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Biomedical subjects

A L Oliveira

Publications and source records attributed to A L Oliveira.

At least 19 recordsLinked to original sources

A comparative analysis of scaffold material modifications for load-bearing applications in bone tissue engineering.

To facilitate optimal application of appropriate scaffold architectures for clinical trials, there is a need to compare different scaffold modifications under similar experimental conditions. In this study was assessed the effectiveness of poly-e-caprolactone (PCL) scaffolds fabricated by fused deposition modelling (FDM), with varying material modifications, for the purposes of bone tissue engineering. The incorporation of hydroxyapatite (HA) in PCL scaffolds, as well as precalcification through immersion in a simulated body fluid (SBF) to produce a biomimetic apatite coating on the scaffolds, was assessed. A series of in vitro studies spanning 3 weeks as well as in vivo studies utilizing a subcutaneous nude mouse model were carried out. PCL and HA-PCL scaffolds demonstrated increasing tissue growth extending throughout the implants, as well as superior mechanical strength and mineralization, as evidenced by X-ray imaging after 14 weeks in vivo. No significant difference was found between PCL and HA-PCL scaffolds. Precalcification with SBF did not result in increased osteoconductivity and cell proliferation as previously reported. Conversely, tensile forces exerted by tissue sheets bridging adjacent struts of the PCL scaffold caused flaking of the apatite coating that resulted in impaired cell attachment, growth and mineralization. The results suggest that scaffolds fabricated by FDM may have load-bearing applications.

Animals↗

Study of the influence of beta-radiation on the properties and mineralization of different starch-based biomaterials.

In this work, the effects of beta-radiation are assessed, for the first time, on starch-based biodegradable polymers, with the aim of using it as an alternative sterilization process to the previously studied sterilization methods. Different doses of radiation were used in order to investigate the possibility of using this sterilization technique as a treatment to tailor the surface and bulk properties (namely mechanical) of these polymers. The as-treated substrates were characterized by water-uptake measurements and contact angle (theta) measurements. The mechanical properties of the materials were characterized by tensile tests by means of ultimate tensile strength (UTS) and strain at break (epsilon). The fracture of the surfaces was observed by scanning electron microscopy (SEM). Dynamic mechanical analysis (DMA) was also used to characterize the viscolelastic behavior of the irradiated materials. The main effect of sterilization with beta-radiation over the starch-based polymers seems to be a surface modification by an increase of the hydrophilicity. Nevertheless, because beta-radiation did not significantly affect the mechanical properties, it can be regarded as an effective way of modifying the surface for applications were more hydrophilic surfaces are desirable.

Beta Particles↗

Development and validation of a LC-method for determination of catechin and epicatechin in aqueous extractives from leaves of Maytenus ilicifolia.

A reverse phase-LC method was developed and validated for separation and quantification of catechin and epicatechin in aqueous extractives from leaves of Maytenus ilicifolia. The analysis was performed using a C(18) column with acetic acid-acetonitrile gradient elution. The detection was carried out by UV at 280 nm and the peak identification was based on the retention times and by co-chromatography with reference substances. High coefficients of determination were achieved for both catechin and epicatechin peaks from the standard solutions (0.9996 and 0.9999), as well as from extractives (0.9981 and 0.9982, respectively). The method showed good repeatability (R.S.D. <1.5%), reproducibility (R.S.D. <5%) and good accuracy for both catechin and epicatechin peaks (101.4%, R.S.D. = 1.18% and 100.6%, R.S.D. = 2.07%, respectively).

Catechin↗

Pre-mineralisation of starch/polycrapolactone bone tissue engineering scaffolds by a calcium-silicate-based process.

This work describes a new methodology to produce bioactive coatings on the surface of starch-based biodegradable polymers or other degradable polymeric biomaterials. As an alternative to the more typical bioactive glass precursors, a calcium silicate gel is being employed as a nucleating agent, for inducing the biomimetic formation of a calcium-phosphate (Ca-P) layer. The method has the advantage of being able to coat efficiently both compact materials and porous 3-D architectures aimed at being used on tissue replacement applications and as bone tissue engineering scaffolds. This treatment is also very effective in reducing the incubation periods, being possible to observe the formation of an apatite-like layer, only after 12 h of immersion in a simulated body fluid (SBF). The apatite coatings formed on the compact surfaces or along the fibres of a fibre mesh scaffold structure made from a starch/polycrapolactone blend (SPCL) were analysed and compared in terms of morphology, chemical composition and structure. After the first days of SBF immersion, the apatite-like films exhibit the typical cauliflower like morphology. With increasing immersion times, these films exhibited a partially amorphous nature and the Ca/P ratios became very closer to the value attributed to hydroxyapatite (1.67). It was possible to fully pre-mineralise the SPCL scaffolds and simultaneously to keep the porous morphology of the fibre-bonded scaffold.

Absorption↗

Sodium silicate gel as a precursor for the in vitro nucleation and growth of a bone-like apatite coating in compact and porous polymeric structures.

In the present work, a new methodology to produce bioactive coatings on the surface of starch-based biodegradable polymers or other polymeric biomaterials is proposed. A sodium silicate gel is employed as an alternative nucleating agent to the more typical bioactive glasses for inducing the formation of a calcium-phosphate (Ca-P) layer. The method has the advantage of being able to coat efficiently both compact materials and porous 3D architectures aimed at being used on tissue replacement applications and as tissue engineering scaffolds. By means of this treatment, it is possible to observe the formation of an apatite-like layer, only after 6 hours of simulated body fluid immersion. For the porous materials, this layer could also be observed inside the pores, clearly covering the cell walls. Furthermore, an increase of the surface hydrophilicity (higher amount of polar groups in the surface) might contribute to the formation of silanol groups that also act as apatite inductors. After 30 days of SBF immersion, the apatite-like films exhibit a partially amorphous nature and the Ca/P ratios became much closer to the value attributed to hydroxyapatite (1.67). The obtained results are very promising for the development of cancellous bone replacement materials and for pre-calcifying bone tissue engineering scaffolds.

Apatites↗

Bi-composite sandwich moldings: processing, mechanical performance and bioactive behavior.

Two composite systems composed of high-density polyethylene (HDPE) filled with hydroxyapatite (HA) and carbon fiber (C fiber) were compounded in a co-rotating twin screw extruder and subsequently molded in a two component injection molding machine in order to produce test bars with a sandwich-like morphology. These moldings are based on a HDPE/HA composite outer layer and a HDPE/C fiber composite core. The mechanical performance of the obtained specimens was assessed by tensile and impact testing. The fracture surfaces were observed by scanning electron microscopy (SEM) and optical reflectance microscopy was used to characterize the morphology within the moldings. In order to study the bioactivity of the molded specimens, the samples were immersed for different periods of time up to 30 days in a simulated-body fluid (SBF) with an ion composition similar to human blood plasma. After each immersion period, the surfaces of the specimens were characterized by SEM. The chemical composition and the structure of the deposited films were studied by electron dispersive spectroscopy (EDS) and thin-film X-ray diffraction (TF-XRD). The evolution of the elemental concentrations in the SBF solution was determined by induced coupled plasma emission (ICP) spectroscopy. Bi-composite moldings featuring a sandwich-like morphology were successfully produced. These moldings present a high stiffness as a result of the C fiber reinforcement present in the molding core. Furthermore, as a result of the HA loading, the sandwich moldings exhibit a clear in vitro bioactive behavior under simulated physiological conditions, which indicates that an in vivo bone-bonding behavior can be expected for these materials.

Journal Article↗

Cell adhesion and proliferation on biomimetic calcium-phosphate coatings produced by a sodium silicate gel methodology.

The present study describes a methodology to produce bioactive coatings on the surface of starch based biodegradable polymers or other polymeric biomaterials. As an alternative to the more typical bioactive glass percursors, a sodium silicate gel is being employed as a nucleating agent, for inducing the formation of a calcium-phosphate (Ca-P) layer. The method has the advantage of being able to coat efficiently both compact materials and porous 3D architectures aimed at being used on tissue replacement applications and as bone tissue engineering scaffolds. This treatment is also very effective in reducing the incubation periods, being possible to observe the formation of an apatite-like layer, only after 6 h of immersion in a simulated body fluid (SBF). The influence of the SBF concentration on the formation of the apatite coating was also studied. The apatite coatings formed under different conditions were analyzed and compared in terms of morphology, chemical composition and structure. After the first days of SBF immersion, the apatite-like films exhibit the typical cauliflower like morphology. With increasing immersion times, these films exhibited a partially amorphous nature and the Ca/P ratios became very closer to the value attributed to hydroxyapatite (1.67). The obtained results are very promising for pre-calcifying bone tissue engineering scaffolds. Therefore, in order to study cell behavior and response to these apatite coatings, adhesion, morphology, and proliferation of a human osteoblast cell line (SaOS-2) was also analyzed after being cultured in the coatings formed after 15 days of immersion in SBF. Results indicate a good correlation between crystallinity of the apatite like coatings formed in these conditions and respective cell spreading and morphology. In general, higher cell proliferation was observed for higher crystalline Ca-P coatings.

Journal Article↗

Population genetics of four PKLR intragenic polymorphisms in Portugal and São Tomé e Príncipe (Gulf of Guinea).

Four intragenic PKLR polymorphisms [1705A/C, 1738C/T. T10/19, and (ATT)n microsatellite] were studied in normal population samples of Central Portugal and São Tomé e Príncipe, a small archipelago located in the Gulf of Guinea, West Africa. For all loci, the observed genotype distributions do not deviate from Hardy-Weinberg equilibrium. The allele frequencies found in the Portuguese population are similar to those previously described in Caucasian populations. Mother-child pair analysis for the (ATT)n microsatellite does not show deviations to the Mendelian rules. In São Tomé e Príncipe the biallelic polymorphisms 1705A/C, 1738C/T, and T10/19 presented inverse allelic frequencies when compared with the Portuguese population. Two new alleles were found at the (ATT)n microsatellite. Significant statistical differences were found between both populations. The results showed that São Tomeans had higher haplotype diversity and lower linkage disequilibrium among the polymorphic sites. The PKLR intragenic polymorphisms, commonly used in haplotype analysis with the gene mutations in PK-deficient patients, can thus be successfully employed in anthropological genetics.

Atlantic Islands↗

Apoptosis of sensory neurons and satellite cells after sciatic nerve transection in C57BL/6J mice.

The rate of axonal regeneration, after sciatic nerve lesion in adult C57BL/6J mice, is reduced when compared to other isogenic strains. It was observed that such low regeneration was not due just to a delay, since neuronal death was observed. Two general mechanisms of cell death, apoptosis and necrosis, may be involved. By using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) technique, we demonstrated that a large number of sensory neurons, as well as satellite cells found in the dorsal root ganglia, were intensely labeled, thus indicating that apoptotic mechanisms were involved in the death process. Although almost no labeled neurons or satellite cells were observed one week after transection, a more than ten-fold increase in TUNEL labeling was detected after two weeks. The results obtained with the C57BL/6J strain were compared with those of the A/J strain, which has a much higher peripheral nerve regeneration potential. In A/J mice, almost no labeling of sensory neurons or satellite cells was observed after one or two weeks, indicating the absence of neuronal loss. Our data confirm previous observations that approximately 40% of C57BL/6J sensory neurons die after sciatic nerve transection, and indicate that apoptotic events are involved. Also, our observations reinforce the hypothesis that the low rate of axonal regeneration occurring in C57BL/6J mice may be the result of a mismatch in the timing of the neurons need for neurotrophic substances, and production of the latter by non-neuronal cells in the distal stump.

Animals↗

GM-1 ganglioside treatment reduces motoneuron death after ventral root avulsion in adult rats.

After ventral root avulsion, a large percentage of the wounded spinal motoneurons die within 2 weeks. Neuronal death is preceded by a series of morphological and physiological changes probably as a result of both apoptotic and necrotic mechanisms. Associated with neuronal death, inflammation develops in the wounded area, resulting in additional neuronal loss as well as the degeneration of fibers from different tracts of the ventral and lateral funiculi. In the present study we tested the neuroprotective and local anti-inflammatory effects of monosialoganglioside (GM-1) after an initial 150 mg/kg body weight dose, followed by the daily administration of 100 mg/kg body weight for 2 weeks. The results showed a statistically significant enhancement of surviving motoneurons which showed good morphological preservation. Also, GM-1 treatment reduced by less then a half, the number of degenerating fibers into the ventral and lateral funiculi. Taken together, our results indicate that the administration of GM-1 in high doses during the critical period of motoneuron death after avulsion is neuroprotective, and diminishes local inflammation.

Animals↗

Differential schwann cell migration in adult and old mice: an in vitro study.

The influence of aging on Schwann cell (SC) proliferation, migration and viability was studied in vitro. SCs were cultured in Ham F-10 medium enriched with 20% fetal calf serum (FCS), 40% FCS or collagen I gel plus 20% FCS. The migration of adult mice derived SCs was stimulated with FCS and collagen. With aging, SC migration, multiplication and viability decreased, indicating that ideal culturing conditions should be adjusted.

Age Factors↗

[Infantile spasms: experience in 13 cases].

Infantile spasms are typical crisis found in the childhood being a serious pathology with an obscure prognosis. We present the experience of the Service of Neuropediatrics in the Pediatric General Center, Belo Horizonte - Brasil, during the year of 1997, with a review of the literature about. After appropriate propaedeutics, were found 12 cases considered symptomatics and 1 case cryptogenic. All these cases were treated with ACTH during 6 weeks associated with oral antiepileptic drugs in mono or polytherapy for maintenance. We had excellent results using ACTH at the beginning of the treatment, obtaining good results in all of the cases with some side effects that were not important enough to interrupt the treatment. However, there was a recurrence rate of 55% and vigabatrin was used as second option, controlling 4 of the 5 cases. All the cases showed retardation of the neuropsychomotor development.

Adrenocorticotropic Hormone↗

Non-neuronal cells are not the limiting factor for the low axonal regeneration in C57BL/6J mice.

Peripheral axonal regeneration was investigated in adult male mice of the C57BL/6J (C), BALB/cJ (B) and A/J (A) strains and in their F1 descendants using a predegenerated nerve transplantation model. Four types of transplants were performed: 1) isotransplants between animals of the C, B and A strains; 2) donors of the C strain and recipients of the C x B and C x A breeding; 3) donors of the B strain and recipients of the C x B breeding, and 4) donors of the A strain and recipients of the C x A breeding. Donors had the left sciatic nerve transected and two weeks later a segment of the distal stump was transplanted into the recipient. Four weeks after transplantation the regenerated nerves were used to determine the total number of regenerated myelinated fibers (TMF), diameter of myelinated fibers (FD) and myelin thickness (MT). The highest TMF values were obtained in the groups where C57BL/6J mice were the donors (C to F1 (C x B) = 4658 +/- 304; C to F1 (C x A) = 3899 +/- 198). Also, A/J grafts led to a significantly higher TMF (A to F1 (C x A) = 3933 +/- 565). Additionally, isotransplant experiments showed that when the nerve is previously degenerated, C57BL/6J mice display the largest number of myelinated fibers (C to C = 3136 +/- 287; B to B = 2759 +/- 170, and A to A = 2835 +/- 239). We also observed that when C57BL/6J was the graft donor, FD was the highest and MT did not differ significantly when compared with the other groups. These morphometric results reinforce the idea that Schwann cells and the nerve environment of C57BL/6J provide enough support to the regenerative process. In this respect, the present results support the hypothesis that the non-neuronal cells, mainly Schwann cells, present in the sciatic nerve of C57BL/6J mice are not the main limiting factor responsible for low axonal regeneration.

Animals↗

Surface modification tailors the characteristics of biomimetic coatings nucleated on starch-based polymers.

This work describes the influence of surface pretreatments over the nucleation and growth of an apatite layer, formed by a biomimetic process, on which a bioactive glass is used as a precursor of the calcium-phosphate (Ca-P) formation on the materials surface. SEVA-C, a corn starch-based biodegradable blend, was used as substrate. The surfaces were pretreated during various periods by: (i) physical methods, namely ultraviolet radiation (u.v.), and over exposure to ethylene oxide sterilization (EtO); and (ii) chemical methods, namely potassium hydroxide (KOH) and acetic anhydride (CH3CO)2 etchings. The surface modifications, performed before the production of the biomimetic coatings, resulted in a faster formation of Ca-P nuclei during the first stages of SBF immersion, particularly in the case of the KOH etching. In this case, it was possible to observe a decrease in the average surface roughness, as measured by laser profilometry, and an increase of the hydrophilicity of the material, which was evident from a clear increment in the water-uptake ability and quantified by contact angle measurements. With this treatment it was possible not only to reduce the induction period for the formation of a well defined and dense apatite-like layer, as observed by scanning electron microscopy (SEM), but also to improve the adhesion of the Ca-P layer to the substrate, as confirmed by the adhesion strength tests. For all the studied pre-treatments, the composition of the films, analyzed by energy dispersive spectroscopy (EDS) and identified by thin-film X-ray diffraction (TF-XRD), seems to be very similar to that of human bone apatites.

Journal Article↗

Protective effect of bifidus milk on the experimental infection with Salmonella enteritidis subsp. typhimurium in conventional and gnotobiotic mice.

The ability of Bifidobacterium bifidum from a commercial bifidus milk to antagonize Salmonella enteritidis subsp. typhimurium in vivo, and to reduce the pathological consequences for the host, was determined using conventional and gnotobiotic mice. Conventional animals received daily, by gavage, 0.1 ml bifidus milk containing about 10(9) cfu B. bifidum and germ-free animals received a single 0.1 ml dose. The conventional and gnotobiotic groups were challenged orally with 10(2) cfu of the pathogenic bacteria 5 and/or 10 d after the beginning of treatment. Control groups were treated with milk. Bifidus milk protected both animal models against the challenge with the pathogenic bacteria, as demonstrated by survival and histopathological data. However, to obtain the protective effect in gnotobiotic animals, the treatment had to be initiated 10 d before the challenge. In experimental and control gnotobiotic mice, Salm. enteritidis subsp. typhimurium became similarly established at levels ranging from 10(8) to 10(9) viable cells g-1 of faeces and remained at these high levels until the animals died or were sacrificed. It was concluded that the protection against Salm. enteritidis subsp. typhimurium observed in conventional and gnotobiotic mice treated with bifidus milk was not due to the reduction of the intestinal populations of the pathogenic bacteria.

Animals↗

[Risk factors for perinatal mortality in Pelotas, a southern city of Brazil, 1993].

INTRODUCTION: Although there was a considerable reduction in infant mortality in Pelotas, Rio Grande do Sul in the last decade, its perinatal causes were reduced only by 28%. The associated factors of these causes were analysed. MATERIAL AND METHOD: All hospital births and perinatal deaths were assessed by daily visits to all the maternity hospitals in the city, throughout 1993 and including the first week of 1994. RESULTS: The perinatal mortality rate was 22.1 per thousand births. The multivariate analysis showed the following risk factors: low socioeconomic level, male sex and maternal age above 35 years. Among multigravidae women, the fetal mortality rate was significantly increased for mothers with a previously low birthweight and a previous stillbirth. For early neonatal mortality the risk was significantly increased by a smaller number of antenatal visits than 5 and low birthweight. CONCLUSIONS: Main risk factors for perinatal mortality: low socioeconomic level, maternal age above 35 years and male sex. For early neonatal mortality the risk was significantly increased by a smaller number of antenatal visits than 5 and low birthweight.

Adult↗

Cutaneous scars in American tegumentary leishmaniasis patients: a site of Leishmania (Viannia) braziliensis persistence and viability eleven years after antimonial therapy and clinical cure.

Two former patients treated for the cutaneous form of American tegumentary leishmaniasis were reviewed eight and 11 years, respectively, following clinical cure. We were able to isolate Leishmania parasites in a culture of material from the two scar biopsies, and in one of them the parasite was characterized as Leishmania (Viannia) braziliensis. In both cases, the histopathology revealed discreet hyperceratosis and a slight infiltrate of mononuclear cells surrounding and on the walls of the surface and deep dermal vessels. No amastigotes were seen on immunohistochemical or histopathologic examination. The Montenegro skin test result and the in vitro lymphoproliferative response to Leishmania antigen were positive, but no specific IgG and IgM antibodies were detected. Otorhinolaryngologic examination showed no macroscopic alteration in the mucosae. These findings are important for the evaluation and criteria of post-treatment cure.

Adult↗