Incorporating pTGF-β1/calcium phosphate nanoparticles with fibronectin into 3-dimensional collagen/chitosan scaffolds: Efficient, sustained gene delivery to stem cells for chondrogenic differentiation

Abstract

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-1 concentrations in MSC cultures revealed that the MSCs transfected with the 3D-NGDS showed remarkably high levels of TGF-1 over long periods, retaining a concentration of TGF-1 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 signifi cantly 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.

Tissue engineering and regenerative approaches to improving the healing of large bone defects

“…The free amine groups on the chitosan molecule allow it to be crosslinked with the same agents as used for protein matrices (Reves et al, 2013;Wang and Stegemann, 2011), which can increase its mechanical strength and resistance to degradation. In addition, the positive charge on the chitosan molecule allows the material to be used for drug and gene delivery directly from the scaffold (Cao et al, 2012b;Goncalves et al, 2012).…”

The effects of graphene nanostructures on mesenchymal stem cells

“…Nanoparticles have been employed to deliver growth factors/genes into MSCs to manipulate their differentiation [17, 18]. They have also been used as contrast agents/nanoprobes for stem cell tracking [19, 20] to locate the site of stem cell activity, and determine the efficacy of the therapy.…”

Enhanced efficiency in isolation and expansion of hAMSCs via dual enzyme digestion and micro-carrier

“…The above results indicate that the CMs had a slow degradation rate and poor adhesiveness to hAMSCs, which was not beneficial to cell proliferation leading to a weakness in biological activity. The results also suggested that CCMs and GCMs were favorable to the development of the cells and played a certain role in promoting hAM-SCs growth [50]. From the above results it is possible to state that the proliferation rate of hAMSCs grown on the CCMs and GCMs are better compared with the cells grown on Petri dishes.…”