Skeletal myotube formation enhanced by electrospun polyurethane carbon nanotube scaffolds

Sirinrath Sirivisoot, Benjamin S Harrison Wake Forest Institute for Regenerative Medicine, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC, USA Background: This study examined the effects of electrically conductive materials made from electrospun single- or multiwalled carbon nanotubes with polyurethane to promote myoblast differentiation into myotubes in the presence and absence of electrical stimulation. Methods and results: After electrical stimulation, the number of multinucleated myotubes on the electrospun polyurethane carbon nanotube scaffolds was significantly larger than that on nonconductive electrospun polyurethane scaffolds (5% and 10% w/v polyurethane). In the absence of electrical stimulation, myoblasts also differentiated on the electrospun polyurethane carbon nanotube scaffolds, as evidenced by expression of Myf-5 and myosin heavy chains. The myotube number and length were significantly greater on the electrospun carbon nanotubes with 10% w/v polyurethane than on those with 5% w/v polyurethane. The results suggest that, in the absence of electrical stimulation, skeletal myotube formation is dependent on the morphology of the electrospun scaffolds, while with electrical stimulation it is dependent on the electrical conductivity of the scaffolds. Conclusion: This study indicates that electrospun polyurethane carbon nanotubes can be used to modulate skeletal myotube formation with or without application of electrical stimulation. Keywords: myoblasts, electrical field, single-walled, multiwalled, carbon nanotubes, nanocomposites, musculoskeletal, tissue engineerin

Ionic liquids-based processing of electrically conducting chitin nanocomposite scaffolds for stem cell growth

In the present study, we have successfully combined the biocompatible properties of chitin with the high electrical conductivity of carbon nanotubes (CNTs) by mixing them using an imidazolium-based ionic liquid as a common solvent/dispersion medium. The resulting nanocomposites demonstrated uniform distribution of CNTs, as shown by scanning electron microscopy (SEM) and optical microscopy. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction confirmed the α-crystal structure of chitin in the regenerated chitin nanocomposite scaffolds. Increased CNT concentration in the chitin matrix resulted in higher conductivity of the scaffolds. Human mesenchymal stem cells adhered to, and proliferated on, chitin/CNT nanocomposites with different ratios. Cell growth in the first 3 days was similar on all composites at a range of (0.01 to 0.07) mass fraction of CNT. However, composites at 0.1 mass fraction of CNT showed reduced cell attachment. There was a significant increase in cell proliferation using 0.07 mass fraction CNT composites suggesting a stem cell enhancing function for CNTs at this concentration. In conclusion, ionic liquid allowed the uniform dispersion of CNTs and dissolution of chitin to create a biocompatible, electrically conducting scaffold permissive for mesenchymal stem cell function. This method will enable the fabrication of chitin- based advanced multifunctional biocompatible scaffolds where electrical conduction is critical for tissue function

Magnetically stimulated ciprofloxacin release from polymeric microspheres entrapping iron oxide nanoparticles

Sirinrath Sirivisoot,1 Benjamin S Harrison2 1Biological Engineering Program, Faculty of Engineering, King Mongkut’s University of Technology Thonburi, Bangkok, Thailand; 2Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Wake Forest University Health Sciences, Winston-Salem, NC, USA Abstract: To extend the external control capability of drug release, iron oxide nanoparticles (NPs) encapsulated into polymeric microspheres were used as magnetic media to stimulate drug release using an alternating magnetic field. Chemically synthesized iron oxide NPs, maghemite or hematite, and the antibiotic ciprofloxacin were encapsulated together within polycaprolactone microspheres. The polycaprolactone microspheres entrapping ciprofloxacin and magnetic NPs could be triggered for immediate drug release by magnetic stimulation at a maximum value of 40%. Moreover, the microspheres were cytocompatible with fibroblasts in vitro with a cell viability percentage of more than 100% relative to a nontreated control after 24 hours of culture. Macrophage cell cultures showed no signs of increased inflammatory responses after in vitro incubation for 56 hours. Treatment of Staphylococcus aureus with the magnetic microspheres under an alternating (isolating) magnetic field increased bacterial inhibition further after 2 days and 5 days in a broth inhibition assay. The findings of the present study indicate that iron oxide NPs, maghemite and hematite, can be used as media for stimulation by an external magnetic energy to activate immediate drug release. Keywords: antibacterial, maghemite, hematite, Staphylococcus aureu

New method for arbuscular mycorrhizal fungus spore separation using a microfluidic device based on manual temporary flow diversion.

Arbuscular mycorrhizal fungi are beneficial components often included in biofertilizers. Studies of the biology and utilization of these fungi are key to their successful use in the biofertilizer industry. The acquisition of isolated spores is a required step in these studies; however, spore quality control and spore separation are bottlenecks. Filtered and centrifuged spores have to be hand-picked under a microscope. The conventional procedure is skill-demanding, labor-intensive, and time-consuming. Here, we developed a microfluidic device to aid manual separation of spores from a filtered and centrifuged suspension. The device is a single spore streamer equipped with a manual temporary flow diversion (MTFD) mechanism to select single spores. Users can press a switch to generate MTFD when the spore arrives at the selection site. The targeted spore flows in a stream to the collection chamber via temporary cross flow. Using the device, spore purity, the percentage of spore numbers against the total number of particles counted in the collecting chamber reached 96.62% (median, n = 10) which is greater than the spore purity obtained from the conventional method (88.89% (median, n = 10))