title: Evaluation of Adenovirus-Mediated Down-Regulation of Connective Tissue Growth Factor on Postoperative Wound Healing After Experimental Glaucoma Surgery.
abstract: This study was aimed to determine whether adenovirus-mediated down-regulation of connective tissue growth factor (CTGF) can modulate postoperative scarring in a rabbit receiving simplified glaucoma surgery.
  
  
  
  In vitro studies were performed using a replication-deficient recombinant adenovirus that transcribes a small interfering RNA (siRNA) specific to the CTGF gene under the control of the modified CMV promoter. Primary tenon cells from a New Zealand White rabbit were transduced with 10-100 plaque-forming units (pfu) per cell of the viral vector. Seventy-two hours later, CTGF expression was analyzed by Western blot analysis. In vivo studies were conducted using 10 New Zealand White rabbits, which underwent simplified glaucoma surgery and received a postoperative subconjunctival injection of 5 µl suspension of adenovirus carrying shRNA for CTGF (2 × 10(11) pfu/ml) in the right eye, and the same amount of null virus in the left eye. Eyes were enucleated 5 d after the surgery, and immunohistochemical and histological examinations of the surgical outcome were performed.
  
  
  
  Western blot analysis showed that CTGF was depleted to less than 10% of its original level in cells transduced with the adenovirus expressing CTGF-specific siRNA. This demonstrates RNA interference (RNAi)-mediated CTGF inactivation in vitro. Immunohistochemical analysis also showed that CTGF was significantly depleted in eyes transduced with the adenovirus expressing CTGF siRNA. This demonstrates RNAi-mediated CTGF inactivation in vivo. In addition, less scar tissue was observed on histological evaluation in the transduced eyes, demonstrating that inhibition of CTGF expression can modulate the wound healing process after surgery.
  
  
  
  Down-regulation of CTGF is effective in inhibiting postoperative scarring in vivo. This suggests that RNAi with CTGF siRNA may potentially pave the road for a novel therapeutic strategy to improve glaucoma surgery results.
authors.affiliation: a Department of Ophthalmology, Samsung Medical Center , Sungkyunkwan University School of Medicine , Seoul.
authors.forename: Dong Hui
authors.initials: DH
authors.lastname: Lim
authors.affiliation: b Center for Clinical Research , Samsung Biomedical Research Institute, Samsung Medical Center , Seoul , Korea.
authors.forename: Tae Eun
authors.initials: TE
authors.lastname: Kim
authors.affiliation: a Department of Ophthalmology, Samsung Medical Center , Sungkyunkwan University School of Medicine , Seoul.|b Center for Clinical Research , Samsung Biomedical Research Institute, Samsung Medical Center , Seoul , Korea.
authors.forename: Changwon
authors.initials: C
authors.lastname: Kee
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D055513:Connective Tissue Growth Factor / Q000096:biosynthesis / Q000235:genetics*; D004195:Disease Models, Animal; D015536:Down-Regulation; D018463:Filtering Surgery; D005901:Glaucoma / Q000378:metabolism / Q000601:surgery*; D007150:Immunohistochemistry; D034741:RNA, Small Interfering / Q000235:genetics*; D011817:Rabbits; D014162:Transfection; D014945:Wound Healing / Q000502:physiology*
chemical_list: D034741:RNA, Small Interfering; D055513:Connective Tissue Growth Factor
keywords: Adenovirus; connective tissue growth factor; fibrosis; gene therapy; glaucoma; glaucoma surgery; wound healing