Mineralization of Phosphorylated Fish Skin Collagen/Mangosteen Scaffolds as Potential Materials for Bone Tissue Regeneration.

In this study, a potential hard tissue substitute was mimicked using collagen/mangosteen porous scaffolds. Collagen was extracted from Tilapia fish skin and mangosteen from the waste peel of the respective fruit. Sodium trimetaphosphate was used for the phosphorylation of these scaffolds to improve the nucleation sites for the mineralization process. Phosphate groups were incorporated in the collagen structure as confirmed by their attenuated total reflection Fourier transform infrared (ATR-FTIR) bands. The phosphorylation and mangosteen addition increased the thermal stability of the collagen triple helix structure, as demonstrated by differential scanning calorimetry (DSC) and thermogravimetry (TGA) characterizations. Mineralization was successfully achieved, and the presence of calcium phosphate was visualized by scanning electron microscopy (SEM). Nevertheless, the porous structure was maintained, which is an essential characteristic for the desired application. The deposited mineral was amorphous calcium phosphate, as confirmed by energy dispersive X-ray spectroscopy (EDX) results.

Influence of blend ratio and mangosteen extract in chitosan/collagen gels and scaffolds: Rheological and release studies.

This study proposes the incorporation of mangosteen peel extract in chitosan and collagen gels and scaffolds, at different ratios, for fabricating materials with potential wound dressing applications. The extract addition increases the thermal stability of the collagen while decreasing to about one-fifth the swelling capability of its scaffolds. Oppositely, it enables chitosan and its blends to withstand high swelling percentages. Release studies showed an extract delivery of 30%, indicating that the formulation does not affect this property. Additionally, the models of Weibull and the Korsmeyer-Peppas adequately fitted the release curves, in which the last one suggested a faster release regarding extract concentration. In contrast, rheology profiling demonstrated distinct behavior associated with the formulations. Even though all the samples exhibit a shear-thinning characteristic, changes in the blend ratio increased the extension of the Newtonian plateau range. The applied Cross mathematical model showed an increase in interactions between the components.

Effects of Mangosteen Peel Phenolic Compounds on Tilapia Skin Collagen-Based Mineralized Scaffold Properties.

A proper valorization of biological waste sources for an effective conversion into composites for tissue engineering is discussed in this study. Hence, the collagen and the phenolic compound applied in this investigation were extracted from waste sources, respectively, fish industry rejects and the peels of the mangosteen fruit. Porous scaffolds were prepared by combining both components at different compositions and mineralized at different temperatures to evaluate the modifications in the biomimetic formation of apatite. The inclusion of mangosteen extract showed the advantage of increasing the collagen denaturation temperature, improving the stability of its triple helix. Moreover, the extract provided antioxidant activity due to its phenolic composition, as confirmed by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) antioxidant assays. Mineralization was successfully achieved as indicated by thermogravimetry and scanning electron microscopy. A higher temperature and a lower extract concentration reduced the calcium phosphate deposits. The extract also affected the pore size, particularly at a lower concentration. The X-ray diffraction pattern identified a low degree of crystallization. A high mineralization temperature induced the formation of smaller crystallites ranging from 18.9 to 25.4 nm. Although the deposited hydroxyapatite showed low crystallinity, the scaffolds are suitable for bone tissue applications and may be effective in controlling the resorbability rate in tissue regeneration.

1,4-Dioxane enhances properties and biocompatibility of polyanionic collagen for tissue engineering applications.

Polyanionic collagen obtained from bovine pericardial tissue submitted to alkaline hydrolysis is an acellular matrix with strong potential in tissue engineering. However, increasing the carboxyl content reduces fibril formation and thermal stability compared to the native tissues. In the present work, we propose a chemical protocol based on the association of alkaline hydrolysis with 1,4-dioxane treatment to either attenuate or revert the drastic structural modifications promoted by alkaline treatments. For the characterization of the polyanionic membranes treated with 1,4-dioxane, we found that (1) scanning electron microscopy (SEM) shows a stronger reorientation and aggregation of collagen microfibrils; (2) histological evaluation reveals recovering of the alignment of collagen fibers and reassociation with elastic fibers; (3) differential scanning calorimetry (DSC) shows an increase in thermal stability; and (4) in biocompatibility assays there is a normal attachment, morphology and proliferation associated with high survival of the mouse fibroblast cell line NIH3T3 in reconstituted membranes, which behave as native membranes. Our conclusions reinforce the ability of 1,4-dioxane to enhance the properties of negatively charged polyanionic collagen associated with its potential use as biomaterials for grafting, cationic drug- or cell-delivery systems and for the coating of cardiovascular devices.

Rheological and antioxidant properties of chitosan/gelatin-based materials functionalized by pomegranate peel extract.

Biopolymer-based materials are potential candidates for food coatings application. In this study, pomegranate (Punica granatum L.) peel extract (PPE) at different concentrations was incorporated to chitosan/gelatin gels and the rheological, antioxidant and structural properties were evaluated. Due to its high phenolic content, PPE enhanced the antioxidant capacity of chitosan/gelatin mixtures. PPE addition extended linear viscoelastic range and enabled the samples to easily flow under the applied shear rate. Rheological properties indicated that both viscosity and activation energy of materials containing natural compounds are highly dependent on temperature. Scanning electron microscopy (SEM) images revealed the influence of PPE concentration in the scaffolds pores size. Findings of this study proved that PPE was capable to improve the functional characteristics of chitosan/gelatin-based materials enhancing the desired properties for their potential application as food coatings.

Development and evaluation of calcium hydroxide-coated, pericardium-based biomembranes for direct pulp capping.

AIM: The aim of the present study was to develop a bovine pericardium biomembrane (BPB) and to evaluate pulp response in vivo. METHODS: A double-layer bovine BPB/chitosan was manufactured, and the porous chitosan side was coated with calcium hydroxide. The microstructure of the matrices was evaluated with electron microscopy. To test pulp response, cavities were prepared on the occlusal surface of Wistar rats' mandibular left first molars and capped with matrices, followed by appropriate adhesives/composite restorations. The animals were divided into three groups: group 1, calcium hydroxide alone; group 2, BPB without calcium hydroxide; and group 3, BPB coated with calcium hydroxide. Specimens were processed and histologically evaluated at 7, 14, and 30 days, postoperatively. RESULTS: Electron microscopy showed porous chitosan surface and a cohesive calcium hydroxide layer. Histological analysis showed that groups 1 and 3 had mild odontoblast layer disorganization, but normal pulp tissue appearance at 7, 14, and 30 days. At the same time points, group 2 showed a loss of general pulp tissue, pulp necrosis, and periapical abscess in some teeth. CONCLUSION: Coated bovine pericardium-based biomembranes resulted in favorable outcomes in cases of pulp exposure after a 30-day observation period, and might protect against injuries caused by adhesive systems and composites.

Porous poly (D,L-lactide-co-glycolide) acid/biosilicate® composite scaffolds for bone tissue engineering.

This study evaluated the effects of the Biosilicate® and poly (D,L-lactic-co-glycolic) acid composites on bone repair in a tibial bone defect model in rats by means of using histological evaluation (histopathological and morphometric analysis) and gene expression analysis. Eighty male Wistar rats (12 weeks old, weighing ±300 g) were randomly divided into two groups: Biosilicate® group (BG) and Biosilicate® /PLGA group (BG/PLGA). Each group was euthanized at 3, 7, 14, and 21 days after surgery (n = 10 animals per time point). The main findings showed that the incorporation of PLGA into BG had a significant effect on the morphological structure of the material, accelerating mass loss, decreasing the pH and increasing the calcium release. Furthermore, histologic analysis revealed that the BG/PLGA showed increased material degradation, accompanied by higher bone formation compared to BG, after 21 days of implantation. In addition, qRT-PCR analysis showed that BG/PLGA induced an upregulation of the osteogenic genes related to BMP4, Runx2, ALP, and OC. These results show that the present BG/PLGA composite may be used as a bone graft for inducing bone repair. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 63-71, 2017.

Modifications on collagen structures promoted by 1,4-dioxane improve thermal and biological properties of bovine pericardium as a biomaterial.

Collagen is a widely used raw material for biomaterial manufacture, which generally depends on chemical modifications of this fibrillar protein with cross-linking agents to improve biocompatibility and mechanical properties. However, cross-linking reduces the natural properties of collagen, such as low immune response, low toxicity as well as the ability to promote cellular growth and attachment. In this work, the modifications promoted by 1,4-dioxane solvent on the collagen present in native bovine pericardium (NBP) matrix routinely used in bioprosthesis manufacture, with or without subsequent cross-linking by glutaraldehyde, has been studied. The structural changes of NBP evaluated by scanning electron microscopy show that 1,4-dioxane induces a more homogeneous material by increasing aggregation of collagen fibers, while transmission scanning electron microscopy shows that natural collagen fibril arrangement, integrity, and the D-periodicity pattern are maintained by solvent treatments. Measurements of thermal stability and resistance to collagenase enzymatic digestion of NBP matrices treated with 1,4-dioxane show an increase in melting temperature and decrease in biodegradability, as compared to native pericardium. Cross-linking with glutaraldehyde improves all the analyzed NBP properties, which are not impaired by previous treatment with 1,4-dioxane. Histological evaluation of NBP submitted to 1,4-dioxane treatment shows lower lipid and cell contents and improvement in other morphologic characteristics compared to native pericardium. Altogether, these results suggest the use of 1,4-dioxane organic solvent as an alternative non-cross-linking treatment for direct utilization on rich collagen matrices, resulting in materials with improved biocompatibility and physicochemical properties suitable for tissue engineering.

The effect of the polymerization initiator and light source on the elution of residual Bis-GMA and TEGDMA monomers: A study using liquid chromatography with UV detection.

A method for the extraction and quantification of two residual monomers, bisphenol glycidyl dimethacrylate (Bis-GMA) and triethylene glycol dimethacrylate (TEGDMA), that were evaluated using high efficiency liquid chromatography with UV detection was developed and validated in this study. Three types of solvents were applied in the extraction of the monomers (methanol, ethanol and acetonitrile), where the highest extraction efficiency was obtained using acetonitrile. The different resins were prepared by photoactivation of Bis-GMA and TEGDMA monomers. Additionally, the effects of the addition of two photoinitiators (camphorquinone (CQ) and phenyl propanodione (PPD) and that of a co-initiator (N,N-dimethyl-p-toluidine) were also analyzed. When only the CQ photoinitiator was used, a smaller amount of residual monomers was obtained, whereas a larger amount was obtained with PPD. When the two photoinitiators were used in the same matrix, however, no significant changes were observed in relation to the amount of residual TEGDMA monomers. For the addition of the co-initiator, there were no large changes in the extraction of residual monomers. The effect of the two photoactivation sources (halogen lamp and LED) led to small differences in the elution of the two monomers, although all of the resins differed significantly when photoactivated with a LED. Quantum chemical calculations using Density Functional Theory were carried out to characterize several molecular properties of each monomer.

Hidrólise seletiva de grupos carboxamidos de matrizes colagênicas: um novo biomaterial / Selective hydrolysis of carboxamides groups of collagenics matrix: a new biomaterial

Este trabalho estudou a hidrólise alcalina seletiva dos resíduos de glutamina e asparagina de colágeno para obtenção de colágeno aniônico. As caracterizações foram realizadas por titulação potenciométrica e calorimetria exploratória diferencial, mostraram que este tratamento hidrolisou seletivamente os grupos carboxamidos presentes no colágeno, sem prejuízo das ligações peptidicas ou estrutura secundária da proteína. Estes materiais mostraram um coeficiente piroelétrico de 1.16 x 10(-4) Cm(-2), maior que o encontrado para o PVDF polarizado, de 7X10(-6 Cm(-2).

Abstract: This work studied the selective alkaline hydrolysis of glutamine and asparagine residues of collagen in order to produce anionic collagen. Characterization of the hydrolysed material by potenciometric titration and differential scanning calorimetry, showed that the selective hydrolysis was achieved, without unpairing both, the peptide bond or the native protein structure. The piroelectric coefficient determined for this material was l. l 6x 1 o-" Cm-2 · is in comparison to 7x 10-6 Cm-2 for polarized PVDF