Li Jingtang, Liao Guanxiang, Long Zhisheng, Qiu Peng, Ding Linghua, Xiong Long
Department of Orthopedics, Jiangxi Provincial People's Hospital Affiliated to Nanchang University, China.
Adv Clin Exp Med. 2020 Apr;29(4):431-440. doi: 10.17219/acem/116752.
Autogenous or allogenic bone transplantation is the main treatment for bone defects and nonunions. However, the shortcomings of autogenous or allogenic bone transplantation limit its wide application in clinical use.
This study investigated the effect of poly(lactic-co-glycolic acid) (PLGA) microspheres loaded with pOsterix (pOsx)/polyethylenimine (PEI) nanoparticles in repairing bone defects and explored its mechanism.
Poly(lactic-co-glycolic acid) microspheres loaded with pOsx/PEI nanoparticles were constructed. The Osx transfection effect was detected by fluorescence quantitative PCR and western blotting methods. 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-diphenytetrazoliumromide (MTT) and flow cytometry methods were used to detect cell proliferation. The collagen I (Col-1), osteopontin (OPN) and osteocalcin (OC) expression levels were detected using real-time polymerase chain reaction (RT-PCR) and western blotting methods. Bone defect model was constructed. Bone repair was detected using X-ray, hematoxylin and eosin (H&E) staining, and Mason staining methods.
PLGA@pOsx/PEI has transfection effect both in vitro and in vivo, does not affect cell proliferation and is safe for cells. PLGA@pOsx/PEI could promote the expression of Col-1, OPN and OC in vitro and in vivo. PLGA@pOsx/PEI could promote osteogenesis in vivo.
PLGA@pOsx/PEI with high Osx expression could promote the expression of OC, OPN, and COL-I. PLGA@pOsx/PEI can be used as a material for repairing bone defects and can promote bone formation. These results provide a theoretical and practical basis for its further clinical application.
自体或异体骨移植是治疗骨缺损和骨不连的主要方法。然而，自体或异体骨移植的缺点限制了其在临床中的广泛应用。
本研究探讨负载osterix（Osx）/聚乙烯亚胺（PEI）纳米颗粒的聚乳酸-羟基乙酸共聚物（PLGA）微球修复骨缺损的效果并探究其机制。
构建负载Osx/PEI纳米颗粒的PLGA微球。采用荧光定量PCR和蛋白质免疫印迹法检测Osx转染效果。采用3-(4,5)-二甲基噻唑-2,5-二苯基四氮唑溴盐（MTT）法和流式细胞术检测细胞增殖。采用实时聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测I型胶原（Col-1）、骨桥蛋白（OPN）和骨钙素（OC）的表达水平。构建骨缺损模型。采用X射线、苏木精-伊红（H&E）染色和Mason染色法检测骨修复情况。
PLGA@pOsx/PEI在体内外均具有转染效果，不影响细胞增殖且对细胞安全。PLGA@pOsx/PEI在体内外均可促进Col-1、OPN和OC的表达。PLGA@pOsx/PEI在体内可促进成骨。
高表达Osx的PLGA@pOsx/PEI可促进OC、OPN和COL-I的表达。PLGA@pOsx/PEI可作为修复骨缺损的材料并促进骨形成。这些结果为其进一步临床应用提供了理论和实践依据。
