Gravity spun polycaprolactone fibers for applications in vascular tissue engineering:proliferation and function of human vascular endothelial cells

Poly(ε-caprolactone) (PCL) fibers produced by wet spinning from solutions in acetone under low-shear (gravity-flow) conditions resulted in fiber strength of 8 MPa and stiffness of 0.08 Gpa. Cold drawing to an extension of 500% resulted in an increase in fiber strength to 43 MPa and stiffness to 0.3 GPa. The growth rate of human umbilical vein endothelial cells (HUVECs) (seeded at a density of 5 × 104 cells/mL) on as-spun fibers was consistently lower than that measured on tissue culture plastic (TCP) beyond day 2. Cell proliferation was similar on gelatin-coated fibers and TCP over 7 days and higher by a factor of 1.9 on 500% cold-drawn PCL fibers relative to TCP up to 4 days. Cell growth on PCL fibers exceeded that on Dacron monofilament by at least a factor of 3.7 at 9 days. Scanning electron microscopy revealed formation of a cell layer on samples of cold-drawn and gelatin-coated fibers after 24 hours in culture. Similar levels of ICAM-1 expression by HUVECs attached to PCL fibers and TCP were measured using RT-PCR and flow cytometry, indicative of low levels of immune activation. Retention of a specific function of HUVECs attached to PCL fibers was demonstrated by measuring their immune response to lipopolysaccharide. Levels of ICAM-1 expression increased by approximately 11% in cells attached to PCL fibers and TCP. The high fiber compliance, favorable endothelial cell proliferation rates, and retention of an important immune response of attached HUVECS support the use of gravity spun PCL fibers for three-dimensional scaffold production in vascular tissue engineering. © Mary Ann Liebert, Inc.

Fiber Bragg grating structures inscribed using 800nm femtosecond laser in single- and multi-core mid-IR glass fibers with their spectral, thermal and strain properties

Single- and multi-core passive and active germanate and tellurite glass fibers represent a new class of fiber host for in-fiber photonics devices and applications in mid-IR wavelength range, which are in increasing demand. Fiber Bragg grating (FBG) structures have been proven as one of the most functional in-fiber devices and have been mass-produced in silicate fibers by UV-inscription for almost countless laser and sensor applications. However, because of the strong UV absorption in germanate and tellurite fibers, FBG structures cannot be produced by UVinscription. In recent years femtosecond (fs) lasers have been developed for laser machining and microstructuring in a variety of glass fibers and planar substrates. A number of papers have been reported on fabrication of FBGs and long-period gratings in optical fibers and also on the photosensitivity mechanism using 800nm fs lasers. In this paper, we demonstrate for the first time the fabrication of FBG structures created in passive and active single- and three-core germanate and tellurite glass fibers by using 800nm fs-inscription and phase mask technique. With a fs peak power intensity in the order of 1011W/cm2, the FBG spectra with 2nd and 3rd order resonances at 1540nm and 1033nm in a single-core germanate glass fiber and 2nd order resonances between ~1694nm and ~1677nm with strengths up to 14dB in all three cores of three-core passive and active tellurite fibers were observed. Thermal and strain properties of the FBGs made in these mid-IR glass fibers were characterized, showing an average temperature responsivity of ~20pm/°C and a strain sensitivity of 1.219±0.003pm/µe.

Continuous-wave ultraviolet light induced fiber Bragg gratings in few- and single-mode microstructured polymer optical fibers

Reported are observations and measurements of the inscription of fibre Bragg gratings in two different types of microstructured polymer optical fibre: few-moded and endlessly single mode. Contrary to FBG inscription in silica microstructured fibre, where high energy laser pulses are a prerequisite, we have successfully used a low power CW laser source operating at 325nm to produce 1-cm long gratings with a reflection peak at 1570 nm. Peak reflectivities of more than 10% have been observed.

Measurements of stress-optic coefficient in polymer optical fibers

We have systematically measured the differential stress-optic coefficient, ?C, in a number of poly(methyl methacrylate) (PMMA) fibers drawn with different stress, ranging from 2 up to 27 MPa. ?C was determined in transverse illumination by measuring the dependence of birefringence on additional axial stress applied to the fiber. Our results show that ?C in PMMA fibers has a negative sign and ranges from -4.5 to -4.5×10-12 Pa-1, depending on the drawing stress. Increase of the drawing stress results in greater initial fiber birefringence and lower ?C.

Thermal response of Bragg gratings in PMMA microstructured optical fibers

We report on the thermal characteristics of Bragg gratings fabricated in polymer optical fibers. We have observed a permanent shift in the grating wavelength at room temperature which occurs when the grating has been heated above a threshold temperature. This threshold temperature is dependent on the thermal history of the grating, and we attribute the effect to a shrinking of the fiber. This effect can be avoided by annealing the fiber before grating inscription, resulting in a linear response with temperature and an increased linear operating temperature range of the grating.

Fabrication of micro-channels in optical fibers using femtosecond laser pulses and selective chemical etching

The fabrication of micro-channels in single-mode optical fibers is demonstrated using focused femtosecond laser processing and chemical etching. Straight line micro-channels are achieved based on a simple technique which overcomes limitations imposed by the fiber curved surface.

Magnetic drive system for a new centrifugal rotary blood pump

The purpose of this investigation was to design a novel magnetic drive and bearing system for a new centrifugal rotary blood pump (CRBP). The drive system consists of two components: (i) permanent magnets within the impeller of the CRBP; and (ii) the driving electromagnets. Orientation of the magnets varies from axial through to 60° included out-lean (conical configuration). Permanent magnets replace the electromagnet drive to allow easier characterization. The performance characteristics tested were the axial force of attraction between the stator and rotor at angles of rotational alignment, Ø, and the corresponding torque at those angles. The drive components were tested for various magnetic cone angles, ?. The test was repeated for three backing conditions: (i) non-backed; (ii) steel-cupped; and (iii) steel plate back-iron, performed on an Instron tensile testing machine. Experimental results were expanded upon through finite element and boundary element analysis (BEM). The force/torque characteristics were maximal for a 12-magnet configuration at 0° cone angle with steel-back iron (axial force = 60 N, torque = 0.375 Nm). BEM showed how introducing a cone angle increases the radial restoring force threefold while not compromising axial bearing force. Magnets in the drive system may be orientated not only to provide adequate coupling to drive the CRBP, but to provide significant axial and radial bearing forces capable of withstanding over 100 m/s2 shock excitation on the impeller. Although the 12 magnet 0° (?) configuration yielded the greatest force/torque characteristic, this was seen as potentially unattractive as this magnetic cone angle yielded poor radial restoring force characteristics.

Fiber Bragg gratings inscribed using 800nm femtosecond laser and a phase mask in singleand multi-core mid-IR glass fibers

For the first time, Fiber Bragg grating (FBG) structures have been inscribed in single-core passive germanate and three-core passive and active tellurite glass fibers using 800nm femtosecond (fs) laser and phase mask technique. With fs peak power intensity in the order of 1011W/cm2, the FBG spectra with 2nd and 3rd order resonances at 1540 and 1033nm in the germanate glass fiber and 2nd order resonances at ~1694 and ~1677nm with strengths up to 14dB in all three cores in the tellurite fiber were observed. Thermal responsivities of the FBGs made in these mid-IR glass fibers were characterized, showing average temperature responsivity ~20pm/°C. Strain responsivities of the FBGs in germanate glass fiber were measured to be 1.219pm/µe.

Enzymatic production of oligosaccharides in centrifugal fields

The aims of this work have been to identify an enzymatic reaction system suitable to investigate and develop the high-speed centrifuge as a novel reaction system for performing such reactions. The production of galacto-oligosaccharides by the trans-galactosyl activity of the enzyme β-galactosidase on lactose monohydrate was identified as a model enzymatic system to elucidate the principles of this type of process. Galacto-oligosaccharides have attracted considerable commercial interest as food additives which have been shown to be beneficial to the health of the human gastrointestinal tract. The development of a single unit operation capable of controlling the biosynthesis of galacto-oligosaccharides whilst simultaneously separating the enzyme from the reaction products would reduce downstream processing costs. This thesis shows for the first time that by using a combination of (a) immobilised or insolubilised β-galactosidase , (b) a rate-zonal centrifugation technique, and (c) various applied centrifugal fields, that a high-speed centrifuge could be used to control the formation of galacto-oligosaccharides whilst removing the enzyme from the reaction products. By layering a suspension of insolubilised β-galactosidase on top of a lactose monohydrate density gradient and centrifuging, the applied centrifugal fields generated produced sedimentation of the enzyme particles through the substrate. The higher sedimentation rate of the enzyme compared to those of the reaction products allowed for separation to take place. Complete sedimentation, or pelleting of the enzyme permits the possible recovery and re-use. Insolubilisation of the enzyme allowed it to be sedimented through the substrate gradient using much lower applied centrifugal fields than that required to sediment free soluble enzyme and this allowed for less expensive centrifugation equipment to be used. Using free soluble and insolubilised β-galactosidase stirred-batch reactions were performed to investigate the kinetics of lactose monohydrate hydrolysis and galacto-oligosaccharide formation. Based on these results a preliminary mathematical model based on Michaelis-Menten kinetics was produced. It was found that the enzyme insolubilisation process using a chemical cross-linking agent did not affect the process of galacto-oligosaccharide formation. Centrifugation experiments were performed and it was found that by varying the applied centrifugal fields that the yield of galacto-oligosaccharides could be controlled. The higher the applied centrifugal fields the lower the yield of galacto-oligosaccharides. By increasing the applied centrifugal fields the 'contact time' between the sedimenting enzyme and the substrate was reduced, which produced lower yields. A novel technique involving pulsing the insolubilised enzyme through the substrate gradient was developed and this was found to produce higher yields of galacto-oligosaccharide compared to using a single enzyme loading equivalent to the total combined activity of the pulses. Comparison of the galacto-oligosaccharide yields between stirred-batch and centrifugation reactions showed that the applied centrifugal fields did not adversely affect the transgalactosyl activity of the insolubilised enzyme.

Combined bioreaction and separation in centrifugal fields

The aim of this work has been to investigate the principle of combined centrifugal bioreaction-separation. The production of dextran and fructose by the action of the enzyme dextransucrase on sucrose was employed to elucidate some of the principles of this type of process. Dextran is a valuable pharmaceutical product used mainly as a blood volume expander and blood flow improver whilst fructose is an important dietary product. The development of a single step process capable of the simultaneous biosynthesis of dextran and the separation of the fructose by-product should improve dextran yields whilst reducing capital and processing costs. This thesis shows for the first time that it is possible to conduct successful bioreaction-separations using a rate-zonal centrifugation technique. By layering thin zones of dextrasucrase enzyme onto sucrose gradients and centrifuging, very high molecular weight (MW) dextran-enzyme complexes were formed that rapidly sedimented through the sucrose substrate gradients under the influence of the applied centrifugal field. The low MW fructose by-product sedimented at reduced rates and was thus separated from the enzyme and dextran during the reaction. The MW distribution of dextran recovered from the centrifugal bioreactor was compared with that from a conventional batch bioreactor. The results indicated that the centrifugal bioreactor produced up to 100% more clinical dextran with MWs of between 12 000 and 98 000 at 20% w/w sucrose concentrations than conventional bioreactors. This was due to the removal of acceptor fructose molecules from the sedimenting reaction zone by the action of the centrifugal field. Higher proportions of unwanted lower MW dextran were found in the conventional bioreactor than in the centrifugal bioreactor-separator. The process was studied on a number of alternative centrifugal systems. A zonal rotor fitted with a reorienting gradient core proved most successful for the evaluation of bioreactor performance. Results indicated that viscosity build-up in the reactor must be minimised in order to increase the yields of dextran per unit time and improve product separation. A preliminary attempt at modelling the process has also been made.

New developments in the study of optical parabolic pulses in normally dispersive fibers

We report two recent studies dealing with the evolution of parabolic pulses in normally dispersive fibres. On the one hand, the nonlinear reshaping from a Gaussian intensity profile towards the asymptotic parabolic shape is experimentally investigated in a Raman amplifier. On the other hand, the significant impact of the fourth order dispersion on a passive propagation is theoretically discussed: we numerically demonstrate flat-top, coherent supercontinuum generation in an all-normal dispersion-flattened photonic crystal fiber. This shape is associated to a strong reshaping of the temporal profile what becomes triangular.

Dissipative nonlinear structures in optical fiber systems II : self-similar parabolic pulses in active fibers

In this second talk on dissipative structures in fiber applications, we overview theoretical aspects of the generation, evolution and characterization of self-similar parabolic-shaped pulses in fiber amplifier media. In particular, we present a perturbation analysis that describes the structural changes induced by third-order fiber dispersion on the parabolic pulse solution of the nonlinear Schrödinger equation with gain. Promising applications of parabolic pulses in optical signal post-processing and regeneration in communication systems are also discussed.