Textile structures made of biocompatible, osteoconductive and resorbable chitosan-filaments provide excellent preconditions as scaffolds for Bone Tissue Engineering applications. The novel Net Shape Nonwoven (NSN) technique that enables short fibers to be processed into three-dimensional net-shaped nonwoven structures with adjustable pore size distributions is described. NSN scaffolds made of pure chitosan fibers were fabricated. NSN hybrid scaffolds for improved initial cell adhesion were realized by combining the NSN technique with electrospinning and dip-coating with collagen, respectively. Scanning electron microscopy and liquid displacement porosimetry revealed an interconnecting open porous scaffold structure. The novel chitosan-hybrid scaffolds provide proper conditions for adhesion, proliferation and differentiation of the seeded human bone marrow stromal cells, proving that they are suitable for usage in hard-tissue regeneration.

Nowadays, exceptional advantages of silk fibroin over synthetic and natural polymers have impelled the scientists to application of this biomaterial for tissue engineering purposes. Recently, we showed that embedding natural degummed silk fibers in regenerated Bombyx mori silk-based scaffold significantly increases the mechanical stiffness, while the porosity of the scaffolds remains the same. In the present study, we evaluated degradation rate, biocompatibility and regenerative properties of the regenerated 2% and 4% wt silk-based composite scaffolds with or without embedded natural degummed silk fibers within 90 days in both athymic nude and wild-type C57BL/6 mice through subcutaneous implantation. In all scaffolds, a suitable interconnected porous structure for cell penetration was seen under scanning electron microscopy. Compressive tests revealed a functional relationship between fiber reinforcement and compressive modulus. In addition, the fiber/fibroin composite scaffolds support cell attachment and proliferation. On days 30 to 90 after subcutaneous implantation, the retrieved tissues were examined via gross morphology, histopathology, immunofluorescence staining and reverse transcription-polymerase chain reaction as shown in Figure 1. Results showed that embedding the silk fibers within the matrix enhances the biodegradability of the matrix resulting in replacement of the composite scaffolds with the fresh connective tissue. Fortification of the composites with degummed fibers not only regulates the degradation profile but also increases the mechanical performance of the scaffolds. This report also confirmed that pore size and structure play an important role in the degradation rate. In conclusion, the findings of the present study narrate key role of additional surface area in improving in vitro and in vivo biological properties of the scaffolds and suggest the potential ability of these fabricated composite scaffolds for connective tissue regeneration.