Chitosan-Based Biomaterials: Insights into Chemistry, Properties, Devices, and Their Biomedical Applications.

Chitosan is a marine-origin polysaccharide obtained from the deacetylation of chitin, the main component of crustaceans' exoskeleton, and the second most abundant in nature. Although this biopolymer has received limited attention for several decades right after its discovery, since the new millennium chitosan has emerged owing to its physicochemical, structural and biological properties, multifunctionalities and applications in several sectors. This review aims at providing an overview of chitosan properties, chemical functionalization, and the innovative biomaterials obtained thereof. Firstly, the chemical functionalization of chitosan backbone in the amino and hydroxyl groups will be addressed. Then, the review will focus on the bottom-up strategies to process a wide array of chitosan-based biomaterials. In particular, the preparation of chitosan-based hydrogels, organic-inorganic hybrids, layer-by-layer assemblies, (bio)inks and their use in the biomedical field will be covered aiming to elucidate and inspire the community to keep on exploring the unique features and properties imparted by chitosan to develop advanced biomedical devices. Given the wide body of literature that has appeared in past years, this review is far from being exhaustive. Selected works in the last 10 years will be considered.

Bioactive Glass Modified with Zirconium Incorporation for Dental Implant Applications: Fabrication, Structural, Electrical, and Biological Analysis.

Implantology is crucial for restoring aesthetics and masticatory function in oral rehabilitation. Despite its advantages, certain issues, such as bacterial infection, may still arise that hinder osseointegration and result in implant rejection. This work aims to address these challenges by developing a biomaterial for dental implant coating based on 45S5 Bioglass® modified by zirconium insertion. The structural characterization of the glasses, by XRD, showed that the introduction of zirconium in the Bioglass network at a concentration higher than 2 mol% promotes phase separation, with crystal phase formation. Impedance spectroscopy was used, in the frequency range of 102-106 Hz and the temperature range of 200-400 K, to investigate the electrical properties of these Bioglasses, due to their ability to store electrical charges and therefore enhance the osseointegration capacity. The electrical study showed that the presence of crystal phases, in the glass ceramic with 8 mol% of zirconium, led to a significant increase in conductivity. In terms of biological properties, the Bioglasses exhibited an antibacterial effect against Gram-positive and Gram-negative bacteria and did not show cytotoxicity for the Saos-2 cell line at extract concentrations up to 25 mg/mL. Furthermore, the results of the bioactivity test revealed that within 24 h, a CaP-rich layer began to form on the surface of all the samples. According to our results, the incorporation of 2 mol% of ZrO2 into the Bioglass significantly improves its potential as a coating material for dental implants, enhancing both its antibacterial and osteointegration properties.

Extensive Investigation on the Effect of Niobium Insertion on the Physical and Biological Properties of 45S5 Bioactive Glass for Dental Implant.

Dental implants have emerged as one of the most consistent and predictable treatments in the oral surgery field. However, the placement of the implant is sometimes associated with bacterial infection leading to its loss. In this work, we intend to solve this problem through the development of a biomaterial for implant coatings based on 45S5 Bioglass® modified with different amounts of niobium pentoxide (Nb2O5). The structural feature of the glasses, assessed by XRD and FTIR, did not change in spite of Nb2O5 incorporation. The Raman spectra reveal the Nb2O5 incorporation related to the appearance of NbO4 and NbO6 structural units. Since the electrical characteristics of these biomaterials influence their osseointegration ability, AC and DC electrical conductivity were studied by impedance spectroscopy, in the frequency range of 102-106 Hz and temperature range of 200-400 K. The cytotoxicity of glasses was evaluated using the osteosarcoma Saos-2 cells line. The in vitro bioactivity studies and the antibacterial tests against Gram-positive and Gram-negative bacteria revealed that the samples loaded with 2 mol% Nb2O5 had the highest bioactivity and greatest antibacterial effect. Overall, the results showed that the modified 45S5 bioactive glasses can be used as an antibacterial coating material for implants, with high bioactivity, being also non-cytotoxic to mammalian cells.

Preparation of Well-Dispersed Chitosan/Alginate Hollow Multilayered Microcapsules for Enhanced Cellular Internalization.

Hollow multilayered capsules have shown massive potential for being used in the biomedical and biotechnology fields, in applications such as cellular internalization, intracellular trafficking, drug delivery, or tissue engineering. In particular, hollow microcapsules, developed by resorting to porous calcium carbonate sacrificial templates, natural-origin building blocks and the prominent Layer-by-Layer (LbL) technology, have attracted increasing attention owing to their key features. However, these microcapsules revealed a great tendency to aggregate, which represents a major hurdle when aiming for cellular internalization and intracellular therapeutics delivery. Herein, we report the preparation of well-dispersed polysaccharide-based hollow multilayered microcapsules by combining the LbL technique with an optimized purification process. Cationic chitosan (CHT) and anionic alginate (ALG) were chosen as the marine origin polysaccharides due to their biocompatibility and structural similarity to the extracellular matrices of living tissues. Moreover, the inexpensive and highly versatile LbL technology was used to fabricate core-shell microparticles and hollow multilayered microcapsules, with precise control over their composition and physicochemical properties, by repeating the alternate deposition of both materials. The microcapsules' synthesis procedure was optimized to extensively reduce their natural aggregation tendency, as shown by the morphological analysis monitored by advanced microscopy techniques. The well-dispersed microcapsules showed an enhanced uptake by fibroblasts, opening new perspectives for cellular internalization.

Unraveling the effect of the hydration level on the molecular mobility of nanolayered polymeric systems.

This work investigates the influence of the hydration level on the molecular mobility and glass transition dynamics of freestanding chitosan/alginate (CHT/ALG) nanolayered systems. Nonconventional dynamic mechanical analysis identifies two relaxation processes assigned to the α-relaxation of the two biopolymers, respectively, CHT and ALG, when immersed in water/ethanol mixtures. This phenomenon explains the shape memory properties of the multilayered systems induced by hydration, thus constituting promising smart materials that would be of paramount importance in a plethora of research fields, including in the biomedical and biotechnological fields.

An alternative hybrid lipid nanosystem combining cytotoxic and magnetic properties as a tool to potentiate antitumor effect of 5-fluorouracil.

AIMS: Colorectal cancer is the third most frequent type of cancer and the second leading cause of cancer-related deaths worldwide. The majority of cases are diagnosed at a later stage, leading to the need for more aggressive treatments such as chemotherapy. 5-Fluorouracil (5-FU), known for its high cytotoxic properties has emerged as a chemotherapeutic agent. However, it presents several drawbacks such as lack of specificity and short half-life. To reduce these drawbacks, several strategies have been designed namely chemical modification or association to drug delivery systems. MATERIALS AND METHODS: Current research was focused on the design, physicochemical characterization and in vitro evaluation of a lipid-based system loaded with 5-FU. Furthermore, aiming to maximize preferential targeting and release at tumour sites, a hybrid lipid-based system, combining both therapeutic and magnetic properties was developed and validated. For this purpose, liposomes co-loaded with 5-FU and iron oxide (II, III) nanoparticles were accomplished. KEY FINDINGS: The characterization of the developed nanoformulation was performed in terms of incorporation parameters, mean size and surface charge. In vitro studies assessed in a murine colon cancer cell line confirmed that 5-FU antiproliferative activity was preserved after incorporation in liposomes. In same model, iron oxide (II, III) nanoparticles did not exhibit cytotoxic properties. Additionally, the presence of these nanoparticles was shown to confer magnetic properties to the liposomes, allowing them to respond to external magnetic fields. SIGNIFICANCE: Overall, a lipid nanosystem loading a chemotherapeutic agent displaying magnetic characteristics was successfully designed and physicochemically characterized, for further in vivo applications.

Recent Developments in Layer-by-Layer Assembly for Drug Delivery and Tissue Engineering Applications.

Surfaces with biological functionalities are of great interest for biomaterials, tissue engineering, biophysics, and for controlling biological processes. The layer-by-layer (LbL) assembly is a highly versatile methodology introduced 30 years ago, which consists of assembling complementary polyelectrolytes or biomolecules in a stepwise manner to form thin self-assembled films. In view of its simplicity, compatibility with biological molecules, and adaptability to any kind of supporting material carrier, this technology has undergone major developments over the past decades. Specific applications have emerged in different biomedical fields owing to the possibility to load or immobilize biomolecules with preserved bioactivity, to use an extremely broad range of biomolecules and supporting carriers, and to modify the film's mechanical properties via crosslinking. In this review, the focus is on the recent developments regarding LbL films formed as 2D or 3D objects for applications in drug delivery and tissue engineering. Possible applications in the fields of vaccinology, 3D biomimetic tissue models, as well as bone and cardiovascular tissue engineering are highlighted. In addition, the most recent technological developments in the field of film construction, such as high-content liquid handling or machine learning, which are expected to open new perspectives in the future developments of LbL, are presented.

Natural Polymer-Polyphenol Bioadhesive Coacervate with Stable Wet Adhesion, Antibacterial Activity, and On-Demand Detachment.

Medical adhesives are emerging as an important clinical tool as adjuvants for sutures and staples in wound closure and healing and in the achievement of hemostasis. However, clinical adhesives combining cytocompatibility, as well as strong and stable adhesion in physiological conditions, are still in demand. Herein, a mussel-inspired strategy is explored to produce adhesive coacervates using tannic acid (TA) and methacrylate pullulan (PUL-MA). TA|PUL-MA coacervates mainly comprise van der Waals forces and hydrophobic interactions. The methacrylic groups in the PUL backbone increase the number of interactions in the adhesives matrix, resulting in enhanced cohesion and adhesion strength (72.7 Jm-2), compared to the non-methacrylated coacervate. The adhesive properties are kept in physiologic-mimetic solutions (72.8 Jm-2) for 72 h. The photopolymerization of TA|PUL-MA enables the on-demand detachment of the adhesive. The poor cytocompatibility associated with the use of phenolic groups is here circumvented by mixing reactive oxygen species-degrading enzyme in the adhesive coacervate. This addition does not hamper the adhesive character of the materials, nor their anti-microbial or hemostatic properties. This affordable and straightforward methodology, together with the tailorable adhesivity even in wet environments, high cytocompatibility, and anti-bacterial activity, enables foresee TA|PUL-MA as a promising ready-to-use bioadhesive for biomedical applications.

Fabrication, Structural and Biological Characterization of Zinc-Containing Bioactive Glasses and Their Use in Membranes for Guided Bone Regeneration.

Polymeric membranes are widely used in guided bone regeneration (GBR), particularly in dentistry. In addition, bioactive glasses can be added to the polymers in order to develop a matrix that is osteoconductive and osteoinductive, increasing cell adhesion and proliferation. The bioactive glasses allow the insertion into its network of therapeutic ions in order to add specific biological properties. The addition of zinc into bioactive glasses can promote antibacterial activity and induce the differentiation and proliferation of the bone cells. In this study, bioactive glasses containing zinc (0.25, 0.5, 1 and 2 mol%) were developed and structurally and biologically characterized. The biological results show that the Zn-containing bioactive glasses do not present significant antibacterial activity, but the addition of zinc at the highest concentration does not compromise the bioactivity and promotes the viability of Saos-2 cells. The cell culture assays in the membranes (PCL, PCL:BG and PCL:BGZn2) showed that zinc addition promotes cell viability and an increase in alkaline phosphatase (ALP) production.

Bioactivity Enhancement of Plasma-Sprayed Hydroxyapatite Coatings through Non-Contact Corona Electrical Charging.

Atmospheric plasma spray (APS) remains the only certified industrial process to produce hydroxyapatite (Hap) coatings on orthopaedic and dental implants intended for commercialization. Despite the established clinical success of Hap-coated implants, such as hip and knee arthroplasties, a concern is being raised regarding the failure and revision rates in younger patients, which are increasing rapidly worldwide. The lifetime risk of replacement for patients in the 50-60 age interval is about 35%, which is significantly higher than 5% for patients aged 70 or older. Improved implants targeted at younger patients are a necessity that experts have been alerted to. One approach is to enhance their bioactivity. For this purpose, the method with the most outstanding biological results is the electrical polarization of Hap, which remarkably accelerates implant osteointegration. There is, however, the technical challenge of charging the coatings. Although this is straightforward on bulk samples with planar faces, it is not easy on coatings, and there are several problems regarding the application of electrodes. To the best of our knowledge, this study demonstrates, for the first time, the electrical charging of APS Hap coatings using a non-contact, electrode-free method: corona charging. Bioactivity enhancement is observed, establishing the promising potential of corona charging in orthopedics and dental implantology. It is found that the coatings can store charge at the surface and bulk levels up to high surface potentials (>1000 V). The biological in vitro results show higher Ca2+ and P5+ intakes in charged coatings compared to non-charged coatings. Moreover, a higher osteoblastic cellular proliferation is promoted in the charged coatings, indicating the promising potential of corona-charged coatings when applied in orthopedics and dental implantology.

Engineering dual-stimuli responsive poly(vinyl alcohol) nanofibrous membranes for cancer treatment by magnetic hyperthermia.

The development of new cancer treatment options, such as multifunctional devices, allows for a more personalized treatment, avoiding the known severe side effects of conventional options. In this context, on-demand drug delivery systems can actively control the rate of drug release offering a precise control of treatment. Magnetically and thermally controlled drug delivery systems have been explored as on-demand devices to treat chronic diseases and cancer tumors. In the present work, dual-stimuli responsive systems were developed by incorporating Fe3O4 magnetic nanoparticles (NPs) and poly(N-isopropylacrylamide) (PNIPAAm) microgels into electrospun polymeric fibers for application in cancer treatment. First, Fe3O4 NPs with an average diameter of 8 nm were synthesized by chemical precipitation technique and stabilized with dimercaptosuccinic acid (DMSA) or oleic acid (OA). PNIPAAm microgels were synthesized by surfactant-free emulsion polymerization (SFEP). Poly(vinyl alcohol) (PVA) was used as a fiber template originating fibers with an average diameter of 179 ± 14 nm. Stress tests of the membranes showed that incorporating both microgels and Fe3O4 NPs in electrospun fibers increases their Young's modulus. Swelling assays indicate that PVA membranes have a swelling ratio of around 3.4 (g/g) and that the presence of microgels does not affect its swelling ability. However, with the incorporation of Fe3O4 NPs, the swelling ratio of the membranes decreases. Magnetic hyperthermia assays show that a higher concentration of NPs leads to a higher heating ability. The composite membrane with the most promising results is the one incorporated with DMSA-coated NPs, since it shows the highest temperature variation, 5.1 °C. To assess the membranes biocompatibility and ability to promote cell proliferation, indirect and direct contact cell viability assays were performed, as well as cell adhesion assays. Following an extract method viability assay, all membrane designs did not reveal cytotoxic effects on dermal fibroblasts and melanoma cancer cells, after 48 h exposure and support long-term viability. The present work demonstrates the potential of dual-stimuli composite membranes for magnetic hyperthermia and may in the future be used as an alternative cancer treatment particularly in anatomically reachable solid tumors.

Marine-origin polysaccharides-based free-standing multilayered membranes as sustainable nanoreservoirs for controlled drug delivery.

The layer-by-layer (LbL) assembly technology has been widely used to functionalise surfaces and precisely engineer robust multilayered bioarchitectures with tunable structures, compositions, properties, and functions at the nanoscale by resorting to a myriad of building blocks exhibiting complementary interactions. Among them, marine-origin polysaccharides are a sustainable renewable resource for the fabrication of nanostructured biomaterials for biomedical applications owing to their wide bioavailability, biocompatibility, biodegradability, non-cytotoxicity, and non-immunogenic properties. Chitosan (CHT) and alginate (ALG) have been widely employed as LbL ingredients to shape a wide repertoire of size- and shape-tunable electrostatic-driven multilayered assemblies by exploring their opposite charge nature. However, the insolubility of CHT in physiological conditions intrinsically limits the range of bioapplications of the as-developed CHT-based LbL structures. Herein, we report the preparation of free-standing (FS) multilayered membranes made of water-soluble quaternised CHT and ALG biopolymers for controlled release of model drug molecules. The influence of the film structure in the drug release rate is studied by assembling two distinct set-ups of FS membranes, having the model hydrophilic drug fluorescein isothiocyanate-labelled bovine serum albumin (FITC-BSA) either as an intrinsic building block or added as an outer layer after the LbL assembly process. Both FS membranes are characterised for their thickness, morphology, in vitro cytocompatibility, and release profile, with those having FITC-BSA as an intrinsic LbL ingredient denoting a more sustained release rate. This work opens up new avenues for the design and development of a wide array of CHT-based devices for biomedical applications, overcoming the limitations associated with the insolubility of native CHT under physiological conditions.

Bioactive Glasses Containing Strontium or Magnesium Ions to Enhance the Biological Response in Bone Regeneration.

The non-surgical treatments are being required to reconstruct damaged tissue, prioritizing our body's natural healing process. Thus, the use of bioactive materials such as bioactive glass has been studied to support the repair and restoration of hard and soft tissue. Thus, in this work Bioglass 45S5 was developed, adding 1 and 2%mol of SrO or MgO and the physical and biological properties were evaluated. The addition of MgO and SrO at the studied concentrations promoted the slight increase in non-bridging oxygens number, observed through the temperature shift in phase transitions to lower values compared to Bioglass 45S5. The insertion of the ions also showed a positive effect on Saos-2 cell viability, decreasing the cytotoxic of Bioglass 45S5. Besides the Ca/P ratio on the pellets surface demonstrating no evidence of higher reactivity between Bioglass 45S5 and Bioglass with Sr and Mg, micrographs show that at 24 h the Ca/P rich layer is denser than in Bioglass 45S5 after the contact with simulated body fluid. The samples with Sr and Mg show a higher antibacterial effect compared to Bioglass 45S5. The addition of the studied ions may benefit the biological response of Bioglass 45S5 in dental applications as scaffolds or coatings.

Supramolecular presentation of bioinstructive peptides on soft multilayered nanobiomaterials stimulates neurite outgrowth.

Peptide amphiphiles (PAs) have emerged as effective molecular building blocks for creating self-assembling nanobiomaterials for multiple biomedical applications. Herein, we report a straightforward approach to assemble soft bioinstructive platforms to recreate the native neural extracellular matrix (ECM) aiming for neuronal regeneration based on the electrostatic-driven supramolecular presentation of laminin-derived IKVAV-containing self-assembling PA (IKVAV-PA) on biocompatible multilayered nanoassemblies. Spectroscopic and microscopic techniques show that the co-assembly of positively charged low-molecular-weight IKVAV-PA with oppositely charged high-molecular-weight hyaluronic acid (HA) triggers the formation of ordered ß-sheet structures denoting a one-dimensional nanofibrous network. The successful functionalization of poly(L-lysine)/HA layer-by-layer nanofilms with an outer positively charged layer of self-assembling IKVAV-PA is demonstrated by the quartz crystal microbalance with dissipation monitoring and the nanofibrous morphological properties revealed by atomic force microscopy. The bioactive ECM-mimetic supramolecular nanofilms promote the enhancement of primary neuronal cells' adhesion, viability, and morphology when compared to the PA without the IKVAV sequence and PA-free biopolymeric multilayered nanofilms, and stimulate neurite outgrowth. The nanofilms hold great promise as bioinstructive platforms for enabling the assembly of customized and robust multicomponent supramolecular biomaterials for neural tissue regeneration.

Retrievable metal ceramic implant-supported fixed prostheses with milled titanium frameworks and all-ceramic crowns: retrospective clinical study with up to 10 years of follow-up.

PURPOSE: The purpose of this study was to report on the outcome of metal ceramic implant-supported fixed prostheses with milled titanium frameworks and all-ceramic crowns. MATERIALS AND METHODS: The clinical study included 108 patients (67 women, 41 men), mean age of 58.6 years (range: 34-82), followed between 9 months and 10 years (post occlusal loading). The mean follow-up time for all patients in the study was 5 years. A total of 125 prostheses were fabricated. The data were divided into 2 groups. Development group (DG): 52 patients with 66 prostheses (28 maxillary, 38 mandibular) fabricated with individual Procera crowns (Alumina copings, Nobel Biocare AB) and Allceram ceramics (Ducera Dental GmbH) cemented onto a CAD/CAM fabricated Ti framework (Nobel Biocare AB) with pink ceramic (Duceram, Ducera Dental GmbH) that replicated the missing gingival tissues. Routine group (RG): 56 patients with 59 prostheses (49 maxillary, 10 mandibular) fabricated with individual Procera crowns (Zirconia copings and Nobel Rondo Zirconia Ceramic; Nobel Biocare AB) cemented onto a CAD/CAM fabricated Ti framework (Nobel Biocare AB) with pink acrylic resin (PallaXpress Ultra, Heraeus Kulzer GmbH) that replicated the missing gingival tissues. Primary outcome measures were prosthetic survival and mechanical complications. Secondary outcome measures were biological complications testing the retrievability characteristic of the prosthesis. Survival estimates were calculated on the patient level with the Kaplan-Meier product limit estimator (95% confidence intervals [CI]). Data were analyzed with descriptive and inferential analyses. RESULTS: The cumulative survival rates for the implant-supported fixed prostheses were 92.4% for the DG at 10 years and 100% for the RG at 5 years (overall 96%) (Kaplan-Meier). Mechanical complications occurred in 44 patients (DG: 29 patients, 36 prostheses; RG: 15 patients, 16 prostheses); the large majority were crown fractures, occurring in 48 patients (DG: 33 patients, 36 prostheses; RG: 15 patients, 16 prostheses). In the DG, univariate analysis of logistic regression disclosed the presence of a metal ceramic implant-supported fixed prosthesis opposing dentition as a risk factor for crown fracture (OR = 1.97). Biological complications occurred in 33 patients (DG: 18 patients; RG: 15 patients), the majority being peri-implant pathologies in 19 patients (DG: 9 patients, RG: 10 patients). All situations were resolved except one in the DG that led to fixture and prosthesis loss. CONCLUSIONS: The results of this study indicated that, within the limitations of this study, the CAD/CAM protocol is acceptable for definitive prosthetic rehabilitation. This protocol provided these patients with a good prognosis on a middle- to long-term basis (5 years).

Biocompatibility, Bioactivity, and Antibacterial Behaviour of Cerium-Containing Bioglass®.

The main reason for the increased use of dental implants in clinical practice is associated with aesthetic parameters. Implants are also presented as the only technique that conserves and stimulates natural bone. However, there are several problems associated with infections, such as peri-implantitis. This disease reveals a progressive inflammatory action that affects the hard and soft tissues surrounding the implant, leading to implant loss. To prevent the onset of this disease, coating the implant with bioactive glasses has been suggested. In addition to its intrinsic function of promoting bone regeneration, it is also possible to insert therapeutic ions, such as cerium. Cerium has several advantages when the aim is to improve osseointegration and prevent infectious problems with dental implant placement. It promotes increased growth and the differentiation of osteoblasts, improves the mechanical properties of bone, and prevents bacterial adhesion and proliferation that may occur on the implant surface. This antibacterial effect is due to its ability to disrupt the cell wall and membrane of bacteria, thus interfering with vital metabolic functions such as respiration. In addition, its antioxidant effect reverses oxidative stress after implantation in bone. In this work, Bioglass 45S5 with CeO2 with different percentages (0.25, 0.5, 1, and 2 mol%) was developed by the melt-quenching method. The materials were analyzed in terms of morphological, structural, and biological (cytotoxicity, bioactivity, and antibacterial activity) properties. The addition of cerium did not promote structural changes to the bioactive glass, which shows no cytotoxicity for the Saos-2 cell line up to 25 mg/mL of extract concentration for all cerium contents. For the maximum cerium concentration (2 mol%) the bioactive glass shows an evident inhibitory effect for Escherichia coli and Streptococcus mutans bacteria. Furthermore, all samples showed the beginning of the deposition of a CaP-rich layer on the surface of the material after 24 h.

Erysipelas in a stranded common bottlenose dolphin: first report in a South American odontocete.

Erysipelas is a zoonotic disease caused by Erysipelothrix rhusiopathiae. In cetaceans, this disease has two main clinical forms: a cutaneous one, grossly characterized by rhomboid lesions, and a septicemic and often fatal form. Erysipelas is considered an important cause of morbidity and mortality in captive cetaceans; however, information in free-ranging cetaceans is limited. An adult common bottlenose dolphin (Tursiops truncatus) was found dead and in advanced autolysis in Paraíba state, northeastern Brazil, on July 19th, 2020. Upon gross examination, 80% of the body surface presented disseminated rhomboid cutaneous lesions ranging from 4 to 6 cm-width, characterized by well-defined edges and occasional ulceration, consistent with erysipelas. Additionally, anthropic-made postmortem linear cuts and partial mechanical removal of the flank musculature were noted. Skin samples were collected for histopathologic and molecular analyses. Microscopically, it was possible to observe multifocal dermatitis with vasculitis. Erysipelothrix sp. was detected by PCR. Despite previous reports of human consumption of cetacean meat in northeastern Brazil, the observed marks and advanced carcass autolysis suggested that the animal was most likely used as bait for fishing instead of human intake. This case highlights the value of postmortem examination and PCR even in poorly preserved cadavers and contributes to the understanding of the epidemiology of cutaneous erysipelas in free-ranging cetaceans (first report in an odontocete from the Southern Hemisphere). Due to the zoonotic potential of certain Erysipelothrix species (i.e., E. rhusiopathiae), active public health policies are required to inform field professionals and the general public about the health threats associated with marine mammal manipulation and consumption.

Advanced Bottom-Up Engineering of Living Architectures.

Bottom-up tissue engineering is a promising approach for designing modular biomimetic structures that aim to recapitulate the intricate hierarchy and biofunctionality of native human tissues. In recent years, this field has seen exciting progress driven by an increasing knowledge of biological systems and their rational deconstruction into key core components. Relevant advances in the bottom-up assembly of unitary living blocks toward the creation of higher order bioarchitectures based on multicellular-rich structures or multicomponent cell-biomaterial synergies are described. An up-to-date critical overview of long-term existing and rapidly emerging technologies for integrative bottom-up tissue engineering is provided, including discussion of their practical challenges and required advances. It is envisioned that a combination of cell-biomaterial constructs with bioadaptable features and biospecific 3D designs will contribute to the development of more robust and functional humanized tissues for therapies and disease models, as well as tools for fundamental biological studies.

Electrospun biodegradable chitosan based-poly(urethane urea) scaffolds for soft tissue engineering.

The composition and architecture of a scaffold determine its supportive role in tissue regeneration. In this work, we demonstrate the feasibility of obtaining a porous electrospun fibrous structure from biodegradable polyurethanes (Pus) synthesized using polycaprolactone-diol as soft segment and, as chain extenders, chitosan (CS) and/or dimethylol propionic acid. Fourier transform infrared spectroscopy and proton nuclear magnetic resonance confirmed the syntheses. Fibre mats' properties were analysed and compared with those of solvent cast films. Scanning electron microscopy images of the electrospun scaffolds revealed fibres with diameters around 1 µm. From tensile tests, we found that Young's modulus increases with CS content and is higher for films (2.5 MPa to 6.5 MPa) than for the corresponding fibre mats (0.8 MPa to 3.2 MPa). The use of CS as the only chain extender improves recovery ratio and resilience. From X-ray diffraction, a higher crystalline degree was identified in fibre mats than in the corresponding films. Films' wettability was enhanced by the presence of CS as shown by the decrease of water contact angle. X-ray photoelectron spectroscopy revealed that while ester groups are predominant at the films' surface, ester and urethanes are present in similar concentrations at fibres' surface, favouring the interaction with water molecules. Both films and fibres undergo hydrolytic degradation. In vitro evaluation was performed with human dermal fibroblasts. No PU sample revealed cytotoxicity. Cells adhered to fibre mats better than to films and proliferation was observed only for samples of CS-containing PUs. Results suggest that electrospun fibres of CS-based polyurethanes are good candidate scaffolds for soft tissue engineering.