Abstract:

BACKGROUND: Gene interference can promote certain gene expressions, change the cell matrix synthesis condition by regulating the cell biological activity and stimulate the matrix synthesis which is benefit for the regeneration of the intervertebral discs to achieve therapeutic purposes.
OBJECTIVE: To construct a recombinant adenovirus vector containing transforming growth factor-β3 gene and to detect its effects on the proliferation of degenerated nucleus pulposus cells.
METHODS: Intervertebral disc degenerated model was constructed to gain the primary degenerated nucleus pulposus cells. Total RNA was extracted. Then the cDNA of the transforming growth factor-β3 was cloned into the shuttle plasmid pAdTrack-CMV. The recombinant adenovirus vector containing transforming growth factor-β3 was constructed by homogenous recombination in bacteria using PAdEasy system. The correct recombinant was packaged by liposome and transfected into 293A cells to amplify.
RESULTS AND CONCLUSION: The adenovirus vector containing transforming growth factor-β3 was successfully constructed with high infection; the titer was 1010 pfu/L. MTT assay showed that the adenovirus vector containing transforming growth factor-β3 significantly enhanced the activity of degenerated nucleus pulposus cells; it provides experimental and theoretical basis for research on the mechanism of transforming growth factor-β3 and its application on the gene therapy for intervertebral disc degeneration.