Collagen gel delivery of TGF-β3 non-viral plasmid DNA in rat osteoblast and calvarial culture

S. Premaraj, B. Mundy, J. Parker-Barnes, P. L. Winnard, Amr Moursi

Research output: Contribution to journalArticle

Abstract

Authors -Premaraj S, Mundy B, Parker-Barnes J, Winnard PL, Moursi AM Different forms of collagen as a carrier for naked plasmid DNA have shown potential as vehicles for therapeutic gene delivery and tissue engineering. The objective of this study was to determine the suitability of a dense collagen gel as a vehicle for sustained delivery of plasmid DNA in cell and organ culture. Plasmid DNA encoding Tgf-b3 was combined with collagen gel. DNA released into the media was measured by Pico-Green spectrophotometry. Results showed that DNA was released from the collagen gel at a gradual rate for up to 14 days. To evaluate collagen-mediated transfection in tissue, calvariae were exposed to collagen containing plasmid encoding GFP or DsRed. Transfection was visualized by fluorescence localized to tissue adjacent to the vehicle. To evaluate protein production, fetal rat calvarial osteoblasts were cultured with a collagen/Tgf-β3 plasmid mixture or in media containing plasmid alone. Media was collected at various time points to measure Tgf-β3 protein production. ELISA assays showed that collagentransfected osteoblasts demonstrated an elevated Tgf-β3 protein production for up to 14 days. Therefore, collagen delivery of viable plasmid DNA created a sustained transient transfection of calvarial osteoblasts resulting in prolonged and elevated growth factor production. Together, these results suggest that use of collagen gel as a vehicle may provide a strategy to achieve localized and controlled, non-viral gene delivery in vivo.

Original languageEnglish (US)
Pages (from-to)320-322
Number of pages3
JournalOrthodontics and Craniofacial Research
Volume8
Issue number4
DOIs
StatePublished - 2005

Fingerprint

Osteoblasts
Plasmids
Collagen
Gels
DNA
Transfection
Fetal Proteins
Spectrophotometry
Organ Culture Techniques
Tissue Engineering
Skull
Genes
Intercellular Signaling Peptides and Proteins
Proteins
Cell Culture Techniques
Fluorescence
Enzyme-Linked Immunosorbent Assay

Keywords

  • Collagen gel
  • Cranial suture
  • Craniosynostosis
  • Nonviral plasmid delivery
  • Osteoblast
  • Tgf-β3

ASJC Scopus subject areas

  • Orthodontics
  • Oral Surgery
  • Otorhinolaryngology
  • Surgery

Cite this

Collagen gel delivery of TGF-β3 non-viral plasmid DNA in rat osteoblast and calvarial culture. / Premaraj, S.; Mundy, B.; Parker-Barnes, J.; Winnard, P. L.; Moursi, Amr.

In: Orthodontics and Craniofacial Research, Vol. 8, No. 4, 2005, p. 320-322.

Research output: Contribution to journalArticle

Premaraj, S. ; Mundy, B. ; Parker-Barnes, J. ; Winnard, P. L. ; Moursi, Amr. / Collagen gel delivery of TGF-β3 non-viral plasmid DNA in rat osteoblast and calvarial culture. In: Orthodontics and Craniofacial Research. 2005 ; Vol. 8, No. 4. pp. 320-322.
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