The basement membrane (BM) is crucial in regulating the physical and biological activities of esophageal epithelial cells which attach to the underlying BM. In order to simulate the natural construction of BM, we prepared the fibrous scaffolds using biodegradable polylactide (PLA) and silk fibroin (SF) as the materials via electrospinning technology. BM's proteins containing collagen (IV), laminin, entactin and proteoglycan were extracted from porcine esophagus and coated on the eletrospun fibers. Morphology, mechanical strength, biodegradability and cytocompatibility of the coated and uncoated scaffolds were tested and evaluated using scanning electron micrography, mechanical test system, immunofluorescence assay and western blotting with CK14 as the primary antibody. The fibrous scaffold PLA or PLA/SF, generated from the present protocol had good formation and mechanical and biodegradable properties. After coating with BM's proteins, the scaffold could enhance the growth and differentiation of esophageal epithelial cells, which would contribute to remodel and regenerate the tissue engineered epithelium and further contribute to engineer the whole esophagus in future.
Absorbable Implants ; Basement Membrane ; Biocompatible Materials ; chemistry ; Epithelium ; Esophagus ; physiology ; Fibroins ; chemistry ; Humans ; Nanostructures ; chemistry ; Polyesters ; chemistry ; Regeneration ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

BACKGROUND:We have found that oriented fibers can guide the alignment of smooth muscle cells in our previous experiments. Thus, we designed the experiment to prepare wel aligned polymeric fibers using electrospinning technology, aiming at guiding the growth of esophageal smooth muscle cells to maintain cellmorphology and biological function. OBJECTIVE:Using electrospinning technology, to fabricate isotropic and directed nano-fibrous scaffolds made of polycaprolacton, gelatin and silk fibroin. METHODS:Polycaprolacton/silk fibroin fibers at a ratio of 4:1 were prepared with proper parameters, including solution concentration, voltage and injection speed, under the self-made spinning system. The polycaprolacton/gelatin sheets with mass ratio of 2:1, 1:1 and 1:2, respectively, were also fabricated under suitable process parameters. Using the rol er col ector instead of the metal plate, polycaprolacton/gelatin nano-fibrous scaffold with good alignment of fibers was manufactured. RESULTS AND CONCLUSION:The isotropic polycaprolacton/silk fibroin scaffold with fiber diameter of (535.9±126.7) nm was prepared under conditions of solution concentration (0.08 g/mL), injection speed (1.6 mL/h) and voltage (22.5 kV), and these fibers were uniform with no beads. The isotropic polycaprolacton/gelatin scaffold with fiber diameter of (257.9±117.8) nm was prepared under conditions of solution concentration (0.10 g/mL), injection speed (0.8 mL/h) and voltage (22.5 kV). Using the rol er col ector instead of the previous metal plate, polycaprolacton/gelatin (w:w, 1:2) nano-fibrous scaffold with good alignment of fibers was manufactured. The process parameters were 3 000 r/min of rol ing speed, 0.8 mL/h of injection speed and 15 kV of voltage.