Compositions and Methods for Making and Using Laminin Nanofibers

University Of Virginia ((UVA) Charlottesville, VA) scientists developed methodologies and parameters for the formation of nanofibrous (to microfibrous) laminin via electrospinning. 

Inventors Roy Clinton Ogle, an Associate Professor of Cell Biology, and Edward A Botchwey, III, , an  Assistant Professor of Biomedical Engineering and Orthopaedic Surgery, in  U.S. Patent Application 20100120115, disclose conditions and appropriate parameters to synthesize laminin fibers from a diameter of about 10 nM to a diameter of over 1,000 nM via electrospinning.

The application covers a method of preparing electrospun laminin, said method comprising obtaining purified laminin, dissolving said purified laminin in HFP, loading said dissolved laminin into a dispensing container comprising a positive lead, subjecting said lead to driving voltage from a power supply, pumping said laminin dissolved in HFP through an opening in said dispensing container, and collecting said laminin dissolved in HFP on a substrate placed on a grounded collector.

Laminins are a family of large extracellular matrix (ECM) proteins found primarily in basement membranes associated with all epithelial, endothelial, muscle, fat and Schwann cells. The laminins serve critical functions in cell attachment, growth, migration, and differentiation of many cell types. Laminin I is the first extracellular matrix protein to appear during embryonic development, where it surrounds the inner cell mass of the compacted blastocyst. 


Studies of laminin I purified from the Engelbreth-Holm-Swarm (EHS) tumor established that laminin is required for cell attachment and growth, and many studies confirm the importance of laminins in development and survival. Laminin interacts with cells through a variety of integrins, the dystroglycan receptor, syndecan, and other type receptors broadly expressed on many cell types  

Extracellular matrix (ECM) provides the extracellular environment for almost all mammalian cell types. It is composed of structural proteins such as collagen and elastin, proteoglycans, and proteins such as fibrin, fibronectin, and laminin. One of the over-reaching goals of cell biology and tissue engineering is to recreate the extracellular environment a cell experiences in vivo, and attaining the appropriate ECM components in appropriate morphological and physical characteristics is of the utmost importance. 

If it is to possess the biological properties of a natural basement membrane, the laminin nanofiber (LNF) mesh should be a favorable substrate for cell attachment and growth in a wide variety of tissue engineering applications. Laminin is particularly relevant for nervous system tissue engineering, as laminin has been shown to encourage neurite extension.

There is a long felt need in the art to recreate the extracellular environment a cell experiences in vivo and to attain the appropriate ECM components in appropriate morphological and physical characteristics is of the utmost importance. The UVA invention helps to satisfy these needs

FIG. 1, comprising FIGS. 1A to 1F, represents images of scanning electron micrographs of laminin electrospun at 20 kV driving voltage and 1.5 mL/hr flow rate. Concentrations (wt/vol) in HFP are shown across the top (3%, 5%, and 8%), and collecting distance is shown along the left side (12.5 cm; upper panels; 25 cm; lower panels). An increase in fiber diameter and decrease in bead area density are correlated with increasing weight percent laminin in HFP of the original solution. White arrows indicate matrisome morphology.