title: Characterization and cytocompatibility of thermosensitive hydrogel embedded with chitosan nanoparticles for delivery of bone morphogenetic protein-2 plasmid DNA.
abstract: A novel injectable chitosan thermosensitive hydrogel was designed as a target multi-effect scaffold for endogenous repair of the periodontium. The hydrogel complex was designed by embedding chitosan nanoparticles (CSn) loaded with bone morphogenetic protein-2 plasmid DNA (pDNA-BMP2) into a chitosan (CS)-based hydrogel with α,β-glycerophosphate (α,β-GP), termed CS/CSn(pDNA-BMP2)-GP. Characterization, the in vitro release profile for pDNA-BMP2, and cytocompatibility to human periodontal ligament cells (HPDLCs), were then conducted. The average diameter of the CSn(pDNA-BMP2) was 270.1 nm with a polydispersity index (PDI) of 0.486 and zeta potential of +27.0 mv. A DNase I protection assay showed that CSn could protect the pDNA-BMP2 from nuclease degradation. Encapsulation efficiency and loading capacity of CSn(pDNA-BMP2) were more than 80 and 30 %, respectively. The sol-gel transition time was only 3 min when CSn(pDNA-BMP2) was added into the CS/α,β-GP system. Scanning electron microscopy showed that CSn(pDNA-BMP2) was randomly dispersed in a network with regular holes and a porous structure. Weighting method showed the swelling ratio and degradation was faster in medium of pH 4.0 than pH 6.8. An in vitro pDNA-BMP2 release test showed that the cumulative release rate of pDNA-BMP2 was much slower from CS/CSn-GP than from CSn in identical release media. In release media with different pH, pDNA-BMP2 release was much slower at pH 6.8 than at pH 4.0. Three-dimensional culture with HPDLCs showed good cell proliferation and the Cell-Counting Kit-8 assay indicated improved cell growth with the addition of CSn(pDNA-BMP2) to CS/α,β-GP. In summary, the CS/CSn(pDNA-BMP2)-GP complex system exhibited excellent biological properties and cytocompatibility, indicating great potential as a gene delivery carrier and tissue regeneration scaffold for endogenous repair of the periodontium.
authors.affiliation: Stomatological Hospital of Zhengzhou, Zhengzhou, 450000, Henan Province, China.|The Affiliated Hospital of Qingdao University, Qingdao, 266001, Shandong Province, China.|School of Stomatology, Qingdao University, Qingdao, 266001, Shandong Province, China.
authors.forename: Dan-Dan
authors.initials: DD
authors.lastname: Li
authors.affiliation: The Arrail Dental Hospital of Beijing, Beijing, 100107, Chaoyang Province, China.
authors.forename: Jian-Feng
authors.initials: JF
authors.lastname: Pan
authors.affiliation: The Affiliated Hospital of Qingdao University, Qingdao, 266001, Shandong Province, China. jiqx@qduhospital.cn.|School of Stomatology, Qingdao University, Qingdao, 266001, Shandong Province, China. jiqx@qduhospital.cn.
authors.forename: Qiu-Xia
authors.initials: QX
authors.lastname: Ji
authors.affiliation: The Affiliated Hospital of Qingdao University, Qingdao, 266001, Shandong Province, China.|School of Stomatology, Qingdao University, Qingdao, 266001, Shandong Province, China.
authors.forename: Xin-Bo
authors.initials: XB
authors.lastname: Yu
authors.affiliation: The Affiliated Hospital of Qingdao University, Qingdao, 266001, Shandong Province, China.|School of Stomatology, Qingdao University, Qingdao, 266001, Shandong Province, China.
authors.forename: Ling-Shuang
authors.initials: LS
authors.lastname: Liu
authors.affiliation: The Affiliated Hospital of Qingdao University, Qingdao, 266001, Shandong Province, China.|School of Stomatology, Qingdao University, Qingdao, 266001, Shandong Province, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: The Affiliated Hospital of Qingdao University, Qingdao, 266001, Shandong Province, China.|School of Stomatology, Qingdao University, Qingdao, 266001, Shandong Province, China.
authors.forename: Xiao-Ju
authors.initials: XJ
authors.lastname: Jiao
authors.affiliation: The Affiliated Hospital of Qingdao University, Qingdao, 266001, Shandong Province, China.|School of Stomatology, Qingdao University, Qingdao, 266001, Shandong Province, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
mesh_terms: D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D048271:Chitosan / Q000737:chemistry*; D003470:Culture Media; D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D005994:Glycerophosphates / Q000737:chemistry; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry; D010513:Periodontal Ligament / Q000166:cytology / Q000502:physiology*; D010957:Plasmids / Q000737:chemistry*; D012038:Regeneration; D054457:Tissue Scaffolds
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D003470:Culture Media; D005994:Glycerophosphates; D020100:Hydrogels; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D004247:DNA; D048271:Chitosan; C029620:alpha-glycerophosphoric acid; C031463:beta-glycerophosphoric acid