Disseny i fabricació de scaffolds de Poli(Ɛ-caprolactona) (PCL) per al cultiu 3D de fibroblasts

Abstract

Skull base neurosurgery means an important challenge due to the difficulty to reach this area without affecting nearby neural and vascular structures and the barrier between the cerebrospinal fluid and the external environment. Traditional surgery is an invasive technique, which consists in drilling a hole in the cranium (craniotomy), being dangerous as it can compromise some important structures. For this reason, endoscopic techniques have emerged. They use a minimally invasive procedure, taking advantage of the natural openings, as the nose and the mouth, to reach the skull base, by drilling a hole in the clivus. The background of this study is Poly(ε-caprolactone) prostheses' design and manufacturing that can be used to close the drilled hole in the clivus, by tissue engineering. In this way, the current project is based on PCL scaffolds' design and manufacturing for three-dimensional fibroblast culture. In the connective-tissue cell family, fibroblasts are the easiest to grow in culture and the most versatile to differentiate into other members of the family. Scaffolds have been designed with different deposition angles (90, 60 and 45°) and different distances between filaments (0.5, 0.7 and 0.9 mm). They have been produced using the RepRap BCN3D+ printer in order to evaluate the optimal parameters for the three-dimensional cell culture. The different designs have been tested in cell culture with the fibroblastic line NIH/3T3. The scaffolds with a deposition angle of 90° are those which have showed the largest cell viability, followed by 60° scaffolds and, finally, by 45° ones. On the other hand, it has been observed a higher number of cells attached to filaments in scaffolds with a distance between filaments of 0.5 mm than in the other ones. No differences have been appreciated between 0.7 and 0.9 mm scaffolds. The obtained results confirm the ease of fibroblasts to grow under any culture condition. In this way, three-dimensional fibroblast culture with PCL scaffolds manufactured with RepRap BCN3D+ printer could be useful to generate constructs that by tissue engineering could close the drilled hole in the clivus, which serves as a door in neurosurgery