Optimal construction and delivery of dual-functioning lentiviral vectors for type I collagen-suppressed chondrogenesis in synovium-derived mesenchymal stem cells.
Pharm Res
; 28(6): 1338-48, 2011 Jun.
Article
in En
| MEDLINE
| ID: mdl-21057858
ABSTRACT
PURPOSE:
This study aims to deliver both transforming growth factor ß3 (TGF-ß3) and shRNA targeting type I collagen (Col I) by optimal construction and application of various dual-functioning lentiviral vectors to induce Col I-suppressed chondrogenesis in synovium-derived mesenchymal stem cells (SMSCs).METHODS:
We constructed four lentiviral vectors (LV-1, LV-2, LV-3 and LV-4) with various arrangements of the two expression cassettes in different positions and orientations. Col I inhibition efficiency and chondrogenic markers were assessed with qPCR, ELISA and staining techniques. Among the four vectors, LV-1 has two distant and reversely oriented cassettes, LV-2 has two distant and same-oriented cassettes, LV-3 has two proximal and reversely oriented cassettes, and LV-4 has two proximal and same-oriented cassettes. Col I and chondrogenic markers, including type II collagen (Col II), aggrecan and glycosaminoglycan (GAG), were examined in SMSCs cultured in 3-D alginate hydrogel.RESULTS:
All of the four vectors showed distinct effects in Col I level as well as diverse inductive efficiencies in upregulation of the cartilaginous markers. Based on real-time PCR results, LV-1 was optimal towards Col I-suppressed chondrogenesis.CONCLUSION:
LV-1 vector is competent to promote Col I-suppressed chondrogenesis in SMSCs.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Lentivirus
/
Chondrogenesis
/
Collagen Type I
/
RNA, Small Interfering
/
Mesenchymal Stem Cells
/
Genetic Vectors
Limits:
Animals
Language:
En
Journal:
Pharm Res
Year:
2011
Document type:
Article