title: Gene transfection of human mesenchymal stem cells with a nano-hydroxyapatite-collagen scaffold containing DNA-functionalized calcium phosphate nanoparticles.
abstract: This study aimed to fabricate a growth factor-releasing biodegradable scaffold for tissue regeneration. We prepared multishell calcium phosphate (CaP) nanoparticles functionalized with DNA, polyethyleneimine (PEI), protamine and octa-arginine (R8) and compared their respective transfection activity and cell viability measures using human mesenchymal stem cells. DNA-protamine complexes improved the transfection efficiency of CaP nanoparticles with the exception of those functionalized with R8. These complexes also greatly reduced the cytotoxicity of PEI. In addition, we also fabricated DNA-protamine-functionalized CaP nanoparticle-loaded nano-hydroxyapatite-collagen scaffolds and investigated their gene transfection efficiencies. These experiments showed that the scaffolds were associated with moderate hMSC cell viability and were capable of releasing the BMP-2 protein into hMSCs following gene transfection. In particular, the scaffold loaded with protamine-containing CaP nanoparticles showed the highest cell viability and transfection efficiency in hMSCs; thus, it might be suitable to serve as an efficient growth factor-releasing scaffold.
authors.affiliation: Liaison Center for Innovative Dentistry, Tohoku University Graduate School of Dentistry, 4-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.
authors.forename: Taichi
authors.identifier: http://orcid.org/0000-0003-2237-1040
authors.initials: T
authors.lastname: Tenkumo
authors.affiliation: Division of Advanced Prosthetic Dentistry, Tohoku University Graduate school of Dentistry, 4-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.
authors.forename: Juan Ramón
authors.initials: JR
authors.lastname: Vanegas Sáenz
authors.affiliation: Division of Dental Biomaterials, Tohoku University Graduate School of Dentistry, 4-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.
authors.forename: Yukyo
authors.initials: Y
authors.lastname: Takada
authors.affiliation: Division of Dental Biomaterials, Tohoku University Graduate School of Dentistry, 4-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Institute of Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitätsstraße 5-7, Essen, D-45117, Germany.
authors.forename: Olga
authors.initials: O
authors.lastname: Rotan
authors.affiliation: Institute of Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitätsstraße 5-7, Essen, D-45117, Germany.
authors.forename: Viktoriya
authors.initials: V
authors.lastname: Sokolova
authors.affiliation: Institute of Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitätsstraße 5-7, Essen, D-45117, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Epple
authors.affiliation: Liaison Center for Innovative Dentistry, Tohoku University Graduate School of Dentistry, 4-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.|Division of Advanced Prosthetic Dentistry, Tohoku University Graduate school of Dentistry, 4-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Sasaki
mesh_terms: D001854:Bone Marrow Cells / Q000187:drug effects; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics; D002130:Calcium Phosphates / Q000737:chemistry; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen / Q000737:chemistry / Q000627:therapeutic use*; D017886:Durapatite / Q000737:chemistry / Q000627:therapeutic use*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry* / Q000627:therapeutic use; D010957:Plasmids / Q000235:genetics; D012038:Regeneration; D054457:Tissue Scaffolds; D014162:Transfection
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D002130:Calcium Phosphates; C509317:nano-hydroxyapatite-collagen; D049452:Green Fluorescent Proteins; D003094:Collagen; D017886:Durapatite; C020243:calcium phosphate