TY - JOUR
T1 - Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming
AU - Wang, Xiao
AU - Qiu, Yiwei
AU - Carr, Andrew J.
AU - Triffitt, James T.
AU - Sabokbar, Afsie
AU - Xia, Zhidao
PY - 2011/6
Y1 - 2011/6
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=79960085495&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79960085495&partnerID=8YFLogxK
U2 - 10.1088/1748-6041/6/3/035010
DO - 10.1088/1748-6041/6/3/035010
M3 - Article
C2 - 21555843
AN - SCOPUS:79960085495
VL - 6
JO - Biomedical Materials
JF - Biomedical Materials
SN - 1748-6041
IS - 3
M1 - 035010
ER -