Mouse growth and differentiation factor-5 protein and DNA therapy potentiates intervertebral disc cell aggregation and chondrogenic gene expression.

摘要: Abstract Background context Growth and differentiation factor-5 (GDF-5)–deficient mice showed abnormalities in intervertebral disc (IVD) structure extracellular matrix. Adenovirus-mediated GDF-5 delivery can promote the growth of rabbit cells. Purpose The aim present study was to investigate effect recombinant protein complementary DNA (cDNA) on metabolism IVD Study design effects cDNA mouse cells will be evaluated vitro. Methods Mouse vitro were treated with protein. cloned into an expression vector used transfect Therapy assessed by measuring cell proliferation, proteoglycan production, matrix gene expression. Results Biochemical assays revealed elevated sulfated glycosaminoglycan (GAG)/DNA ratio that cultured presence various concentrations GDF-5(mGDF-5) Real-time reverse transcription-polymerase chain reaction (RT-PCR) demonstrated treating increased collagen Type II aggrecan genes a dose-dependent manner but decreased metalloproteinase (MMP)-3 Immunohistochemistry increase aggregation mGDF-5 culture compared control group. successfully plasmid vector, production confirmed Western blot analysis. significantly transfected nucleofection plasmid. Conclusions This is first report cloning use method transfer data suggest both forms for proteins Future attempts at therapy treat degenerative disease novel ex vivo technique are needed develop would alleviate condition patients clinically relevant axial spine pain.