title: Mechanical stimulation increases collagen type I and collagen type III gene expression of stem cell-collagen sponge constructs for patellar tendon repair.
abstract: Our group has shown that mechanical stimulation increases the stiffness of stem cell-collagen sponge constructs at 14 days in culture and subsequent rabbit patellar tendon repairs at 12 weeks postsurgery. What remains unclear is which genes might be responsible for this increase in stiffness. Therefore, the objective of this study was to determine how a tensile stimulus affects the gene expression of stem cell-collagen sponge constructs used to repair rabbit central patellar tendon defects. Tissue-engineered constructs were created by seeding mesenchymal stem cells (MSCs) from 10 adult rabbits at 0.14 x 10(6) cells/construct in type I collagen sponges. Half of the constructs were mechanically stimulated once every 5 min for 8 h/d to a peak strain of 2.4% for 2 weeks. The other half remained in an incubator without mechanical stimulation for 2 weeks. After 14 days in culture, half of the stimulated and nonstimulated constructs were prepared to determine the expression of collagen type I, collagen type III, decorin, fibronectin, and glyceraldehyde-3-phosphate dehydrogenase genes using real-time quantitative reverse transcriptase polymerase chain reaction. The remaining constructs were mechanically tested to determine their mechanical properties. Two weeks of in vitro mechanical stimulation significantly increased collagen type I and collagen type III gene expression of the stem cell-collagen sponge constructs. Stimulated constructs showed 3 and 4 times greater collagen type I (p = 0.0001) and collagen type III gene expression (p = 0.001) than nonstimulated controls. Stimulated constructs also had 2.5 times the linear stiffness and 4 times the linear modulus of nonstimulated constructs. However, mechanical stimulation did not significantly increase decorin or fibronectin gene expression (p = 0.2) after 14 days in culture. This study shows that mechanical stimulation of cell-sponge constructs produces similar increases in the expression of 2 structural genes, as well as linear stiffness and linear modulus.
authors.affiliation: Department of Biomedical Engineering, University of Cincinnati, Cincinnati, Ohio 45221-0048, USA. juncosln@email.uc.edu
authors.forename: Natalia
authors.initials: N
authors.lastname: Juncosa-Melvin
authors.forename: Karl S
authors.initials: KS
authors.lastname: Matlin
authors.forename: Robert W
authors.initials: RW
authors.lastname: Holdcraft
authors.forename: Victor S
authors.initials: VS
authors.lastname: Nirmalanandhan
authors.forename: David L
authors.initials: DL
authors.lastname: Butler
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D024042:Collagen Type I / Q000737:chemistry / Q000378:metabolism*; D024043:Collagen Type II / Q000378:metabolism*; D004548:Elasticity; D005260:Female; D015870:Gene Expression / Q000502:physiology; D066298:In Vitro Techniques; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology*; D017847:Patellar Ligament / Q000293:injuries / Q000473:pathology / Q000601:surgery; D010812:Physical Stimulation; D011817:Rabbits; D013314:Stress, Mechanical; D013718:Tensile Strength
chemical_list: D024042:Collagen Type I; D024043:Collagen Type II