Extracellular Matrix Of Soft Tissues Research Articles

Graphene Hybrid Tough Hydrogels with Nanostructures for Tissue Regeneration.

Over the past few decades, hydrogels have attracted considerable attention as promising biomedical materials. However, conventional hydrogels require improved mechanical properties, such as brittleness, which significantly limits their widespread use. Recently, hydrogels with remarkably improved toughness have been developed; however, their low biocompatibility must be addressed. In this study, we developed a tough graphene hybrid hydrogel with nanostructures. The resultant hydrogel exhibited remarkable mechanical properties while representing an aligned nanostructure that resembled the extracellular matrix of soft tissue. Owing to the synergistic effect of the topographical properties, and the enhanced biochemical properties, the graphene hybrid hydrogel had excellent stretchability, resilience, toughness, and biocompatibility. Furthermore, the hydrogel displayed outstanding tissue regeneration capabilities (e.g., skin and tendons). Overall, the proposed graphene hybrid tough hydrogel may provide significant insights into the application of tough hydrogels in tissue regeneration.

Implantable Biologics for Soft Tissue Surgery Reinforcement.

Soft tissue reinforcement focuses on medical grafts that are designed to support and regenerate soft tissue under or near suture lines. Soft tissue is defined as areas of similarly specialized cells that function to connect, support, and surround other structures and organs of the body. These tissues include skin, subcutaneous tissue, fascia, ligaments, tendons, fibrous tissues, fat, synovial membranes, and muscle. Most of the implantable devices used for this purpose are made of collagen, the most abundant protein in mammals and a key component of the extracellular matrix of soft tissues that allows for tissue repair. For suture line reinforcement, exogenous collagen from various sources is implanted under or in continuity with the suture line to allow for increased strength and better healing. First introduced in the field of breast reconstruction, this practice is now also used in hernia repair, dural repair, vaginal slings, amputation reinforcement, tendon repair reinforcement, and even dental soft tissue regeneration.

Advances in antimicrobial hydrogels for dental tissue engineering: regenerative strategies for endodontics and periodontics.

Dental tissue infections have been affecting millions of patients globally leading to pain, severe tissue damage, or even tooth loss. Commercial sterilizers may not be adequate to prevent frequent dental infections. Antimicrobial hydrogels have been introduced as an effective therapeutic strategy for endodontics and periodontics since they have the capability of imitating the native extracellular matrix of soft tissues. Hydrogel networks are considered excellent drug delivery platforms due to their high-water retention capacity. In this regard, drugs or nanoparticles can be incorporated into the hydrogels to endow antimicrobial properties as well as to improve their regenerative potential, once biocompatibility criteria are met avoiding high dosages. Herein, novel antimicrobial hydrogel formulations were discussed for the first time in the scope of endodontics and periodontics. Such hydrogels seem outstanding candidates especially when designed not only as simple volume fillers but also as smart biomaterials with condition-specific adaptability within the dynamic microenvironment of the defect site. Multifunctional hydrogels play a pivotal role against infections, inflammation, oxidative stress, etc. along the way of dental regeneration. Modern techniques (e.g., 3D and 4D-printing) hold promise to develop the next generation of antimicrobial hydrogels together with their limitations such as infeasibility of implantation.

Extracellular vesicles present in bone, blood and extracellular matrix have distinctive characteristics and biologic roles

IntroductionExtracellular vesicles (EV) have long been recognized as an important means of cell to cell communication, but current metrics to delineate various subpopulations of EV are limited. Recently, a distinctive subpopulation of EV embedded within the extracellular matrix of soft tissues, termed matrix-bound nanovesicles (MBV), has been described. Although the lipid membrane composition and intravesicular cargo of MBV clearly differ from liquid phase EV (i.e. exosomes), a more comprehensive characterization of the physical and biologic properties of MBV vs. exosomes and those of a separate subpopulation of EV, specifically bone matrix vesicles, would contribute to our understanding of the biogenesis and physiologic role of these three EV subpopulations. ResultsThe physical characteristics, protein and miRNA cargo profiling, vesicle membrane lipidomics, and immunomodulatory activity were used to compare skeletal muscle-derived MBV, liquid phase plasma exosomes, and mineralization-competent matrix vesicles of provisional bone matrix. We show that despite similar physical characteristics, these three preparations of EV represent distinct entities with different biologic activity. ConclusionThese results inform metrics for the categorization of EV and provide tools for the isolation of EV for potential diagnostic and therapeutic applications.

Nonlinear Mechanical Properties of Prestressed Branched Fibrous Networks

Random fiber networks constitute the solid skeleton of many biological materials such as the cytoskeleton of cells and extracellular matrix of soft tissues. These random networks show unique mechanical properties such as nonlinear shear strain-stiffening and strain softening when subjected to preextension and precompression, respectively. In this study, we perform numerical simulations to characterize the influence of axial prestress on the nonlinear mechanical response of random network structures as a function of their micromechanical and geometrical properties. We build our numerical network models using the microstructure of disordered hexagonal lattices and quantify their nonlinear shear response as a function of uniaxial prestress strain. We consider three different material models for individual fibers and fully characterize their influence on the mechanical response of prestressed networks. Moreover, we investigate both the influence of geometric disorder keeping the network connectivity constant and the influence of the randomness in the stiffness of individual fibers keeping their mean stiffness constant. The effects of network connectivity and bending rigidity of fibers are also determined. Several important conclusions are made, including that the tensile and compressive prestress strains, respectively, increase and decrease the initial network shear stiffness but have no effect on the maximal shear modulus. We discuss the findings in terms of microstructural properties such as the local strain energy distribution.

Heparan Sulfate: A Potential Candidate for the Development of Biomimetic Immunomodulatory Membranes.

Clinical trials have demonstrated that heparan sulfate (HS) could be used as a therapeutic agent for the treatment of inflammatory diseases. Its anti-inflammatory effect makes it suitable for the development of biomimetic innovative strategies aiming at modulating stem cells behavior toward a pro-regenerative phenotype in case of injury or inflammation. Here, we propose collagen type I meshes fabricated by solvent casting and further crosslinked with HS (HS-Col) to create a biomimetic environment resembling the extracellular matrix of soft tissue. HS-Col meshes were tested for their capability to provide physical support to stem cells’ growth, maintain their phenotypes and immunosuppressive potential following inflammation. HS-Col effect on stem cells was investigated in standard conditions as well as in an inflammatory environment recapitulated in vitro through a mix of pro-inflammatory cytokines (tumor necrosis factor-α and interferon-gamma; 20 ng/ml). A significant increase in the production of molecules associated with immunosuppression was demonstrated in response to the material and when cells were grown in presence of pro-inflammatory stimuli, compared to bare collagen membranes (Col), leading to a greater inhibitory potential when mesenchymal stem cells were exposed to stimulated peripheral blood mononuclear cells. Our data suggest that the presence of HS is able to activate the molecular machinery responsible for the release of anti-inflammatory cytokines, potentially leading to a faster resolution of inflammation.

Probing Micromechanical Properties of the Extracellular Matrix of Soft Tissues by Atomic Force Microscopy.

The extracellular matrix (ECM) determines 3D tissue architecture and provides structural support and chemical and mechanical cues to the cells. Atomic force microscopy (AFM) has unique capabilities to measure ECM mechanics at the scale at which cells probe the mechanical features of their microenvironment. Moreover, AFM measurements can be readily combined with bright field and fluorescence microscopy. Performing reliable mechanical measurements with AFM requires accurate calibration of the device and correct computation of the mechanical parameters. A suitable approach to isolate ECM mechanics from cell contribution is removing the cells by means of an effective decellularization process that preserves the composition, structure and mechanical properties of the ECM. AFM measurement of ECM micromechanics provides important insights into organ biofabrication, cell-matrix mechanical crosstalk and disease-induced tissue stiffness alterations. J. Cell. Physiol. 232: 19-26, 2017. © 2016 Wiley Periodicals, Inc.

Is the use of STABHA™ for supplementation of damaged extracellular matrix of soft tissues in the musculoskeletal system an effective treatment of acute injuries and tendinopathies?

Viscosupplementation, or the intra articular administration of hyaluronic acid in order to stabilise synovial fluid chemistry and improve its functional quality, is now a popular therapeutic method whose efficacy, based on numerous published studies, makes it not only a form of symptomatic treatment, but also, to a considerable extent, a cause-oriented treatment. However, a possibly controversial aspect of this therapy is the use of “intra articular hyaluronate” in the treatment of post-traumatic or inflammatory soft-tissue lesions in in the musculoskeletal system. Inappropriate administration of this dosage form to the area of the injured soft tissue (Achilles tendon, periarticular tendon of tarsal joint or knee, tennis elbow, tendinopathy within the rotator cuff, etc.) may not only lead to a failure to achieve the desired therapeutic effect but can even increase the severity of the symptoms, including a rupture of the frayed tendon. The role and importance of hyaluronate in the process of natural regeneration of damaged soft tissue has been demonstrated unequivocally and beyond any doubt. Subsequent research aimed to produce forms of hyaluronic acid that would be characterised by a greater influence and support of the regeneration processes in musculoskeletal soft tissue after an acute or chronic injury, as well as to develop the technology to produce a formulation which would be biocompatible, efficient and adapted to the treatment of ligament and tendon injuries. Following administration, such formulation would also need to be identified by the body as a naturally produced hyaluronate, which plays an essential role in the repair of damaged tissue, beginning with the bleeding phase and involving in all phases of the healing process, as has been demonstrated in numerous scientific studies. The development of a technology for producing hyaluronic acid known as STABHA™ (Soft Tissue Adapted Biocompatible Hyaluronic Acid) in 2008 proved to be a significant achievement in this area. The therapeutic efficacy of STABHA™-containing preparations has been documented in many clinical trials with patients treated for common musculoskeletal disorders and injuries, such as talocrural sprain, so called tennis elbow or rotator cuff tendinopathy. This paper, based on the available literature and the authors’ clinical experience, reviews the therapeutic efficacy of preparations containing STABHA™ used in the treatment of acute and chronic musculoskeletal soft-tissue injuries to enable the reader to make decisions regarding the use of this form of treatment in clinical practice.

From variome to phenome: Pathogenesis, diagnosis and management of ectopic mineralization disorders.

Ectopic mineralization - inappropriate biomineralization in soft tissues - is a frequent finding in physiological aging processes and several common disorders, which can be associated with significant morbidity and mortality. Further, pathologic mineralization is seen in several rare genetic disorders, which often present life-threatening phenotypes. These disorders are classified based on the mechanisms through which the mineralization occurs: metastatic or dystrophic calcification or ectopic ossification. Underlying mechanisms have been extensively studied, which resulted in several hypotheses regarding the etiology of mineralization in the extracellular matrix of soft tissue. These hypotheses include intracellular and extracellular mechanisms, such as the formation of matrix vesicles, aberrant osteogenic and chondrogenic signaling, apoptosis and oxidative stress. Though coherence between the different findings is not always clear, current insights have led to improvement of the diagnosis and management of ectopic mineralization patients, thus translating pathogenetic knowledge (variome) to the phenotype (phenome). In this review, we will focus on the clinical presentation, pathogenesis and management of primary genetic soft tissue mineralization disorders. As examples of dystrophic calcification disorders Pseudoxanthoma elasticum, Generalized arterial calcification of infancy, Keutel syndrome, Idiopathic basal ganglia calcification and Arterial calcification due to CD73 (NT5E) deficiency will be discussed. Hyperphosphatemic familial tumoral calcinosis will be reviewed as an example of mineralization disorders caused by metastatic calcification.

Engineering cell-adhesive gellan gum spongy-like hydrogels for regenerative medicine purposes

The similarity between the extracellular matrix of soft tissue and hydrogels, characterized by high-water-content viscoelastic polymeric networks, has been sustaining the advancement of hydrogels for tissue engineering and regenerative medicine (TERM) purposes. Current research on hydrogels has focused on introducing cell-adhesive peptides to promote cell adhesion and spreading, a critical applicability limitation. Here we report the development of gellan gum (GG) spongy-like hydrogels with ameliorated mechanical performance and flexibility in relation to hydrogels, using a simple and cost-effective method. Most importantly, these materials allow the entrapment of different cell types representing mesenchymal, epidermal and osteoblastic phenotypes that spread within the three-dimensional microstructure. This effect was associated with microstructural rearrangements characterized by pore wall thickening and pore size augmentation, and lower water content than precursor hydrogels. These properties significantly affected protein adsorption once cell adhesion was inhibited in the absence of serum. Spongy-like hydrogels are not adhesive for endothelial cells; however, this issue was surpassed by a pre-incubation with a cell-adhesive protein, as demonstrated for other substrates but not for traditional hydrogels. The proposed cell-compatible GG-based structures avoid time-consuming and expensive strategies that have been used to include cell-adhesive features in traditional hydrogels. This, associated with their off-the-shelf availability in an intermediary dried state, represents unique and highly relevant features for diverse TERM applications.

Multiscale Model Predicts Tissue-Level Failure From Collagen Fiber-Level Damage

Excessive tissue-level forces communicated to the microstructure and extracellular matrix of soft tissues can lead to damage and failure through poorly understood physical processes that are multiscale in nature. In this work, we propose a multiscale mechanical model for the failure of collagenous soft tissues that incorporates spatial heterogeneity in the microstructure and links the failure of discrete collagen fibers to the material response of the tissue. The model, which is based on experimental failure data derived from different collagen gel geometries, was able to predict the mechanical response and failure of type I collagen gels, and it demonstrated that a fiber-based rule (at the micrometer scale) for discrete failure can strongly shape the macroscale failure response of the gel (at the millimeter scale). The model may be a useful tool in predicting the macroscale failure conditions for soft tissues and engineered tissue analogs. In addition, the multiscale model provides a framework for the study of failure in complex fiber-based mechanical systems in general.

In situ forming hydrogels of new amino hyaluronic acid/benzoyl-cysteine derivatives as potential scaffolds for cartilage regeneration

A new chemical strategy is described to link ethylenediamino (EDA) groups to primary hydroxyl groups of hyaluronic acid (HA) and the obtained derivatives have been characterized by 1H-NMR and 13C-NMR analyses. Such HA–EDA derivatives have been exploited to control the functionalization degree in benzoyl-cysteine (BC) groups, chosen as moieties able to allow both self-assembling in aqueous media and an oxidative crosslinking. In particular, the kinetics of oxidation of thiol groups in HA–EDA–BC derivatives has been studied in Dulbecco's Phosphate Buffer Solution (DPBS) pH 7.4 by colorimetric assays and rheological measurements. Mechanical properties of chemical hydrogels obtained after oxidative crosslinking have been evaluated by using two different concentration values (1 and 1.5% w/v) of HA–EDA–BC. These hydrogels show a dense interconnected fibrillar structure similar to the extracellular matrix of soft tissue and a resistance to chemical and enzymatic hydrolysis. Therefore, the potential suitability of HA–EDA–BC hydrogels as scaffolds for cartilage regeneration has been preliminarily assessed using primary human chondrocytes and evaluating their viability and ability to produce extracellular collagen.