Effects of transgenic silk materials that incorporate FGF‐7 protein microcrystals on the proliferation and differentiation of human keratinocytes

Abstract

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A Bioengineering Approach for the Development of Fibroblast Growth Factor-7-Functionalized Sericin Biomaterial Applicable for the Cultivation of Keratinocytes

“…To obtain silkworm cocoons containing FGF-7 in the sericin layer, we generated transgenic silkworm line S1sp-F7 carrying FGF-7 cDNA-fused with the polyhedra-encapsulation signal helix-1 sequence (H1/FGF-7) and regulated by the sericin-1 promoter ( Figure 1 A). The fusion with the H1 peptide facilitated the encapsulation of foreign proteins using microcrystals of cypovirus polyhedrin (also known as polyhedra) and showed no adverse effects on the biological activities of the fused proteins, including FGF-7 [ 11 , 28 ]. The stabilization effect of cypovirus polyhedra on encapsulated proteins against the hostile environment [ 11 , 12 , 28 ] drove us to investigate the stability of H1/FGF-7 as a combined effect of encapsulation by polyhedra and incorporation into sericin cocoons.…”

“…The fusion with the H1 peptide facilitated the encapsulation of foreign proteins using microcrystals of cypovirus polyhedrin (also known as polyhedra) and showed no adverse effects on the biological activities of the fused proteins, including FGF-7 [ 11 , 28 ]. The stabilization effect of cypovirus polyhedra on encapsulated proteins against the hostile environment [ 11 , 12 , 28 ] drove us to investigate the stability of H1/FGF-7 as a combined effect of encapsulation by polyhedra and incorporation into sericin cocoons. Therefore, transgenic silkworm line S1sp-F7/poly producing cocoons with sericin layers incorporated with co-expressed H1/FGF-7 and polyhedrin was generated as described in the Supplementary Materials .…”

“…Transgenic silkworm line S1sp-F7 was created to produce cocoons shells incorporating H1/FGF-7 in the sericin layer of silkworm cocoons ( Figure 1 A). Microinjection was performed as previously described [ 12 , 28 , 50 , 51 ] to generate S1sp-F7. In brief, the constructed vector, pBacMCS (hr3-Ser1sp-H1/FGF-7; 3 × P3-EGFP-SV40) ( Figure S2(Ai) ), was co-injected with a helper plasmid that encodes the piggyBac transposase into pre-blastodermic w1-pnd embryos [ 52 ].…”

Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications

“…Fibroin-based materials are useful for biomedical applications, such as in fine sutures [6], electrospun nanofiber film-based dressings [7], hydrogels for drug delivery and tissue engineering [8,9], biodegradable carriers for cell growth factors [10,11], and sponge scaffolds for cell growth [12][13][14]. Fibroin-based biomaterials, which are biocompatible, biodegradable, and have low immunostimulatory properties, have been used for revascularization [15], bone tissue regeneration [16], and cutaneous wound healing [17,18].…”

“…Fibroin-based biomaterials, which are biocompatible, biodegradable, and have low immunostimulatory properties, have been used for revascularization [15], bone tissue regeneration [16], and cutaneous wound healing [17,18]. Our previous study showed that fibroin powders incorporated with protein microcrystalencapsulated cytokines processed from transgenic silk glands effectively controlled NIH-3T3 proliferation [10] and facilitated the formation of a three-dimensional (3D) epidermis model [11]. These examples highlight the potential of fibroin-based biomaterials that can be generated through the bioengineering of silkworms for biomedical applications, particularly for tissue engineering.…”