Bioinspired Design of Novel Microscaffolds for Fibroblast Guidance toward In Vitro Tissue Building

Porous microscaffolds (μ-scaffs) play a crucial role in modular tissue engineering as they control cell functions and guide hierarchical tissue formation toward building new functional tissue analogues. In the present study, we developed a new route to prepare porous polycaprolactone (PCL) μ-scaffs with a bioinspired trabecular structure that supported in vitro adhesion, growth, and biosynthesis of human dermal fibroblasts (HDFs). The method involved the use of poly(ethylene oxide) (PEO) as a biocompatible porogen and a fluidic emulsion/porogen leaching/particle coagulation process to obtain spherical μ-scaffs with controllable diameter and full pore interconnectivity. To achieve this objective, we investigated the effect of PEO concentration and the temperature of the coagulation bath on the μ-scaff architecture, while we modulated the μ-scaff diameter distribution by varying the PCL–PEO amount in the starting solution and changing the flow rate of the continuous phase (QCP). μ-Scaff morphology, pore architecture, and diameter distribution were assessed using scanning electron microscopy (SEM) analysis, microcomputed tomography (microCT), and Image analysis. We reported that the selection of 60 wt % PEO concentration, together with a 4 °C coagulation bath temperature and ultrasound postprocessing, allowed for the design and fabrication of μ-scaff with porosity up to 80% and fully interconnected pores on both the μ-scaff surface and the core. Furthermore, μ-scaff diameter distributions were finely tuned in the 100–600 μm range with the coefficient of variation lower than 5% by selecting the PCL–PEO concentration in the 1–10% w/v range and QCP of either 8 or 18 mL/min. Finally, we investigated the capability of the HDF-seeded PCL μ-scaff to form hybrid (biological/synthetic) tissue in vitro. Cell culture tests demonstrated that PCL μ-scaff enabled HDF adhesion, proliferation, colonization, and collagen biosynthesis within inter- and intraparticle spaces and guided the formation of a large (centimeter-sized) viable tissue construct.