Effects of antisense transforming growth factor-beta1 gene transfer on the biological activities of tendon sheath fibroblasts.
Recent studies have shown the importance of transforming growth factor-beta (TGF-beta) in flexor tendon wound healing. Decreased adhesion formation and increased range of motion after the administration of TGF-beta antibodies after tendon repair have been shown. But TGF-beta antibodies have a short biologic half-life, and continuous supplementation of exogenous TGF-beta antibodies is not practical. Transfer of growth factor genes to tenocytes provides an alternative to protein therapeutics, and a gene therapy approach will prolong the availability of therapeutic proteins.We investigated the biological activities effects of rabbit tendon sheath fibroblasts transfected by antisense TGF-beta1 gene. Tendon sheath fibroblasts were isolated from New Zealand white rabbits and transfected by antisense TGF-beta1 gene with Lipofectin (Invitrogen, Carlsbad, California). Reverse transcription polymerase chain reaction was used to measure collagen I, collagen III, and TGF-beta1 expression, and Western blot was used to measure collagen protein I expression in tendon sheath fibroblasts after being transfected by antisense TGF-beta1 gene. Reverse transcription polymerase chain reaction displayed that tendon sheath fibroblasts transfected with antisense TGF-beta1 gene showed marked decrease collagen I, collagen III, and TGF-beta1 mRNA expression. Western blot showed that tendon sheath fibroblasts transfected with antisense TGF-beta1 gene showed marked decrease expression of collagen I protein, and there was significant difference compared with the untransfected and empty transfected groups (P<.01). Tendon sheath fibroblasts can transfect with antisense TGF-beta1 gene successfully and can decrease production of collagen I, collagen III, and TGF-beta1, which were factors of tendon adhere formation.
Xia
Changsuo
C
Department of Orthopedics, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, China. xcs009@163.com
Ding
Changrong
C
Yang
Xuanying
X
Sun
Kang
K
Tian
Shaoqi
S
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D024061:Collagen Type III / Q000096:biosynthesis / Q000235:genetics; D004195:Disease Models, Animal; D005346:Fibroblast Growth Factors / Q000235:genetics / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D008297:Male; D012313:RNA / Q000235:genetics; D011817:Rabbits; D013708:Tendon Injuries / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D013710:Tendons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics; D014945:Wound Healing / Q000187:drug effects*
D024042:Collagen Type I; D024061:Collagen Type III; D016212:Transforming Growth Factor beta; C108111:fibroblast growth factor 13; D005346:Fibroblast Growth Factors; D012313:RNA