Home › Microbiology › Peptides from Phage Display Library Modulate Gene Expression in Mesenchymal Cells and Potentiate Osteogenesis in Unicortical Bone Defects
Microbiology JoVE (Open Access) Citable · DOI

Peptides from Phage Display Library Modulate Gene Expression in Mesenchymal Cells and Potentiate Osteogenesis in Unicortical Bone Defects

DOI: 10.3791/2362-v
What you'll learn
  • ✓ Perform phage display biopanning to identify bone-targeting peptide sequences
  • ✓ Assess peptide binding to mesenchymal cells using immunofluorescence staining
  • ✓ Prepare and implant peptide-loaded gel foam composites in bone defects
  • ✓ Evaluate osteogenic effects of phage-display peptides in vitro and in vivo
Protocol

A phage display library was used to identify peptide sequences that target bone. The objective was to investigate the effect of these peptides on mesenchymal cell differentiation and to determine their effect on bone regeneration.

Difficulty
advanced
Total time
~3-4 weeks (biopanning ~1 week; cell culture differentiation ~2 weeks; surgical healing/histology ~2 weeks)
Model organism
Mouse (surgical bone defect model)
Biosafety
BSL-1

Steps

1
Screen phage display library via iterative biopanning

Perform multiple rounds of biopanning to enrich phage clones that bind bone tissue. Select and amplify phages displaying bone-targeting peptide sequences.

▶ 01:44
2
Stain peptides on cultured mesenchymal cells

Incubate mesenchymal cells with identified peptides and perform immunofluorescence staining to confirm peptide binding and cellular localization.

▶ 03:39
3
Prepare peptide-gel foam composite and implant

Combine peptides with gel foam scaffold material and surgically implant into unicortical bone defects in mice.

▶ 04:42
4
Process tissue samples for histological analysis

Harvest implanted bone defect sites and prepare histological sections to evaluate tissue organization and bone regeneration.

▶ 05:25
5
Compare osteogenic effects in vitro and in vivo

Analyze mesenchymal cell differentiation markers in culture and quantify bone formation in implanted defects to assess peptide efficacy.

▶ 05:40
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