title: Incorporation of osteogenic and angiogenic small interfering RNAs into chitosan sponge for bone tissue engineering.
abstract: Engineered bone substitutes are being extensively explored in response to growing demand. However, the angiogenesis that occurs during bone formation is often overlooked in scaffold design. In this novel study, we incorporated two small interfering RNAs (siRNAs), ie, small interfering RNA targets casein kinase 2 interaction protein 1 (siCkip-1) and small interfering RNA targets soluble VEGF receptor 1 (siFlt-1), which can promote osteogenesis and angiogenesis, into a chitosan sponge. This scaffold could maintain siRNAs for over 2 weeks in neutral phosphate-buffered saline and degraded rapidly in the presence of lysozyme. The chitosan sponge with siCkip-1 and siFlt-1 in vitro bioactivity was investigated using mesenchymal stem cells. Target genes were significantly suppressed, and osteocalcin, alkaline phosphatase, and vascular endothelial growth factor were significantly upregulated. Alizarin Red staining revealed that mineralization of the extracellular matrix was markedly enhanced by dual transfection. Further analysis by immunofluorescence confirmed that the siRNA-modified scaffold simultaneously improved the expression of osteocalcin and von Willebrand factor. In vivo testing in a skull critical-size defect model showed marked bone regeneration in rats treated with siCkip-1 and siFlt-1. In conclusion, chitosan sponge containing osteogenic and angiogenic siRNAs may be used as a scaffold for bone regeneration. The dual siRNA concept may also be useful in the biofunctionalization of other materials.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Sen
authors.initials: S
authors.lastname: Jia
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Xinjie
authors.initials: X
authors.lastname: Yang
authors.affiliation: Department of Prosthetic Dentistry, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Wen
authors.initials: W
authors.lastname: Song
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Implant Dentistry, School of Stomatology, State Key Laboratory of Military Stomatology, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Kaixiu
authors.initials: K
authors.lastname: Fang
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China ; Department of Otorhinolaryngology, No 113 Hospital of People's Liberation Army, Ningbo, People's Republic of China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Zihui
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Chun
authors.initials: C
authors.lastname: Shan
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Delin
authors.initials: D
authors.lastname: Lei
authors.affiliation: Department of Oral and Maxillofacial Surgery, Fourth Military Medical University, Xi'an People's Republic of China.
authors.forename: Bin
authors.initials: B
authors.lastname: Lu
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells; D001861:Bone Regeneration / Q000187:drug effects*; D002448:Cell Adhesion; D049109:Cell Proliferation; D048271:Chitosan / Q000737:chemistry*; D008297:Male; D059630:Mesenchymal Stem Cells; D010012:Osteogenesis / Q000187:drug effects* / Q000235:genetics*; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012886:Skull / Q000187:drug effects / Q000293:injuries; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D034741:RNA, Small Interfering; D048271:Chitosan
keywords: angiogenesis; chitosan sponge; osteogenesis; small interfering RNA