Multilayer composite scaffolds with mechanical properties similar to small intestinal submucosa

Use of biodegradable scaffolds to engineer new tissues has become an attractive option in various transplantation protocols. In particular, small intestinal submucosa (SIS) has generated immense interest in various tissue engineering applications because of its diverse favorable properties. However, it is a natural matrix, which leads to problems in large-scale preparations and contains sample to sample heterogeneity. In this study, we explored the formation of synthetic matrix mimicking the characteristics of the SIS. Three-dimensional composite structures were developed by sandwiching 50:50 PLGA film between porous chitosan matrices. The outer chitosan layers provide biological activity while the inner PLGA layer provides mechanical strength. PLGA films were initially perforated at 1 cm distance, and the porous chitosan matrix was formed sequentially on each side by controlled rate freezing and lyophilization technique at -80 degrees C. Scanning electron microscopy analysis showed a layered microarchitecture with chitosan filling the perforations of PLGA membrane. Urea permeability studies confirmed that the perforations were filled (negligible urea transfer across composite over 8 h). Tensile strength analysis showed that the matrices formed using 160 kDa PLGA had sufficient break stress ( approximately 4.5 MPa). Degradation analysis over 8 weeks in the presence of 10 mg/L lysozyme showed a 50% decrease in total weight and an 80% decrease in PLGA molecular weight. When cellular adhesion and actin distribution of mouse embryonic fibroblasts were evaluated, for 7 days, cells showed their typical spindle shape and redistribution of actin fibers on composite matrices. Viability studies and MMP-2/MMP-9 activity showed that the cells were viable and functional, similar to tissue culture plastic. Further, canine bladder smooth muscle cells also showed similar cell adhesion and spreading on the composite matrix. In summary, composite structures mimicking SIS were constructed and show potential as a tissue engineering material.