Although several major progresses have been introduced in the field of bone regenerative medicine during the years, current therapies, such as bone grafts, still have many limitations. Moreover, and in spite of the fact that material science technology has resulted in clear improvements in the field of bone substitution medicine, no adequate bone substitute has been developed and hence large bone defects/injuries still represent a major challenge for orthopaedic and reconstructive surgeons. It is in this context that TE has been emerging as a valid approach to the current therapies for bone regeneration/substitution. In contrast to classic biomaterial approach, TE is based on the understanding of tissue formation and regeneration, and aims to induce new functional tissues, rather than just to implant new spare parts. The present review pretends to give an exhaustive overview on all components needed for making bone tissue engineering a successful therapy. It begins by giving the reader a brief background on bone biology, followed by an exhaustive description of all the relevant components on bone TE, going from materials to scaffolds and from cells to tissue engineering strategies, that will lead to "engineered" bone. Scaffolds processed by using a methodology based on extrusion with blowing agents.

Colorectal carcinoma (CRC) is one of the most common causes of cancer-related mortality. Short-chain fatty acids secreted by dietary propionibacteria from the intestine, such as acetate, induce apoptosis in CRC cells and may therefore be relevant in CRC prevention and therapy. We previously reported that acetic acid-induced apoptosis in Saccharomyces cerevisiae cells involves partial vacuole permeabilization and release of Pep4p, the yeast cathepsin D (CatD), which has a protective role in this process. In cancer cells, lysosomes have emerged as key players in apoptosis through selective lysosomal membrane permeabilization (LMP) and release of cathepsins. However, the role of CatD in CRC survival is controversial and has not been assessed in response to acetate. We aimed to ascertain whether LMP and CatD are involved in acetate-induced apoptosis in CRC cells. We showed that acetate per se inhibits proliferation and induces apoptosis. More importantly, we uncovered that acetate triggers LMP and CatD release to the cytosol. Pepstatin A (a CatD inhibitor) but not E64d (a cathepsin B and L inhibitor) increased acetate-induced apoptosis of CRC cells, suggesting that CatD has a protective role in this process. Our data indicate that acetate induces LMP and subsequent release of CatD in CRC cells undergoing apoptosis, and suggest exploiting novel strategies using acetate as a prevention/therapeutic agent in CRC, through simultaneous treatment with CatD inhibitors.

Starch-based biomaterials and scaffolds have been proposed for several biomedical applications. In the present work new scaffolds based on a 50/50 (wt%) blend of corn starch/ethylene-vinyl alcohol (SEVA-C) were studied. These scaffolds were processed by a melt-based technology, which has been used before with other starch-based materials but never with SEVA-C. Scanning electron microscopy (SEM) observation showed that the developed porous structures were 60% porous with pore size between 200 and 900 mm and a reasonable degree of interconnectivity. Moreover, scaffolds presented a compressive modulus of 117.50 6 3.7 MPa and a compressive strength of 20.8 6 2.4 MPa. Cytotoxicity evaluation was performed according to ISO/EN 10993 part 5 guidelines, and revealed that the developed scaffolds were nontoxic and did not inhibit cell growth. Direct contact assays were also carried out by use of a cell line of human osteoblast-like cells (SaOS-2). Cells were seeded (3 3 10 5 per scaffold) and allowed to grow for 4 weeks at 37°C, in a humidified atmosphere containing 5% CO 2. Total protein assay showed that the cells were able to grow for the 4 weeks of the experiment. These data were further confirmed by SEM. Moreover, a cell viability assay (MTS test) demonstrated that cells were perfectly viable after the 4 weeks of culture, showing the adequacy of the developed structure in supporting them. Finally, Western blot analysis revealed that osteopontin was being actively expressed by the cells, which, in association with collagen deposition observed by SEM, seems to indicate that bone extracellular matrix was being deposited. Consequently it is believed that starch-based scaffolds should be considered as an alternative for bone tissueengineering applications in the near future.

Blends of polysaccharides and proteins are a source for the development of novel materials with interesting and tailorable properties, with potential to be used in a range of biomedical applications. in this work a series of blended membranes composed by chitosan and soy protein isolate was prepared by solvent casting methodology. in addition, cross-linking was performed in situ with glutaraldehyde solutions in the range 5x10(-3)-0.1 M. Furthermore, the influence of the composition and cross-linking on the degradation behaviour, water uptake and cell adhesion was investigated. The obtained results showed that the incorporation of chitosan, associated to network formation by cross linking, promoted a slight decrease of water absorption and a slower degradability of the membranes. Moreover, direct contact biocompatibility studies, with L929 cells, indicate that the cross-linking enhances the capability of the material to support cell growth.

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