title: Using poly(lactic-co-glycolic acid) microspheres to encapsulate plasmid of bone morphogenetic protein 2/polyethylenimine nanoparticles to promote bone formation in vitro and in vivo.
abstract: Repair of large bone defects is a major challenge, requiring sustained stimulation to continually promote bone formation locally. Bone morphogenetic protein 2 (BMP-2) plays an important role in bone development. In an attempt to overcome this difficulty of bone repair, we created a delivery system to slowly release human BMP-2 cDNA plasmid locally, efficiently transfecting local target cells and secreting functional human BMP-2 protein. For transfection, we used polyethylenimine (PEI) to create pBMP-2/PEI nanoparticles, and to ensure slow release we used poly(lactic-co-glycolic acid) (PLGA) to create microsphere encapsulated pBMP-2/PEI nanoparticles, PLGA@pBMP-2/PEI. We demonstrated that pBMP-2/PEI nanoparticles could slowly release from the PLGA@pBMP-2/PEI microspheres for a long period of time. The 3-15 μm diameter of the PLGA@pBMP-2/PEI further supported this slow release ability of the PLGA@pBMP-2/PEI. In vitro transfection assays demonstrated that pBMP-2/PEI released from PLGA@pBMP-2/PEI could efficiently transfect MC3T3-E1 cells, causing MC3T3-E1 cells to secrete human BMP-2 protein, increase calcium deposition and gene expressions of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), SP7 and I type collagen (COLL I), and finally induce MC3T3-E1 cell differentiation. Importantly, in vivo data from micro-computed tomography (micro-CT) and histological staining demonstrated that the human BMP-2 released from PLGA@pBMP-2/PEI had a long-term effect locally and efficiently promoted bone formation in the bone defect area compared to control animals. All our data suggest that our PLGA-nanoparticle delivery system efficiently and functionally delivers the human BMP-2 cDNA and has potential clinical application in the future after further modification.
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.forename: Kai
authors.initials: K
authors.lastname: Zhang
authors.forename: Han
authors.initials: H
authors.lastname: Jin
authors.forename: Leiying
authors.initials: L
authors.lastname: Miao
authors.forename: Ce
authors.initials: C
authors.lastname: Shi
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.forename: Anliang
authors.initials: A
authors.lastname: Yuan
authors.forename: Jinzhong
authors.initials: J
authors.lastname: Liu
authors.forename: Daowei
authors.initials: D
authors.lastname: Li
authors.forename: Changyu
authors.initials: C
authors.lastname: Zheng
authors.forename: Guirong
authors.initials: G
authors.lastname: Zhang
authors.forename: Xiangwei
authors.initials: X
authors.lastname: Li
authors.forename: Bai
authors.initials: B
authors.lastname: Yang
authors.forename: Hongchen
authors.initials: H
authors.lastname: Sun
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D016503:Drug Delivery Systems; D015870:Gene Expression / Q000187:drug effects; D015316:Genetic Therapy; D007150:Immunohistochemistry; D019344:Lactic Acid / Q000737:chemistry* / Q000494:pharmacology; D008297:Male; D051379:Mice; D008863:Microspheres; D053758:Nanoparticles / Q000737:chemistry*; D010012:Osteogenesis / Q000187:drug effects*; D010316:Particle Size; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry* / Q000494:pharmacology; D011100:Polyglycolic Acid / Q000737:chemistry* / Q000494:pharmacology; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D051381:Rats; D017208:Rats, Wistar; D012886:Skull / Q000187:drug effects / Q000293:injuries / Q000473:pathology; D014162:Transfection
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D011094:Polyethyleneimine
keywords: biodegradable polymer; bone regeneration; gene therapy; human BMP-2