title: Chemically modified RNA activated matrices enhance bone regeneration.
abstract: There exists a dire need for improved therapeutics to achieve predictable bone regeneration. Gene therapy using non-viral vectors that are safe and efficient at transfecting target cells is a promising approach to overcoming the drawbacks of protein delivery of growth factors. Here, we investigated the transfection efficiency, cytotoxicity, osteogenic potential and in vivo bone regenerative capacity of chemically modified ribonucleic acid (cmRNA) (encoding BMP-2) complexed with polyethylenimine (PEI) and made comparisons with PEI complexed with conventional plasmid DNA (encoding BMP-2). The polyplexes were fabricated at an amine (N) to phosphate (P) ratio of 10 and characterized for transfection efficiency using human bone marrow stromal cells (BMSCs). The osteogenic potential of BMSCs treated with these polyplexes was validated by determining the expression of bone-specific genes, osteocalcin and alkaline phosphatase as well as through the detection of bone matrix deposition. Using a calvarial bone defect model in rats, it was shown that PEI-cmRNA (encoding BMP-2)-activated matrices promoted significantly enhanced bone regeneration compared to PEI-plasmid DNA (BMP-2)-activated matrices. Our proof of concept study suggests that scaffolds loaded with non-viral vectors harboring cmRNA encoding osteogenic proteins may be a powerful tool for stimulating bone regeneration with significant potential for clinical translation.
authors.affiliation: Department of Periodontics, University of Iowa College of Dentistry, Iowa City, IA, United States. Electronic address: satheesh-elangovan@uiowa.edu.
authors.forename: Satheesh
authors.initials: S
authors.lastname: Elangovan
authors.affiliation: Division of Pharmaceutics and Translational Therapeutics, University of Iowa College of Pharmacy, Iowa City, IA, United States.
authors.forename: Behnoush
authors.initials: B
authors.lastname: Khorsand
authors.affiliation: Division of Pharmaceutics and Translational Therapeutics, University of Iowa College of Pharmacy, Iowa City, IA, United States.
authors.forename: Anh-Vu
authors.initials: AV
authors.lastname: Do
authors.affiliation: Department of Prosthodontics, University of Iowa College of Dentistry, Iowa City, IA, United States.
authors.forename: Liu
authors.initials: L
authors.lastname: Hong
authors.affiliation: Department of Pediatrics I-Pediatric Infectiology and Immunology, Translational Genomics and Gene Therapy, University of Tübingen, Wilhelstr. 56, 72074 Tübingen, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Dewerth
authors.affiliation: Department of Pediatrics I-Pediatric Infectiology and Immunology, Translational Genomics and Gene Therapy, University of Tübingen, Wilhelstr. 56, 72074 Tübingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Kormann
authors.affiliation: Department of Anatomy and Cell Biology, Rush Medical College, Chicago, IL, United States.
authors.forename: Ryan D
authors.initials: RD
authors.lastname: Ross
authors.affiliation: Department of Anatomy and Cell Biology, Rush Medical College, Chicago, IL, United States.
authors.forename: D Rick
authors.initials: DR
authors.lastname: Sumner
authors.affiliation: Central Microscopy Research Facility, University of Iowa, Iowa City, IA, United States.
authors.forename: Chantal
authors.initials: C
authors.lastname: Allamargot
authors.affiliation: Department of Periodontics, University of Iowa College of Dentistry, Iowa City, IA, United States; Division of Pharmaceutics and Translational Therapeutics, University of Iowa College of Pharmacy, Iowa City, IA, United States. Electronic address: aliasger-salem@uiowa.edu.
authors.forename: Aliasger K
authors.initials: AK
authors.lastname: Salem
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D001861:Bone Regeneration; D002470:Cell Survival; D006259:Craniocerebral Trauma / Q000628:therapy; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry; D015316:Genetic Therapy; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D015675:Osteocalcin / Q000235:genetics; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry*; D012313:RNA / Q000008:administration & dosage* / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use; D011916:Rats, Inbred F344; D017154:Stromal Cells / Q000378:metabolism
chemical_list: D055396:Bone Morphogenetic Protein 2; D015675:Osteocalcin; D012313:RNA; D011094:Polyethyleneimine; D004247:DNA; D000469:Alkaline Phosphatase
keywords: Bone morphogenetic protein-2; Bone regeneration; Chemically modified RNA; Gene delivery; Plasmid DNA; Polyethylenimine