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1.
Mater Today Bio ; 9: 100093, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33665602

ABSTRACT

Spinal cord injury is extremely debilitating, both at physiological and psychological levels, changing completely the patient's lifestyle. The introduction of biomaterials has opened a new window to develop a therapeutic approach to induce regeneration after injury due to similarities with extracellular matrix. Particularly, hydrogels have the ability to support axonal growth and endogenous regeneration. Moreover, they can also act as potential matrixes in which to load and deliver therapeutic agents at injury site. In this review, we highlight some important characteristics to be considered when designing hydrogels as delivery systems (DS), such as rheology, mesh size, swelling, degradation, gelation temperature and surface charge. Additionally, affinity-based release systems, incorporation of nanoparticles, or ion-mediated interactions are also pondered. Overall, hydrogel DS aim to promote a sustained, controlled and prolonged release at injury site, allowing a targeted oriented action of the therapeutic agent that will be used.

2.
J Tissue Eng Regen Med ; 11(7): 1949-1962, 2017 07.
Article in English | MEDLINE | ID: mdl-26510640

ABSTRACT

Osteochondral defects of the ankle are common lesions affecting the talar cartilage and subchondral bone. Current treatments include cell-based therapies but are frequently associated with donor-site morbidity. Our objective is to characterize the posterior process of the talus (SP) and the os trigonum (OT) tissues and investigate their potential as a new source of viable cells for application in tissue engineering and regenerative medicine. SP and OT tissues obtained from six patients were characterized by micro-computed tomography and histological, histomorphometric and immunohistochemical analyses. Proliferation and viability of isolated cells were evaluated by MTS assay, DNA quantification and live/dead staining. The TUNEL assay was performed to evaluate cell death by apoptosis. Moreover, the production of extracellular matrix was evaluated by toluidine blue staining, whereas cells phenotype was investigated by flow cytometry. Characterization of ankle explants showed the presence of a cartilage tissue layer in both SP and OT tissues, which represented at least 20%, on average, of the explant. The presence of type II collagen was detected in the extracellular matrix. Isolated cells presented a round morphology typical of chondrocytes. In in vitro studies, cells were viable and proliferating for up to 21 days of culture. No signs of apoptosis were detected. Flow-cytometry analysis revealed that isolated cells maintained the expression of several chondrocytic markers during culture. The results indicated that the SP and OT tissues were a reliable source of viable chondrocytes, which could find promising applications in ACI/MACI strategies with minimal concerns regarding donor zone complications. Copyright © 2015 John Wiley & Sons, Ltd.


Subject(s)
Cartilage , Cell Proliferation , Talus/cytology , Talus/metabolism , Tissue Engineering/methods , Humans
3.
ACS Biomater Sci Eng ; 1(9): 726-739, 2015 Sep 14.
Article in English | MEDLINE | ID: mdl-33445249

ABSTRACT

This review analyzes the outcomes and technical aspects of in vivo studies published in the past decade using gels and hydrogels for cartilage repair. Using PubMed search engine, original research publications during the period of 2002/01/01 to 2015/04/30 identified 115 published papers. Of these, 3 studies failed to find a statistically significant improvement of treatment group as compared to control and 18 studies did not clearly identify hyaline-like cartilage formation in the treated groups. The most frequent repaired lesion was the rabbit acute full thickness trochlear defect, using a scaffold combining a gel or hydrogel and other material. One third of the scaffolds were cell-free (35%) and the majority of the studies did not use growth factors (71%). The present review may constitute a useful tool in design of future studies, as limitations of study designs are pointed and results in terms of translation to human application is discussed.

4.
Biomed Microdevices ; 16(1): 69-78, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24122322

ABSTRACT

The use of bottom-up approaches in tissue engineering applications is advantageous since they enable the combination of various layers that could be made from different materials and/or incorporate different biochemical cues. Regarding the complex structure and the vascular system of the bone tissue, the aim of this study was to develop an innovative bottom-up approach that allows the construction of 3D biodegradable scaffolds from 2D microfabricated membranes with precise shape, pore size and porosity. For that purpose, poly (caprolactone) (PCL) and starch ­ poly (caprolactone) (SPCL (30 % starch)) blended sheets were used as substrates to produce the microfabricated membranes using micro hotembossing. The use of this micro fabrication process allowed accurately imprinting micropillars and microholes in reproducible way. The assembling of the microfabricated membranes was performed using an easy, highly reproducible and inexpensive approach based on its successive stacking. Additionaly, the suitability of the microfabricated membranes to support the attachment and the cytoskeletal organization of human bone marrow stem cells (hBMSCs), macrovascular endothelial cells and osteoblasts derived from hBMSCs was demonstrated. Furthermore, hBMSCs proliferated and maintained the expression of the stromal progenitor marker STRO-1 when cultured on both PCL and SPCL microfabricated membranes. The proposed methodology constitutes a promising alternative to the traditional processing methods used to prepare tissue engineering scaffolds.


Subject(s)
Bone and Bones/chemistry , Microtechnology/methods , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Cell Adhesion , Cell Differentiation , Cells, Cultured , Human Umbilical Vein Endothelial Cells , Humans , Immunohistochemistry , Mesenchymal Stem Cells/chemistry , Microscopy, Electron, Scanning , Osteoblasts/chemistry , Polymers/chemistry , Porosity
5.
J Tissue Eng Regen Med ; 7(9): 687-96, 2013 Sep.
Article in English | MEDLINE | ID: mdl-22396311

ABSTRACT

The hypothesis of this study was that the extent of bone regeneration could be enhanced by using scaffolds with appropriate geometry, and that such an effect could be further increased by mimicking the natural timing of appearance of bone morphogenetic proteins BMP-2 and BMP-7 after fracture. Bioplotted poly(ε-caprolactone) (PCL) disks with four different fibre organizations were used to study the effect of 3D scaffold architecture on the healing of bone defects in a rat pelvis model. Moreover, one PCL construct was further modified by introducing a nanoparticulate sequential BMP-2/BMP-7 delivery system into this scaffold. Scaffolds and functionalized construct along with free nanocapsules were implanted using a rat iliac crest defect model. Six weeks post-implantation, the defects were evaluated by CT scan and histology. Analysis revealed that the basic architecture, having the highest pore volume for tissue ingrowth, presented the highest bone formation as determined by the bone mineral density (BMD) within the defect (144.2 ± 7.1); about four-fold higher than that of the empty defect (34.9 ± 10.7). It also showed the highest histological analysis scores with a high amount of bone formation within the defect, within the scaffold pores and along the outer surfaces of the scaffold. The basic scaffold carrying the BMP-2/BMP-7 delivery system showed significantly higher bone formation than the growth factor-free basic scaffold at 6 weeks (BMD 206.8 ± 15.7). Histological analysis also revealed new bone formation in close to or in direct contact with the construct interface. This study indicates the importance of open and interconnecting pore geometry on the better healing of bone defects, and that this effect could be further increased by supplying growth factors, as is the case in nature.


Subject(s)
Biocompatible Materials/chemistry , Bone Regeneration/drug effects , Bone and Bones/pathology , Osteogenesis/drug effects , Polyesters/chemistry , Tissue Scaffolds/chemistry , Animals , Bone Density , Bone Morphogenetic Protein 2/administration & dosage , Bone Morphogenetic Protein 2/chemistry , Bone Morphogenetic Protein 7/administration & dosage , Bone Morphogenetic Protein 7/chemistry , Humans , Imaging, Three-Dimensional , Intercellular Signaling Peptides and Proteins , Nanocapsules/chemistry , Pelvis/pathology , Rats , Rats, Sprague-Dawley , Tomography, X-Ray Computed , Wound Healing
6.
Work ; 41 Suppl 1: 1403-8, 2012.
Article in English | MEDLINE | ID: mdl-22316914

ABSTRACT

Public space is endowed with undeniable social relevance, thus becoming a defining element of integration and interaction among its users. Aware of this importance the Universidade Federal da Paraíba (UFPB), linked to the Ministério da Educação e Cultura (MEC), develops the project "UFPB para todos: eliminando barreiras'' (UFPB for all: removing barriers) that aims to conceive an architectural design of an accessible rout to UFPB's campus I, and execute a pilot stretch of this route. This article aims to subsidize the preprojectual phase by understanding the needs of this campus' users, through the concepts of ergonomics and universal design. Was performed out direct observation of the space, and the methods of interviews and observation of user behavior were applied to a group of students with disabilities, through the techniques of assisted walking, photographic and video recording. Based on those, projective guidelines have been defined, which will contribute to the final project's quality, so that this is not a simple application of the rule, but works free of segregating barriers.


Subject(s)
Architectural Accessibility , Disabled Persons , Environment Design , Ergonomics , Universities , Brazil , City Planning , Humans , Interviews as Topic , Needs Assessment , Observation
7.
J Tissue Eng Regen Med ; 5(6): e97-107, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21604382

ABSTRACT

Intervertebral disc (IVD) degeneration is a challenging clinical problem that urgently demands viable nucleus pulposus (NP) implant materials. The best suited biomaterial for NP regeneration has yet to be identified, but it is believed that biodegradable hydrogel-based materials are promising candidates. In this work, we have developed ionic- and photo-crosslinked methacrylated gellan gum (GG-MA) hydrogels to be used in acellular and cellular tissue-engineering strategies for the regeneration of IVDs. The physicochemical properties of the developed hydrogels were investigated by Fourier-transform infrared spectroscopy, (1) H nuclear magnetic resonance and differential scanning calorimetry. The swelling ability and degradation rate of hydrogels were also analysed in phosphate-buffered saline solution at physiological pH for a period of 30 days. Additionally, the morphology and mechanical properties of the hydrogels were assessed under a scanning electron microscope and dynamic compression, respectively. An in vitro study was carried out to screen possible cytotoxicity of the gellan gum-based hydrogels by culturing rat lung fibroblasts (L929 cells) with hydrogel leachables up to 7 days. The results demonstrated that gellan gum was successfully methacrylated. We observed that the produced GG-MA hydrogels possess improved mechanical properties and lower water uptake ability and degradation rate as compared to gellan gum. This work also revealed that GG-MA hydrogels are non-cytotoxic in vitro, thus being promising biomaterials to be used in IVD tissue-engineering strategies.


Subject(s)
Hydrogels/pharmacology , Intervertebral Disc/drug effects , Intervertebral Disc/physiology , Polysaccharides, Bacterial/pharmacology , Tissue Engineering/methods , Animals , Calorimetry, Differential Scanning , Cell Death/drug effects , Cells, Cultured , Fibroblasts/cytology , Fibroblasts/drug effects , Freeze Drying , Hydrogels/chemical synthesis , Hydrogels/chemistry , Magnetic Resonance Spectroscopy , Materials Testing , Mechanical Phenomena/drug effects , Molecular Weight , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/ultrastructure , Rats , Spectroscopy, Fourier Transform Infrared , Water
8.
J Biomed Mater Res A ; 93(3): 852-63, 2010 Jun 01.
Article in English | MEDLINE | ID: mdl-19658177

ABSTRACT

Gellan gum is a polysaccharide manufactured by microbial fermentation of the Sphingomonas paucimobilis microorganism, being commonly used in the food and pharmaceutical industry. It can be dissolved in water, and when heated and mixed with mono or divalent cations, forms a gel upon lowering the temperature under mild conditions. In this work, gellan gum hydrogels were analyzed as cells supports in the context of cartilage regeneration. Gellan gum hydrogel discs were characterized in terms of mechanical and structural properties. Transmissionelectron microscopy revealed a quite homogeneous chain arrangement within the hydrogels matrix, and dynamic mechanical analysis allowed to characterize the hydrogels discs viscoelastic properties upon compression solicitation, being the compressive storage and loss modulus of approximately 40 kPa and 3 kPa, respectively, at a frequency of 1 Hz. Rheological measurements determined the sol-gel transition started to occur at approximately 36 degrees C, exhibiting a gelation time of approximately 11 s. Evaluation of the gellan gum hydrogels biological performance was performed using a standard MTS cytotoxicity test, which showed that the leachables released are not deleterious to the cells and hence were noncytotoxic. Gellan gum hydrogels were afterwards used to encapsulate human nasal chondrocytes (1 x 10(6) cells/mL) and culture them for total periods of 2 weeks. Cells viability was confirmed using confocal calcein AM staining. Histological observations revealed normal chondrocytes morphology and the obtained data supports the claim that this new biomaterial has the potential to serve as a cell support in the field of cartilage regeneration.


Subject(s)
Biocompatible Materials/pharmacology , Cartilage/drug effects , Cartilage/physiology , Polysaccharides, Bacterial/pharmacology , Tissue Engineering/methods , Adult , Aged , Animals , Cell Death/drug effects , Cell Line , Cell Survival/drug effects , Chondrocytes/cytology , Chondrocytes/drug effects , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Mechanical Phenomena/drug effects , Membranes, Artificial , Middle Aged , Phase Transition/drug effects , Polysaccharides, Bacterial/ultrastructure , Rats , Rheology/drug effects , Sepharose/pharmacology , Staining and Labeling , Tissue Scaffolds
9.
Acta Biomater ; 5(5): 1626-38, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19188103

ABSTRACT

Apatite layers were grown on the surface of newly developed starch/polycaprolactone (SPCL)-based scaffolds by a 3D plotting technology. To produce the biomimetic coatings, a sodium silicate gel was used as nucleating agent, followed by immersion in a simulated body fluid (SBF) solution. After growing a stable apatite layer for 7 days, the scaffolds were placed in SBF under static, agitated (80 strokes min(-1)) and circulating flow perfusion (Q=4 ml min(-1); t(R)=15s) for up to 14 days. The materials were characterized by scanning electron microscopy/energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy and thin-film X-ray diffraction. Cross-sections were obtained and the coating thickness was measured. The elemental composition of solution and coatings was monitored by inductively coupled plasma spectroscopy. After only 6 h of immersion in SBF it was possible to observe the formation of small nuclei of an amorphous calcium phosphate (ACP) layer. After subsequent SBF immersion from 7 to 14 days under static, agitated and circulating flow perfusion conditions, these layers grew into bone-like nanocrystalline carbonated apatites covering each scaffold fiber without compromising its initial morphology. No differences in the apatite composition/chemical structure were detectable between the coating conditions. In case of flow perfusion, the coating thickness was significantly higher. This condition, besides mimicking better the biological milieu, allowed for the coating of complex architectures at higher rates, which can greatly reduce the coating step.


Subject(s)
Apatites/chemistry , Apatites/chemical synthesis , Coated Materials, Biocompatible/chemistry , Coated Materials, Biocompatible/chemical synthesis , Polyesters/chemistry , Starch/chemistry , Tissue Scaffolds/chemistry , Bioreactors , Elements , Microscopy, Electron, Scanning , Silicates/chemistry , Solutions , Spectroscopy, Fourier Transform Infrared , X-Ray Diffraction
10.
Electromyogr Clin Neurophysiol ; 47(4-5): 243-50, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17711042

ABSTRACT

AIM: To assess the effect of acupuncture on the temporal and masseter muscles activity employing surface electromyography. METHODS: Thirty volunteers were evaluated according to three groups of acupuncture application: selected local points, selected points at distance and association of local and distant points. Bipolar surface electrodes were positioned bilaterally on the anterior portion of temporal muscle, as well as in the median region of masseter muscle. An electromyograph connected to a computer and a specific software registered the muscular activity before and after acupuncture, in the following experimental conditions: 1- Rest Position (RP); 2- Maximum Intercuspation Clenching (MIC); 3- Bilateral Molar Clenching with Cotton Rolls (BMCCR). The mean values obtained for the activities of the studied muscles were submitted to Analysis of Variance and Tukey complementary test. RESULTS: The electromyographic activity of the studied muscles was lower after the application of the methods of acupuncture in the Rest, and higher after the application of the acupuncture in the Maximum Intercuspation Clenching; the electromyographic activity of the temporal muscle was higher than the masseter muscle in the Rest and lower in the Bilateral molar clenching with cotton rolls; the electromyographic activity showed to be modified after the three used methods of acupuncture in the Maximum Intercuspation Clenching. In this clinical condition, the method of long-distance acupuncture was higher than the application of local needles and the associated method. CONCLUSION: Acupuncture provided alterations in the activity of the studied muscles, favoring conditions of rest and muscular tightness.


Subject(s)
Acupuncture , Electromyography , Masseter Muscle/physiology , Temporal Muscle/physiology , Acupuncture Points , Adult , Bite Force , Female , Humans , Isometric Contraction/physiology , Male
11.
J R Soc Interface ; 4(17): 999-1030, 2007 Dec 22.
Article in English | MEDLINE | ID: mdl-17412675

ABSTRACT

The fields of tissue engineering and regenerative medicine aim at promoting the regeneration of tissues or replacing failing or malfunctioning organs, by means of combining a scaffold/support material, adequate cells and bioactive molecules. Different materials have been proposed to be used as both three-dimensional porous scaffolds and hydrogel matrices for distinct tissue engineering strategies. Among them, polymers of natural origin are one of the most attractive options, mainly due to their similarities with the extracellular matrix (ECM), chemical versatility as well as typically good biological performance. In this review, the most studied and promising and recently proposed naturally derived polymers that have been suggested for tissue engineering applications are described. Different classes of such type of polymers and their blends with synthetic polymers are analysed, with special focus on polysaccharides and proteins, the systems that are more inspired by the ECM. The adaptation of conventional methods or non-conventional processing techniques for processing scaffolds from natural origin based polymers is reviewed. The use of particles, membranes and injectable systems from such kind of materials is also overviewed, especially what concerns the present status of the research that should lead towards their final application. Finally, the biological performance of tissue engineering constructs based on natural-based polymers is discussed, using several examples for different clinically relevant applications.


Subject(s)
Biocompatible Materials , Regenerative Medicine , Tissue Engineering
12.
J Mater Sci Mater Med ; 18(2): 211-23, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17323152

ABSTRACT

This work studies the influence of dynamic biomimetic coating procedures on the growth of bone-like apatite layers at the surface of starch/polycaprolactone (SPCL) scaffolds produced by a 3D-plotting technology. These systems are newly proposed for bone Tissue Engineering applications. After generating stable apatite layers through a sodium silicate-based biomimetic methodology the scaffolds were immersed in Simulated Body Fluid solutions (SBF) under static, agitation and circulating flow perfusion conditions, for different time periods. Besides the typical characterization techniques, Micro-Computed Tomography analysis (micro-CT) was used to assess scaffold porosity and as a new tool for mapping apatite content. 2D histomorphometric analysis was performed and 3D virtual models were created using specific softwares for CT reconstruction. By the proposed biomimetic routes apatite layers were produced covering the interior of the scaffolds, without compromising their overall morphology and interconnectivity. Dynamic conditions allowed for the production of thicker apatite layers as consequence of higher mineralizing rates, when comparing with static conditions. micro-CT analysis clearly demonstrated that flow perfusion was the most effective condition in order to obtain well-defined apatite layers in the inner parts of the scaffolds. Together with SEM, this technique was a useful complementary tool for assessing the apatite content in a non-destructive way.


Subject(s)
Apatites/chemistry , Biomimetic Materials/chemistry , Coated Materials, Biocompatible/chemistry , Imaging, Three-Dimensional/methods , Polyesters/chemistry , Tissue Engineering/methods , Tomography, X-Ray Computed/methods , Bone Substitutes/chemistry , Cell Culture Techniques/methods , Crystallization/methods , Extracellular Matrix/chemistry , Materials Testing/methods , Particle Size , Porosity , Starch/chemistry , Surface Properties , Tomography, X-Ray Computed/instrumentation
13.
J Mater Sci Mater Med ; 14(6): 475-87, 2003 Jun.
Article in English | MEDLINE | ID: mdl-15348431

ABSTRACT

Several coupling treatments based on silane chemicals were investigated for the development of high density (HDPE)/hydroxyapatite (HA) composites. Two HA powders, sintered HA (HAs) and non sintered HA (HAns), were studied in combination with five silanes, namely y-methacryloxy propyltrimethoxy silane (MEMO), 3-(2-aminoethyl)aminopropyltrimethoxy silane (DAMO), vinyltrimethoxy silane (VTMO), 3-aminopropyltriethoxy silane (AMEO) and trimethoxypropyl silane (PTMO). The HA particles were treated by a dipping in method or by spraying with silane solutions. After drying, the treated powders were compounded with HDPE or HDPE with acrylic acid and/or organic peroxide and subsequently compression molded. The tensile test specimens obtained from the molded plates were tensile tested and their fracture surfaces were observed by scanning electron microscopy (SEM). For the sintered HA (HAs) composites, the most effective coupling treatments concerning stiffness are those based on MEMO and AMEO. The low influence of these coupling procedures on strength is believed to be associated to the low volume fraction and the relatively smooth surface of the used HA particles. For the non-sintered HA (HAns) composites, it was possible to improve significantly both the stiffness and the strength. Amino silanes demonstrated to be highly efficient concerning strength enhancement. The higher effectiveness of the coupling treatments for HAns filled composites is attributed to their higher particle surface area, smaller particle size distribution and expected higher chemical reactivity. For both cases, the improvement in mechanical performance after the coupling treatment is consistent with the enhancement in interfacial adhesion observed by SEM.

14.
J Mater Sci Mater Med ; 14(5): 385-97, 2003 May.
Article in English | MEDLINE | ID: mdl-15348441

ABSTRACT

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.

15.
J Mater Sci Mater Med ; 13(10): 939-45, 2002 Oct.
Article in English | MEDLINE | ID: mdl-15348187

ABSTRACT

The present research aims to evaluate the possibility of creating new degradable, stiff and highly bioactive composites based on a biodegradable thermoplastic starch-based polymeric blend and a Bioglass filler. Such combination should allow for the development of bioactive and degradable composites with a great potential for a range of temporary applications. A blend of starch with ethylene-vinyl alcohol copolymer (SEVA-C) was reinforced with a 45S5 Bioglass powder presenting a granulometric distribution between 38 and 53 microm. Composites with 10 and 40 wt % of 45S5 Bioglass were compounded by twin-screw extrusion (TSE) and subsequently injection molded under optimized conditions. The mechanical properties of the composites were evaluated by tensile testing, and their bioactivity assessed by immersion in a simulated body fluid (SBF) for different periods of time. The biodegradability of these composites was also monitored after several immersion periods in an isotonic saline solution. The tensile tests results obtained indicated that SEVA-C/Bioglass composites present a slightly higher stiffness and strength (a modulus of 3.8 GPa and UTS of 38.6 MPa) than previously developed SEVA-C/Hydroxylapatite (HA) composites. The bioactivity of SEVA-C composites becomes relevant for 45S5 amounts of only 10 wt %. This was observed by scanning electron microscopy (SEM) and confirmed for immersion periods up to 30 days by both thin-film X-ray diffraction (TF-XRD) (where HA typical peaks are clearly observed) and induced coupled plasma emission (ICP) spectroscopy used to follow the elemental composition of the SBF as function of time. Additionally, it was observed that the composites are biodegradable being the results correlated with the correspondent materials composition.

16.
J Biochem ; 130(2): 227-33, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11481039

ABSTRACT

There is growing evidence in the literature emphasizing the significance of the post-translational modification of cysteine thiols to sulfenic acids (SOH), which have been found in a number of proteins. Crystallographic and mass spectrometric evidence has shown the presence of this group in an inactive form of the industrially important enzyme nitrile hydratase (NHase). This oxidized cysteine is unique in that it forms part of the coordination sphere of the low-spin iron III at the active site of the enzyme. The presence of this unstable sulfenic group in the active form of NHase is the subject of some controversy. To try to detect this function in NHase, we have studied the inhibitory effect on nitrile hydration of reagents known to react with sulfenic acids. Two NHases were studied, namely, Rhodococcus rhodochrous R312 NHase and Comamonas testosteroni NI1 NHase, and the reagents used were meta-chlorocarbonyldicyano-phenylhydrazone (m-ClCP), 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl), and 2-nitro-5-thiocyanato-benzoic acid (NTBA). Following this approach we report three novel inhibitors of NHases. In addition, we report thiocyanate reagents that can be used to monitor NHase activity spectroscopically.


Subject(s)
4-Chloro-7-nitrobenzofurazan/pharmacology , Enzyme Inhibitors/pharmacology , Hydro-Lyases/antagonists & inhibitors , Iron/metabolism , Sulfenic Acids/metabolism , Thiocyanates/pharmacology , 4-Chloro-7-nitrobenzofurazan/metabolism , Catalytic Domain , Enzyme Inhibitors/metabolism , Hydrazones/metabolism , Hydrazones/pharmacology , Hydro-Lyases/chemistry , Hydro-Lyases/metabolism , Molecular Structure , Oxidation-Reduction , Spectrum Analysis , Thiocyanates/metabolism
17.
J Inorg Biochem ; 84(3-4): 207-13, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11374583

ABSTRACT

A new [Co(N2(SO2)2)(CNtBu)2](Et4N) complex 6 was prepared from N,N'-(3-mercapto-3-methyl-butyryl)-o-phenylenediamine and completely characterized. While the starting square planar complex [Co(N2S2)](Et4N) 4 was destroyed by dioxirane, the Co ligated thiolates of the six-coordinate intermediate [Co(N2S2)(CNtBu)2](Et4N) complex 5 was readily oxidized to sulfinates with a stoichiometric amount of this oxidant. The resulting complex 6 crystallizes with an octahedral structure. The SO bonds of the SO2 groups are almost equivalent (approximately 1.483 and approximately 1.453 A). The isonitrile is linearly bonded to the cobalt with a Co-C-N angle of 177.5 degrees and a very short C-N(tBu) distance of 1.13 A, which has a triple bond character. As expected for six-coordinate CoIII complexes, 5 and 6 are diamagnetic in agreement with their 1H and 13C NMR spectra. The SO2 IR bands are located at 1210 cm(-1) (v(as)SO2) and 1070 cm(-1) (v(s)SO2), while the CN vibration of the isonitrile is observed at 2170 cm(-1) in 5 and 2210 cm(-1) in 6. Very recently, it has been reported in the literature that oxidation of the coordinated thiolates was required for activity of both Fe and Co nitrile hydratases. Complex 6, with two oxidized thiolates trans to two deprotonated carboxamido nitrogens, is the first to have an in-plane closely related to that of the Co-NHase active site.


Subject(s)
Cobalt , Sulfinic Acids/chemical synthesis , Catalytic Domain , Hydro-Lyases , Molecular Structure , Organometallic Compounds/chemistry , Oxidation-Reduction , Spectrophotometry, Infrared
19.
Acta Med Port ; 4 Suppl 1: 59S-63S, 1991 Dec.
Article in Portuguese | MEDLINE | ID: mdl-1805535

ABSTRACT

In the first part of this article some structural and biological aspects of the HIV viruses are presented, in our opinion among the most interesting ones, connected with the AIDS viruses. Viral infection and its evolution, particularly related with infection by HIV-2, will be presented later, in the light of our experience, obtained over several years work with African people infected by the virus. The AIDS viruses are complex retroviruses, with their own identity, but also with marked structural and biological resemblances to other retroviruses, equally pathogenical for animals. The lentivirinae subfamily to which the AIDS viruses belong includes other agents, usually classified according to the host they infect. In this way, the lentiviruses of the primates contain in the same group, besides those of HIV-1 and HIV-2, viruses that infect monkeys such as SIVMAC, SIVAGM, SIVSMM, etc. The comparative study of molecular genetics and biology of human and animal retroviruses in recent years has permitted significant progress in the understanding of the possible mechanisms that lead to the Immunodepressive Acquired Syndrome that characterizes AIDS. The presence of a gene that deactivates the activated lymphocytes only present in the lentiviruses of the primates, as well as the known tropism of those viruses to CD4 lymphocytes, and not found in the other groups, are biological aspects that are pointed out. We also refer to other characteristics of HIVs such as the cytolytic and sincicial capacity of these viruses in lymphocyte culture. Finally we present an analysis of what we were able to observe in individuals infected by HIV-2 in their own and African habitat.


Subject(s)
HIV-1 , HIV-2 , Acquired Immunodeficiency Syndrome/microbiology , Africa, Western , Animals , HIV-1/genetics , HIV-1/physiology , HIV-1/ultrastructure , HIV-2/genetics , HIV-2/physiology , HIV-2/ultrastructure , Humans
20.
Acta Med Port ; 4 Suppl 1: 64S-66S, 1991 Dec.
Article in Portuguese | MEDLINE | ID: mdl-1805536

ABSTRACT

In the geographic distribution of HIV-2, it is known that this infection is most prevalent in West Africa. Since 1986 we have studied seropositive and seronegative clusters, in Guinea-Bissau with follow-ups in 1988, 1989, 1990 and 1991. Analysis of the results show the high incidence of this infection. 8.51% of the 4,372 people of the general population studied were seropositive, showing the high predominance of HIV-2 infection. Only 4 cases were exclusively reactive to HIV-1 and a slow evolution of HIV-1 infections. In the seroconversions of HIV-2 infections the antibodies appeared first to the core components and secondly to the surface glycoproteins. Some of the laboratory parameters affected in the evolution of the infection include a gradual increase in immunoglobulins and a decrease in CD4 lymphocytes and in the CD4/CD8 ratio. A comparison of these variations in HIV-2 infected people, with or without cross-reactivity to HIV-1, reveals that they are much more evident in exclusively HIV-2 positive people. This fact can indicate that the variants responsible for the cross-reactions are less pathogenic and phylogenetically less developed.


PIP: In 1986, a total of 4372 persons were included in an HIV-2 seroepidemiological study covering the whole of Guinea-Bissau. An 8.5% incidence of HIV-2 infection was revealed by the enzyme-linked immunosorbent assay (ELISA) confirmed by Western Blot and/or RIPA. Only 4 cases of HIV-1 seropositivity were found. Annual follow-up of 78 seropositive (e died of AIDS) and 320 seronegative individuals in 1988, 1989, 1990, and 1991 was initiated. In the present investigation, a total of 70 individuals were included: 19 were seropositive (6 with double reactivity) and 51 were seronegative. During this period there were 3 seroconversion cases among seronegative persons, and the appearance of double reactivity in 3 previously HIV-2 positive cases. The reexamination of sera before seroconversion indicated reactivity to the core proteins and in 3 cases to GP-160 or GP-140 surface glycoproteins. The average values of immunoglobulins increased, but the beta-2 microglobulin, and cytomegalovirus antibodies (CMV) did not differ in the sera of 1250 decreased patients compared with seronegative persons. In seropositives, a progressive diminution of the CD4 lymphocyte values and of the T4/T8 (CD4/CD8) ratio was observed which was much more evident in HIV-2 monoreactive patients than in double reactive patients. The interpretation of HIV-1 and HIV-2 double reactivity whether the result of a double infection or of an intermediate variant of the virus between HIV-1 and HIV-2 has not been reconciled with these results. These findings indicate that the variants responsible for double reactivity are less pathogenic, less adapted to the human host, and less developed phylogenetically.


Subject(s)
HIV Seropositivity/immunology , HIV-2/immunology , Africa, Western , Follow-Up Studies , Humans
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