Patricia L. Moreira

Embedding techniques were first developed in the mid 1800s in response to the significant improvements in light microscopy. As the resolution of microscopy increased, so did the need for improved quality of the tissue specimens to be analyzed. The specimens needed to be cut in much thinner slices, which could only be done if embedded in a suitable medium supporting the material and providing the hardness required for thinner sectioning. Edwin Klebs introduced the paraffin-embedding methodology in 1869.18 However, at that time, the material was not completely infiltrated with paraffin and the sections were cut freehand with a razor.

To overcome the current lack of in vitro models to specifically reproduce hormonal skin ageing in women, and in search of active ingredients with innovative efficacy claim for cosmetic skin care, we developed a cell culture-based model by simulating menopause's hormonal decline and assessed several parameters of collagen metabolism. Human dermal fibroblasts were incubated with media containing 17β-oestradiol, progesterone, dehydroepiandrosterone, growth hormone and insulin-like growth factor-1 at concentrations corresponding to those of non-menopausal women's sera and then of menopausal women's sera. We measured cell proliferation [by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT)], matrix metalloproteinase-1 and metalloproteinase-3 (MMPs) release (by enzyme-linked immunosorbent assay - ELISA), total collagen deposition (by Sirius red staining), types I and III collagen deposition (by ELISA), and types I and III procollagen gene expression (by real-ti...

Oxidative stress occurs when there is an over production of free radicals and cells are not able to neutralize them by their own antioxidant mechanisms. These excess of free radicals will attack cellular macromolecules leading to cell damage, function impairment or death. Because of that, antioxidant substances have been largely used in products to offer complementary protection. In this study a new mixture of three known antioxidants (cocoa, green tea and alpha-tocopherol) was evaluated and its antioxidant protection was assessed focusing on its capacity to protect main cell macromolecules. Results have shown that it has a high antioxidant capacity by protecting lipids, DNA and proteins against oxidative damage. The antioxidant effect of the mixture on cells was also investigated and it was able to reduce oxidative stress generated by lipopolisacharide in human fibroblasts. Finally, as the mixture has proved to be highly antioxidant, its effect on cell senescence was evaluated, and...

The aim of this work was to perform a systematic study about the effects induced by chitosan solution concentration and by chitin or glycerol incorporation on dense chitosan membranes with potential use as burn dressings. The membrane properties analyzed were total raw material cost, thickness, morphology, swelling ratio, tensile strength, percentage of strain at break, crystallinity, in vitro enzymatic degradation with lysozyme, and in vitro Vero cells adhesion. While the use of the most concentrated chitosan solution (2.5% w/w) increased membrane cost, it also improved the biomaterial mechanical resistance and ductility, as well as reduced membrane degradation when exposed for 2 months to lysozyme. The remaining evaluated properties were not affected by initial chitosan solution concentration. Chitin incorporation, on the other hand, reduced the membranes cost, swelling ratio, mechanical properties, and crystallinity, resulting in thicker biomaterials with irregular surface more easily degradable when exposed to lysozyme. Glycerol incorporation also reduced the membranes cost and crystallinity and increased membranes degradability after exposure to lysozyme. Strong Vero cells adhesion was not observed in any of the tested membrane formulations. The overall results indicate that the majority of the prepared membranes meet the performance requirements of temporary nonbiodegradable burn dressings (e.g. adequate values of mechanical resistance and ductility, low values of in vitro cellular adhesion on their surfaces, low extent of degradation when exposed to lysozyme solution, and high stability in aqueous solutions).

The biggest challenge to improve extracorporeal circulation (ECC) circuits lays on avoiding platelet adhesion to their surfaces, because this contributes to thrombus formation, resulting in the activation of blood coagulation. One approach to minimize this effect is to improve the biocompatibility of ECC circuits by modifying their surfaces. This can be achieved by coating them with heparin or phospholipids. The present study investigated the adhesion and morphology characteristics of fibroblastic and blood cells cultured on uncoated poly (vinyl) chloride PVC tubes as well as on heparin, phosphatidylcholine (DMPC), and phosphatidylethanolamine (DMPE) -coated tubing. The results showed the importance of uniform coating regardless of the substance used, because the coatings cover the grooves on PVC surfaces, which favor cell adhesion. The comparison among the three different coatings showed the best biocompatibility results for the PVC tubes coated with heparin, followed by the coating with DMPE and with DMPC. For all coated tubes, cells did not spread on the PVC surfaces and, consequently, did not adhere to their surfaces, increasing the overall biocompatibility of PVC tubes. However, possible DMPE's alkylation, caused by sterilization, resulted in increased material hydrophobicity, which explains the decrease in fibroblastic adhesion. Furthermore, sterilization of DMPC-PVC improves its hydrophilic character, also decreasing adhesion. Based on these results, coating PVC with the phospholipids DMPC and DMPE seems to be a promising technique to improve the biocompatibility of PVC tubes, and is worthy of further investigation.

Collagen has been extensively described as a beneficial material in bone tissue engineering due to its biocompatibility, biodegradability, low antigenicity, and high tensile strength. However, collagen scaffolds in their pure form have some drawbacks and improvements in the physical, chemical, and biologic properties of collagen are necessary to overcome those inadequacies. Recently, the selective hydrolysis of carboxyamides of asparagine and glutamine residues of collagen has been employed to increase the number of negative sites and enhance the piezoelectric properties of collagen. Anionic collagen scaffolds were prepared by use of a hydrolysis treatment for either 24 h [bovine pericardium (BP 24)] or 48 h (BP 48). Bovine osteoblasts were cultured on them and on native matrices to understand the cellular interactions responsible for the good osteoconductivity and biocompatibility reported with in vivo tests. Based on the data obtained on cell adhesion, alkaline phosphatase (ALP) and extracellular matrix macromolecule production, and cellular proliferation through histological analysis, we may conclude that the materials tested reveal sufficient biocompatibility level for bone repair. Further, the evidence of some connection between ALP activity and the mineralization process should be emphasized. BP 48 presented the most promising results stimulating in vitro mineralization, ALP production, and possible osteoblast differentiation.

... Texto completo. experimental, Documentos relacionados. Id: 355073. Autor: Estacia, Paulo; Rodrigues Junior, Arnaldo Santos; Moreira, Patrícia da Luz; Genari, Selma Calendária. Título: The cytotoxicity in vero cells of a perfluorocarbon used in vitreoretinal surgery. Fonte: Braz. ...