Fabrication and Characterization of Three Dimensional Electrospun Cortical Bone Scaffolds

Bone is a composite tissue composed of an organic matrix, inorganic mineral matrix and water. Structurally, bone is organized into two distinct types: trabecular (or cancellous) and cortical (or compact) bone. Cortical bone is highly organized, dense and composed of tightly packed units or osteons whereas trabecular bone is highly porous and usually found within the confines of cortical bone. Osteons, the subunits of cortical bone, consist of concentric layers of mineralized collagen fibers. While many scaffold fabrication techniques have sought to replicate the structure and organization of trabecular bone, very little research focuses on mimicking the organization of native cortical bone. In this study we fabricated three-dimensional electrospun cortical scaffolds by heat sintering individual osteon-like scaffolds. The scaffolds contained a system of channels running parallel to the length of the scaffolds, as found naturally in the haversian systems of bone tissue. The purpose of the studies discussed in this paper was to develop a mechanically enhanced biomimetic electrospun cortical scaffold. To that end we investigated the appropriate mineralization and cross-linking methods for these structures and to evaluate the mechanical properties of scaffolds with varying fiber angles. Cross-linking the gelatin in the scaffolds prior to the mineralization of the scaffolds proved to help prevent channels of the osteons from collapsing during fabrication. Premineralization, before larger scaffold formation and mineralization, increased mineral deposition between the electrospun layers of the scaffolds. A combination of cross-linking and premineralization significantly increased the compressive moduli of the individual scaffolds. Furthermore, scaffolds with fibers orientation ranging between 15° and 45° yielded the highest compressive moduli and yield strength.

Laminin- and basement membranepolycaprolactone blend nanofibers as a scaffold for regenerative medicine

Mimicking one or more components of the basement membrane (BM) holds great promise for overcoming insufficiencies in tissue engineering therapies. We have electrospun laminin nanofibers (NFs) isolated from the murine Engelbreth-Holm Swarm (EHS) tumor and evaluated them as a scaffold for embryonic stem cell culture. Seeded human embryonic stem cells were found to better maintain their undifferentiated, colony environment when cultured on laminin NFs compared to laminin mats, with 75% remaining undifferentiated on NFs. Mouse embryonic stem cells cultured on 10% laminin-polycaprolactone (PCL) NFs maintained their colony formation for twice as long without passage compared to those on PCL or gelatin substrates. In addition, we have established a protocol for electrospinning reconstituted basement membrane aligned (RBM)-PCL NFs within 10° of angular deviation. Neuron-like PC12 cells show significantly greater attachment (p < 0.001) and percentage of neuriteextending cells