This study developed a solvent merging/particulate leaching method for preparing three-dimensional porous scaffolds. Poly(L-lactic-co-glycolic acid) (PLGA) and sodium chloride particles were dry-mixed and cast into a special mold, through which a liquid could pass due to a pressure difference. An organic solvent was then poured into the mold to dissolve and merge the PLGA particles under negative pressure. A nonsolvent was conducted into the PLGA/salt composite to solidify and precipitate the merged PLGA matrix. Finally, a large amount of water was passed through the mold to leach out the salt particles so as to create a porous structure. The results revealed that a highly porous three-dimensional scaffold (>85 vol %) with a well interconnected porous structure could be achieved by this process. Porosity and the pore size of the scaffold were controlled using the ratio and the particle size of the added salt particles. A larger-volume scaffold was produced using a larger mold. This work provides a continuous and simple procedure for fabricating a bulk three-dimensional porous scaffold for tissue engineering.
The research goals were to fabricate a crosslinked hyaluronate (HA)/collagen membrane and study its efficacy in preventing peridural adhesion. Different weight ratios of HA/collagen membranes crosslinked with carbodiimide were tested for biocompatibility and biodegradability first. Forty-eight adult New Zealand white rabbits underwent an L6 laminectomy. Sixteen rabbits each received a weight ratio of HA/collagen = 60/40 membrane (Membrane A) or a 40/60 membrane (Membrane B) on the exposed dura. The last 16 rabbits received no treatment and served as controls. No adverse reaction of the membranes was noted. Magnetic resonance images revealed a hyposignal space between the dura and the scar tissues at Membrane B-treated laminectomy site 3 months after surgery. Histological examination showed that the amount of scarring decreased with time in all groups. Amount of scarring decreased significantly at laminectomy sites treated with either membrane. Compared with the control group, the extent of peridural adhesion decreased significantly in the Membrane B-treated group at 3 months after surgery; while it decreased substantially, but not significantly, in the Membrane A-treated group. The carbodiimide-crosslinked HA/collagen membrane is a safe and effective antiadhesive material in vivo. When placed onto the laminectomy site, the membrane with a weight ratio of HA/collagen = 40/60 appears to reduce peridural scar adhesion.
We established histopathologic and neurophysiologic approaches to examine whether different designs of polycaprolactone-engineered nerve conduits (hollow vs. laminated) could promote nerve regeneration as autologous grafts after transection of sciatic nerves. The assessments included morphometric analysis at the level of sciatic nerve, neuromuscular junction (NMJ) and gastrocnemius muscle, and nerve conduction studies on sciatic nerves. Six months after nerve grafting, the nerve fiber density in the hollow-conduit group was similar to that in the autologous-graft group; the laminated-conduit group only achieved approximately 20% of these values. The consequences of these differences were reflected in nerve growth into muscular targets; this was demonstrated by combined cholinesterase histochemistry for NMJ and immunohistochemistry for nerve fibers innervating NMJ with an axonal marker, protein gene product 9.5. Hollow conduits had similar index of NMJ innervation as autologous grafts; the values were higher than those of laminated conduits. Among the 3 groups there were same patterns of differences in the cross-sectional area of muscle fibers and amplitudes of compound muscle action potential. These results indicate that hollow conduits were as efficient as autologous grafts to facilitate nerve regeneration, and provide a multidisciplinary approach to quantitatively evaluate muscular reinnervation after nerve injury.
The influence of polyurethane compositions and fabrication conditions on the pore morphology and mechanical properties of microporous segmented polyetherurethane (SPEU) grafts, which were produced by the coagulation technique, were carefully investigated in this article. SPEU resins based on polytetramethylene oxide (PTMO) were synthesized by the solution polymerization method. Different types of coagulant were adapted to examine the feasibility of producing a microporous SPEU graft with good structural regularity. The experimental results indicate that a microporous SPEU graft with a uniform pore structure can be fabricated quite conveniently by using a proper concentration of water and ethanol mixed coagulant. Tensile tests demonstrated that the fabricated microporous SPEU grafts possess high mechanical strengths and satisfy the requirements as vessel replacements. The burst strength test also revealed that the SPEU graft can sustain extremely high internal pressure. Furthermore, a high compliant SPEU (high porosity) graft can be obtained by blending a proper amount of "soluble filler" (i.e., free PTMO polyol in this study) into the SPEU resin.
Hyaluronic acid (HA) is a high‐molecular‐weight glycosaminoglycan and extracellular matrix component that promotes cell proliferation. This study aimed to evaluate the effects of HA on alkali‐injured human corneal epithelial cells in vitro, and to elucidate the mechanisms by which HA mediates corneal cell protection. A human corneal epithelial cell line (HCE‐2) was treated with sodium hydroxide before incubation with low‐molecular‐weight HA (LMW‐HA, 127 kDa) or high‐molecular‐weight HA (HMW‐HA, 1525 kDa). A global proteomic analysis was then performed. Our data indicated that HA treatment protects corneal epithelial cells from alkali injury, and that the molecular weight of HA is a crucial factor in determining its effects. Only HMW‐HA reduced NaOH‐induced cytotoxic effects in corneal cells significantly and increased their migratory and wound healing ability. Results from 2D‐DIGE and MALDI‐TOF/TOF MS analyses indicated that HMW‐HA modulates biosynthetic pathways, cell migration, cell outgrowth, and protein degradation to stimulate wound healing and prevent cell death. To our knowledge, our study is the first to report the possible mechanisms by which HMW‐HA promotes repair in alkali‐injured human corneal epithelial cells.
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