As a potential alternative to currently available skin substitutes and wound dressings, we explored the use of bioactive scaffolds made of plant-derived proteins. We hypothesized that 'green' materials, derived from renewable and biodegradable natural sources, may confer bioactive properties to enhance wound healing and tissue regeneration. We optimized and characterized fibrous scaffolds electrospun from soy protein isolate (SPI) with addition of 0.05% poly(ethylene oxide) (PEO) dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol, and from corn zein dissolved in glacial acetic acid. Fibrous mats electrospun from either of these plant proteins remained intact without further cross-linking, possessing a skin-like pliability. Soy-derived scaffolds supported the adhesion and proliferation of cultured primary human dermal fibroblasts. Using targeted PCR arrays and qPCR validation, we found similar gene expression profiles of fibroblasts cultured for 2 and 24 h on SPI substrates and on collagen type I at both time points. On both substrates there was a pronounced time-dependent upregulation of several genes related to ECM deposition remodelling, including MMP-10, MMP-1, collagen VII, integrin-α2 and laminin-β3, indicating that both plant- and animal-derived materials induce similar responses from the cells after initial adhesion, degrading substrate proteins and depositing extracellular matrix in a 'normal' remodelling process. These results suggest that 'green' proteins, such as soy and zein, are promising as a platform for organotypic skin equivalent culture, as well as implantable scaffolds for skin regeneration. Future studies will determine specific mechanisms of their interaction with skin cells and their efficacy in wound-healing applications.
Electrospinning is an increasingly popular technique to generate 3D fibrous tissue scaffolds that mimic the submicron sized fibers of extracellular matrices. A major drawback of electrospun scaffolds is the small interfibrillar pore size, which normally prevents cellular penetration in between fibers. In this study, we introduced a novel process, based on electrospinning, to manufacture a unique gradient porous fibrous (GPF) scaffold from soy protein isolate (SPI). The pore sizes in the GPF scaffolds gradually increase from one side of the scaffold to the other, ranging from 7.8 ± 2.5 μm in the small pore side, 21.4 ± 10.3 μm in the mid layer to 58.0 ± 23.6 μm in the large pore side. The smallest pores of the GPF scaffolds appeared to be somewhat larger than those in conventionally electrospun SPI scaffolds (4.2 ± 1.3 μm). Hydrated GPF scaffolds exhibited J-shaped stress-strain curves, reminiscent of those for soft biological scaffolds. Attachment, spreading, and proliferation of human dermal fibroblasts (HDFB) were supported on both the small and the large pore sides of the GPF scaffolds. Cultured HDFB and murine RAW 264.7 macrophages penetrated significantly deeper (98.7 ± 24.2 μm and 53.3 ± 9.6 μm, respectively) into the larger pores than when seeded onto the small pore side of GPF scaffolds (22.8 ± 6.2 μm and 25.7 ± 7.3 μm) and control SPI scaffolds. (11.3 ± 3.8 μm and 15.3 ± 3.1 μm). This study introduces a novel fabrication technique, which, by convergence of several biofabrication technologies, produces scaffolds with enhanced cellular penetration.
We have generated a model for custom 3-D synthesis of incudal prostheses. While current 3-D printing in biocompatible materials at the size required is limited, the technology is rapidly advancing, and 3-D printing of incudal replacements with polylactic acid (PLA) is of the correct size and shape.
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