In vitro evaluation of crosslinked bovine pericardium as potential scaffold for the oral cavity.

BACKGROUND: Bovine pericardium (BP) is a scaffold widely used in soft tissues regeneration; however, its calcification in contact with glutaraldehyde, represent an opportunity for its application in hard tissues, such as bone in the oral cavity. OBJECTIVE: To develop and to characterize decellularized and glutaraldehyde-crosslinked bovine pericardium (GC-BP) as a potential scaffold for guided bone regeneration GBR. METHODS: BP samples from healthy animals of the bovine zebu breed were decellularized and crosslinked by digestion with detergents and glutaraldehyde respectively. The resulting cell-free scaffold was physical, chemical, mechanical, and biologically characterized thought hematoxylin and eosin staining, DNA quantification, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), uniaxial tensile test, cell viability and live and dead assay in cultures of dental pulp stem cells (DPSCs). RESULTS: The decellularization and crosslinking of BP appeared to induce conformational changes of the CLG molecules, which led to lower mechanical properties at the GC-BP scaffold, at the same time that promoted cell adhesion and viability of DPSCs. CONCLUSION: This study suggests that the decellularized and GC-BP is a scaffold with the potential to be used promoting DPSCs recruitment, which has a great impact on the dental area.

Comparative physicochemical analysis of chitin quality of the two tagmata in the exoskeleton of the horseshoe crab Limulus polyphemus.

Recently, chitin biopolymer has received much attention as a wide variety of biomedical application for this and its derivatives have been reported, in fact, the study of non-conventional species as alternative sources of them compounds has taken particular interest. Here, we present a comparative physicochemical survey of the two tagmata in the exoskeleton of the horseshoe crab Limulus polyphemus: the prosoma and the opisthosoma, collected in Yucatán, Mexico. The characterization included CHNSO analysis, FTIR, TGA, DSC, XRD, and SEM. The CHNSO analysis revealed that C is present in the highest proportion (∼45 %) and that chemical composition did not show significant differences (P < 0.05) between the two tagmata. FTIR spectra of two tagmata presented a wide characteristic band of the chitin between 3600 and 3000 cm-1, confirming the presence of this biopolymer in the exoskeleton studied. TGA and DTGA profiles resulted very similar for both tagmata being the residual mass at 650 °C of around 30 % for both samples; these values were associated to the presence of minerals. SEM micrographs showed a porous matrix with infinite large number of irregularly shaped particles. Results show that both tagmata are made up of chitin, and they seem to have a high mineral content.

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.

Influence of Molecular Weight and Grafting Density of PEG on the Surface Properties of Polyurethanes and Their Effect on the Viability and Morphology of Fibroblasts and Osteoblasts.

Grafting polyethylene glycol (PEG) onto a polymer's surface is widely used to improve biocompatibility by reducing protein and cell adhesion. Although PEG is considered to be bioinert, its incorporation onto biomaterials has shown to improve cell viability depending on the amount and molecular weight (MW) used. This phenomenon was studied here by grafting PEG of three MW onto polyurethane (PU) substrata at three molar concentrations to assess their effect on PU surface properties and on the viability of osteoblasts and fibroblasts. PEG formed a covering on the substrata which increased the hydrophilicity and surface energy of PUs. Among the results, it was observed that osteoblast viability increased for all MW and grafting densities of PEG employed compared with unmodified PU. However, fibroblast viability only increased at certain combinations of MW and grafting densities of PEG, suggesting an optimal level of these parameters. PEG grafting also promoted a more spread cell morphology than that exhibited by unmodified PU; nevertheless, cells became apoptotic-like as PEG MW and grafting density were increased. These effects on cells could be due to PEG affecting culture medium pH, which became more alkaline at higher MW and concentrations of PEG. Results support the hypothesis that surface energy of PU substrates can be tuned by controlling the MW and grafting density of PEG, but these parameters should be optimized to promote cell viability without inducing apoptotic-like behavior.

Physicochemical, Mechanical, and Structural Properties of Bio-Active Films Based on Biological-Chemical Chitosan, a Novel Ramon (Brosimum alicastrum) Starch, and Quercetin.

The properties of biological-chemical chitosan (BCh) films from marine-industrial waste and a non-conventional Ramon starch (RS) (Brosimum alicastrum) were investigated. Blended films of BCh/RS were prepared to a volume ratio of 4:1 and 1:4, named (BChRS-80+q, biological-chemical chitosan 80% v/v and Ramon starch, BChRS-20+q, biological-chemical chitosan 20% v/v and Ramon starch, both with quercetin), Films from commercial chitosan (CCh) and corn starch (CS), alone or blended (CChCS-80+q, commercial chitosan 80% v/v and corn starch, CChCS-20+q commercial chitosan 20% v/v and corn starch, both with quercetin) were also prepared for comparison purposes. Films were investigated for their physicochemical characteristics such as thickness, moisture, swelling, water-vapor permeability, and water solubility. In addition, their mechanical and structural properties were studied using Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetric analysis (TGA) and Scanning Electron Microscopy (SEM) techniques. Antioxidant activity was evaluated as radical scavenging, and antimicrobial effect was also determined. The BCh and RS films presented similar tensile strength values compared with commercial biopolymers. Only films with chitosan presented antioxidant and antimicrobial activity. The FTIR spectra confirmed the interactions between functional groups of the biopolymers. Although, BChRS-80+q and BChRS-20+q films exhibited poor mechanical performance compared to their commercial counterparts, they showed good thermal stability, and improved antioxidant and antimicrobial activity in the presence of quercetin. BChRS-80+q and BChRS-20+q films have promising applications due to their biological activity and mechanical properties, based on a novel material that has been underutilized (Ramon starch) that does not compete with materials for human feeding and may be used as a coating for food products.

Antiviral capacity of polypropylene/(1-Hexadecyl) trimethyl-ammonium bromide composites against COVID-19.

During the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, scientists from different areas are looking for alternatives to fight it. SARS-CoV-2, the cause of the infectious respiratory disease COVID-19, is mainly transmitted through direct or indirect contact with infected respiratory droplets. The integrity of the virus structure is crucial for its viability to attack human cells. Quaternary ammonium salts are characterized by having antiviral capabilities which alter or destroy the structure of the viral capsid. In this work, polypropylene (PP)/(1-Hexadecyl) trimethyl-ammonium bromide (CTAB) composites have been prepared in order to create an antiviral material. The composites were melt processed and blown to produce thin films. The CTAB content on the antiviral effect was evaluated using antibodies and serum from infected patients with the SARS-CoV-2 virus. In addition, the mechanical and thermal properties of blown films were investigated, and CTAB release kinetics from the films was followed by UV-Vis. The results indicate that the virus tends to remain less on the polymer surface by increasing the amount of CTAB in the PP matrix.

Kinetic, Thermodynamic, Physicochemical, and Economical Characterization of Pectin from Mangifera indica L. cv. Haden Residues.

The effect of temperature (60, 70, 80, and 90 °C) and time (30, 45, 60, 75, and 90 min) on citric acid extraction of Haden mango (Mangifera indica L. cv. Haden) peel pectin was evaluated in the present study. In order to obtain a better understanding of both the extraction process and the characteristics of the pectin (obtained from an agro-industrial waste) for a future scaling process, the following characterizations were performed: (1) Kinetic, with the maximum extraction times and yields at all evaluated temperatures; (2) thermodynamic, obtaining activation energies, enthalpies, entropies, and Gibbs free energies for each stage of the process; (3) physicochemical (chemical analysis, monosaccharide composition, degree of esterification, galacturonic acid content, free acidity, Fourier-transform infrared spectroscopy, thermogravimetric and derivative thermogravimetric analyses); and (4) economical, of the pectin with the highest yield. The Haden mango peel pectin was found to be characterized by a high-esterified degree (81.81 ± 0.00%), regular galacturonic acid content (71.57 ± 1.26%), low protein (0.83 ± 0.05%) and high ash (3.53 ± 0.02%) content, low mean viscometric molecular weight (55.91 kDa), and high equivalent weight (3657.55 ± 8.41), which makes it potentially useful for food applications.

Biocompatibility studies of polyurethane electrospun membranes based on arginine as chain extender.

Electrospun polymers are an example of multi-functional biomaterials that improve the material-cellular interaction and aimed at enhancing wound healing. The main objective of this work is to fabricate electrospun polyurethane membranes using arginine as chain extender (PUUR) in order to test the fibroblasts affinity and adhesion on the material and the polymer toxicity. Polyurethane membranes were prepared in two steps: (i) the polyurethane synthesis, and ii) the electrospinning process. The membranes were characterized by scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy, gel permeation chromatography, and differential scanning calorimetry techniques. The evaluation of PUUR as a scaffolding biomaterial for growing and developing of cells on the material was realized by LIVE/DEAD staining. The results show that the fluorescent surface area of human fibroblasts (hFB), was greater in control dense membranes made from Tecoflex than in electrospun and dense PUUR. From SEM analysis, the electrospun membranes show relatively uniform attachment of cells with a well-spread shape, while Tecoflex dense membranes show a non-proliferating round shape, which is attributed to the fiber's structure in electrospun membranes. The cell morphology and the cell attachment assay results reveal the well spreading of hFB cells on the surface of electrospun PUUR membranes which indicates a good response related to cell adhesion.

Ceiba pentandra cellulose crosslinked with citric acid for drug release systems.

An alternative for the production of drug delivery system is proposed based on the Ceiba pentandra milkweed. The kapok cellulose was chemically crosslinked with citric acid (CA) at different CA proportions, and loaded with chlorhexidine diacetate (CHX) at different concentrations. Cellulose crosslinking was followed with FTIR and XPS analysis, and the CHX loading was determined using elemental analysis. In vitro studies showed a burst release within the first 2-3 h and the drug release kinetics was described with several models. In addition, the crosslinked Ceiba pentandra fibers did not exhibit a cytotoxic effect on human dermic fibroblasts. Results indicate that the crosslinked Ceiba pentandra fibers are a feasible material for the production of systems for drug release applications.

The Effect of PEGDE Concentration and Temperature on Physicochemical and Biological Properties of Chitosan.

Chitosan (CHT) is a polysaccharide with multiple claimed properties and outstanding biocompatibility, generally attributed to the presence of protonable amino groups rendering a cationic natural polymer. However, the effect of changes in CHT structure due to hydration is not considered in its performance. This study compares the effects on biocompatibility after drying at 25 °C and 150 °C scaffolds of chitosan, polyethylene glycol diglycidyl ether (PEGDE) crosslinked CHT (low, medium and high concentration) and glutaraldehyde (GA) crosslinked CHT. PEGDE crosslinked CHT showed a reduction in free amino groups and the amide I/II ratio, which exhaustive drying reduced further. In X-ray diffraction (DRX) analysis, PEGDE crosslinked CHT showed multiple peaks, whereas the crystallinity percentage was reduced with an increase in PEGDE concentration and thermal treatments at 150 °C. In a direct contact cell assay, high osteoblast viability was achieved at low and medium PEDGE concentrations, which was improved when the crosslinked scaffolds were thermally treated at 150 °C. This was attributed to its partial hydrophilicity, low crystallinity and low surface roughness; this in spite of the small reduction in the amount of free amino groups on the surface induced during drying at 150 °C. Furthermore, PEGDE crosslinked CHT scaffolds showed strong vinculin and integrin 1ß expression, which render them suitable for bone contact applications.

Titanium - castor oil based polyurethane composite foams for bone tissue engineering.

Polyurethanes (PU) foams with titanium particles (Ti) were prepared with castor oil (CO) and isophorone diisocyanate (IPDI) as polymeric matrix, and 1, 3 and 5 wt.% of Ti. Composites were physicochemically and mechanically characterized and their biocompatibility assessed using human dental pulp stem cells (HDPSC). PU synthesis was confirmed by FTIR, but the presence of Ti was detected by RAMAN, X-ray diffraction (peak at 2θ = 40.2°) and by EDX-mapping. Materials showed three decomposition temperatures between 300 °C and 500 °C and their decomposition were not catalyzed by Ti particles. Compressive modulus (164-846 kPa), compressive strength (12.9-116.7 kPa) and density (128-240 kg/m3) tend to increase with Ti concentration but porosity was reduced (87% to 80%). Composites' foams were fully degraded in acid and oxidative media while remained stable in distilled water. HDPSC viability on all composites was higher than 80% up to 14 days while proliferation dropped up to 60% at 21 days. Overall, these results suggest that these foams can be used as scaffolds for bone tissue regeneration.

Preparation and characterization of titanium-segmented polyurethane composites for bone tissue engineering.

Segmented polyurethanes were prepared with polycaprolactone diol as soft segment and 4,4-methylene-bis cyclohexyl diisocyanate and l-glutamine as the rigid segment. These polyurethanes were filled with 1 wt.% to 5 wt.% titanium particles (Ti), physicochemically characterized and their biocompatibility assessed using human dental pulp stem cells and mice osteoblasts. Physicochemical characterization showed that composites retained the properties of the semicrystalline polyurethane as they exhibited a glass transition temperature (Tg) between -35°C and -45°C, melting temperature (Tm) at 52°C and crystallinity close to 40% as determined by differential scanning calorimetry. In agreement with this, X-ray diffraction showed reflections at 21.3° and 23.6° for polycaprolactone diol and reflections at 35.1°, 38.4°, and 40.2° for Ti particles suggesting that these particles are not acting as nucleating sites. The addition of up to 5 wt.% of Ti reduced both, tensile strength and maximum strain from 1.9 MPa to 1.2 MPa, and from 670% to 172% for pristine and filled polyurethane, respectively. Although there were differences between composites at low strain rates, no significant differences in mechanical behavior were observed at higher strain rate where a tensile stress of 8.5 MPa and strain of 223% were observed for 5 wt.% composites. The addition to titanium particles had a beneficial effect on both human dental pulp stem cells and osteoblasts viability, as it increased with the amount of titanium in composites up to 10 days of incubation.