Textile-based technologies are considered as potential routes for the production of 3D porous architectures for tissue engineering applications. We describe the use of two polymers, namely polybutylene succinate (PBS) and silk fibroin (SF) to produce fiber-based finely tuned porous architectures by weft and warp knitting. The obtained knitted constructs are described in terms of their morphology, mechanical properties, swelling ability, degradation behaviour and cytotoxicity. Each type of polymer fibers allow for the processing of a very reproducible intra-architectural scaffold geometry, with distinct characteristics in terms of the surface physicochemistry, mechanical performance and degradation capability, which has an impact on the resulting cell behaviour at the surface of the respective biotextiles. Preliminary cytotoxicity screening shows that both materials can support cell adhesion and proliferation. Furthermore, different surface modifications were performed (acid/alkaline treatment, UV radiation and plasma) for modulating cell behavior. An increase of cell-material interactions were observed, indicating the important role of materials surface in the first hours of culturing. Human Adipose-derived Stem Cells (hASCs) became an emerging possibility for regenerative medicine and tissue replacement therapies. The potential of the recently developed silk-based biotextile structures to promote hASCs adhesion, proliferation and differentiation is also evaluated. The obtained results validate the developed constructs as viable matrices for TE applications. Given the processing efficacy and versatility of the knitting technology, and the interesting structural and surface properties of the proposed polymer fibers, it is foreseen that our developed systems can be attractive for the functional engineering of tissues such as bone, skin, ligaments or cartilage and also for develop more complex systems for further industrialization of TE products.