Novel porous aortic elastin and collagen scaffolds for tissue engineering

Decellularized vascular matrices are used as scaffolds in cardiovascular tissue engineering because they retain their natural biological composition and three-dimensional (3-D) architecture suitable for cell adhesion and proliferation. However, cell infiltration and subsequent repopulation of these scaffolds was shown to be unsatisfactory due to their dense collagen and elastic fiber networks. In an attempt to create more porous structures for cell repopulation, we selectively removed matrix components from decellularized porcine aorta to obtain two types of scaffolds, namely elastin and collagen scaffolds. Histology and scanning electron microscopy examination of the two scaffolds revealed a well-oriented porous decellularized structure that maintained natural architecture of the aorta. Quantitative DNA analysis confirmed that both scaffolds were completely decellularized. Stress-strain analysis demonstrated adequate mechanical properties for both elastin and collagen scaffolds. In vitro enzyme digestion of the scaffolds suggested that they were highly biodegradable. Furthermore, the biodegradability of collagen scaffolds could be controlled by crosslinking with carbodiimides. Cell culture studies showed that fibroblasts adhered to and proliferated on the scaffold surfaces with excellent cell viability. Fibroblasts infiltrated about 120 microm into elastin scaffolds and about 40 microm into collagen scaffolds after 4 weeks of rotary cell culture. These results indicated that our novel aortic elastin and collagen matrices have the potential to serve as scaffolds for cardiovascular tissue engineering.     

Solution properties of ultrahigh-molecular-weight polymers, 16. Unperturbed dimensions of poly(isobutyl methacrylate)

Three techniques, i. e., the Lenka extrapolation method, the Stockmayer-Fixman method, and the extrapolation of the Mark-Houwink constants to a = 0,5 were applied to determine the unperturbed dimensions of ultrahigh-molecular-weight poly(isobutyl methacrylate). A correction term, depending on the exponent a of the Mark-Houwink relation, is suggested to be introduced into the Stockmayer-Fixman-Burchard equation.     

The cerebral microvasculature of the rat: structure and luminal surface properties during early development

The development of the cerebral microvasculature of the rat was studied during three successive postnatal periods, namely: 1) neonatal period, i.e., 1 to 9 days after birth (capillary sprouting period); 2) myelinization period, i.e., 10 to 20 days; and 3) young adult period, i.e., 2 to 3 months. The survey covered structural aspects and distribution of binding sites for anionic or cationic probes and for albumin-gold complexes on the luminal surface of the microvascular endothelium. The salient results are: a) an extensive development of the endoplasmic reticulum of endothelial cells during the first period (presumably in relation with the production of basement membrane components); b) the high surface density of coated pits and coated vesicles that peaks during the myelinization period; c) the paucity of plasmalemmal vesicle and their differential distribution (their volume density is higher in the endothelium of arterioles than in that of capillaries and venules); d) the existence of an extensive smooth surface tubular system in the cytoplasm of endothelial cells, whose structural connections and functional significance remains to be established; and e) the presence of pericytes with elaborate interactions with endothelia in the early developmental periods. Labeling by perfused tracers indicates an uneven patchy distribution of binding sites for cationic ferritin (generally limited to the plasmalemma proper) and a more even distribution of binding sites for cationic and anionic hemeundecapeptides. Binding patterns did not change during the developmental periods studied. No binding sites were detected for albumin-gold complexes.     

Changes in the organization of membrane lipids during human platelet activation. Study by fluorescent and freeze-fracture cytochemistry

Modifications in the membrane lipid organization of human platelets activated with different agents (adenosine 5'-diphosphate, thrombin, collagen type I, and monosaccharides such as fucose, mannose, and galactose) were analyzed in vitro by using three lipid markers. Cholesterol was detected upon interaction with filipin, the anionic phospholipids were reacted with polymyxin B, and alterations in the degree of lipid packing were evaluated with the lipophilic fluorescent probe merocyanine 540, which reportedly inserts into bilayer domains whose lipids are more disordered. Filipin-sterol complexes and polymyxin B-anionic phospholipid complexes form characteristic membrane deformations which were examined in freeze-fracture preparations, whereas the merocyanine 540 binding to platelet membrane was recorded by fluorescent microscopy. In contrast to the resting cells, thrombin-stimulated platelets displayed an uneven distribution of filipin-sterol complexes which occurred in much higher density on the cell body than on pseudopods: on the latter, apparently cholesterol-free domains were very common. Unlike the non-stimulated cells, the platelets aggregated with the various agents employed showed characteristic polymixin B-anionic phospholipid complexes deformations of plasmalemma suggesting the appearance in uneven concentration of anionic phospholipids in the outer membrane leaflet. Incubation with merocyanine 540 did not result in staining of resting platelets when these were maintained in plasma, but a slight fluorescence was observed when platelets were kept in Tyrode buffer. However, platelets stimulated with thrombin, collagen type I, and monosaccharides bound very heavily the fluorescent dye; platelets aggregated with adenosine-5'-diphosphate bound only small amounts of merocyanine 540. The results showed that, during activation by different agents, modifications in lipid membrane organization include alterations in cholesterol and anionic phospholipid distribution, transbilayer movement of anionic phospholipids accompanied by more disordered membrane.     

Mechano-chemischer abbau bei der schwingmahlung von poly(ε-caprolactam) in gegenwart von sebacoylchlorid und nachfolgende polykondensation mit aromatischen diaminen

Mechano-chemical degradation of poly(ε-caprolactam) {poly[imino(1-oxohexamethylene)]} in the presence of sebacoyl chloride, carried out by vibratory milling, gave polyamide acid chlorides. Subsequently, these products, also by vibratory milling, were condensed with p-phenylenediamine to obtain polymers with aromatic amino groups which by diazotation and coupling reaction were transformed into coloured polymers. The use of some specially selected coupling components (i.e. containing itself primary amino groups) allowed to regulate the colour intensity of the final product when the diazotation and coupling reaction was carried out gradually.     

Lysine-enhanced glutaraldehyde crosslinking of collagenous biomaterials.

Crosslinking of collagenous biomaterials currently employs the use of glutaraldehyde. The putative enhancement of glutaraldehyde crosslinking by lysine was investigated in three model systems: bovine pericardium, collagen membranes, and bovine serum albumin. Repetitive sequential treatment of bovine pericardium with glutaraldehyde and lysine and finally with formaldehyde produced a matrix which, by the two criteria used (shrinkage temperature and urea/SDS soluble collagen), was shown to be more highly crosslinked than pericardium fixed in glutaraldehyde alone. Essentially the same results were obtained when membranes prepared from pepsin-soluble pericardial collagen were subjected to sequential glutaraldehyde and lysine treatments, reaching shrinkage temperatures of more than 90 degrees C. Heart valves prepared from lysine-enhanced glutaraldehyde crosslinked bovine pericardium were tested in vitro in an accelerated fatigue tester and have been shown to behave satisfactorily after 300 million cycles. These additional crosslinks proved to be stable in saline at 37 degrees C. Studies on bovine serum albumin attempted to get an insight into the mechanisms of lysine enhancement of glutaraldehyde crosslinking by treating sequentially albumin with glutaraldehyde and lysine and analysis of the products by gel filtration and SDS-PAGE. These studies suggest that free amino groups exposed by proteins are initially reacted with glutaraldehyde and then bridged by the diamino compound (lysine) producing more extensive intermolecular crosslinking than glutaraldehyde alone.     

Growing skull fractures of childhood. A particular form of posttraumatic encephalopathy

Summary The authors describe the findings in 12 cases of growing skull fracture of childhood as revealed by clinical and paraclinical investigations. Based on these findings the authors regard this pathological entity as a separate form of posttraumatic encephalopathy.

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Zusammenfassung Die Autoren beschreiben die klinischen und paraklinischen Befunde bei 12 Fällen von wachsender Schädelfraktur des Kindesalters. Auf Grund dieser Befunde betrachten die Autoren dieses Krankheitsbild als eine besondere Form der posttraumatischen Enzephalopathie.

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Resumen Los autores describen los hallazgos obtenidos en 12 casos de fractura de craneo en niños con el hueso craneal en crecimiento, como revelan sus estudios clínicos y paraclínieos. Estos hallazgos permiten al autor considerar esta entidad patológica como una forma particular de la encefalopatía postraumética.

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Résumé Les auteurs exposent leurs constatations dans douze cas de fracture progressive du crâne chez l'enfant, ceci à la lumière d'investigations cliniques et para-cliniques. Il leur est ainsi possible de considérer cette entité pathologique comme une forme particulière d'encéphalopathie post-traumatique.

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Riassunto Gli AA. descrivono i risultati di indagini cliniche e paracliniche di 12 casi si accrescimento delle fratture craniche infantili. Essi sono d'avviso che detta entità patologica debba essere considerata corne una forma particolare di encefalopatia posttraumatica.

     

Effect of SO2 Dry Deposition on Porous Dolomitic Limestones

The present study is concerned with the assessment of the relative resistance of a monumental dolomitic limestone (Laspra – Spain) used as building material in stone monuments and submitted to artificial ageing by SO₂ dry deposition in the presence of humidity. To investigate the protection efficiency of different polymeric coatings, three commercially available siloxane-based oligomers (Lotexan-N, Silres BS 290 and Tegosivin HL 100) and a newly synthesized hybrid nanocomposite with silsesquioxane units (TMSPMA) were used. A comparative assessment of the data obtained in this study underlines that a better limestone protection was obtained when treated with the hybrid nanocomposite with silsesquioxane units.