Vascular Tissue Engineering: Recent Advances in Small Diameter Blood Vessel Regeneration

Cardiovascular diseases are the leading cause of mortality around the globe. The development of a functional and appropriate substitute for small diameter blood vessel replacement is still a challenge to overcome the main drawbacks of autografts and the inadequate performances of synthetic prostheses made of polyethylene terephthalate (PET, Dacron) and expanded polytetrafluoroethylene (ePTFE, Goretex). Therefore, vascular tissue engineering has become a promising approach for small diameter blood vessel regeneration as demonstrated by the increasing interest dedicated to this field. This review is focused on the most relevant and recent studies concerning vascular tissue engineering for small diameter blood vessel applications. Specifically, the present work reviews research on the development of tissue-engineered vascular grafts made of decellularized matrices and natural and/or biodegradable synthetic polymers and their realization without scaffold

Composite material comprising pectin and calcium phosphate and method for its realisation

A method for obtaining a composite material including an aqueous solution of pectin and a suspension/solution of calcium phosphate mixed together, wherein said solution of pectin cross-links with a portion of the calcium obtained from the solution of calcium phosphate and wherein a portion of the calcium phosphate in suspension remains as inorganic phase and composite materials obtained by this method

Phytoestrogens/insoluble fibers and colonic estrogen receptor β: Randomized, double-blind, placebo-controlled study

AIM: To assess the safety and effect of the supplementation of a patented blend of dietary phytoestrogens and insoluble fibers on estrogen receptor (ER)-β and biological parameters in sporadic colonic adenomas. METHODS: A randomized, double-blind placebo-controlled trial was performed. Patients scheduled to undergo surveillance colonoscopy for previous sporadic colonic adenomas were identified, and 60 eligible patients were randomized to placebo or active dietary intervention (ADI) twice a day, for 60 d before surveillance colonoscopy. ADI was a mixture of 175 mg milk thistle extract, 20 mg secoisolariciresinol and 750 mg oat fiber extract. ER-β and ER-α expression, apoptosis and proliferation (Ki-67 LI) were assessed in colon samples. RESULTS: No adverse event related to ADI was recorded. ADI administration showed a significant increases in ER-β protein (0.822 ± 0.08 vs 0.768 ± 0.10, P = 0.04) and a general trend to an increase in ER-β LI (39.222 ± 2.69 vs 37.708 ± 5.31, P = 0.06), ER-β/ER-α LI ratio (6.564 ± 10.04 vs 2.437 ± 1.53, P = 0.06), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (35.592 ± 14.97 vs 31.541 ± 11.54, P = 0.07) and Ki-67 (53.923 ± 20.91 vs 44.833 ± 10.38, P = 0.07) approximating statistical significance. A significant increase of ER-β protein (0.805 ± 0.13 vs 0.773 ± 0.13, P = 0.04), mRNA (2.278 ± 1.19 vs 1.105 ± 1.07, P < 0.02) and LI (47.533 ± 15.47 vs 34.875 ± 16.67, P < 0.05) and a decrease of ER-α protein (0.423 ± 0.06 vs 0.532 ± 0.11, P < 0.02) as well as a trend to increase of ER-β/ER-α protein in ADI vs placebo group were observed in patients without polyps (1.734 ± 0.20 vs 1.571 ± 0.42, P = 0.07). CONCLUSION: The role of ER-β on the control of apoptosis, and its amenability to dietary intervention, are supported in our study. © 2013 Baishideng. All rights reserved

Detection and genotyping of human Papillomavirus in urine samples from unvaccinated male and female adolescents in Italy

The introduction of vaccination against Human Papillomavirus (HPV) in adolescent girls in 2006 has focused virological surveillance on this age group. As few studies have evaluated HPV infections in young populations, further data are needed in order to improve and extend prophylactic policy and to monitor epidemiological changes. The present study aimed at evaluating overall and type-specific HPV prevalence in both female and male adolescents in Italy. HPV DNA detection and genotyping was performed on urine samples collected from 870 unvaccinated adolescents (369 females, 501 males, 11-18 years of age) in five cities in Italy. Following DNA extraction by means of a commercial kit (NucliSENS®-miniMAG®, bioMérieux), the L1 gene fragment was PCR amplified and genotyped by restriction fragment length polymorphism analysis. HPV DNA was detected in 1.5% of all samples, and in 3% and 0.4% of samples from females and males, respectively. In approximately 70% of HPV DNA positive adolescents, the infection was due to a single genotype, with 88.9% of genotypes belonging to the HR-clade. The only two HPV-positive boys (14 and 18 years old) had HPV-70 genotype. Only one of the 11 HPV-infected girls was in the 11-14 age-group. HPV prevalence was 4.2% in girls aged 15-18 years and 60% of infections were due to vaccine types HPV-16 or HPV-6/-11. This is one of the few studies, the first conducted in Italy, on HPV infection in adolescents. Urine testing is the easier way of detecting HPV infection in younger populations. Our data revealed a very low HPV prevalence, and no infections were observed in the 12-year-old vaccine target population. The majority of infections were seen in females aged 15-18 years. Overall, more than 50% and 30% of the potentially persistent HPV infections detected in this group could have been prevented by the quadrivalent and the bivalent vaccines, respectively

Functionalization of PU Foams via Inorganic and Organic Coatings to Improve Cell and Tissue Interactions

In this work an innovative method to obtain hybrid bio-functional scaffolds has been developed. Polyether urethane (PU) foam scaffolds were synthetized by one-step gas foaming process. PU foams were coated with crosslinked gelatin hydrogel to promote cell adhesion and proliferation for the regeneration of soft tissues (e.g., adipose tissue). PU foams were coated with inorganic coating (i.e., CaPs) to improve the interaction with osteoblasts for bone tissue regeneration. The functionalized 3D PU porous scaffolds have been characterized investigating morphological properties by SEM and microCT, water uptake and coating stability, and compressive mechanical properties. Adipose tissue derived stem cells (ADSCs), endothelial cells (MS1), amnion mesenchymal cells (AMCs) and chorion mesenchymal cells (CMCs) isolated from human placenta were in vitro cultured on the hybrid functionalized 3D scaffolds. Mechanical properties showed elastic modulus ranging between 15.75 ± 2.14 and 22.9 ± 3.1 kPa; in vitro biological studies showed good cell adhesion, proliferation, and differentiation. In particular, compared to the results with uncoated PU, when cells where differentiated into adipocytes, Oil red O staining confirmed a higher presence of lipid droplets; in case of osteoblasts differentiation, inorganic extracellular matrix deposition was evidenced on CaPs coated PU. The obtained results suggest the important role of an adequate coating on the scaffold to stimulate a better interaction with cells, promoting the differentiation into different cells phenotypes

2D and 3D Electrospun Silk Fibroin Gelatin Coatings to Improve Scaffold Performances in Cardiovascular Applications

A coating with biomacromolecules can improve the performance of electrospun 3D scaffolds and 2D matrices that show morphology similar to that of native ECM, but their mechanical and biological properties are often inadequate, particularly in applications in contact with blood. In this work, a gelatin coating was applied to electrospun silk fibroin (ESF) mats and tubes intended for the regeneration of cardiovascular tissues. The crosslinking reaction used is based on a Michael-type addition in water that promotes the formation of covalent bonds between gelatin amino groups and β-carbons of N-N′-methylene bis-acrylamide (MBA). ESF mats and tubes were coated with gelatin MBA crosslinked in situ by loading or dipping the ESF samples with the crosslinking solution by use of static or dynamic homemade systems. SEM analysis on coated samples showed a homogeneous coating with gelatin penetrating the whole thickness of the SF matrix (≈120 µm for mats and ≈212 µm for tubes), with an increase of thickness of about 40% in wet conditions. Water uptake tests indicated for coated samples a faster and higher swelling (1600% after 14 days) than not coated ones (500%), due to the presence of gelatin. Tensile mechanical tests showed higher values of ultimate stress and elastic modulus for silk fibroin samples (b = 2.4, E = 1.82 MPa) compared to gelatin-coated ones (b = 1.2, E = 0.58 MPa), with not significant differences in the ultimate deformation (≈150%). Indirect cytocompatibility tests, performed by culturing L929 cells in the presence of eluates obtained by immersing coated and uncoated samples up to 7 days in culture medium, demonstrated a cell viability higher than the control. For direct contact tests, primary human umbilical vein endothelial cell (HUVEC), obtained by enzymatic digestion, and cell adhesion and growth on the MBA-crosslinked gelatin was analyzed by OM after fixing with formalin and staining with toluidine blue. HUVEC were seeded onto ESF and ESF-coated samples and cultured under standard tissue culture conditions. After 7 days from seeding, cell proliferation was evaluated from the quantitation of total proteins in cell lysates (BCA protein assay) and the obtained results indicated a significantly higher (p < 0.05) cell growth on gelatin-coated ESF samples. Overall, these results point out that the described gelatin coating allows for the production of a structure with adequate mechanical properties for cardio-vascular applications and biological characteristics even better than those of silk fibroin