In vivo evaluation of Vivere bovine pericardium valvular bioprosthesis with a new anti-calcifying treatment.

The objective of this study was to evaluate the effect of chemical treatment with glutamic acid to avoid calcification of biological cardiac valves. The bovine pericardium (BP) tissues were fixed with 0.5% glutaraldehyde (BP/GA), followed by treatment with glutamic acid (BP/GA + Glu) for neutralization of the free aldehyde groups. Microscopic analysis showed that the wavy structure of collagen fibrils was preserved, but changes in elastin's integrity occurred. However, the treatment did not promote undesirable changes in the thermal and mechanical properties of the modified BPs. These samples were systematically studied in rat subcutaneous tissue: control (BP/GA) and anticalcificant (BP/GA + Glu). After 60 days, both groups induced similar inflammatory reactions. In terms of calcification, BP/GA + Glu remained more stable with a lower index (3.1 ± 0.2 µg Ca2+ /mg dry tissue), whereas for BP/GA it was 5.7 ± 1.3 µg Ca2+ /mg dry tissue. Bioprostheses made from BP/GA + Glu were implanted in the pulmonary position in sheep, and in vivo echocardiographic analyses revealed maintenance of valvar function after 180 days, with low gradients and minimal valve insufficiency. The explanted tissues of the BP/GA + Glu group had a lower average calcium content 3.8 ± 3.0 µg Ca2+ /mg dry tissue. The results indicated high anticalcification efficiency of BP/GA + Glu in both subcutaneous implant in rats and in the experimental sheep model, which is an advantage that should encourage the industrial application of these materials for the manufacture of bioprostheses.

The calcium paradox - what should we have to fear?

The calcium paradox was first mentioned in 1966 by Zimmerman et al. Thereafter gained great interest from the scientific community due to the fact of the absence of calcium ions in heart muscle cells produce damage similar to ischemia-reperfusion. Although not all known mechanisms involved in cellular injury in the calcium paradox intercellular connection maintained only by nexus seems to have a key role in cellular fragmentation. The addition of small concentrations of calcium, calcium channel blockers, and hyponatraemia hypothermia are important to prevent any cellular damage during reperfusion solutions with physiological concentration of calcium.

Paradoxo do cálcio - o que temos a temer? / The calcium paradox - What should we have to fear?

O paradoxo do cálcio foi pela primeira vez citado em 1966 por Zimmerman et al. A partir daí, ganhou grande interesse por parte da comunidade científica internacional devido ao fato da ausência do íon cálcio produzir na célula muscular cardíaca dano semelhante à lesão de isquemia-reperfusão. Apesar de não serem conhecidos todos os mecanismos envolvidos no processo da lesão celular no paradoxo do cálcio, a conexão intercelular mantida somente pelo nexus parece ter papel chave na fragmentação celular. A adição de pequenas concentrações de cálcio, bloqueadores de canal de cálcio, hiponatremia ou hipotermia são importantes para evitar que haja lesão celular no momento da reperfusão com soluções com concentração fisiológica de cálcio.

The calcium paradox was first mentioned in 1966 by Zimmerman et al. Thereafter gained great interest from the scientific community due to the fact of the absence of calcium ions in heart muscle cells produce damage similar to ischemia-reperfusion. Although not all known mechanisms involved in cellular injury in the calcium paradox intercellular connection maintained only by nexus seems to have a key role in cellular fragmentation. The addition of small concentrations of calcium, calcium channel blockers, and hyponatraemia hypothermia are important to prevent any cellular damage during reperfusion solutions with physiological concentration of calcium.

Regeneration of critical bone defects with anionic collagen matrix as scaffolds.

The aim of this study was to make a histomorphometric evaluation of the osteogenic potential of anionic collagen matrix as scaffolds; either crosslinked in glutaraldehyde or not cross-linked and, implanted in critical bone defects in rat calvaria. Seventy-two rats were randomly distributed in three groups: anionic collagen scaffolds treated for 24 h of selective hydrolysis (ACSH); anionic collagen scaffolds treated for 24 h of selective hydrolysis and 5 min of crosslinking in glutaraldehyde 0.05% (ACSHGA); empty bone defect (Control), evaluated at the biological points of 15, 45, 90 and 120 days. The results showed that the biomaterials implanted were biocompatible and showed a high osteogenic potential. These biomaterials presented a speed of biodegradation compatible with bone neoformation, which was shown to be associated with angiogenesis inside the scaffolds at all biological points. The percentage of mineralization of ACSH (87%) differed statistically from that found in ACSHGA (66%). It was concluded that the regeneration of critical bone defect was more evident in anionic collagen without crosslinking (ACSH).

Bovine osteoblasts cultured on polyanionic collagen scaffolds: an ultrastructural and immunocytochemical study.

Collagen is the most abundant protein in the body and is also the most important component of the extracellular matrix. Collagen has several advantages as a biomaterial such as lack of toxicity, biocompatibility, biodegradability, and easy reabsorption. In this study, we examined bovine osteoblasts cultured on native or anionic collagen scaffolds prepared from bovine pericardium after selective hydrolysis of glutamine and asparagine side chain amides for periods from 24 (BP24) and 48 h (BP48). The cells were cultured in control and mineralization medium at 37 °C in the presence of 5% CO(2). Transmission and scanning electron microscopy, energy dispersive spectroscopy, and an immunocytochemical marker were used for analysis. Cells with an irregular morphology forming a confluent multilayer were observed on matrices kept in control medium. Most of these cells presented a polygonal or elongated flattened morphology. Several spherical deposits of calcium crystal associated with phosphorus were observed on the native and BP48 matrices. Similar results were observed in samples kept in control medium except with lower calcium/phosphorus ratio. Vesicles actively expelled from the cell membrane were also seen (do this vesicles corresponds to calcium/phosphorus deposits). Osteocalcin was clearly visible on matrices kept in mineralization medium and was more expression on the surface of BP48 matrices. The results showed that anionic collagen is able to support osteoblastic differentiation, regardless of the medium used. Finally, the BP48 matrix promoted better osteoblast differentiation than the native matrix.

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.

Preparation and characterization of an acellular bovine pericardium intended for manufacture of valve bioprostheses.

Major problems with biological heart valves post-implantation are associated with progressive structural deterioration and calcification attributed to glutaraldehyde processing, dead cells, and cell fragments present in the native tissue. In spite of these problems, glutaraldehyde still is the reagent of choice. The results with acellular matrix xenograft usually prepared by detergent treatment in association with enzymes are rather conflicting because while preserving mechanical properties, tissue morphology and collagen structure are process dependent. This work describes a chemical approach for the preparation of an acellular bovine pericardium matrix intended for the manufacture of heart valve bioprostheses. Cell removal was performed by an alkaline extraction in the presence of calcium salts for periods ranging from 6 to 48 h. The results showed that cell removal was achieved after 12 h, with swelling and negative charge increasing with processing time. Nevertheless, collagen fibril structure, ability to form fibrils, and stability to collagenase were progressive after 24-h processing. There was no denaturation of the collagen matrix. A process is described for the preparation of acellular bovine pericardium matrices with preserved fibril structure and morphology for the manufacture of cardiac valve bioprostheses and may be used in other applications for tissue reconstruction.

In vitro properties and performance of glutaraldehyde-crosslinked bovine pericardial bioprostheses treated with glutamic acid.

Calcification is one of the major causes of failure of heart valve bioprostheses (HVBs) derived from glutaraldehyde (GA)-processed bovine pericardium (BP) or porcine aortic valves. New crosslinking reagent procedures are still far from giving satisfactory results, and this is the main reason why GA is still the reagent of choice for the fixation of native tissue intended for HVB manufacture. Nevertheless, two new findings with respect to GA processing may significantly improve HVB performance postimplantation: the finding that increasing concentrations of GA result in a decrease in calcification; the blocking of free aldehyde usually by nucleophyles or the treatment of processed material at low pH. This work investigates the in vitro properties of BP fixed with GA followed by the treatment with glutamic acid under alkaline conditions in order to prepare BP materials with lower calcification potential postimplantation. In comparison to conventional processing, except for the tensile strength that was slightly lower, elongation and toughness were higher than the accepted values. No significant differences were observed in the performance indexes (mean pressure gradient, mean effective area, regurgitant fraction, performance and efficiency indexes) with wear resistance over 150 × 106 cycles. These results indicate that the processing of BP described in this work may be of potential use in the manufacture of HVBs.

Compósito de colágeno com silicato e hidroxiapatita como material para endodontia: preparação e caracterização / Collagen composite with silicate and hydroxyapatite as endodontic material: preparation and characterization

Objetivo: Este trabalho teve por objetivo preparar e caracterizar um novo compósito de colágeno com silicato e hidroxiapatita que possa vir a ser utilizado como material endodôntico em capeamento pulpar direto e em pulpotomia. Metodologia: Os materiais utilizados como matéria-prima foram colágeno do tipo I na forma aniônica, silicato (cimento Portland Branco) e hidroxiapatita sintetizada. Estes materiais foram misturados em proporções variadas de modo a obter um material compósito na forma de pasta. Os materiais obtidos foram caracterizados por técnicas de calorimetria exploratória diferencial, termogravimetria, microscopia eletrônica de varredura (MEV), espectrometria de dispersão de energia de Raios-X e espectroscopia na região do infravermelho.Resultados: A presença de colágeno no compósito foi demonstrada por transições térmicas típicas daquelas encontradas para o colágeno do tipo I (-48ºC), independentemente da proporção utilizada, e também por imagens de MEV, onde as fibras colagênicas também puderam ser observadas interligando as partículas de silicato. A composição química de todos compósitos de colágeno foi praticamente idêntica à do silicato utilizado.Conclusão: Os resultados mostraram que a estrutura da matriz de colágeno nos compósitos testados não foi alterada. Esta é uma característica importante para a utilização em endodontia do material compósito proposto, objetivando uma reparação tecidual de forma 'mais natural'.

Purpose: This study aimed to prepare and characterize a new collagen composite with silicate and hydroxyapatite as a potential endodontic material for pulp capping and pulpotomy. Methods: Anionic type I collagen was mixed with silicate (White Portland cement) and hydroxyapatite in several proportions. The materials were characterized by standard differential scanning calorimetry, thermogravimetry, scanning electron microscopy (SEM), energy dispersion x-ray, and infrared spectrometry. Results: The presence of collagen in the composite was observed by means of typical thermal transitions for collagen type I (-48ºC), independently from the proportion used. SEM images showed collagen fibers interconnecting silicate particles. The chemical composition of all collagen composites was essentially the same as that of the original silicate. Conclusion: These results showed that the collagen matrix structure of the tested composite was not altered, which is an important feature for a 'more natural' tissue repair in Endodontics.

Implants of polyanionic collagen matrix in bone defects of ovariectomized rats.

In recent years, there has been a great interest in the development of biomaterials that could be used in the repair of bone defects. Collagen matrix (CM) has the advantage that it can be modified chemically to improve its mechanical properties. The aim of the present study was to evaluate the effect of three-dimensional membranes of native or anionic (submitted to alkaline treatment for 48 or 96 h) collagen matrix on the consolidation of osteoporosis bone fractures resulting from the gonadal hormone alterations caused by ovariectomy in rats subjected to hormone replacement therapy. The animals received the implants 4 months after ovariectomy and were sacrificed 8 weeks after implantation of the membranes into 4-mm wide bone defects created in the distal third of the femur with a surgical bur. Macroscopic analysis revealed the absence of pathological alterations in the implanted areas, suggesting that the material was biocompatible. Microscopic analysis showed a lower amount of bone ingrowth in the areas receiving the native membrane compared to the bone defects filled with the anionic membranes. In ovariectomized animals receiving anionic membranes, a delay in bone regeneration was observed mainly in animals not subjected to hormone replacement therapy. We conclude that anionic membranes treated with alkaline solution for 48 and 96 h presented better results in terms of bone ingrowth.

Biocompatibility of anionic collagen matrices and its influence on the orientation of cellular growth / Biocompatibilidade de matrizes de colágeno aniônico e sua influência na orientação do crescimento celular

This study aimed to analyze the biocompatibility of anionic collagen matrices, the local bone response following implantation in surgically-created bone defects, and also the influence of the collagen fiber orientation on the neoformed osseous tissue. Seventy two rats (Rattus norvegicus albinus) were used in this experiment. The animals were divided into four experimental groups: Group 1 (control), without implants; Group 2, pericardium medullar implants 12 hours; Group 3, medullar implants of tendon 24 hours, with the long axis of collagen fibers oriented parallel to the long axis of the tibia; and Group 4, medullar implants of tendon 24 hours, with the long axis of collagen fibers perpendicular to the long axis of the tibia. After the experimental surgery, the evolution of the repair process was microscopically evaluated at 7, 15, and 30 days post-surgery. The results demonstrated that the implanted matrices are biocompatible and act as a scaffold inducing bone formation, mainly in the Group 4 animals. At first, cellularity follows the arrangement of collagen fibers, later obtaining a multidirectional growth.

Este estudo apresentou como objetivo, examinar a biocompatibilidade de matrizes de colágeno aniônico, sua respostaóssea local após implantação em defeitos ósseos criados cirurgicamente e a influência da orientação das fibras colágenasno tecido ósseo neoformado. Foram utilizados 72 ratos (Rattus norvegicus albinus) divididos em 4 grupos experimentaisde 6 animais cada: Grupo 1 (Controle), sem implante; Grupo 2, implante medular de pericárdio 12 horas; Grupo 3,implante medular de tendão 24 horas, com o maior eixo de suas fibras colágenas direcionados paralelamente ao maioreixo da tíbia e, Grupo 4, implante medular de tendão 24 horas, com o maior eixo de suas fibras colágenas direcionados perpendicularmente ao maior eixo da tíbia. A evolução do processo de reparo, após a cirurgia experimental, foi avaliada microscopicamente com 7, 15 e 30 dias pós-operatórios. Os resultados demonstraram que as matrizes implantadas são biocompatíveis e funcionam como uma matriz tridimensional induzindo a formação de osso, maior no grupo 4. A celularidade, inicialmente, acompanha o arranjo das fibras colágenas, adquirindo um arranjo multidirecional crescente.

Rheological behavior of anionic collagen injectable gels in the presence of rhamsan for plastic surgery applications.

The present paper describes the rheological properties of anionic collagen gels and anionic collagen:rhamsan composites gels in the concentration of 0.7, 4 and 6%, estimated to be used as injectable biomaterials for plastic reconstruction. Rheological studies of these gels showed that independently of pH, composition and concentration the viscoelastic behavior was dependent on the frequency, with the storage modulus always greater than the loss modulus (G' > G'' and delta < 45 degrees ). Creep experiments showed that anionic collagen:rhamsan composites equilibrated at pH 7.4 were less elastic and more susceptible to deformation in comparison to gels equilibrated at pH 3.5. Flow experiments indicated that the force needed for the extrusion of anionic collagen:rhamsan composites, in comparison to anionic collagen, was significantly smaller and with a smoother flow, suggesting the association with rhamsan may be a good alternative in the replacement of glutaraldehyde to stabilize the microfibril assembly of commercial collagen gel preparations. Finally, on the basis of dynamic viscosity profiles found for different preparations, some of these composites are potential candidates to be utilized in laryngology.

Morphological assessment of the behavior of three-dimensional anionic collagen matrices in bone regeneration in rats.

The osteogenic behavior of three-dimensional collagenic and anionic matrices (M3DGA) implanted in critical bone defects of 8 mm diameter was morphologically assessed. The defects were performed in the calvaria of 48 adult male (Wistar) rats, and observed at days 15, 30, and 60. The animals were distributed in four groups of equal number: GA1 (M3DGA with 60 min of cross-linking in glutaraldehyde [GA]); GA2 (M3DGA with 30 min of cross-linking in GA); GA3 (M3DGA with 15 min of cross-linking in GA); and G4 (control group, without any implanted biomaterial). The M3DGAs were biocompatible, with mild and regressive chronic granulomatous inflammation, associated with the degradation of their fibers. Furthermore, tissue neovascularization and neomineralization was noted, with statistically significant differences in these amounts among the groups (GA1, GA2, and GA3) and G4. In G4 neoformation was limited to the edges of the defect as well as interstitial fibrosis. Under the experimental conditions and for the parameters analyzed, the M3DGAs had osteogenic behavior, which was more evident in M3DGA 15 min.

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.

Injectable gels of anionic collagen:rhamsan composites for plastic correction: preparation, characterization, and rheological properties.

The present article describes the preparation and characterization of anionic collagen gels obtained from porcine intestinal submucosa after 72 h of alkaline treatment and in the form of rhamsan composites to develop injectable biomaterials for plastic reconstruction. All materials were characterized by SDS/polyacrylamide gel electrophoresis, infrared spectroscopy, thermal stability, potentiometric titration, rheological properties, and fluidity tests. Biocompatibility was appraised after the injection of anionic collagen: rhamsan composites at 2.5% in 60 North Folk rabbits. Independently of processing, the collagen's secondary structure was preserved in all cases, and after 72 h of hydrolysis the collagen was characterized by a carboxyl group content of 346+/-9, which, at physiological pH, corresponds to an increase of 106+/-17 negative charges, in comparison to native collagen, due to the selective hydrolysis of asparagine and glutamine carboxyamide side chain. Rheological studies of composites at pH 7.4 in concentrations of 2, 4, and 6% (in proportions of 75:1 and 50:1) showed a viscoelastic behavior dependent on the frequency, which is independent of concentration and proportion. In both, the concentration of the storage modulus always predominated over the loss modulus (G'>G'' and delta<45 degrees ). The results from creep experiments confirmed this behavior and showed that anionic collagen:rhamsan composites at pH 7.4 in the proportion of 50:1 are less elastic and more susceptible to deformation in comparison to gels in the proportion of 75:1, independent of concentration. This was further confirmed by flow experiments, indicating that the necessary force for the extrusion of anionic collagen:rhamsan composites, in comparison to anionic collagen, was significantly smaller and with a smooth flow. Biocompatibility studies showed that the tissue reaction of anionic collagen:rhamsan composites at 2.5% in the proportion of 75:1 was compatible with the application of these gels in plastic reconstruction. These results suggest that the association of collagen with rhamsan may be a good alternative in the replacement of glutaraldehyde to stabilize the microfibril assembly of commercial collagen gel preparations.

Comparação entre o pericárdio bovino decelularizado e o pericárdio bovino convencional utilizado na confecção de biopróteses valvares cardíacas / Comparison between the decellularized bovine pericardium and the conventional bovine pericardium used in the manufacture of cardiac bioprostheses

Objetivo: Neste estudo, tivemos como objetivo comparar a resistência mecânica do pericárdio decelularizado com o pericárdio convencional, assim como avaliar sua capacidade de induzir resposta inflamatória em modelo experimental com ratos.Método: Dividimos os pericárdios em: GrupoI - pericárdio submetido a tratamento convencional. Após o processamento químico, as amostras do GrupoII foram histologicamente avaliadas para confirmar a eficácia da decelularização. A seguir, apenas para análise da resistência mecânica por testes de tração e de desnaturação térmica, os pericárdios foram divididos em: grupo 1 (pericárdio convencional com critérios de aprovação), grupo 2(pericárdio convencional com critérios de reprovação) e grupo 3(pericárdio decelularizado). A capacidade de induzir resposta inflamatória foi avaliada em estudo experimental em 50 ratos Wistar, os quais foram submetidos a implante subcutâneo d fragmentos dos pericárdios. Nossa terceira etapa de avaliação consistiu em confeccionar três biopróteses com o pericárdio decelularizado e que foram submetidas à avaliação hidrodinâmica, juntamente com uma bioprótese convencional de teste.Resultados: A análise histológica inicial demostrou decelularização completa. A resistência mecânica mostrou diferença significativa com relação às variáveis "tensão de ruptura" e "índice de tenacidade". Não encontramos diferença quanto a atividade inflamatória em modelo experimental com ratos. O desempenho hidrodinâmico foi semelhante e todas biopróteses atingiram a marca de 150 milhões de ciclos. A avaliação histológica ao fim da ciclagem mostrou padrão microscópico habitual, não havendo ruptura ou fragmentação anormal induzida por estresse mecânico.Conclusão: A decelularização mantém a resistência física do pericárdio, além de não induzir resposta inflamatória diferente daquela habitualmente encontrada no pericárdio convencional

A study on the biocompatibility and integration of acellular polyanionic collagen: elastin matrices by soft tissue / Um estudo sobre biocompatibilidade e integração de matrizes acelulares polianiônicas de colágeno:elastina com tecidos moles

Uma alternativa para o transplante de tecido e órgãos ou para a substituição de funções biológicas é a engenharia de tecido, baseada no crescimento de células isoladas em matrizes tridimensionais biodegradáveis. Este trabalho descreve a preparação, a caracterização e o comportamento de matrizes polianiônicas de colágeno:elastina preparadas a partir de pericárdio bovino nativo e implantadas no subcutâneo de ratos. Os resultados mostraram que estes materiais correspondem a matrizes acelulares polianiônicas de colágeno:elastina formadas por fibras colágenas preservadas. Em comparação ao pericárdio nativo, a resposta biológica foi caracterizada pelo decréscimo progressivo da resposta inflamatória, e, praticamente, não existindo para o material com elevados grupos de carboxila, sugerindo alto grau de biocompatibilidade destas matrizes acelulares de colágeno:elastina. Os resultados mostram também evidências da incorporação do material pelo tecido adjacente, sugerindo que as matrizes polianiônicas de colágeno:elastina podem ser de uso potencial na engenharia de tecido

Biomimetic mineralization of charged collagen matrices: in vitro and in vivo study.

Polyanionic collagen matrix prepared by hydrolysis side chain amides of asparagine and glutamine was mineralized in vivo, without inflammatory response, biodegradation, or resorption, with calcium phosphate deposited in close resemblance to the D-periodicity of collagen fibrils assembly. In vitro results with the same material produced mineralized collagen fibers with a similar morphology and chemical characteristics, suggesting that amide hydrolysis may have introduced into this matrix, signs for the controlled mineralization of collagen fiber. TEM indicated that amide hydrolysis occurred near the OVERLAP and GAP zones, as suggested by the significant reduction in inter-band distances in these regions. The lack of an inflammatory response associated to the similar mineralization pattern observed in vivo and in vitro suggests not only the biomimetic behavior of polyanionic collagen matrix, but also its potential uses as scaffold for bone tissue reconstruction. Based on these results, a model for the in vitro mineralization was also proposed.

Tissue response to polyanionic collagen: elastin matrices implanted in rat calvaria.

The tissue response to polyanionic collagen matrices, prepared from bovine pericardium and implanted subperiosteally in rat calvaria, was studied. The materials were implanted in 72 male rats (Rattus norvegicus, albinus, Holtzman), randomly divided into four groups: GI-MBP hydrolyzed for 24 h; GII-MBP hydrolyzed for 36 h; GIII-MBP hydrolyzed for 48 h; GIV-native MBP. The materials were explanted after 15, 30 and 60 days and analyzed by routine histological procedures. Except for group IV (native bovine pericardium), polyanionic collagen from groups GI, GII and GIII showed low inflammatory reaction associated with bone formation, partially or completely integrated to the cranial bone; group GIV was characterized by an intense inflammatory reaction with occasional dystrophic mineralization and with occasional bone formation at 60 days when there was a decrease in the inflammatory reaction. Thus, the MBP from groups I, II and III were biologically compatible, enhancing bone formation with a slight delay at 60 days in GII.

Compósitos de colágeno aniônico: ramsana como géis injetáveis para correções plásticas: preparação, caracterização e propriedades reológicas / Anionic collagen:rhamsan composites as injectable gels for plastics reconstruction: preparation, characterization and rheological properties

Este trabalho descreve a preparação e a caracterização de compósitos de colágeno aniônico:ramsana, com o propósito de desenvolver géis injetáveis para correções plásticas. Os materiais foram caracterizados por eletroforese, espectroscopia no infravermelho, estabilidade térmica, titulação potenciométrica, propriedades reológicas e ensaios de fluidez. Independentemente do processamento, os materiais estudados são formados por colágeno não desnaturado, com um total de grupos carboxílicos de 345,9 ± 9/mol de colágeno, o que representa a pH fisiológico um excedente de 106 ± 17 cargas negativas em relação ao colágeno nativo. O estudo reológico mostrou que: independentemente do pH, géis de colágeno aniônico e seus compósitos com ramsana apresentaram o módulo de armazenamento sempre maior que o módulo de perda (G' > G" e delta <45 graus); géis colégeno aniônico:ramsana equilibrados a pH 7,4, embora menos elásticos que os géis equilibrados a pH 3,5; géis contendo ramsana foram mais susceptíveis à deformação; a força necessária para o escoamento de géis contendo ramsana foi sempre significativamente menor que aquela determinada para o colágeno aniônico e com um perfil mais limpo em relação à presença de picos de contrapressão, sugerindo que a associação colágeno aniônico:ramsana pode substituir com vantagens o glutaraldeído na estabilização das preparações comerciais do gel. Os resultados de biocompatibilidade de géis de CAR injetados na membrana vesical de coelho mostraram que estes compósitos são compatíveis com a sua utilização em correções plásticas.