title: In vitro degradation of silk fibroin.
abstract: A significant need exists for long-term degradable biomaterials which can slowly and predictably transfer a load-bearing burden to developing biological tissue. In this study Bombyx mori silk fibroin yarns were incubated in 1mg/ml Protease XIV at 37 degrees C to create an in vitro model system of proteolytic degradation. Samples were harvested at designated time points up to 12 weeks and (1) prepared for scanning electron microscopy (SEM), (2) lyophilized and weighed, (3) mechanical properties determined using a servohydraulic Instron 8511, (4) dissolved and run on a SDS-PAGE gel, and (5) characterized with Fourier transform infrared spectroscopy. Control samples were incubated in phosphate-buffered saline. Fibroin was shown to proteolytically degrade with predictable rates of change in fibroin diameter, failure strength, cycles to failure, and mass. SEM indicated increasing fragmentation of individual fibroin filaments from protease-digested samples with time of exposure to the enzyme; particulate debris was present within 7 days of incubation. Gel electrophoresis indicated a decreasing amount of the silk 25 kDa light chain and a shift in the molecular weight of the heavy chain with increasing incubation time in protease. Results support that silk is a mechanically robust biomaterial with predictable long-term degradation characteristics.
authors.affiliation: Biomedical Engineering Department, Tufts University, 4 Colby Street, Room 153, Medford, MA 02155, USA.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Horan
authors.forename: Kathryn
authors.initials: K
authors.lastname: Antle
authors.forename: Adam L
authors.initials: AL
authors.lastname: Collette
authors.forename: Yongzhong
authors.initials: Y
authors.lastname: Wang
authors.forename: Jia
authors.initials: J
authors.lastname: Huang
authors.forename: Jodie E
authors.initials: JE
authors.lastname: Moreau
authors.forename: Vladimir
authors.initials: V
authors.lastname: Volloch
authors.forename: David L
authors.initials: DL
authors.lastname: Kaplan
authors.forename: Gregory H
authors.initials: GH
authors.lastname: Altman
mesh_terms: D020341:Absorbable Implants; D001672:Biocompatible Materials / Q000032:analysis / Q000737:chemistry*; D004548:Elasticity; D005349:Fibroins / Q000032:analysis / Q000737:chemistry* / Q000648:ultrastructure*; D008422:Materials Testing; D011402:Pronase / Q000737:chemistry*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013718:Tensile Strength
chemical_list: D001672:Biocompatible Materials; C482288:fibroin, silkworm; D005349:Fibroins; D011402:Pronase