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Biomedical subjects

A P Marques

Publications and source records attributed to A P Marques.

At least 19 recordsLinked to original sources

Effect of starch-based biomaterials on the in vitro proliferation and viability of osteoblast-like cells.

The cytotoxicity of starch-based polymers was investigated using different methodologies. Poly-L-lactic acid (PLLA) was used as a control for comparison purposes. Extracts of four different starch-based blends (corn starch and ethylene vinyl alcohol (SEVA-C), corn starch and cellulose acetate (SCA), corn starch and polycaprolactone (SPCL) and starch and poly-lactic acid (SPLA70) were prepared in culture medium and their toxicity was analysed. Osteoblast-like cells (SaOs-2) were incubated with the extracts and cell viability was assessed using the MTT test and a lactate dehydrogenase (LDH) assay. In addition DNA and total protein were quantified in order to evaluate cell proliferation. Cells were also cultured in direct contact with the polymers for 3 and 7 days and observed in light and scanning electron microscopy (SEM). LDH and DNA quantification revealed to be the most sensitive tests to assess respectively cell viability and cell proliferation after incubation with starch-based materials and PLLA. SCA was the starch blend with higher cytotoxicity index although similar to PLLA polymer. Cell adhesion tests confirmed the worst performance of the blend of starch with cellulose acetate but also showed that SPCL does not perform as well as it could be expected. All the other materials were shown to present a comparable behaviour in terms of cell adhesion showing slight differences in morphology that seem to disappear for longer culture times. The results of this study suggest that not only the extract of the materials but also their three-dimensional form has to be biologically tested in order to analyse material-associated parameters that are not possible to consider within the degradation extract. In this study, the majority of the starch-based biomaterials presented very promising results in terms of cytotoxicity, comparable to the currently used biodegradable PLLA which might lead the biocompatibility evaluation of those novel biomaterials to other studies.

Biocompatible Materials↗

The effect of starch-based biomaterials on leukocyte adhesion and activation in vitro.

Leukocyte adhesion to biomaterials has long been recognised as a key element to determine their inflammatory potential. Results regarding leukocyte adhesion and activation are contradictory in some aspects of the material's effect in determining these events. It is clear that together with the wettability or hydrophilicity/hydrophobicity, the roughness of a substrate has a major effect on leukocyte adhesion. Both the chemical and physical properties of a material influence the adsorbed proteins layer which in turn determines the adhesion of cells. In this work polymorphonuclear (PMN) cells and a mixed population of monocytes/macrophages and lymphocytes (mononuclear cells) were cultured separately with a range of starch-based materials and composites with hydroxyapatite (HA). A combination of both reflected light microscopy and scanning electron microscopy (SEM) was used in order to study the leukocyte morphology. The quantification of the enzyme lactate dehydrogenase (LDH) was used to determine the number of viable cells adhered to the polymers. Cell adhesion and activation was characterised by immunocytochemistry based on the expression of several adhesion molecules, crucial in the progress of an inflammatory response. This work supports previous in vitro studies with PMN and monocytes/macrophages, which demonstrated that there are several properties of the materials that can influence and determine their biological response. From our study, monocytes/macrophages and lymphocytes adhere in similar amounts to more hydrophobic (SPCL) and to moderately hydrophilic (SEVA-C) surfaces and do not preferentially adhere to rougher substrates (SCA). Contrarily, more hydrophilic surfaces (SCA) induced higher PMN adhesion and lower activation. In addition, the hydroxyapatite reinforcement induces changes in cell behaviour for some materials but not for others. The observed response to starch-based biodegradable polymers was not significantly different from the control materials. Thus, the results reported herein indicate the low potential of the starch-based biodegradable polymers to induce inflammation especially the HA reinforced composite materials.

Biocompatible Materials↗

Surface modification of starch based blends using potassium permanganate-nitric acid system and its effect on the adhesion and proliferation of osteoblast-like cells.

The surface modification of three starch based polymeric biomaterials, using a KMnO4/HNO3 oxidizing system, and the effect of that modification on the osteoblastic cell adhesion has been investigated. The rationale of this work is as follows--starch based polymers have been proposed for use as tissue engineering scaffolds in several publications. It is known that in biodegradable systems it is quite difficult to have both cell adhesion and proliferation. Starch based polymers have shown to perform better than poly-lactic acid based materials but there is still room for improvement. This particular work is aimed at enhancing cell adhesion and proliferation on the surface of several starch based polymer blends that are being proposed as tissue engineering scaffolds. The surface of the polymeric biomaterials was chemically modified using a KMnO4/HNO3 system. This treatment resulted in more hydrophilic surfaces, which was confirmed by contact angle measurements. The effect of the treatment on the bioactivity of the surface modified biomaterials was also studied. The bioactivity tests, performed in simulated body fluid after biomimetic coating, showed that a dense film of calcium phosphate was formed after 30 days. Finally, human osteoblast-like cells were cultured on unmodified (control) and modified materials in order to observe the effect of the presence of higher numbers of polar groups on the adhesion and proliferation of those cells. Two of the modified polymers presented changes in the adhesion behavior and a significant increase in the proliferation rate kinetics when compared to the unmodified controls.

Body Fluids↗

Quantifying pain threshold and quality of life of fibromyalgia patients.

The most typical symptom of fibromyalgia (FM) is diffuse pain, and pain at specific points-tender points-is crucial for its diagnosis. By comparing healthy individuals and FM patients, this study was aimed at assessing pain and quality of life of Brazilian females with FM, while seeking for a correlation between pain threshold and quality of life. A total of 178 women were evaluated: 124 were FM patients and 54 were healthy women. Pain threshold at tender points was quantified by dolorimetry, and diffuse pain by means of the visual analogue scale (VAS); the Fibromyalgia Impact Questionnaire (FIQ) was used to evaluate quality of life. Statistical treatment of the data allowed for proposing two indexes: a pain threshold index (PT) and a quality of life one (QOL). PT is the lowest value among all pain thresholds measured at the 18 tender points; QOL is the mean of responses to the FIQ and VAS. Both indexes were tested and showed significant differences between the test and control groups. By pairing pain threshold values of each tender point in the test and control groups, it was found that the most sensitive points matched between the two groups, that is, the most sensitive anatomic spots in a healthy individual are also likely to be the most sensitive points in a person with FM. This suggests that a stimulus that provokes slight discomfort to a healthy person may produce more pain in FM patients--which may bear implications for FM clinical treatment. In this sample of Brazilian women, FM patients had both lower pain threshold and worse quality of life than healthy women.

Adult↗

Evaluation of the potential of starch-based biodegradable polymers in the activation of human inflammatory cells.

The inflammatory response resulting from the implantation of a medical device may compromise its performance and efficiency leading, in certain cases, to the failure of the implant. Thus, the assessment of the behavior of inflammatory cells in vitro, constitutes a key feature in the evaluation of the adverse potential, or not, of new promising biomaterials. The objectives of this study were to determine whether starch-based polymers and composites activated human neutrophils. Blends of starch with ethylene-vinyl alcohol, with cellulose acetate and polycaprolactone, as well as composites based on all these materials filled with hydroxyapatite have been studied. A lysozyme assay was adapted to examine enzyme secretion from human neutrophils incubated with different starch-based materials. Changes in the free radical and degranulation activity of the neutrophil were also determined by measuring the luminescent response of Pholasin, a photoprotein that emits light after excitation by reactive oxygen species. The amount of lysozyme secreted by neutrophils incubated with the polymers did not exhibit significant differences between the tested materials. Results were in all cases similar to those obtained for the control (polypropylene) except for one of the starch blends (corn starch with polycaprolactone reinforced with 30% (w/w) of HA). The chemiluminescence experiments showed that polymers reduce the signal produced by activated neutrophils. Furthermore, for some polymers it was demonstrated that the phenomenon was due to an effect of the surface of the materials in cell adhesion or a simultaneous competition for the photoprotein in solution, which results in the decrease of the intensity of light emitted and detected.

Journal Article↗

The biocompatibility of novel starch-based polymers and composites: in vitro studies.

Studies with biodegradable starch-based polymers have recently demonstrated that these materials have a range of properties. which make them suitable for use in several biomedical applications, ranging from bone plates and screws to drug delivery carriers and tissue engineering scaffolds. The aim of this study was to screen the cytotoxicity and evaluate starch-based polymers and composites as potential biomaterials. The biocompatibility of two different blends of corn-starch, starch ethylene vinyl alcohol (SEVA-C) and starch cellulose acetate (SCA) and their respective composites with hydroxyapatite (HA) was assessed by cytotoxicity and cell adhesion tests. The MTT assay was performed with the extracts of the materials in order to evaluate the short-term effect of the degradation products. The cell morphology of L929 mouse fibroblast cell line was also analysed after direct contact with polymers and composites for different time periods and the number of cells adhered to the surface of the polymers was determined by quantification of the cytosolic lactate dehydrogenase (LDH) activity. Both types of starch-based polymers exhibit a cytocompatibility that might allow for their use as biomaterials. SEVA-C blends were found to be the less cytotoxic for the tested cell line, although cells adhere better to SCA surface. The cytotoxicity test also revealed that SCA and SEVA-C composites have a similar response to the one obtained for SCA polymer. Scanning electron microscopy (SEM) analysis showed that cells were much more spread on the SCA polymer and LDH measurements showed a higher number of cells on this surface.

Animals↗

A comparative study for results obtained using biomonitors and PM10 collectors in Sado Estuary.

In 1996 a program was started, financed by the Environmental Ministry of Portugal and IAEA, aiming to study the inorganic atmospheric pollutant dispersion in Sado Estuary. Gent PM10 air samplers were used for air particulate matter sampling. Three sampling sites were chosen, forming a triangle around the fuel power station of Setúbal. Transplants of Parmelia sulcata Taylor were suspended in nylon bags within a rectangle 15 km wide and 25 km long on a 2.5 x 2.5 km grid. Two sets of four transplants were hung in each of the 47 locations, one set facing the wind and the other set opposing the wind. The transplants were suspended in December 1997 for a 1-year period; every 3 months, one transplant of each set was collected. Both lichen transplants and PM10 filters were analysed by INAA and PIXE. A comparative study of results obtained for the two sampling procedures is presented in this work.

Air Pollutants↗

Growth deficiency, mental retardation and unusual facies.

An 8-year-old girl with growth deficiency, mental retardation, unusual facies and hypertrichosis is described. The case is compared with one described by Wiedemann et al. [(1989) Atlas of Clinical Syndromes, 2nd ed. London: Wolfe Publishing pp. 198-199].

Child↗

Environmental assessment in an industrial area of Portugal.

The region of Lisbon and south of Lisbon (Sado estuary) is densely industrialized, and, therefore, air pollution should be studied in a more detailed scale there. The topography of the Sado estuary region and the predominant wind direction from the northwest contribute to the influence in this region of the industries located in the north. The region selected includes a fuel-fired power station. Transplants of the lichen Parmelia sulcata Taylor were suspended in nylon bags within a rectangle 15 km wide and 25 km long on a grid 2.5 km x 2.5 km, centered in the power station. In each of the 47 sites, 2 sets of 4 transplants each were hung. Care was taken (1) in covering the two sets with a polyethylene roof to prevent leaching of elements in the lichen, (2) in building a hanging system that could rotate according to the wind direction, and (3) in orienting one set toward the wind and the other set opposite the wind. For a 1-yr period and every 3 mo, one transplant of each set is collected. In this work, the results of the first campaign (after 3 mo suspension) obtained by instrumental neutron activation analysis and proton-induced X-ray emission are shown. Some elemental contents are mapped and discussed.

Air Pollutants↗

Comparison between enzyme-linked immunosorbent assay, indirect fluorescent antibody and rapid conglutination tests in detecting antibodies against Babesia bovis.

The performance of an enzyme-linked immunosorbent assay (ELISA), an indirect fluorescent antibody test (IFAT) and a rapid conglutination test (RCT) for the detection of antibodies against Babesia bovis, was evaluated with 462 cattle sera from Bahia State; Brazil. The results showed a concordance of 96.6% between the ELISA and IFAT, 90.5% between the ELISA and RCT, and 91.8% between the IFAT and RCT. Although the prevalence rates determined by ELISA (97.2%) and IFAT (96.8%) were higher than that indicated by the RCT (92.9%), performances of the three serological tests were very similar and characterized the region studied as enzootically stable to B. bovis.

Animals↗

Erythrophagocytosis in the caprine trophoblast.

The mechanism of iron transfer by extravasation of maternal blood in the maternal-fetal interface and the subsequent phagocytosis of the erythrocytes by the trophoblast cells was described in ovine, bovine and other species. This research was performed due to the absence of studies on this process in the goat. Fragments of placentomes were obtained from 9 adult goats of an unspecified breed and were separated into Groups A, B and C, based on 90, 120 and 150 d of gestation, respectively. Fragments of 3 placentomes were obtained from each animal of all groups. The first of them was removed from the base, the second from the middle part and the third from the end of the gestational uterine horn. The fragments were fixed in Bouin solution and histologically processed according to the usual paraffin inclusion techniques. The slides were stained by hematoxilin-eosin and potassium ferrocyanide and examined under light microscopy The erythrophagocytosis (EP) process of the trophoblast was identified in all the examined histological sections, throughout the gestation and for each placentome. An accumulation of spots of blood was observed only in the maternal-fetal interface located in the arcade zone of the placentome. The main cells of the trophoblast of these areas presented an intense phagocytic activity and the observation of erythrocytes with hemossiderin pigments in their citoplasm was possible. The trophoblast epithelium of these areas was thicker and with a columnar aspect than the cubic aspect observed in other areas. The trophoblast binucleate cells do not seem to participate in EP. Our findings indicate that the phagocitosis process of the maternal erythrocites by the trophoblast also occurs in caprine species, indicating that this could be a mechanism of transplacentary transfer of iron in this species.

Journal Article↗

Steroid synthesis by the fetal part of the bovine placenta of late pregnancy in vitro: Effect of a low dose of dexamethasone in vivo.

Pregnant cows were treated with 5 mg dexamethasone (DEX) (n = 5) or 0.9% saline (n = 3) on Day 255 of gestation. Placentomes were removed on Day 250, Day 260 by laparotomy, and immediately after parturition. Fetal placental tissue (500 mg) was homogenized and frozen for the determination of initial tissue content of progesterone (P(4)) and estrone (E(1)) by radioimmunoassay (RIA). Tissue samples (500 mg) were incubated for 1, 5, or 9 h in the presence of 0, 10, or 100 ng/ml androstenedione (A) or pregnenolone (P(5)). After incubation, tissue was homogenized in the incubation medium (HOM) and P(4) and E(1) content assessed by RIA. Finally, tissue was incubated in medium containing (3)H-A or (3)H-P(5) with or without cortisol (70 ng/ml) to measure conversion of precursors into estrogens (E). Overall initial tissue concentration of P(4) decreased as pregnancy progressed (P < 0.05). An interaction between DEX treatment and stage of gestation (P < 0.05) indicated that P(4) decreased only in tissue from DEX-treated cows, whereas P(4) in tissue from control cows remained constant. Tissue concentrations of E(1) increased from Day 250 to parturition (P < 0.05) and were not influenced by in vivo DEX treatment. Progesterone concentration in HOM increased during incubation indicating P(4) synthesis (P < 0.001). Addition of 100 ng/ml P(5) stimulated P(4) production (P < 0.05). There was a larger increase in P(4) in HOM from DEX-treated cows than in HOM from control cows (P < 0.05). Concentration of E(1) decreased during incubation (P < 0.0001) and was not influenced by the addition of A or P(5). Estrogen synthesis was indicated by the conversion of A or P(5) into E (pmol/g/h). The conversion of precursors (A or P(5)) into E increased during gestation and was not influenced by in vivo DEX treatment. Cortisol application in vitro did not affect E synthesis. It is concluded that treatment with 5 mg DEX in vivo on Day 255 of pregnancy does not seem to influence placental E synthesis in vitro. However, the decrease in initial P(4) tissue concentration and the increase in P(4) synthesis by fetal placental tissue from DEX-treated cows in vitro suggest that DEX treatment in vivo stimulates P(4) synthesis with increased metabolism of P(4) into other metabolites.

Journal Article↗

Pain evaluation of patients with fibromyalgia, osteoarthritis, and low back pain.

UNLABELLED: The purpose of this study was to evaluate and compare pain as reported by outpatients with fibromyalgia, osteoarthritis, and low back pain, in view of designing more adequate physical therapy treatment. PATIENTS AND METHODS: A Portuguese version of the McGill Pain Questionnaire - where subjects are asked to choose, from lists of pre-categorized words, one or none that best describes what they feel - was used to assess pain intensity and quality of 64 patients, of which 24 had fibromyalgia, 22 had osteoarthritis, and 18 had low back pain. The pre-categorized words were organized into 4 major classes - sensory, affective, evaluative, and miscellaneous. RESULTS: Patients with fibromyalgia reported, comparatively, more intense pain through their choice of pain descriptors, both sensory and affective; they also chose a higher number of words from these classes than patients in the other groups and were the only ones to choose specific affective descriptors such as "vicious", "wretched", "exhausting", "blinding". CONCLUSION: Assuming that each disease presents unique qualities of pain experience, and that these can be pointed out by means of this questionnaire by patients' choice of specific groups of words, the findings suggest that fibromyalgia include not only a physical component, but also a psycho-emotional component, indicating that they require both emotional/affective and physical care.

Confidence Intervals↗

Nutrient intake and dental caries in the primary dentition.

An extensive literature has assessed the influence of diet on dental caries, but to date dietary influences on caries of the primary dentition have not been studied widely. This study examined the role of specific nutrients in the caries experience of preschool children. A total of 628 children ages 2 to 6 years received a dental examination, parent interview, and a seven-day diet diary. The diary was completed adequately for 269 children (43%). The average daily intakes of 15 nutrients were computed and compared with recommended dietary allowances (RDA); total sugar consumption and Ca/P ratio were calculated. Each child's gender, age, fluoride history, previous dental experience, and nutrient intakes were examined as independent variables. Stepwise logistic regression showed the combination of categories most predictive of caries was older age, suboptimal fluoride consumption, and recall dental visit. No association was apparent between dental caries and the intake of specific nutrients, total sugar consumption, or Ca/P ratio.

Age Factors↗