Nanostructured biomaterials from electrospun demineralized bone matrix: a survey of processing and crosslinking strategies.

In the design of scaffolds for tissue engineering biochemical function and nanoscale features are of particular interest. Natural polymers provide a wealth of biochemical function, but do not have the processability of synthetic polymers, limiting their ability to mimic the hierarchy of structures in the natural extracellular matrix. Thus, they are often combined with synthetic carrier polymers to enable processing. Demineralized bone matrix (DBM), a natural polymer, is allograft bone with inorganic material removed. DBM contains the protein components of bone, which includes adhesion ligands and osteoinductive signals, such as important growth factors. Herein we describe a novel method for tuning the nanostructure of DBM through electrospinning without the use of a carrier polymer. This work surveys solvents and solvent blends for electrospinning DBM. Blends of hexafluoroisopropanol and trifluoroacetic acid are studied in detail. The effects of DBM concentration and dissolution time on solution viscosity are also reported and correlated to observed differences in electrospun fiber morphology. We also present a survey of techniques to stabilize the resultant fibers with respect to aqueous environments. Glutaraldehyde vapor treatment is successful at maintaining both macroscopic and microscopic structure of the electrospun DBM fibers. Finally, we report results from tensile testing of stabilized DBM nanofiber mats, and preliminary evaluation of their cytocompatibility. The DBM nanofiber mats exhibit good cytocompatibility toward human dermal fibroblasts (HDF) in a 4-day culture; neither the electrospun solvents nor the cross-linking results in any measurable residual cytotoxicity toward HDF.