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

Silk fibroin exhibits high biocompatibility and biodegradability, making it a versatile biomaterial for medical applications. However, contaminated silkworm-derived substances in remnant sericin from the filature and degumming process can result in undesired immune reactions and silk allergy, limiti...

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Autores principales: Yamano, Mana, Hirose, Ryoko, Lye, Ping Ying, Takaki, Keiko, Maruta, Rina, On Liew, Mervyn Wing, Sakurai, Shinichi, Mori, Hajime, Kotani, Eiji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9267247/
https://www.ncbi.nlm.nih.gov/pubmed/35806440
http://dx.doi.org/10.3390/ijms23137433
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author Yamano, Mana
Hirose, Ryoko
Lye, Ping Ying
Takaki, Keiko
Maruta, Rina
On Liew, Mervyn Wing
Sakurai, Shinichi
Mori, Hajime
Kotani, Eiji
author_facet Yamano, Mana
Hirose, Ryoko
Lye, Ping Ying
Takaki, Keiko
Maruta, Rina
On Liew, Mervyn Wing
Sakurai, Shinichi
Mori, Hajime
Kotani, Eiji
author_sort Yamano, Mana
collection PubMed
description Silk fibroin exhibits high biocompatibility and biodegradability, making it a versatile biomaterial for medical applications. However, contaminated silkworm-derived substances in remnant sericin from the filature and degumming process can result in undesired immune reactions and silk allergy, limiting the widespread use of fibroin. Here, we established transgenic silkworms with modified middle silk glands, in which sericin expression was repressed by the ectopic expression of cabbage butterfly-derived cytotoxin pierisin-1A, to produce cocoons composed solely of fibroin. Intact, nondegraded fibroin can be prepared from the transgenic cocoons without the need for sericin removal by the filature and degumming steps that cause fibroin degradation. A wide-angle X-ray diffraction analysis revealed low crystallinity in the transgenic cocoons. However, nondegraded fibroin obtained from transgenic cocoons enabled the formation of fibroin sponges with varying densities by using 1–5% (v/v) alcohol. The effective chondrogenic differentiation of ATDC5 cells was induced following their cultivation on substrates coated with intact fibroin. Our results showed that intact, allergen-free fibroin can be obtained from transgenic cocoons without the need for sericin removal, providing a method to produce fibroin-based materials with high biocompatibility for biomedical uses.
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spelling pubmed-92672472022-07-09 Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications Yamano, Mana Hirose, Ryoko Lye, Ping Ying Takaki, Keiko Maruta, Rina On Liew, Mervyn Wing Sakurai, Shinichi Mori, Hajime Kotani, Eiji Int J Mol Sci Article Silk fibroin exhibits high biocompatibility and biodegradability, making it a versatile biomaterial for medical applications. However, contaminated silkworm-derived substances in remnant sericin from the filature and degumming process can result in undesired immune reactions and silk allergy, limiting the widespread use of fibroin. Here, we established transgenic silkworms with modified middle silk glands, in which sericin expression was repressed by the ectopic expression of cabbage butterfly-derived cytotoxin pierisin-1A, to produce cocoons composed solely of fibroin. Intact, nondegraded fibroin can be prepared from the transgenic cocoons without the need for sericin removal by the filature and degumming steps that cause fibroin degradation. A wide-angle X-ray diffraction analysis revealed low crystallinity in the transgenic cocoons. However, nondegraded fibroin obtained from transgenic cocoons enabled the formation of fibroin sponges with varying densities by using 1–5% (v/v) alcohol. The effective chondrogenic differentiation of ATDC5 cells was induced following their cultivation on substrates coated with intact fibroin. Our results showed that intact, allergen-free fibroin can be obtained from transgenic cocoons without the need for sericin removal, providing a method to produce fibroin-based materials with high biocompatibility for biomedical uses. MDPI 2022-07-04 /pmc/articles/PMC9267247/ /pubmed/35806440 http://dx.doi.org/10.3390/ijms23137433 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yamano, Mana
Hirose, Ryoko
Lye, Ping Ying
Takaki, Keiko
Maruta, Rina
On Liew, Mervyn Wing
Sakurai, Shinichi
Mori, Hajime
Kotani, Eiji
Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications
title Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications
title_full Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications
title_fullStr Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications
title_full_unstemmed Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications
title_short Bioengineered Silkworm for Producing Cocoons with High Fibroin Content for Regenerated Fibroin Biomaterial-Based Applications
title_sort bioengineered silkworm for producing cocoons with high fibroin content for regenerated fibroin biomaterial-based applications
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9267247/
https://www.ncbi.nlm.nih.gov/pubmed/35806440
http://dx.doi.org/10.3390/ijms23137433
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