Name: peptides from phage display library modulate gene expression in mesenchymal cells and potentiate osteogenesis in unicortical bone defects
Uploaded: 2010-12-10T17:00:00
Duration: 7 min 53 s
Description: Watch this Scientific Journal Video about Peptides from Phage Display Library Modulate Gene Expression in Mesenchymal Cells and Potentiate Osteogenesis in Unicortical Bone Defects at JoVE.com

National Institutes of Health, AR53579, Osteogenesis: Potentiation with Bone Tropic Peptides 
National Institutes of Health, AR056422, Anabolic Effect of Osteogenic Peptides 
National Institutes of Health, T32 AR050960, Musculoskeletal Tissue Repair and Regeneration 
DOD, PC051219 Nucleolin: A Novel Mediator of Prostate Cancer and Bone Marrow Endothelial Cell Adhesion 
Department of Defense, PC061508, Prostate Cancer Metastasis Cell-Bone Marrow adhesion Mediators

<ol>
	<li>Diduch, D. R., Coe, M. R., Joyner, C., Owen, M. E., Balian, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+cell+lines+from+bone+marrow+that+differ+in+terms+of+collagen+synthesis,+osteogenic+characteristics,+and+matrix+mineralization.">Two cell lines from bone marrow that differ in terms of collagen synthesis, osteogenic characteristics, and matrix mineralization.</a> J. Bone Joint Surg. Am. 75, 92-105 (1993).</li><li>Stoddart, M. J., Furlong, P. I., Simpson, A. E., Davies, C. M., Richards, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viability+in+ex+vivo+cultured+cancellous.">Viability in ex vivo cultured cancellous.</a> Eur. Cells. Mater. 13, 69-70 (2007).</li><li>Lehr, J. E., Pienta, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preferential+adhesion+of+prostate+cancer+cells+to+a+human+bone+marrow+endothelial+cell+line.">Preferential adhesion of prostate cancer cells to a human bone marrow endothelial cell line.</a> J. Natl. Cancer Inst. 90, 118-123 (1998).</li><li>Schuster, N., G&#246;tz, C., Faust, M., Schneider, E., Prowald, A., Jungbluth, A., Montenarh, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wild-type+p53+inhibits+protein+kinase+CK2+activity.">Wild-type p53 inhibits protein kinase CK2 activity.</a> J. Cell Biochem. 81, 172-183 (2001).</li><li>Cwirla, S. E., Peters, E. A., Barrett, R. W., Dower, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peptides+on+phage:+a+vast+library+of+peptides+for+identifying+ligands.">Peptides on phage: a vast library of peptides for identifying ligands.</a> Proc. Natl. Acad. Sci. U. S. A. 87, 6378-6382 (1990).</li><li>Doorbar, J., Winter, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+a+peptide+antagonist+to+the+thrombin+receptor+using+phage+display.">Isolation of a peptide antagonist to the thrombin receptor using phage display.</a> J. Mol. Biol. 244, 361-369 (1994).</li><li>Ellerby, H. M., Arap, W., Ellerby, L. M., Kain, R., Andrusiak, R., Rio, G. D., Krajewski, S., Lombardo, C. R., Rao, R., Ruoslahti, E., Bredesen, D. E., Pasqualini, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-cancer+activity+of+targeted+pro-apoptotic+peptides.">Anti-cancer activity of targeted pro-apoptotic peptides.</a> Nat. Med. 5, 1032-1038 (1999).</li><li>Goodson, R. J., Doyle, M. V., Kaufman, S. E., Rosenberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-affinity+urokinase+receptor+antagonists+identified+with+bacteriophage+peptide+display.">High-affinity urokinase receptor antagonists identified with bacteriophage peptide display.</a> Proc. Natl. Acad. Sci. U. S. A. 91, 7129-7133 (1994).</li><li>Pasqualini, R., Koivunen, E., Ruoslahti, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+peptide+isolated+from+phage+display+libraries+is+a+structural+and+functional+mimic+of+an+RGD-binding+site+on+integrins.">A peptide isolated from phage display libraries is a structural and functional mimic of an RGD-binding site on integrins.</a> J. Cell Biol. 130, 1189-1196 (1995).</li><li>Pasqualini, R., Koivunen, E., Ruoslahti, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha+v+integrins+as+receptors+for+tumor+targeting+by+circulating+ligands.">Alpha v integrins as receptors for tumor targeting by circulating ligands.</a> Nat. Biotechnol. 15, 542-546 (1997).</li><li>Pasqualini, R., Koivunen, E., Kain, R., Lahdenranta, J., Sakamoto, M., Stryhn, A., Ashmun, R. A., Shapiro, L. H., Arap, W., Ruoslahti, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aminopeptidase+N+is+a+receptor+for+tumor-homing+peptides+and+a+target+for+inhibiting+angiogenesis.">Aminopeptidase N is a receptor for tumor-homing peptides and a target for inhibiting angiogenesis.</a> Cancer Res. 60, 722-727 (2000).</li><li>Rajotte, D., Arap, W., Hagedorn, M., Koivunen, E., Pasqualini, R., Ruoslahti, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+heterogeneity+of+the+vascular+endothelium+revealed+by+in+vivo+phage+display.">Molecular heterogeneity of the vascular endothelium revealed by in vivo phage display.</a> J. Clin. Invest. 102, 430-437 (1998).</li><li>Rajotte, D., Ruoslahti, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+dipeptidase+is+the+receptor+for+a+lung-targeting+peptide+identified+by+in+vivo+phage+display.">Membrane dipeptidase is the receptor for a lung-targeting peptide identified by in vivo phage display.</a> J. Biol. Chem. 274, 11593-11598 (1999).</li><li>Wang, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+high-affinity+peptide+antagonists+of+14-3-3+proteins+by+phage+display.">Isolation of high-affinity peptide antagonists of 14-3-3 proteins by phage display.</a> Biochemistry. 38, 12499-12504 (1999).</li><li>Sugahara, K. N., Teesalu, T., Karmali, P. P., Kotamraju, V. R., Agemy, L., Greenwald, D. R., Ruoslahti, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coadministration+of+a+tumor-penetrating+peptide+enhances+the+efficacy+of+cancer+drugs.">Coadministration of a tumor-penetrating peptide enhances the efficacy of cancer drugs.</a> Science. 328, 1031-1035 (2010).</li><li>Sikes, R. A., Cooper, C. R., Beck, G. L., Pruitt, F., Brown, M. L., Balian, G., Meadows, G., Kluwer, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+Stromal+Cells+as+Therapeutics+Targets+in+Osseous+Metastasis.">Bone Stromal Cells as Therapeutics Targets in Osseous Metastasis.</a> Cancer Growth and Progression. "Integration/Interaction of Oncologic Growth". , 369-386 (2005).</li><li>Santos, J. L., Pandita, D., Rodrigues, J., P&#234;go, A. P., Granja, P. L., Tom&#225;s, H., Balian, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Receptor-Mediated+Gene+Delivery+in+Mesenchymal+Stem+Cells+using+PAMAM+Dendrimers+conjugated+with+Osteotropic+Peptides.">Receptor-Mediated Gene Delivery in Mesenchymal Stem Cells using PAMAM Dendrimers conjugated with Osteotropic Peptides.</a> Molecular Pharmaceutics. , (2010).</li></ol>

The production of this video was sponsored by New England Biolabs, Inc, which produces some of the reagents used in this article.

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Phage Display Peptide Library Kit-Ph.D-12</td>
<td>&nbsp;</td>
<td>New England Biolabs</td>
<td>E8110S</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

peptides from phage display library modulate gene expression in mesenchymal cells and potentiate osteogenesis in unicortical bone defects

Two novel synthetic peptides accelerate bone formation and can be delivered using a collagen matrix. The aim of this study was to investigate the effects on bone repair in a unicortical defect model. Treatment of mesenchymal cells produced an increase in alkaline phosphatase activity, showed nodule formation by the cells, and increased the expression of genes for runx2, osterix, bone sialoprotein, and osteocalcin. A collagen sponge soaked with peptide promoted repair of bone defects, whereas the control was less effective. The results from this study demonstrated that mesenchymal cells treated with peptide in vitro differentiate towards osteogenesis, and, that peptides delivered in vivo using a collagen sponge promote the repair of unicortical defects.

Watch this Scientific Journal Video about Peptides from Phage Display Library Modulate Gene Expression in Mesenchymal Cells and Potentiate Osteogenesis in Unicortical Bone Defects at JoVE.com

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

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.

Two novel synthetic peptides accelerate bone formation and can be delivered using a collagen matrix. The aim of this study was to investigate the effects ...

Research

JoVE Journal

Biology

Culture of myeloid dendritic cells from bone marrow precursors

Myeloid dendritic cells (DCs) are frequently used to study the interactions between innate and adaptive immune mechanisms and the early response to ...

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser ...

Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency

Implantation of Ferumoxides Labeled Human Mesenchymal Stem Cells in Cartilage Defects

The field of tissue engineering integrates the principles of engineering, cell biology and medicine towards the regeneration of specific cells and ...

Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells

The use of siRNA mediated gene knockdown is continuing to be an important tool in studies of gene expression.  siRNA studies are being conducted not only ...

In vitro Reconstitution of the Active T. castaneum Telomerase

In vitro Reconstitution of the Active T. castaneum Telomerase

Efforts to isolate the catalytic subunit of telomerase, TERT, in sufficient quantities for structural studies, have been met with limited success for more ...

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides

Brug SecM Arrest Sequence som et redskab til at isolere ribo bundne polypeptider

Extensive research has provided ample evidences suggesting that protein folding in the cell is a co-translational process1-5. However, the exact pathway ...

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits

En protokol for fagvisning og Affinity Selection Brug rekombinant protein Lokkemad

Using  recombinant phage as a scaffold to present various protein portions encoded by  a directionally cloned cDNA library to immobilized bait molecules ...

siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells

siRNA Screening at identificere Ubiquitin og ubiquitinlignende System Regulators Biologisk Pathways i dyrkede pattedyrceller

Post-translational modification of proteins with ubiquitin and ubiquitin-like molecules (UBLs) is emerging as a dynamic cellular signaling network that ...