Bacterial cellulose-collagen nanocomposite for bone tissue engineering

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Elastic system of the rat ventral prostate and its modifications following orchiectomy

BACKGROUND. The extracellular matrix (ECM) has important roles in prostatic development, and marked stromal changes take place in the rat ventral prostate (VP) after androgen deprivation. However, little knowledge exists about individual ECM components.METHODS. The distribution of elastic fibers (EF) and elastic-related fibers (ERF) in the VP of castrated and control rats was investigated, using histochemistry and transmission electron microscopy (TEM).RESULTS. EF are barely detected in the prostatic stroma, but ERF are relatively abundant. Castration results in a relative increase in the number and thickness of ERF. TEM showed an open network of ECM microfibrils throughout the stroma and thin and short EF, which increase in number and thickness after orchiectomy.CONCLUSIONS. The presence of elastic system components in the rat VP warrants the deformability required for the secretion exclusion under the action of smooth muscle cells, and the castration-induced modification may be related to the contraction of the tissue and maintenance of peculiar arrangements of other ECM components. (C) 1997 Wiley-Liss, Inc.

Tissue remodeling in Guinea pig lateral prostate at different ages after estradiol treatment

Estrogen seems to have an essential role in the fibromuscular growth characteristic of benign prostatic hyperplasia (BPH). This paper describes the effects of chronic estradiol treatment on Guinea pig prostatic stroma at different ages. Tissues from experimental animals were studied by histological and histochemical procedures, morphometric-stereological analysis and transmission electron microscopy (TEM). Marked fibromuscular hypertrophy was observed after estradiol treatment in animals of pre-pubertal and adult ages. Increases in the density and thickness of the collagen and elastic fibers were observed by histochemistry. TEM revealed wide distributions of collagen fibrils and large elastic fibers adjacent to the epithelial basal lamina and between the stromal cells, establishing contacts between them. These results indicate that the Guinea pig prostate simulates the stromal modifications observed in BPH in some aged animals after estrogen treatment at different ages, making it a good model for this disease. (c) 2005 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Evaluation of water and sucrose diffusion coefficients in potato tissue during osmotic concentration

The water and sucrose effective diffusion coefficients behavior were studied in potato tubers immersed in aqueous sucrose solution, 50% (w/,A), at 27 degreesC. Water and sucrose concentration profiles were measured as function of the position for 3, 6 and 12 h of immersion. These were adjusted to a mathematical model for three components that take into account the bulk flow in a shrinking tissue and the concentration dependence of the diffusion coefficients.The binary effective coefficients were an order of magnitude lower than those for pure solutions of sucrose. These coefficients show an unusual concentration dependence. Analysis of these coefficients as functions of the concentration and position demonstrates that, cellular tissue promotes high resistance to diffusion in the tuber and also the elastic contraction of material influences the species diffusion. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Acellular dermal tissue study: an ultrastructural evaluation of human and porcine derived tissues in a rat model

The purpose of this study was to evaluate the host response of a human and a porcine derived acellular dermal tissue (ADT) implanted in the subcutaneous tissue of a rat model. Two subcutaneous pockets were surgically created along the dorsal midline of 25 rats (5 rats/group). The human ADT was placed superiorly and the porcine ADT, inferiorly. The animals were sacrificed at 07, 15, 30, 60 and 180 postoperative days (PO) and the ADTs and surrounding soft tissues were assessed for ultrastructural evaluation by transmission electron microscopy. The ultrastructural findings were similar in both materials. Normal collagen and elastic fibers bundles were observed during all experimental moments, as well as macrophages presenting cytoplasmic enlargements digesting cellular portions after 15 PO. From 30 until 180 PO, vacuolar structures filled with an amorphous, electron-transparent substance, were present inside and outside the fibroblasts. Both human and porcine ADT showed similar pattern of ultrastructural response when implanted in the subcutaneous tissue of rats. The porcine ADT appears as a good alternative to be used as a biomaterial.

Ultrasound Transient Shear Wave Elasticity Imaging for Tendon Tissue

Degeneration of tendon tissue is a common cause of tendon dysfunction with the symptoms of repeated episodes of pain and palpable increase of tendon thickness. Tendon mechanical properties are directly related to its physiological composition and the structural organization of the interior collagen fibers which could be altered by tendon degeneration due to overuse or injury. Thus, measuring mechanical properties of tendon tissue may represent a quantitative measurement of pain, reduced function, and tissue health. Ultrasound elasticity imaging has been developed in the last two decades and has proved to be a promising tool for tissue elasticity imaging. To date, however, well established protocols of tendinopathy elasticity imaging for diagnosing tendon degeneration in early stages or late stages do not exist. This thesis describes the re-creation of one dynamic ultrasound elasticity imaging method and the development of an ultrasound transient shear wave elasticity imaging platform for tendon and other musculoskeletal tissue imaging. An experimental mechanical stage with proper supporting systems and accurate translating stages was designed and made. A variety of high-quality tissue-mimicking phantoms were made to simulate homogeneous and heterogeneous soft tissues as well as tendon tissues. A series of data acquisition and data processing programs were developed to collect the displacement data from the phantom and calculate the shear modulus and Young’s modulus of the target. The imaging platform was found to be capable of conducting comparative measurements of the elastic parameters of the phantoms and quantitatively mapping elasticity onto ultrasound B-Mode images. This suggests the system has great potential for not only benefiting individuals with tendinopathy with an earlier detection, intervention and better rehabilitation, but also for providing a medical tool for quantification of musculoskeletal tissue dysfunction in other regions of the body such as the shoulder, elbow and knee.

Preparation and evaluation of the electrospun chitosan/PEO fibers for potential applications in cartilage tissue engineering

Fibrous materials have morphological similarities to natural cartilage extracellular matrix and have been considered as candidate for bone tissue engineering scaffolds. In this study, we have evaluated a novel electrospun chitosan mat composed of oriented sub-micron fibers for its tensile property and biocompatibility with chondrocytes (cell attachment, proliferation and viability). Scanning electronic microscope images showed the fibers in the electrospun chitosan mats were indeed aligned and there was a slight cross-linking between the parent fibers. The electrospun mats have significantly higher elastic modulus (2.25 MPa) than the cast films (1.19 MPa). Viability of cells on the electrospun mat was 69% of the cells on tissue-culture polystyrene (TCP control) after three days in culture, which was slightly higher than that on the cast films (63% of the TCP control). Cells on the electrospun mat grew slowly the first week but the growth rate increased after that. By day 10, cell number on the electrospun mat was almost 82% that of TCP control, which was higher than that of cast films (56% of TCP). The electrospun chitosan mats have a higher Young’s modulus (P <0.01) than cast films and provide good chondrocyte biocompatibility. The electrospun chitosan mats, thus, have the potential to be further processed into three-dimensional scaffolds for cartilage tissue repair.

Bacterial cellulose-collagen nanocomposite for bone tissue engineering

A nanocomposite based on bacterial cellulose (BC) and type I collagen (COL) was evaluated for in vitro bone regeneration. BC membranes were modified by glycine esterification followed by cross-linking of type I collagen employing 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide. Collagen incorporation was studied by spectroscopy analysis. X-Ray diffraction showed changes in the BC crystallinity after collagen incorporation. The elastic modulus and tensile strength for BC-COL decreased, while the strain at failure showed a slight increase, even after sterilization, as compared to pristine BC. Swelling tests and contact angle measurements were also performed. Cell culture experiments performed with osteogenic cells were obtained by enzymatic digestion of newborn rat calvarium revealed similar features of cell morphology for cultures grown on both membranes. Cell viability/proliferation was not different between BC and BC-COL membranes at day 10 and 14. The high total protein content and ALP activity at day 17 in cells cultured on BC-COL indicate that this composite allowed the development of the osteoblastic phenotype in vitro. Thus, BC-COL should be considered as alternative biomaterial for bone tissue engineering.

Cell-specific expression of human HGF by alveolar type II cells induces remodeling of septal wall tissue in the lung: a morphometric study

Hepatocyte growth factor (HGF) is involved in development and regeneration of the lungs. Human HGF, which was expressed specifically by alveolar epithelial type II cells after gene transfer, attenuated the bleomycin-induced pulmonary fibrosis in an animal model. As there are also regions that appear morphologically unaffected in fibrosis, the effects of this gene transfer to normal lungs is of interest. In vitro studies showed that HGF inhibits the formation of the basal lamina by cultured alveolar epithelial cells. Thus we hypothesized that, in the healthy lung, cell-specific expression of HGF induces a remodeling within septal walls. Electroporation of a plasmid of human HGF gene controlled by the surfactant protein C promoter was applied for targeted gene transfer. Using design-based stereology at light and electron microscopic level, structural alterations were analyzed and compared with a control group. HGF gene transfer increased the volume of distal air spaces, as well as the surface area of the alveolar epithelium. The volume of septal walls, as well as the number of alveoli, was unchanged. Volumes per lung of collagen and elastic fibers were unaltered, but a marked reduction of the volume of residual extracellular matrix (all components other than collagen and elastic fibers) and interstitial cells was found. A correlation between the volumes of residual extracellular matrix and distal air spaces, as well as total surface area of alveolar epithelium, could be established. Cell-specific expression of HGF leads to a remodeling of the connective tissue within the septal walls in the healthy lung, which is associated with more pronounced stretching of distal air spaces at a given hydrostatic pressure during instillation fixation.

Role for ephrinB2 in postnatal lung alveolar development and elastic matrix integrity

Alveoli are formed in the lung by the insertion of secondary tissue folds, termed septa, which are subsequently remodeled to form the mature alveolar wall. Secondary septation requires interplay between three cell types: endothelial cells forming capillaries, contractile interstitial myofibroblasts, and epithelial cells. Here, we report that postnatal lung alveolization critically requires ephrinB2, a ligand for Eph receptor tyrosine kinases expressed by the microvasculature. Mice homozygous for the hypomorphic knockin allele ephrinB2DeltaV/DeltaV, encoding mutant ephrinB2 with a disrupted C-terminal PDZ interaction motif, show severe postnatal lung defects including an almost complete absence of lung alveoli and abnormal and disorganized elastic matrix. Lung alveolar formation is not sensitive to loss of ephrinB2 cytoplasmic tyrosine phosphorylation sites. Postnatal day 1 mutant lungs show extracellular matrix alterations without differences in proportions of major distal cell populations. We conclude that lung alveolar formation relies on endothelial ephrinB2 function.

The influence of yield surface shape and damage in the depth-dependent response of bone tissue to nanoindentation using spherical and Berkovich indenters

Prevention and treatment of osteoporosis rely on understanding of the micromechanical behaviour of bone and its influence on fracture toughness and cell-mediated adaptation processes. Postyield properties may be assessed by nonlinear finite element simulations of nanoindentation using elastoplastic and damage models. This computational study aims at determining the influence of yield surface shape and damage on the depth-dependent response of bone to nanoindentation using spherical and conical tips. Yield surface shape and damage were shown to have a major impact on the indentation curves. Their influence on indentation modulus, hardness, their ratio as well as the elastic-to-total work ratio is well described by multilinear regressions for both tip shapes. For conical tips, indentation depth was not statistically significant (p<0.0001). For spherical tips, damage was not a significant parameter (p<0.0001). The gained knowledge can be used for developing an inverse method for identification of postelastic properties of bone from nanoindentation.

A quantitative analysis of microcirculation in sore-prone pressure areas on conventional and pressure relief hospital mattresses using laser Doppler flowmetry and tissue spectrophotometry

BACKGROUND Pressure ulcers are associated with severe impairment for the patients and high economic load. With this study we wanted to gain more insight to the skin perfusion dynamics due to external loading. Furthermore, we evaluated the effect of different types of pressure relief mattresses. METHODS A total of 25 healthy volunteers were enrolled in the study. Perfusion dynamics of the sacral and the heel area were assessed using the O2C-device, which combines a laser light, to determine blood flow, and white light to determine the relative amount of hemoglobin. Three mattresses were evaluated compared to a hard surface: a standard hospital foam mattress bed, a visco-elastic foam mattress, and an air-fluidized bed. RESULTS In the heel area, only the air-fluidized bed was able to maintain the blood circulation (mean blood flow of 13.6 ± 6 versus 3.9 ± 3 AU and mean relative amount of hemoglobin of 44.0 ± 14 versus 32.7 ± 12 AU.) In the sacral area, all used mattresses revealed an improvement of blood circulation compared to the hard surface. CONCLUSION The results of this study form a more precise pattern of perfusion changes due to external loading on various pressure relief mattresses. This knowledge may reduce the incidence of pressure ulcers and may be an influencing factor in pressure relief mattress selection.

Effect of microstructure upon elastic behaviour of human tooth enamel

Tooth enamel is the stiffest tissue in the human body with a well-organized microstructure. Developmental diseases, such as enamel hypomineralisation, have been reported to cause marked reduction in the elastic modulus of enamel and consequently impair dental function. We produce evidence, using site-specific transmission electron microscopy (TEM), of difference in microstructure between sound and hypomineralised enamel. Built upon that, we develop a mechanical model to explore the relationship of the elastic modulus of the mineral-protein composite structure of enamel with the thickness of protein layers and the direction of mechanical loading. We conclude that when subject to complex mechanical loading conditions, sound enamel exhibits consistently high stiffness, which is essential for dental function. A marked decrease in stiffness of hypomineralised enamel is caused primarily by an increase in the thickness of protein layers between apatite crystals and to a lesser extent by an increase in the effective crystal orientation angle. © 2009 Elsevier Ltd. All rights reserved.

Caputo Derivatives in Viscoelasticity: A Non-Linear Finite-Deformation Theory for Tissue

Mathematics Subject Classification: 26A33, 74B20, 74D10, 74L15

Defective Apoptosis In Intestinal And Mesenteric Adipose Tissue Of Crohn's Disease Patients.

Crohn's disease (CD) is associated with complex pathogenic pathways involving defects in apoptosis mechanisms. Recently, mesenteric adipose tissue (MAT) has been associated with CD ethiopathology, since adipose thickening is detected close to the affected intestinal area. However, the potential role of altered apoptosis in MAT of CD has not been addressed. To evaluate apoptosis in the intestinal mucosa and MAT of patients with CD. Samples of intestinal mucosa and MAT from patients with ileocecal CD and from non-inflammatory bowel diseases patients (controls) were studied. Apoptosis was assessed by TUNEL assay and correlated with the adipocytes histological morphometric analysis. The transcriptional and protein analysis of selected genes and proteins related to apoptosis were determined. TUNEL assay showed fewer apoptotic cells in CD, when compared to the control groups, both in the intestinal mucosa and in MAT. In addition, the number of apoptotic cells (TUNEL) correlated significantly with the area and perimeter of the adipose cells in MAT. Transcriptomic and proteomic analysis reveal a significantly lower transcript and protein levels of Bax in the intestinal mucosa of CD, compared to the controls; low protein levels of Bax were found localized in the lamina propria and not in the epithelium of this tissue. Furthermore, higher level of Bcl-2 and low level of Caspase 3 were seen in the MAT of CD patients. The defective apoptosis in MAT may explain the singular morphological characteristics of this tissue in CD, which may be implicated in the pathophysiology of the disease.