Incorporating pTGF-β1/calcium phosphate nanoparticles with fibronectin into 3-dimensional collagen/chitosan scaffolds: efficient, sustained gene delivery to stem cells for chondrogenic differentiation.
The objective of this study was to prepare a 3-dimensional nanoparticle gene delivery system (3D-NGDS) based on collagen/chitosan scaffolds, in which plasmid transforming growth factor beta 1 (TGF-β1)/calcium phosphate nanoparticles mixed with fibronectin (FN) were used to transfect mesenchymal stem cells (MSCs). Scanning electron microscopy was used to characterise the microstructure of 3-dimensional collagen/chitosan scaffolds. An analysis performed to quantify the TGF-b1 concentrations in MSC cultures revealed that the MSCs transfected with the 3D-NGDS showed remarkably high levels of TGF-b1 over long periods, retaining a concentration of TGF-b1 of approximately 10 ng/mL within two weeks, with the highest level (12.6 ng/mL) being observed on the 6th day. An immunohistochemistry analysis for collagen type II revealed that much higher production of collagen II from the 9th to 15th day was observed in the 3D-NGDS-transfected MSCs than that in MSCs transfected by the Lipofectamine 2000 method. The glycosaminoglycan content of the 3D-NGDS was comparable to those treated with TGF-β1 as well as TGF-β1 plus dexamethasone, and was significantly higher than those treated with free plasmid and Lipofectamine 2000. A remarkable type I collagen expression inhibition of the 3D-NGDS at day 21 was observed via ELISA. These results suggested that transfection with the 3D-NGDS could successfully induce MSC chondrogenic differentiation in vitro without dexamethasone. In summary, the 3D-NGDS could be developed into a promising alternative method to transfer exogenous nucleic acid to MSCs in clinical trials.
Department of Pharmaceutics, School of Pharmacy, Jiangsu University, 301 Xuefu Road, Jingkou District, Zhenjiang 212001, RP China.
Xia
X
Cao
Wenwen
W
Deng
Yuan
Y
Wei
Yan
Y
Yang
Weiyan
W
Su
Yawei
Y
Wei
Ximing
X
Xu
Jiangnan
J
Yu
D000818:Animals; D002130:Calcium Phosphates / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism; D020219:Chondrogenesis / Q000235:genetics*; D003094:Collagen / Q000737:chemistry / Q000378:metabolism; D024042:Collagen Type I / Q000096:biosynthesis / Q000187:drug effects; D024043:Collagen Type II / Q000096:biosynthesis / Q000187:drug effects; D045424:Complex Mixtures / Q000008:administration & dosage*; D005353:Fibronectins / Q000378:metabolism; D018014:Gene Transfer Techniques / Q000295:instrumentation*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D054457:Tissue Scaffolds; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*
D002130:Calcium Phosphates; D024042:Collagen Type I; D024043:Collagen Type II; D045424:Complex Mixtures; D005353:Fibronectins; D053773:Transforming Growth Factor beta1; D003094:Collagen; D048271:Chitosan; C020243:calcium phosphate