We have recently shown that it is possible to recombinantly produce a miniature spider silk protein, 4RepCT, that spontaneously self-assembles into mechanically stable macroscopic fibers (Stark, M.; Grip, S.; Rising, A.; Hedhammar, M.; Engstrom, W.; Hjalm, G.; Johansson, J. Macroscopic fibers self-assembled from recombinant miniature spider silk proteins. Biomacromolecules 2007, 8 (5), 1695-1701). When produced as a soluble fusion protein (with thioredoxin) in Escherichia coli , the spider silk protein can be subjected to several purification steps without aggregating. Here, combined purification and endotoxin removal is achieved using a simple cell wash procedure, protein affinity purification, and LPS depletion. No toxic chemicals were included in the process and the protein retained its ability to self-assemble into fibers. With this method, fibers with pyrogenicity corresponding to less than 1 EU/mg could be recovered. Moreover, the fibers could be sterilized through autoclaving with retained morphology, structure, and mechanical properties. This implies that this recombinant silk is suitable for usage as biomaterial, which is further supported by data showing that the fibers allow growth of human primary fibroblasts.

The prevalent challenge facing tissue engineering today is the lack of adequate vascularization to support the growth, function, and viability of tissue substitutes that require blood vessel supply. Researchers rely on the increasing knowledge of angiogenic and vasculogenic processes to stimulate vascular network formation within three-dimensional tissue constructs. These processes are mainly endothelial cell-regulated, although in the context of tissue engineering, specific interactions with scaffold materials, growth factors and other cell types may require in vitro vascularization schemes to be altered accordingly. To better mimic the complete in vivo environment, increasing attention is given to the integration of co-cultures and mechanical conditioning in bioreactors. Such approaches show great promise for the enhancement of the functionality and clinical applicability of tissue engineering constructs. This paper reviews some scaffold materials used in tissue engineering and the effect of their properties on the vascularization process. Also, it specifically addresses the pivotal role of biomaterials vascularization in tissue engineering applications, along with the effect of angiogenic factors and adhesive molecules on angiogenesis. Assays and markers of angiogenesis are also outlined. One section highlights the need for bioreactor cultures and mechanical conditioning in controlling endothelial cell responses. Finally, we conclude with a brief section on the effects of oxygen concentration and hypoxia over microvessel formation.

A series of three biocompatible P(CL-co-LA)-PEG-P(CL-co-LA) copolymers were synthesized using ring-opening polymerization and characterized by 1H-NMR, gel permeation chromatography, DSC, dynamic-mechanical analysis, and X-ray diffraction. The number of monomer units was kept constant, while the D,L-LA fraction was varied so as to constitute 0, 30, or 70% of the end segments. The molecular weights were sufficiently high to eventually permit 3D scaffold preparation. A degradation study was carried out over 26 weeks, and the effect of monomer composition on the rate of degradation as well as on changes in mechanical strength was investigated. Pure polycaprolactone (PCL)-poly(ethylene glycol) (PEG)-PCL copolymer, P(100/0), was a crystalline material displaying no measurable mass loss, a 30% reduction in mean molecular weight (Mn), and only very slight changes in tensile strength. The random incorporation of 30 and 70% D,L-LA into the end sections of the polymer chain, produced more and more amorphous materials, exhibiting increasingly high rates of degradation, mass loss, and loss of tensile strength. Compared with random P(CL-co-LA), the presence of the PEG block was found both to improve hydrophilicity and thus the rate of degradation and to infer a stabilizing quality, thereby pacing the decrease in tensile strength during degradation. The tested copolymers range from materials exhibiting low mechanical strength and high rate of degradation to slow-degrading materials with high mechanical strength suitable, e.g., for three-dimensional scaffolding.