Effects of adeno-associated virus-2-mediated human BMP-7 gene transfection on the phenotype of nucleus pulposus cells.
Bone morphogenetic protein-7 (BMP-7) was found to stimulate the synthesis of proteoglycans (PGs) and collagen type II. To increase the biological function of the nucleus pulposus (NP) cells, the Ad-hBMP-7 vector was also successfully constructed and transfected NP cells. However, the disadvantages of adenovirus limit the usefulness of the Ad-hBMP7 vector for clinical application. The rAAV2 vector has empirical advantages, especially for clinical use, to transfer exogenous genes into cells. The purpose of this study was to first determine whether a rAAV2-hBMP-7 vector could be used to transfect canine NP cells and effect on the biological functions of canine NP cells. The canine NP cells transfected by the rAAV-BMP7 were assessed semi-quantitatively for BMP-7 expression with real-time PCR and westernbloting. Aggrecan and collagens type I and II secreted by the NP cells were qualitatively assessed at 4, 7, and 14 days post-transfection in the transfection and control groups. We found that rAAV2 can successfully transfer the hBMP-7 gene into canine NP cells. NP cells transfected by the rAAV-hBMP-7 vector express hBMP-7 for at least 14 days. At 7 and 14 days, the expressed hBMP-7 promotes a remarkable and significant accumulation of both proteoglycans (42% and 77% higher than non-transfected cells) (p<0.05) and collagen type II (63% and 94% higher than non-transfected cells) (p<0.05). Thus, we could speculate that the rAAV-based gene delivery technique promotes the expression of proteoglycans and collagen type II of nucleus pulposus cells. Moreover, this technique may be applicable for the future treatment of degenerative disc disease.
Department of Orthopaedic Surgery, Navy General Hospital, Beijing, China.
Chaofeng
C
Wang
Di-Ke
DK
Ruan
Chao
C
Zhang
De-Li
DL
Wang
Hongkui
H
Xin
Yan
Y
Zhang
D053674:Aggrecans / Q000378:metabolism; D000818:Animals; D055419:Bone Morphogenetic Protein 7 / Q000235:genetics*; D049109:Cell Proliferation; D024042:Collagen Type I / Q000378:metabolism; D024043:Collagen Type II / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D004285:Dogs; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology / Q000378:metabolism*; D010641:Phenotype; D011509:Proteoglycans / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D053674:Aggrecans; C525819:BMP7 protein, human; D055419:Bone Morphogenetic Protein 7; D024042:Collagen Type I; D024043:Collagen Type II; D011509:Proteoglycans; D012333:RNA, Messenger; D011994:Recombinant Proteins