[Construction of recombinant adenovirus bearing human transforming growth factor beta1 and bone morphogenetic protein 7 genes and its directional inducement effect on marrow stromal cells].
OBJECTIVE
To construct the recombinant adenovirus bearing human transforming growth factor beta1 (TGF-beta1) and bone morphogenetic protein 7 (BMP-7) genes, and investigate its co-expression in the marrow stromal stem cells (MSCs) and bioactivity effect.


METHODS
Using the replication defective adenovirus AdEasy as a carrier, MSCs were infected by the high-titer-level infected recombinant adenovirus taking TGF-beta1 and BMP genes. Immunocytochemistry, in situ hybridization ,reverse transcription-polymerase chain reaction (RT-PCR), and hexuronic acid level test were used to detect the co-expression of the exogenous genes and to analyze their effect transfection on directive differentiation of MSCs.


RESULTS
The immunocytochemistry staining showed that the brown coarse grains were situated in the cytoplasm of the most MSCs 72 h after infection. Procollagen I mRNA in the cells was detected by the in situ hybridization, and the content of hexuronic acid in the culture medium was significantly increased 10 days after infection compared with the level before infection (P<0. 01).


CONCLUSIONS
The recombinant adenovirus bearing human TGF-beeta1 and BMP-7 genes can be constructed, and the exogenous gene can be co-expressed in MSCs, which may offer a novel approach to the local combination gene therapy for repairing joint cartilage defects.
Long
Hua
H
Department of Orthopedics, Tangdu Hospital, Fourth Military Medical University, Xi'an Shaanxi, 710038, PR China.
Yuan
Hua
H
Ma
Baoan
B
D000256:Adenoviridae / Q000235:genetics; D000257:Adenoviridae Infections; D001854:Bone Marrow Cells / Q000166:cytology*; D055419:Bone Morphogenetic Protein 7 / Q000235:genetics*; D002454:Cell Differentiation; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D017154:Stromal Cells; D023822:Tissue Engineering / Q000379:methods; D053773:Transforming Growth Factor beta1 / Q000235:genetics*
D055419:Bone Morphogenetic Protein 7; D053773:Transforming Growth Factor beta1