Single-lumen and multi-lumen poly(ethylene glycol) nerve conduits fabricated by stereolithography for peripheral nerve regeneration in vivo.

BACKGROUND: The use of nerve conduits to facilitate nerve regrowth after peripheral nerve injury is limited to defects less than 3 cm. The purpose of this study is to determine the capability of novel single and multi-lumen poly(ethylene glycol) (PEG) conduits manufactured by stereolithography to promote peripheral nerve regeneration. MATERIALS AND METHODS: Eight Sprague Dawley rats with sharp transection injuries of the sciatic nerve were randomly assigned to receive single-lumen or multi-lumen PEG conduits to bridge a 10-mm gap. Sciatic nerve and conduit samples were harvested after 5 weeks, and axon number, myelin thickness, fiber diameter, and g-ratio were analyzed. The contralateral intact nerve was also harvested for comparison. RESULTS: Partial nerve regeneration was observed in three out of four single-lumen conduits and one out of four multi-lumen conduits. Axon number in the single-lumen regenerated nerve approached that of the contralateral intact nerve at 4,492 ± 2,810.0 and 6,080 ± 627.9 fibers/mm(2), respectively. The percentage of small fibers was greater in the single-lumen conduit compared with the intact nerve, whereas myelin thickness and g-ratio were consistently greater in the autologous nerve. Axon regrowth through the multi-lumen conduits was severely limited. CONCLUSION: Single-lumen stereolithography-manufactured PEG nerve conduits promote nerve regeneration, with regenerating axon numbers approaching that of normal nerve. Multi-lumen conduits demonstrated significantly less nerve regeneration, possibly due to physical properties of the conduit inhibiting growth. Further studies are necessary to compare the efficacy of the two conduits for functional recovery and to elucidate the reasons underlying their differences in nerve regeneration potential.

Fabrication of Off-the-Shelf Multilumen Poly(Ethylene Glycol) Nerve Guidance Conduits Using Stereolithography.

A manufacturing process for fabricating off-the-shelf multilumen poly(ethylene glycol) (PEG)-based nerve guidance conduits (NGCs) was developed that included the use of stereolithography (SL). A rapid fabrication strategy for complex 3D scaffolds incorporated postprocessing with lyophilization and sterilization to preserve the scaffold, creating an implantable product with improved suturability. SL is easily adaptable to changes in scaffold design, is compatible with various materials and cells, and can be expanded for mass manufacture. The fabricated conduits were characterized using optical and scanning electron microscopy, and measurements of swelling ratio, dimensional swelling factor, resistance to compression, and coefficient of friction were performed. Water absorption curves showed that the conduits after lyophilization and sterilization return easily and rapidly to a swollen state when placed in an aqueous solution, successfully maintaining their original overall structure as required for implantation. Postprocessed conduits at the swollen state were less slippery and therefore easier to handle than those without postprocessing. Suture pullout experiments showed that NGCs fabricated with a higher concentration of PEG were better able to resist suture pullout. NGCs having a multilumen design demonstrated a better resistance to compression than a single-lumen design with an equivalent surface area, as well as a greater force required to collapse the design. Conduits fabricated with a higher PEG concentration were shown to have compressive resistances comparable to those of commercially available NGCs. The use of SL with PEG and the manufacturing process developed here shows promise for improving the current state of the art in peripheral nerve repair strategies.

In vitro validation of finite element analysis of blood flow in deformable models.

The purpose of this article is to validate numerical simulations of flow and pressure incorporating deformable walls using in vitro flow phantoms under physiological flow and pressure conditions. We constructed two deformable flow phantoms mimicking a normal and a restricted thoracic aorta, and used a Windkessel model at the outlet boundary. We acquired flow and pressure data in the phantom while it operated under physiological conditions. Next, in silico numerical simulations were performed, and velocities, flows, and pressures in the in silico simulations were compared to those measured in the in vitro phantoms. The experimental measurements and simulated results of pressure and flow waveform shapes and magnitudes compared favorably at all of the different measurement locations in the two deformable phantoms. The average difference between measured and simulated flow and pressure was approximately 3.5 cc/s (13% of mean) and 1.5 mmHg (1.8% of mean), respectively. Velocity patterns also showed good qualitative agreement between experiment and simulation especially in regions with less complex flow patterns. We demonstrated the capabilities of numerical simulations incorporating deformable walls to capture both the vessel wall motion and wave propagation by accurately predicting the changes in the flow and pressure waveforms at various locations down the length of the deformable flow phantoms.

Stereolithography of spatially controlled multi-material bioactive poly(ethylene glycol) scaffolds.

Challenges remain in tissue engineering to control the spatial, mechanical, temporal and biochemical architectures of scaffolds. Unique capabilities of stereolithography (SL) for fabricating multi-material spatially controlled bioactive scaffolds were explored in this work. To accomplish multi-material builds, a mini-vat setup was designed allowing for self-aligning X-Y registration during fabrication. The mini-vat setup allowed the part to be easily removed and rinsed, and different photocrosslinkable solutions to be easily removed and added to the vat. Two photocrosslinkable hydrogel biopolymers, poly(ethylene glycol) dimethacrylate (PEG-dma, MW 1000) and poly(ethylene glycol) diacrylate (PEG-da, MW 3400), were used as the primary scaffold materials. Multi-material scaffolds were fabricated by including controlled concentrations of fluorescently labeled dextran, fluorescently labeled bioactive PEG or bioactive PEG in different regions of the scaffold. The presence of the fluorescent component in specific regions of the scaffold was analyzed with fluorescent microscopy, while human dermal fibroblast cells were seeded on top of the fabricated scaffolds with selective bioactivity and phase contrast microscopy images were used to show specific localization of cells in the regions patterned with bioactive PEG. Multi-material spatial control was successfully demonstrated in features down to 500 microm. In addition, the equilibrium swelling behavior of the two biopolymers after SL fabrication was determined and used to design constructs with the specified dimensions at the swollen state. The use of multi-material SL and the relative ease of conjugating different bioactive ligands or growth factors to PEG allows for the fabrication of tailored three-dimensional constructs with specified spatially controlled bioactivity.

Stereolithography of three-dimensional bioactive poly(ethylene glycol) constructs with encapsulated cells.

Stereolithography (SL) was used to fabricate complex 3-D poly(ethylene glycol) (PEG) hydrogels. Photopolymerization experiments were performed to characterize the solutions for use in SL, where the crosslinked depth (or hydrogel thickness) was measured at different laser energies and photoinitiator (PI) concentrations for two concentrations of PEG-dimethacrylate in solution (20% and 30% (w/v)). Hydrogel thickness was a strong function of PEG concentration, PI type and concentration, and energy dosage, and these results were utilized to successfully fabricate complex hydrogel structures using SL, including structures with internal channels of various orientations and multi-material structures. Additionally, human dermal fibroblasts were encapsulated in bioactive PEG photocrosslinked in SL. Cell viability was at least 87% at 2 and 24 h following fabrication. The results presented here indicate that the use of SL and photocrosslinkable biomaterials, such as photocrosslinkable PEG, appears feasible for fabricating complex bioactive scaffolds with living cells for a variety of important tissue engineering applications.