title: Efficiently engineered cell sheet using a complex of polyethylenimine-alginate nanocomposites plus bone morphogenetic protein 2 gene to promote new bone formation.
abstract: Regeneration of large bone defects is a common clinical problem. Recently, stem cell sheet has been an emerging strategy in bone tissue engineering. To enhance the osteogenic potential of stem cell sheet, we fabricated bone morphogenetic protein 2 (BMP-2) gene-engineered cell sheet using a complex of polyethylenimine-alginate (PEI-al) nanocomposites plus human BMP-2 complementary(c)DNA plasmid, and studied its osteogenesis in vitro and in vivo. PEI-al nanocomposites carrying BMP-2 gene could efficiently transfect bone marrow mesenchymal stem cells. The cell sheet was made by culturing the cells in medium containing vitamin C for 10 days. Assays on the cell culture showed that the genetically engineered cells released the BMP-2 for at least 14 days. The expression of osteogenesis-related gene was increased, which demonstrated that released BMP-2 could effectively induce the cell sheet osteogenic differentiation in vitro. To further test the osteogenic potential of the cell sheet in vivo, enhanced green fluorescent protein or BMP-2-producing cell sheets were treated on the cranial bone defects. The results indicated that the BMP-2-producing cell sheet group was more efficient than other groups in promoting bone formation in the defect area. Our results suggested that PEI-al nanocomposites efficiently deliver the BMP-2 gene to bone marrow mesenchymal stem cells and that BMP-2 gene-engineered cell sheet is an effective way for promoting bone regeneration.
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Han
authors.initials: H
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, People's Republic of China.
authors.forename: Kai
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Qiao
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Anliang
authors.initials: A
authors.lastname: Yuan
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Daowei
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Ce
authors.initials: C
authors.lastname: Shi
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Shilei
authors.initials: S
authors.lastname: Ni
authors.affiliation: Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Changyu
authors.initials: C
authors.lastname: Zheng
authors.affiliation: Department of Biomedical Sciences, Texas A&M University Baylor College of Dentistry, Dallas, TX, USA.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, People's Republic of China.
authors.forename: Bai
authors.initials: B
authors.lastname: Yang
authors.affiliation: Department of Pathology, School of Stomatology, Jilin University, Changchun, People's Republic of China.
authors.forename: Hongchen
authors.initials: H
authors.lastname: Sun
mesh_terms: D000464:Alginates / Q000737:chemistry*; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism; D018786:Bone Substitutes / Q000138:chemical synthesis; D004867:Equipment Design; D019544:Equipment Failure Analysis; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000295:instrumentation*; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D053761:Nanocomposites / Q000737:chemistry / Q000648:ultrastructure; D010012:Osteogenesis / Q000187:drug effects / Q000502:physiology; D011094:Polyethyleneimine / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D012887:Skull Fractures / Q000175:diagnosis / Q000628:therapy*; D023822:Tissue Engineering / Q000295:instrumentation; D054457:Tissue Scaffolds; D016896:Treatment Outcome
chemical_list: D000464:Alginates; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D018786:Bone Substitutes; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D011094:Polyethyleneimine
keywords: bone morphogenetic protein 2; bone regeneration; bone tissue engineering; cell sheet; nanocomposites