Development of biomimetic trilayer fibrous membranes for guided bone regeneration

in order to build fibrous bone tissue scaffolds for guided bone regeneration and to mimic the trilayer structure and the multifunctional properties of the natural periosteum, we fabricated two fibrous trilayer membranes by conjugate electrospinning technology, in which poly(ε-caprolactone) (PCL) fiber was designed as an outer layer, the mixed fibers of PCL and polyurethane (co-PUPCL) as the interlayer, and degradable polyurethane fibers with or without nano-hydroxyapatite (n-HA) as the inner layer (PUHA or PU). The microstructure and characteristics of the trilayer membranes were evaluated and different monolayer fibers were fabricated as the contrast samples. The tensile strength values of each layer increased from the inner layer to the outer layer in the designed structure, while the step-by-step electrospinning method produced good adhesion of different layers. Furthermore, the degradable properties and hydrophilicity of the layers changed with dissymmetric fibrous structures. Cell proliferation assay and cell morphology observation indicated that the PUHA inner fibrous layer exhibited better cell attachment and proliferation than PU. In addition, the osteogenicity of the PUHA fibrous layer has been attested through protein expression by the differentiation of rat mesenchymal stem cells (rMSCs) into the osteogenic lineage. Cell infiltration testing on the two sides of the trilayer membranes in vitro and in vivo showed that the inner layer had good cellular penetration deep into the scaffolds, whereas the cells were barred by the outer layer. We have developed a trilayer structured membrane with different polymer fibers to replicate the natural periosteum by improving functional outcomes, which is a promising fibrous scaffold for clinical use in the repair of destroyed bone.