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Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming

Abstract

Tendon disorders are common clinical conditions. Tendon tissue engineering provides a new approach for tendon repair by integrating engineered substitutes with their native counterparts. Silk is considered to be a promising candidate for tendon engineering because of its biological and mechanical properties. However, a major concern with using silk for biomedical applications is the immune responses generated by sericin, a glue-like protein that coats the silk fibres. This study improves the existing protocols for silk 'degumming' which removes sericin and enables preparation of silk that is suitable for tendon regeneration. Bombyx mori silks were treated by sequential treatments with different proteases. The efficiency of degumming was determined by measuring weight loss, picric acid and carmine staining and scanning electron microscopy. To evaluate the cellular responses after degumming, the growth and differentiation of human tenocytes on silks were examined. The results showed that sequential protease treatment effectively degummed raw silks. The sequentially degummed silks showed enhanced tenocyte proliferation and upregulated mRNA levels of tendon markers. Thick cell multilayers formed on the treated silks, with cells and collagen fibres penetrating into the spaces in individual silk filaments, resulting in a structure resembling human tendon.
Authors:
Xiao, Wang; Yiwei, Qiu; Carr, Andrew J; Triffitt, James T; Sabokbar, Afsie; Xia Zhidao, E-mail: z.xia@swansea.ac.uk [1] 
  1. Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford (United Kingdom)
Publication Date:
Jun 15, 2011
Product Type:
Journal Article
Resource Relation:
Journal Name: Biomedical Materials (Bristol. Online); Journal Volume: 6; Journal Issue: 3; Other Information: DOI: 10.1088/1748-6041/6/3/035010; PII: S1748-6041(11)67983-7
Subject:
60 APPLIED LIFE SCIENCES; COLLAGEN; HUMAN POPULATIONS; IN VITRO; MECHANICAL PROPERTIES; MESSENGER-RNA; PICRIC ACID; SCANNING ELECTRON MICROSCOPY; TENDONS; ANIMAL TISSUES; AROMATICS; BODY; CHEMICAL EXPLOSIVES; CONNECTIVE TISSUE; ELECTRON MICROSCOPY; EXPLOSIVES; HYDROXY COMPOUNDS; MICROSCOPY; NITRO COMPOUNDS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PHENOLS; POPULATIONS; PROTEINS; RNA; SCLEROPROTEINS
OSTI ID:
21536516
Country of Origin:
United Kingdom
Language:
English
Other Identifying Numbers:
Journal ID: ISSN 1748-605X; TRN: GB12O8812015525
Availability:
Available from http://dx.doi.org/10.1088/1748-6041/6/3/035010
Submitting Site:
GBN
Size:
12 pages
Announcement Date:
Mar 29, 2012

Citation Formats

Xiao, Wang, Yiwei, Qiu, Carr, Andrew J, Triffitt, James T, Sabokbar, Afsie, and Xia Zhidao, E-mail: z.xia@swansea.ac.uk. Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming. United Kingdom: N. p., 2011. Web. doi:10.1088/1748-6041/6/3/035010.
Xiao, Wang, Yiwei, Qiu, Carr, Andrew J, Triffitt, James T, Sabokbar, Afsie, & Xia Zhidao, E-mail: z.xia@swansea.ac.uk. Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming. United Kingdom. doi:10.1088/1748-6041/6/3/035010.
Xiao, Wang, Yiwei, Qiu, Carr, Andrew J, Triffitt, James T, Sabokbar, Afsie, and Xia Zhidao, E-mail: z.xia@swansea.ac.uk. 2011. "Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming." United Kingdom. doi:10.1088/1748-6041/6/3/035010. https://www.osti.gov/servlets/purl/10.1088/1748-6041/6/3/035010.
@misc{etde_21536516,
title = {Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming}
author = {Xiao, Wang, Yiwei, Qiu, Carr, Andrew J, Triffitt, James T, Sabokbar, Afsie, and Xia Zhidao, E-mail: z.xia@swansea.ac.uk}
abstractNote = {Tendon disorders are common clinical conditions. Tendon tissue engineering provides a new approach for tendon repair by integrating engineered substitutes with their native counterparts. Silk is considered to be a promising candidate for tendon engineering because of its biological and mechanical properties. However, a major concern with using silk for biomedical applications is the immune responses generated by sericin, a glue-like protein that coats the silk fibres. This study improves the existing protocols for silk 'degumming' which removes sericin and enables preparation of silk that is suitable for tendon regeneration. Bombyx mori silks were treated by sequential treatments with different proteases. The efficiency of degumming was determined by measuring weight loss, picric acid and carmine staining and scanning electron microscopy. To evaluate the cellular responses after degumming, the growth and differentiation of human tenocytes on silks were examined. The results showed that sequential protease treatment effectively degummed raw silks. The sequentially degummed silks showed enhanced tenocyte proliferation and upregulated mRNA levels of tendon markers. Thick cell multilayers formed on the treated silks, with cells and collagen fibres penetrating into the spaces in individual silk filaments, resulting in a structure resembling human tendon.}
doi = {10.1088/1748-6041/6/3/035010}
journal = {Biomedical Materials (Bristol. Online)}
issue = {3}
volume = {6}
place = {United Kingdom}
year = {2011}
month = {Jun}
}