Biomimetic three-layered membranes comprising (poly)-epsilon-caprolactone, collagen and mineralized collagen for guided bone regeneration

The distinct structural properties and osteogenic capacity are important aspects to be taken into account when developing guided bone regeneration membranes. Herein, inspired by the structure and function of natural periosteum, we designed and fabricated using electrospinning a fibrous membrane comprising (poly)-epsilon-caprolactone (PCL), collagen-I (Col) and mineralized Col (MC). The three-layer membranes, having PCL as the outer layer, PCL/Col as the middle layer and PCL/Col/MC in different ratios (5/2.5/2.5 (PCM-1); 3.3/3.3/3.3 (PCM-2); 4/4/4 (PCM-3) (%, w/w/w)) as the inner layer, were produced. The physiochemical properties of the different layers were investigated and a good integration between the layers was observed. The three-layeredmembranes showed tensile properties in the range of those of natural periosteum. Moreover, the membranes exhibited excellent water absorption capability without changes of the thickness. In vitro experiments showed that the inner layer of the membranes supported attachment, proliferation, ingrowth and osteogenic differentiation of human bone marrowderived stromal cells. In particular cells cultured on PCM-2 exhibited a significantly higher expression of osteogenesis-related proteins. The three-layered membranes successfully supported new bone formation inside a critical-size cranial defect in rats, with PCM-3 being the most efficient. The membranes developed here are promising candidates for guided bone regeneration applications.