title: Ultrasound and microbubble-assisted gene delivery in Achilles tendons: long lasting gene expression and restoration of fibromodulin KO phenotype.
abstract: The aim of this study is to deliver genes in Achilles tendons using ultrasound and microbubbles. The rationale is to combine ultrasound-assisted delivery and the stimulation of protein expression induced by US. We found that mice tendons injected with 10 μg of plasmid encoding luciferase gene in the presence of 5×10⁵ BR14 microbubbles, exposed to US at 1 MHz, 200 kPa, 40% duty cycle for 10 min were efficiently transfected without toxicity. The rate of luciferase expression was 100-fold higher than that obtained when plasmid alone was injected. Remarkably, the luciferase transgene was stably expressed for up to 108 days. DNA extracted from these sonoporated tendons was efficient in transforming competent E. coli bacteria, indicating that persistent intact pDNA was responsible for this long lasting gene expression. We used this approach to restore expression of the fibromodulin gene in fibromodulin KO mice. A significant fibromodulin expression was detected by quantitative PCR one week post-injection. Interestingly, ultrastructural analysis of these tendons revealed that collagen fibrils diameter distribution and circularity were similar to that of wild type mice. Our results suggest that this gene delivery method is promising for clinical applications aimed at modulating healing or restoring a degenerative tendon while offering great promise for gene therapy due its safety compared to viral methods.
authors: Centre de Biophysique Moléculaire UPR 4301 CNRS, Rue Charles Sadron 45071 Orléans Cedex 2, France.
authors: Anthony
authors: A
authors: Delalande
authors: Ayache
authors: A
authors: Bouakaz
authors: Gilles
authors: G
authors: Renault
authors: Flore
authors: F
authors: Tabareau
authors: Spiros
authors: S
authors: Kotopoulis
authors: Patrick
authors: P
authors: Midoux
authors: Brigitte
authors: B
authors: Arbeille
authors: Rustem
authors: R
authors: Uzbekov
authors: Shukti
authors: S
authors: Chakravarti
authors: Michiel
authors: M
authors: Postema
authors: Chantal
authors: C
authors: Pichon
mesh_terms: D000125:Achilles Tendon / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D000071235:Fibromodulin; D005786:Gene Expression Regulation; D008156:Luciferases / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D045423:Microbubbles; D010641:Phenotype; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011509:Proteoglycans / Q000235:genetics*; D014162:Transfection; D019076:Transgenes; D014465:Ultrasonics
chemical_list: D016326:Extracellular Matrix Proteins; C000604332:Fmod protein, mouse; D011509:Proteoglycans; D000071235:Fibromodulin; D004247:DNA; D008156:Luciferases