title: Nanoplex-Mediated Codelivery of Fibroblast Growth Factor and Bone Morphogenetic Protein Genes Promotes Osteogenesis in Human Adipocyte-Derived Mesenchymal Stem Cells.
abstract: This study highlights the importance of transfection mediated coordinated bone morphogenetic protein 2 (BMP-2) and fibroblast growth factor 2 (FGF-2) signaling in promoting osteogenesis. We employed plasmids independently encoding BMP-2 and FGF-2 complexed with polyethylenimine (PEI) to transfect human adipose derived mesenchymal stem cells (hADMSCs) in vitro. The nanoplexes were characterized for size, surface charge, in vitro cytotoxicity, and transfection ability in hADMSCs. A significant enhancement in BMP-2 protein secretion was observed on day 7 post-transfection of hADMSCs with PEI nanoplexes loaded with both pFGF-2 and pBMP-2 (PEI/(pFGF-2+pBMP-2)) versus transfection with PEI nanoplexes of either pFGF-2 alone or pBMP-2 alone. Osteogenic differentiation of transfected hADMSCs was determined by measuring osteocalcin and Runx-2 gene expression using real time polymerase chain reactions. A significant increase in the expression of Runx-2 and osteocalcin was observed on day 3 and day 7 post-transfection, respectively, by cells transfected with PEI/(pFGF-2+pBMP-2) compared to cells transfected with nanoplexes containing pFGF-2 or pBMP-2 alone. Alizarin Red staining and atomic absorption spectroscopy revealed elevated levels of calcium deposition in hADMSC cultures on day 14 and day 30 post-transfection with PEI/(pFGF-2+pBMP-2) compared to other treatments. We have shown that codelivery of pFGF-2 and pBMP-2 results in a significant enhancement in osteogenic protein synthesis, osteogenic marker expression, and subsequent mineralization. This research points to a new clinically translatable strategy for achieving efficient bone regeneration.
authors.affiliation: †Division of Pharmaceutics and Translational Therapeutics, College of Pharmacy, The University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Keerthi
authors.initials: K
authors.lastname: Atluri
authors.affiliation: ‡Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Denise
authors.initials: D
authors.lastname: Seabold
authors.affiliation: ‡Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Liu
authors.initials: L
authors.lastname: Hong
authors.affiliation: ‡Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Satheesh
authors.initials: S
authors.lastname: Elangovan
authors.affiliation: †Division of Pharmaceutics and Translational Therapeutics, College of Pharmacy, The University of Iowa, Iowa City, Iowa 52242, United States.|‡Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Aliasger K
authors.initials: AK
authors.lastname: Salem
mesh_terms: D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D001854:Bone Marrow Cells / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001861:Bone Regeneration / Q000502:physiology*; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D010012:Osteogenesis / Q000235:genetics*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry* / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods; D014162:Transfection
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D016222:Fibroblast Growth Factor 2; D011094:Polyethyleneimine; D004247:DNA
keywords: BMP-2; FGF-2; PEI; bone morphogenetic protein 2; fibroblast growth factor 2; human adipose derived mesenchymal stem cells; nanoplex; nonviral gene delivery; osteogenesis; pDNA; plasmid DNA; polyethylenimine