Multifactorial bioengineering in the tendon context to maintain tenocyte phenotype and to direct dermal fibroblasts towards tenogenic lineage

Considering that cells <i>in vivo</i> are simultaneously exposed to multiple signals, multifactorial bioengineering slowly but surely becomes the state of play in controlling cell function <i>in vitro</i>. In this context, herein we assessed the simultaneous effect of three bioinspired factors (i.e. anisotropic architecture, macromolecular crowding and growth factor supplementation) to maintain human tenocyte phenotype and to direct human dermal fibroblasts towards tenogenic lineage. Anisotropic architecture induced bidirectional cell and deposited extracellular matrix deposition in both cell types. Immunofluorescence analysis revealed that when growth factor and macromolecular crowding were combined significantly increased collagen types I, III, IV and V deposition in both cell types. Proteomics analysis made apparent that the combination of anisotropic architecture, macromolecular crowding and growth factor supplementation brought about the highest degree of tenogenic phenotype maintenance (in the case of human tenocytes) and tenogenic induction (in the case of human dermal fibroblasts). This work lays the foundations for utilising multifactorial bioengineering in the development of tendon-like assemblies.