Hydrogel Based on Chitosan/Gelatin/Poly(Vinyl Alcohol) for In Vitro Human Auricular Chondrocyte Culture.

Three-dimensional (3D) hydrogels provide tissue-like complexities and allow for the spatial orientation of cells, leading to more realistic cellular responses in pathophysiological environments. There is a growing interest in developing multifunctional hydrogels using ternary mixtures for biomedical applications. This study examined the biocompatibility and suitability of human auricular chondrocytes from microtia cultured onto steam-sterilized 3D Chitosan/Gelatin/Poly(Vinyl Alcohol) (CS/Gel/PVA) hydrogels as scaffolds for tissue engineering applications. Hydrogels were prepared in a polymer ratio (1:1:1) through freezing/thawing and freeze-drying and were sterilized by autoclaving. The macrostructure of the resulting hydrogels was investigated by scanning electron microscopy (SEM), showing a heterogeneous macroporous structure with a pore size between 50 and 500 µm. Fourier-transform infrared (FTIR) spectra showed that the three polymers interacted through hydrogen bonding between the amino and hydroxyl moieties. The profile of amino acids present in the gelatin and the hydrogel was determined by ultra-performance liquid chromatography (UPLC), suggesting that the majority of amino acids interacted during the formation of the hydrogel. The cytocompatibility, viability, cell growth and formation of extracellular matrix (ECM) proteins were evaluated to demonstrate the suitability and functionality of the 3D hydrogels for the culture of auricular chondrocytes. The cytocompatibility of the 3D hydrogels was confirmed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reaching 100% viability after 72 h. Chondrocyte viability showed a high affinity of chondrocytes for the hydrogel after 14 days, using the Live/Dead assay. The chondrocyte attachment onto the 3D hydrogels and the formation of an ECM were observed using SEM. Immunofluorescence confirmed the expression of elastin, aggrecan and type II collagen, three of the main components found in an elastic cartilage extracellular matrix. These results demonstrate the suitability and functionality of a CS/Gel/PVA hydrogel as a 3D support for the auricular chondrocytes culture, suggesting that these hydrogels are a potential biomaterial for cartilage tissue engineering applications, aimed at the regeneration of elastic cartilage.

Microplastics: challenges of assessment in biological samples and their implication for in vitro and in vivo effects.

Microplastics (MPs) have attracted global interest because they have been recognized as emerging pollutants that require urgent attention. MPs are plastic particles with a size between 1 micron and 5 mm (1 µm-5mm); those measuring less than 1 µm are known as nanoplastics (NPs). MP is distributed in the environment in various physical forms that depend on the degradation process, the erosion factors to which it was subjected, or the original form in which it was intentionally manufactured. Humans may be exposed to these pollutants mainly by ingestion or inhalation, which could adversely affect human health with effects that are still unknown due to limitations that are often dependent on their analytical determination and lack of studies over time, as it is a relatively new topic. Therefore, this review focuses on the challenges currently faced by laboratories for determining MPs in different matrices. We highlight the application of methods and techniques to assess the precise levels of exposure to MPs in biological samples. In addition, exposure pathways, sources, and evidence of adverse effects reported in vitro and in vivo studies are described to generate knowledge about their potential threat to human health.

Chondrogenic Potential of Human Adipose-Derived Mesenchymal Stromal Cells in Steam Sterilized Gelatin/Chitosan/Polyvinyl Alcohol Hydrogels.

Cross-linked polymer blends from natural compounds, namely gelatin (Gel), chitosan (CS), and synthetic poly (vinyl alcohol) (PVA), have received increasing scrutiny because of their versatility, biocompatibility, and ease of use for tissue engineering. Previously, Gel/CS/PVA [1:1:1] hydrogel produced via the freeze-drying process presented enhanced mechanical properties. This study aimed to investigate the biocompatibility and chondrogenic potential of a steam-sterilized Gel/CS/PVA hydrogel using differentiation of human adipose-derived mesenchymal stromal cells (AD-hMSC) and cartilage marker expression. AD-hMSC displayed fibroblast-like morphology, 90% viability, and 69% proliferative potential. Mesenchymal profiles CD73 (98.3%), CD90 (98.6%), CD105 (97.0%), CD34 (1.11%), CD45 (0.27%), HLA-DR (0.24%); as well as multilineage potential, were confirmed. Chondrogenic differentiation of AD-hMSC in monolayer revealed the formation of cartilaginous nodules composed of glycosaminoglycans after 21 days. Compared to nonstimulated cells, hMSC-derived chondrocytes shifted the expression of CD49a from 2.82% to 40.6%, CD49e from 51.4% to 92.2%, CD54 from 9.66 to 37.2%, and CD151 from 45.1% to 75.8%. When cultured onto Gel/CS/PVA hydrogel during chondrogenic stimulation, AD-hMSC changed to polygonal morphology, and chondrogenic nodules increased by day 15, six days earlier than monolayer-differentiated cells. SEM analysis showed that hMSC-derived chondrocytes adhered to the surface with extended filopodia and abundant ECM formation. Chondrogenic nodules were positive for aggrecan and type II collagen, two of the most abundant components in cartilage. This study supports the biocompatibility of AD-hMSC onto steam-sterilized GE/CS/PVA hydrogels and its improved potential for chondrocyte differentiation. Hydrogel properties were not altered after steam sterilization, which is relevant for biosafety and biomedical purposes.

Porous Chitosan Hydrogels Produced by Physical Crosslinking: Physicochemical, Structural, and Cytotoxic Properties.

Chitosan hydrogels are biomaterials with excellent potential for biomedical applications. In this study, chitosan hydrogels were prepared at different concentrations and molecular weights by freeze-drying. The chitosan sponges were physically crosslinked using sodium bicarbonate as a crosslinking agent. The X-ray spectroscopy (XPS and XRD diffraction), equilibrium water content, microstructural morphology (confocal microscopy), rheological properties (temperature sweep test), and cytotoxicity of the chitosan hydrogels (MTT assay) were investigated. XPS analysis confirmed that the chitosan hydrogels obtained were physically crosslinked using sodium bicarbonate. The chitosan samples displayed a semi-crystalline nature and a highly porous structure with mean pore size between 115.7 ± 20.5 and 156.3 ± 21.8 µm. In addition, the chitosan hydrogels exhibited high water absorption, showing equilibrium water content values from 23 to 30 times their mass in PBS buffer and high thermal stability from 5 to 60 °C. Also, chitosan hydrogels were non-cytotoxic, obtaining cell viability values ≥ 100% for the HT29 cells. Thus, physically crosslinked chitosan hydrogels can be great candidates as biomaterials for biomedical applications.

Circulating exosomal MicroRNAs: New non-invasive biomarkers of non-communicable disease.

Exosomes are vesicles, ranging of 30-150 nm in diameter, which are released by different cell types into the extracellular space. Exosomes are capable of transporting several biomolecules such as proteins, lipids, DNA, mRNA, and non-coding RNA, including microRNAs (miRs). miRs signatures have been linked to the development of non-communicable diseases and their classification into various subtypes and/or stages. Interestingly, the miRs contained in exosomes (exomiRs) are suitable candidates as non-invasive biomarkers due to their stability in body fluids and harsh conditions, as well as they are considered critical players involved in intercellular communication; so that they can be a promising diagnostic tool for several diseases. Besides, exomiRs allow discrimination between different stages of the disease and could be a valuable strategy for the early detection of several pathologies in a non-invasive approach. This review aims to describe exomiRs present in biologic fluids that can be used as a tool for the diagnosis and prognosis of non-communicable diseases such as cancer, cardiovascular, kidney, and neurodegenerative disease.

Classical methods and perspectives for manipulating the human gut microbial ecosystem.

A healthy Human Gut Microbial Ecosystem (HGME) is a necessary condition for maintaining the orderly function of the whole body. Major alterations in the normal gut microbial composition, activity and functionality (dysbiosis) by an environmental or host-related disruptive event, can compromise metabolic, inflammatory, and neurological processes, causing disorders such as obesity, inflammatory bowel disease, colorectal cancer, and depressive episodes. The restore or the maintaining of the homeostatic balance of Gut Microbiota (GM) populations (eubiosis) is possible through diet, the use of probiotics, prebiotics, antibiotics, and even Fecal Microbiota Transplantation (FMT). Although these "classic methods" represent an effective and accepted way to modulate GM, the complexity of HGME requires new approaches to control it in a more appropriate way. Among the most promising emergent strategies for modulating GM are the use of engineered nanomaterials (metallic nanoparticles (NP), polymeric-NP, quantum dots, micelles, dendrimers, and liposomes); phagotherapy (i.e., phages linked with the CRISPR/Cas9 system), and the use of antimicrobial peptides, non-antibiotic drugs, vaccines, and immunoglobulins. Here we review the current state of development, implications, advantages, disadvantages, and perspectives of the different approaches for manipulating HGME.

Polybrominated diphenyl ethers (PBDEs) concentration in soil from San Luis Potosi, Mexico: levels and ecological and human health risk characterization.

The aim of this study was to assess the levels of polybrominated diphenyl ethers (PBDEs) in soils from the city of San Luis Potosi in Mexico and perform an ecological and human health risk characterization. In order to confirm the presence of PBDEs, outdoor surface soil samples were collected and the concentrations of PBDEs in urban, industrial, agricultural, and brick kiln industry areas were determined. The mean total PBDEs levels obtained in the study sites were 25.0 ± 39.5 µg/kg (geometric mean ± standard deviation) in the brick kiln industry zone; 34.5 ± 36.0 µg/kg in the urban zone; 8.00 ± 7.10 µg/kg in the industrial zone and 16.6 ± 15.3 µg/kg in the agricultural zone. The ecological and human health risk characterization showed relatively low-hazard quotient values. However, the moderately high PBDEs levels found in soils highlight the necessity to establish a systematic monitoring process for PBDEs in environmental and biological samples.