[Influence of liposome-mediated recombinant plasmid pIRES-hBMP-2-hVEGF165 on osteogenic activity of hBMSCs in vitro]

Guoping Wu, Xiaochuan He, Zhihui Yang, Li Guo
Chinese Journal of Reparative and Reconstructive Surgery 2009, 23 (9): 1124-8

OBJECTIVE: To investigate the effects of the recombinant plasmid pIRES-hBMP-2-hVEGFl65 on differentiation and maturation of hBMSCs in vitro.

METHODS: The co-expressing vector of hBMP-2 and hVEGF165 was constructed. The BMSCs were isolated and cultured from health adult human denoted marrow. By the lipofection method, the reconstructed plasmids pIRES-hBMP-2-hVEGF165, pIRES-hBMP-2, pIRES-hVEGF165 and pIRES neo empty vector, were transfected to hBMSCs (groups A, B, C and D). The untransfected cells were harvested as control group (group E). After 4 weeks of culture, RT-PCR was employed to assay the hBMP-2, hVEGF165 and osteocalcin mRNA expression in hBMSCs. The expressions of hBMP-2 and hVEGF165 of BMSCs were assayed by Western blot. The level of ALP activities of BMSCs was determined. Col I was also determined by immunohistochemical staining.

RESULTS: Compared to group E, the hBMSCs in group A secreted high level of hBMP-2, hVEGF165, Col I and osteocalcin; osteocalcin and Col I expressed at high level in group B, and hVEGF165 expressed at high level in group C. Otherwise, the expression of hVEGF165 in group B and the expressions of hBMP-2 and Col I in group C resemble to that of groups D and E, no expression or few expression was observed. The activities of ALP in groups A, B, C, D and E were 0.91 +/- 0.03, 0.90 +/- 0.02, 0.64 +/- 0.03, 0.67 +/- 0.01 and 0.66 +/- 0.02, respectively. The activity of ALP of groups A and B were significantly increased compared with that of group E (P < 0.05); there was no significant difference among groups C, D and E (P > 0.05).

CONCLUSION: The recombinant plasmid pIRES-hBMP-2-hVEGF165 can be successfully transfected into BMSCs with cation liposome-mediated transfection method, the exogenous hBMP-2 and hVEGF165 genes can be expressed constitutively in the transfected BMSCs, and it can enhance the differentiation abilities of BMSCs.

Full Text Links

Find Full Text Links for this Article


You are not logged in. Sign Up or Log In to join the discussion.

Related Papers

Remove bar
Read by QxMD icon Read

Save your favorite articles in one place with a free QxMD account.


Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"