Fibrous membranes of aligned poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) fibers have been made through electrospinning. A high-speed rotating drum was used as the fiber collector while the electric field was manipulated by using five knife-edged auxiliary electrodes. It was found that a high drum rotating speed of 3000 rpm could lead to a nearly perfect alignment of PHBV fibers during electrospinning. Multilayered fibrous structures with each layer having a different direction of fiber alignment could also be constructed through electrospinning. The electrospun PHBV fibers were further modified by incorporating carbonated hydroxyapatite (HA) nanospheres (up to 20% of HA) in the fibers. The fibrous membranes made of aligned PHBV fibers and made of HA/PHBV composite fibers should be very useful for the tissue engineering of different human body tissues.

Electrospinning is an effective technology for the fabrication of ultrafine fibers, which can be the basic component of a tissue engineering scaffold. In tissue engineering, because cells seeded on fibrous scaffolds with varying fiber diameters and morphologies exhibit different responses, it is critical to control these characteristics of electrospun fibers. The diameter and morphology of electrospun fibers can be influenced by many processing parameters (e.g., electrospinning voltage, needle inner diameter, solution feeding rate, rotational speed of the fiber-collecting cylinder, and working distance) and solution properties (polymer solution concentration and conductivity). In this study, a factorial design approach was used to systematically investigate the degree of influence of each of these parameters on fiber diameter, degree of fiber alignment, and their possible synergetic effects, using a natural biodegradable polymer, poly(hydroxybutyrate-co-hydroxyvalerate), for the electrospinning experiments. It was found that the solution concentration invoked the highest main effect on fiber diameter, whereas both rotational speed of the fiber-collecting cylinder and addition of a conductivityenhancing salt could significantly affect the degree of fiber alignment. By carefully controlling the electrospinning parameters and solution properties, fibrous scaffolds of desired characteristics could be made to meet the requirements of different tissue engineering applications.

Electrospinning of fibrous tissue engineering scaffolds has been traditionally conducted using positive voltages. In the current study, positive voltage (PV) electrospinning and negative voltage (NV) electrospinning were investigated for forming fibrous membranes of poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV). In both PV-electrospinning and NV-electrospinning, the fiber diameter generally increased with increasing needle inner diameter and PHBV concentration but decreased with increasing working distance. The use of a conductivity-enhancing surfactant, benzyl triethylammonium chloride (BTEAC), significantly reduced PHBV fiber diameters from the micron scale to the sub-micron scale. Interestingly, with increasing applied voltage, the fiber diameter increased for PV-electrospinning but decreased for NV-electrospinning. The PV-electrospun fibrous membranes from solutions without BTEAC (PVEfm) and with BTEAC (PVEfm-B) and NV-electrospun membranes from solutions without BTEAC (NVEfm) and with BTEAC (NVEfm-B) were characterized in terms of their structure, wettability, thermal properties and tensile properties. Both PVEfm and NVEfm exhibited similar water contact angles (∼104°) but the contact angle of PVEfm-B or NVEfm-B was not measurable. The elongation at break of PVEfm-B or NVEfm-B was significantly higher than that of PVEfm or NVEfm. Using NV-electrospinning or a combination of NV- and PV-electrospinning may be very useful for developing suitable scaffolds for tissue engineering applications.

A method is developed for encapsulation of bacterial biocatalysts in silica gels formed by silica nanoparticles (SNP) and a silicon alkoxide crosslinker. Formulation of the gel was optimized by changing the SNP size, SNP to crosslinker ratio and crosslinker functionality. Hydrolysis and condensation reactions of silicon alkoxide were controlled by water to alkoxide ratio (r) and pH of the solution. FTIR analysis verified that a reactive and temporally stable silicon alkoxide crosslinker was obtained. As a case study, recombinant Escherichia coli (E. coli) cells expressing the atrazine dechlorinating enzyme AtzA were encapsulated. Synthesized catalytic biomaterials (silica gel encapsulated bacterial biocatalysts) were evaluated based on their gelation time, biocatalytic activity and mechanical strength. Diffusivity assays and SEM were used for characterization of the gel structure. We found that SNP to crosslinker ratio affected all the features of the gel, whereas crosslinker functionality primarily affected the gelation time and SNP size affected the mechanical strength and diffusivity. Based on systematic evaluation, we selected three gel formulations and subjected them to long-term activity measurements in a continuous-flow bioreactor for removing trace levels of atrazine. The effluent atrazine concentration was sustained below 30% of the influent concentration, <3 ppb, for 2 months.

Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) fibers containing carbonated hydroxyapatite (CHA) nanoparticles with different CHA amounts (5, 10 and 15 wt%) were electrospun with the aid of ultrasonic power for dispersing the nanoparticles. Scanning electron microscopy and energy-dispersive x-ray spectroscopy results showed that the distribution of CHA within the CHA/PHBV nanocomposite fibers was homogeneous when the CHA content was 10 wt%. Slight particle agglomeration occurred when the CHA content was 15 wt%. The diameters of the electrospun CHA/PHBV nanocomposite fibers and PHBV polymer fibers were around 3 µm. Fourier transform infrared spectroscopic analysis further confirmed the presence of CHA in CHA/PHBV nanocomposite fibers. Both PHBV and CHA/PHBV fibrous membranes exhibited similar tensile properties. Compared with PHBV solvent-cast film, the PHBV fibrous membrane was hydrophobic but the incorporation of CHA nanoparticles dramatically enhanced its wettability. In vitro studies revealed that both types of electrospun fibrous membranes (PHBV and CHA/PHBV) supported the proliferation of human osteoblastic cells (SaOS-2). The alkaline phosphatase activity of SaOS-2 cells seeded on the CHA/PHBV fibrous membranes was higher than that of the cells seeded on the PHBV fibrous membranes after 14 days of cell culture. The electrospun CHA/PHBV nanocomposite fibrous membranes show promises for bone tissue engineering applications.