic materials which require an overall composite stiffening without increasing the amount of the reinforcing material

Accumulation, localization, and compartmentation of transforming growth factor beta during endochondral bone development.

Endochondral bone formation was induced in postnatal rats by implantation of demineralized rat bone matrix. Corresponding control tissue was generated by implanting inactive extracted bone matrix, which did not induce bone formation. At various times, implants were removed and sequentially extracted with guanidine hydrochloride, and then EDTA and guanidine hydrochloride. Transforming growth factor beta (TGF beta) in the extracts was quantitated by a radioreceptor assay. TGF beta was present in demineralized bone matrix before implantation, and the concentration had decreased by 1 d after implantation. Thereafter, TGF beta was undetectable by radioreceptor assay until day 9. From day 9-21 the TGF beta was extracted only after EDTA demineralization, indicating tight association with the mineralized matrix. During this time, the content of TGF beta per milligram soluble protein rose steadily and remained high through day 21. This increased concentration correlated with the onset of vascularization and calcification of cartilage. TGF beta was detected only between days 3-9 in the controls; i.e., non-bone-forming implants. Immunolocalization of TGF beta in bone-forming implants revealed staining of inflammatory cells at early times, followed later by staining of chondrocytes in calcifying cartilage and staining of osteoblasts. The most intense staining of TGF beta was found in calcified cartilage and mineralized bone matrix, again indicating preferential compartmentalization of TGF beta in the mineral phase. In contrast to the delayed expression of TGF beta protein, northern blot analysis showed TGF beta mRNA in implants throughout the sequence of bone formation. The time-dependent accumulation of TGF beta when cartilage is being replaced by bone in this in vivo model of bone formation suggests that TGF beta may play a role in the regulation of ossification during endochondral bone development

Colocation and role of polyphosphates and alkaline phosphatase in apatite biomineralization of elasmobranch tesserae

AbstractElasmobranchs (e.g. sharks and rays), like all fishes, grow continuously throughout life. Unlike other vertebrates, their skeletons are primarily cartilaginous, comprising a hyaline cartilage-like core, stiffened by a thin outer array of mineralized, abutting and interconnected tiles called tesserae. Tesserae bear active mineralization fronts at all margins and the tesseral layer is thin enough to section without decalcifying, making this a tractable but largely unexamined system for investigating controlled apatite mineralization, while also offering a potential analog for endochondral ossification. The chemical mechanism for tesserae mineralization has not been described, but has been previously attributed to spherical precursors, and alkaline phosphatase (ALP) activity. Here, we use a variety of techniques to elucidate the involvement of phosphorus-containing precursors in the formation of tesserae at their mineralization fronts. Using Raman spectroscopy, fluorescence microscopy and histological methods, we demonstrate that ALP activity is located with inorganic phosphate polymers (polyP) at the tessera–uncalcified cartilage interface, suggesting a potential mechanism for regulated mineralization: inorganic phosphate (Pi) can be cleaved from polyP by ALP, thus making Pi locally available for apatite biomineralization. The application of exogenous ALP to tissue cross-sections resulted in the disappearance of polyP and the appearance of Pi in uncalcified cartilage adjacent to mineralization fronts. We propose that elasmobranch skeletal cells control apatite biomineralization by biochemically controlling polyP and ALP production, placement and activity. Previous identification of polyP and ALP shown previously in mammalian calcifying cartilage supports the hypothesis that this mechanism may be a general regulating feature in the mineralization of vertebrate skeletons

European sea bass (Dicentrarchus labrax) skin and scale transcriptomes

Fish skin and their appendages, the mineralized scales, are important organs for protection and homeostasis, but little is known about their specific transcript or protein repertoire. This study used RNA-seq to generate transcriptome data for skin and scales in the European sea bass (Dicentrarchus labrax), an important species for fisheries and aquaculture. RNA was extracted from the pectoral skin and from scales collected above the midline of immature one-year old sea bass. More than 20 x 10(6) reads were obtained for each tissue, using RNA-seq Illumina technology. De novo assembly resulted in 31,842 transcripts (of 500 base pairs or greater) for skin and 20,423 transcripts for scale. This dataset provides a useful resource for both aquaculture and fish conservation studies and for research into the physiology and molecular biology of fish skin and scales. (C) 2017 Elsevier B.V. All rights reserved.Foundation for Science and Technology of Portugal (FCT) [PTDC/AAG-GLO/4003/2012, CCMAR/Multi/04326/2013, SFRH/BPD/84033/2012

Improved regeneration and de novo bone formation in a diabetic zebrafish model treated with paricalcitol and cinacalcet

Bone changes related to diabetes have been well stablished, but few strategies have been developed to prevent this growing health problem. In our work, we propose to investigate the effects of calcitriol as well as of a vitamin D analog (paricalcitol) and a calcimimetic (cinacalcet), in fin regeneration and de novo mineralization in a zebrafish model of diabetes. Following exposure of diabetic transgenic Tg(ins: nfsb-mCherry) zebrafish to calcitriol, paricalcitol and cinacalcet, caudal fins were amputated to assess their effects on tissue regeneration. Caudal fin mineralized and regenerated areas were quantified by in vivo alizarin red staining. Quantitative real-time PCR was performed using RNA from the vertebral column. Diabetic fish treated with cinacalcet and paricalcitol presented increased regenerated and mineralized areas when compared with non-treated diabetic group, while no significant increase was observed in nondiabetic fish treated with both drugs. Gene expression analysis showed an up-regulation for runt-related transcription factor 2b (runx2b), bone gamma-carboxyglutamic acid-containing protein (bglap), insulin a (insa) and insulin b (insb) and a trend of increase for sp7 transcription factor (sp7) in diabetic groups treated with cinacalcet and paricalcitol. Expression of insra and vdra was up-regulated in both diabetic and nondiabetic fish treated with cinacalcet. In nondiabetic fish treated with paricalcitol and cinacalcet a similar increase in gene expression could be observed but not so pronounced. The increased mineralization and regeneration in diabetic zebrafish treated with cinacalcet and paricalcitol can be explained by increased osteoblastic differentiation and increased insulin expression indicating pro-osteogenic potential of both drugs.European Regional Development Fund (ERDF) through the COMPETE-Operational Competitiveness ProgramFCT-Fundacao para a Ciencia e a Tecnologia [PEst-CCMAR/Multi/04326/2013]info:eu-repo/semantics/publishedVersio

Nature's conveyor belt - the matrix mediated biomineralization of magnetite in chitons (Mollusca)

Chitons are marine molluscs that use a variety of iron and calcium based minerals to harden their teeth, which they use to scrape algae growing upon, and within, rocks. The teeth are mounted on a long ribbon-like organ termed the radula, with immature, unmineralized teeth at the posterior end and the hardened iron-mineralized teeth at the anterior end (Fig. 1). At any one time, up to 80 individual tooth rows can be observed, with each row becoming progressively mineralized as it moves forward in a conveyor belt-like manner. The ability to study the entire mineralization process in a single animal makes these creatures ideal for the study of matrix mediated biomineralization. The chiton’s ability to mineralize iron has inspired researchers who believe that new biomimetic materials and technologies can be developed based on the principles of biomineral formation

Multiscale approach including microfibril scale to assess elastic constants of cortical bone based on neural network computation and homogenization method

The complexity and heterogeneity of bone tissue require a multiscale modelling to understand its mechanical behaviour and its remodelling mechanisms. In this paper, a novel multiscale hierarchical approach including microfibril scale based on hybrid neural network computation and homogenisation equations was developed to link nanoscopic and macroscopic scales to estimate the elastic properties of human cortical bone. The multiscale model is divided into three main phases: (i) in step 0, the elastic constants of collagen-water and mineral-water composites are calculated by averaging the upper and lower Hill bounds; (ii) in step 1, the elastic properties of the collagen microfibril are computed using a trained neural network simulation. Finite element (FE) calculation is performed at nanoscopic levels to provide a database to train an in-house neural network program; (iii) in steps 2 to 10 from fibril to continuum cortical bone tissue, homogenisation equations are used to perform the computation at the higher scales. The neural network outputs (elastic properties of the microfibril) are used as inputs for the homogenisation computation to determine the properties of mineralised collagen fibril. The mechanical and geometrical properties of bone constituents (mineral, collagen and cross-links) as well as the porosity were taken in consideration. This paper aims to predict analytically the effective elastic constants of cortical bone by modelling its elastic response at these different scales, ranging from the nanostructural to mesostructural levels. Our findings of the lowest scale's output were well integrated with the other higher levels and serve as inputs for the next higher scale modelling. Good agreement was obtained between our predicted results and literature data.Comment: 2

Failure of Mineralized Collagen Microfibrils Using Finite Element Simulation Coupled to Mechanical Quasi-brittle Damage

Bone is a multiscale heterogeneous materiel of which principal function is to support the body structure and to resist mechanical loading and fractures. Bone strength does not depend only on the quantity and quality of bone which is characterized by the geometry and the shape of bones but also on the mechanical proprieties of its compounds, which have a significant influence on its deformation and failure. This work aim to use a 3D nano-scale finite element model coupled to the concept of quasi-brittle damage with the behaviour law isotropic elasticity to investigate the fracture behaviour of composite materiel collagen-mineral (mineralized collagen microfibril). Fracture stress-number of cross-links and damping capacity-number of cross-links curves were obtained under tensile loading conditions at different densities of the mineral phase. The obtained results show that number of cross-links as well as the density of mineral has an important influence on the strength of microfibrils which in turn clarify the bone fracture at macro-scale.Comment: 6; http://www.sciencedirect.com/science/article/pii/S187770581100714