Open AccessGene-activated matrix harboring a miR20a-expressing plasmid promotes rat cranial bone augmentation
Author(s) -
Rena Shido,
Yoshinori Sumita,
Masahito Hara,
Mayumi Iwatake,
Shun Narahara,
Mayumi Umebayashi,
Kei-ichiro Miura,
Yukinobu Kodama,
Izumi Asahina
Publication year - 2021
Publication title -
regenerative biomaterials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.166
H-Index - 25
ISSN - 2056-3426
DOI - 10.1093/rb/rbaa060
Subject(s) - mesenchymal stem cell , microbiology and biotechnology , gene delivery , transgene , transfection , downregulation and upregulation , transplantation , chemistry , microrna , in vivo , progenitor cell , matrix (chemical analysis) , genetic enhancement , gene , stem cell , biology , medicine , biochemistry , genetics , chromatography
Gene-activated matrix (GAM) has a potential usefulness in bone engineering as an alternate strategy for the lasting release of osteogenic proteins but efficient methods to generate non-viral GAM remain to be established. In this study, we investigated whether an atelocollagen-based GAM containing naked-plasmid ( p ) DNAs encoding microRNA (miR) 20a, which may promote osteogenesis in vivo via multiple pathways associated with the osteogenic differentiation of mesenchymal stem/progenitor cells (MSCs), facilitates rat cranial bone augmentation. First, we confirmed the osteoblastic differentiation functions of generated p DNA encoding miR20a ( p miR20a) in vitro , and its transfection regulated the expression of several of target genes, such as Bambi1 and PPARγ, in rat bone marrow MSCs and induced the increased expression of BMP4. Then, when GAMs fabricated by mixing 100 μl of 2% bovine atelocollagen, 20 mg β-TCP granules and 0.5 mg (3.3 μg/μl) AcGFP plasmid-vectors encoding miR20a were transplanted to rat cranial bone surface, the promoted vertical bone augmentation was clearly recognized up to 8 weeks after transplantation, as were upregulation of VEGFs and BMP4 expressions at the early stages of transplantation. Thus, GAM-based miR delivery may provide an alternative non-viral approach by improving transgene efficacy via a small sequence that can regulate the multiple pathways.