Combination gene therapy targeting on interleukin-1β and RANKL for wear debris-induced aseptic loosening.
This study investigated the efficacy of a combination gene therapy to repress interleukin-1 (IL-1) and receptor activator of nuclear factor NF-kappa B ligand (RANKL) for the treatment of particulate debris-induced aseptic loosening, and tried to explore the molecular mechanism of the exogenous gene modifications on osteoclastogenesis. RAW cells activated by titanium particles were transduced with DFG-IL-1Ra (retroviral vector encoding IL-1 receptor antagonist) and AAV-OPG (adeno-associated viral vectors-osteoprotegerin) individually or in combination for 4 weeks. Pro-inflammatory cytokines in culture media were determined by enzyme-linked immunosorbent assay, and gene expressions of RANK, IL-1β, c-Fos, TRAF6, JNK1 and CPK were examined using real-time PCR. An established knee-implant-failure mouse model was employed to evaluate the efficacy of the in vivo double-gene therapy. The surgical implantation of a titanium alloy pin into the proximal tibia was followed by monthly challenge with titanium debris. Peri-implant gene transfers of IL-1Ra and OPG (respectively or in combination) were given 3 weeks after surgery. The combination of OPG and IL-1Ra gene transfer exhibited strong synergetic effects in blockage of inflammation and osteoclastogenesis at 8 weeks after gene modification. The combination therapy reversed peri-implant bone resorption and restored implant stability when compared with either single gene transduction. Real-time PCR data indicated that the action of IL-1Ra gene therapy may be mediated via the JNK1 pathway, while the reduction of osteoclastogenesis by OPG gene modification may be regulated by c-Fos expression. In addition, both gene modifications resulted in significant diminishment of TRAF6 expression.
Orthopaedic Research Institute, Via Christi Wichita Hospitals Inc., Wichita, KS 67214, USA.
H
H
Wang
T-H
TH
Jia
N
N
Zacharias
W
W
Gong
H-X
HX
Du
P H
PH
Wooley
S-Y
SY
Yang
D000818:Animals; D001861:Bone Regeneration / Q000235:genetics; D001862:Bone Resorption / Q000628:therapy; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D015316:Genetic Therapy; D020007:Implants, Experimental; D007249:Inflammation / Q000628:therapy*; D053590:Interleukin 1 Receptor Antagonist Protein / Q000235:genetics; D053583:Interleukin-1beta / Q000037:antagonists & inhibitors*; D007720:Knee Prosthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D010010:Osteoclasts / Q000378:metabolism / Q000473:pathology; D053244:Osteoprotegerin / Q000235:genetics; D011475:Prosthesis Failure; D053245:RANK Ligand / Q000037:antagonists & inhibitors*; D014025:Titanium / Q000008:administration & dosage
D053590:Interleukin 1 Receptor Antagonist Protein; D053583:Interleukin-1beta; D053244:Osteoprotegerin; D053245:RANK Ligand; D014025:Titanium