Polysaccharide-based Scaffolds Research Articles

Magnetic resonance imaging tracking of human adipose derived stromal cells within three-dimensional scaffolds for bone tissue engineering

For bone tissue engineering, human Adipose Derived Stem Cells (hADSCs) are proposed to be associated with a scaffold for promoting bone regeneration. After implantation, cellularised scaffolds require a non-invasive method for monitoring their fate in vivo. The purpose of this study was to use Magnetic Resonance Imaging (MRI)-based tracking of these cells, labelled with magnetic agents for in vivo longitudinal assessment. hADSCs were isolated from adipose tissue and labelled with USPIO-rhodamine (Ultrasmall SuperParamagnetic Iron Oxide). USPIO internalisation, absence of toxicity towards hADSCs, and osteogenic differentiation of the labelled cells were evaluated in standard culture conditions. Labelled cells were then seeded within a 3D porous polysaccharide-based scaffold and imaged in vitro using fluorescence microscopy and MRI. Cellularised scaffolds were implanted subcutaneously in nude mice and MRI analyses were performed from 1 to 28 d after implantation. In vitro, no effect of USPIO labelling on cell viability and osteogenic differentiation was found. USPIO were efficiently internalised by hADSCs and generated a high T2* contrast. In vivo MRI revealed that hADSCs remain detectable until 28 d after implantation and could migrate from the scaffold and colonise the area around it. These data suggested that this scaffold might behave as a cell carrier capable of both holding a cell fraction and delivering cells to the site of implantation. In addition, the present findings evidenced that MRI is a reliable technique to validate cell-seeding procedures in 3D porous scaffolds, and to assess the fate of hADSCs transplanted in vivo.

Fabrication of porous polysaccharide-based scaffolds using a combined freeze-drying/cross-linking process

Biocompatible three-dimensional (3-D) porous scaffolds are of great interest for tissue engineering applications. We here present a novel combined freeze-drying/cross-linking process to prepare porous polysaccharide-based scaffolds. This process does not require an organic solvent or porogen agent. We unexpectedly found that cross-linking of biomacromolecules such as pullulan and dextran with sodium trimetaphosphate could be performed during freeze-drying. We have demonstrated that the freeze-drying pressure modulates the degree of porosity. High freeze-drying pressure scaffolds presented pores with a mean diameter of 55 +/- 4 microm and a porosity of 33 +/- 12%, whereas low freeze-drying pressure scaffolds contained larger pores with a mean diameter of 243 +/- 14 microm and a porosity of 68 +/- 3%. Porous scaffolds of the desired shape could be easily obtained and were stable in culture medium for weeks. In vitro viable mesenchymal stem cells were found associated with porous scaffolds in higher proportions than with non-porous scaffolds. Moreover, cells penetrated deeper into scaffolds with larger pores. This novel combined freeze-drying/cross-linking processing of polysaccharides enabled the fabrication of biocompatible scaffolds with controlled porosity and architectures suitable for 3-D in vitro culture and biomedical applications.

The evaluation of a small-diameter polysaccharide-based arterial graft in rats

Cardiovascular diseases may require surgery such as arterial bypasses that are usually performed with synthetic PTFE and Dacron grafts with diameter above 6mm. For smaller diameter replacement, healthy vascular tissue from the patient is not always available to carry out this type of graft. The purpose of this study was to evaluate the effectiveness of a small-diameter polysaccharide-based scaffold as an alternative arterial replacement. Tubular polysaccharide-based grafts of 2 mm internal diameter were prepared and moulded by a cross-linking technique. Fifteen Wistar adult rats underwent infrarenal aortic bypass with these grafts using microsurgical techniques. Grafts withstood aortic blood pressure and exhibited physiological blood flow, as evaluated with ultrasound techniques and angiographies at 4 and 8 weeks post-surgery. Harvested grafts were morphologically evaluated by light microscopy and immunohistochemistry. Neointima formation at 8 weeks was evidenced through collagen deposition and smooth muscle-like cells circumferential growth on the luminal surface without intimal hyperplasia or aneurysm formation. In conclusion, we described a 2mm polysaccharide-based arterial material being investigated in vivo and demonstrating patency for up to 8 weeks post-surgery with neointima formation and absence of intimal hyperplasia.