Adenovirus-mediated gene transfer of growth and differentiation factor-5 into tenocytes and the healing rat Achilles tendon.
Growth and differentiation factor-5 (GDF-5) is known to induce tendon tissue and stimulate tendon healing. The hypothesis was that adenoviral GDF-5 transfer leads to transitory transgene expression and improves Achilles tendon healing. In vitro experiments were first performed with rat tenocytes. Transgene expression was evaluated by RT-PCR, Western blotting and GDF-5-ELISA. In vivo virus dosage and transgene expression were examined by a marker gene transfer (LacZ and luciferase). In the main experiment in 131 rats, adenovirus particles (3 x 10(10)) were injected into transected Achilles tendons. The time course of GDF-5 mRNA expression was assessed by real-time RT-PCR. Histology and biomechanical testing were used to evaluate tendon healing and tensile strength. In vitro GDF-5 was secreted with a maximum after 2 weeks (330 ng GDF-5/10(6) cells per 24 hr). In vivo GDF-5 transgene expression showed a maximum at 4 weeks. At 8 weeks, GDF-5 specimens were thicker (p<0.05) with a trend to higher strength (p=0,064). Histology showed greater cartilage formation in type II collagen stains than in controls. Injection of adenovirus particles successfully can deliver the GDF-5 gene in healing tendons and leads to thicker tendon regenerates after 8 weeks. This technique might become a new approach for nonsurgical treatment of tendon injuries.
Department of Orthopaedic Surgery, University of Heidelberg, Heidelberg, Germany. markus.rickert@ok.uni-heidelburg.de
Markus
M
Rickert
Haili
H
Wang
Peter
P
Wieloch
Helga
H
Lorenz
Eric
E
Steck
Desiderius
D
Sabo
Wiltrud
W
Richter
D000125:Achilles Tendon / Q000166:cytology / Q000293:injuries* / Q000378:metabolism; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis / Q000235:genetics*; D002356:Cartilage / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D024043:Collagen Type II / Q000378:metabolism; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000502:physiology; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000627:therapeutic use*; D055428:Growth Differentiation Factor 5; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012038:Regeneration / Q000502:physiology*; D013708:Tendon Injuries / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D019076:Transgenes / Q000235:genetics; D016896:Treatment Outcome; D014945:Wound Healing / Q000502:physiology*
D019485:Bone Morphogenetic Proteins; D024043:Collagen Type II; D055428:Growth Differentiation Factor 5; D012333:RNA, Messenger