The Osteoprint: a bioinspired two-photon polymerized 3-D structure for the enhancement of bone-like cell differentiation.

The need for a better understanding of cell behavior and for exploiting cell functions in various healthcare applications has driven biomedical research to develop increasingly complex fabrication strategies to reproduce the natural biological microenvironment in vitro. Different approaches have led to the development of refined examples of 2- and 3-D structures able to sustain cellular proliferation, differentiation and functionality very similar to those normally occurring in living organisms. One such approach is two-photon polymerization. In this paper, we present a trabecula-like structure (which we have named "Osteoprint") that resembles to the typical microenvironment of trabecular bone cells. Starting from microtomography images of the trabecular bone, we prepared several Osteoprints through two-photon polymerization and tested the behavior of SaOS-2 bone-like cells cultured on our structures. Interestingly, we found that Osteoprints deeply affect cellular behavior, determining an exit from the cell cycle and an enhancement of osteogenic differentiation. Indeed, we found an up-regulation of the genes involved in SaOS-2 cell maturation and an increase in hydroxyapatite production and accumulation upon SaOS-2 culture on the Osteoprints. The findings we obtained are extremely interesting, and open up new perspectives in "bioinspired" approaches for tissue engineering and regenerative medicine.