Abstract
Purpose: Statin is an effective factor for promoting osteogenesis. The aim of the present study was to evaluate the effect of simvastatin (SIM) and/or HA addition on changes in osteogenesis levels by human DPSCs transferred onto three-dimensional (3D) nanofibrous Poly (ε-caprolactone) (PCL)/Poly lactic acide (PLLA) polymeric scaffolds. Methods: For this purpose, a 3D nanofibrous composite scaffold of PCL/PLLA/HA was prepared by electrospinning method. SIM was added to scaffolds during DPSCs culturing step. Cell proliferation and osteogenic activity levels were assessed by using MTT assay and Alizarin Red assay methods. In addition, the expression of genes responsible for osteogenesis, including BMP2, Osteocalcin, DSPP and RUNX2, were determined before and 2 weeks after incorporation of SIM. Results: The MTT assay showed that PCL/PLLA/HA scaffolds seeded with DPSCs has significant (p<0.05) more proliferative effect than PCL/PLLA or DMEM cultured cells, additionally SIM administration improved this result over the PCL/PLLA/HA scaffolds without SIM treatment. SEM imaging revealed improved adhesion and probably osteogenic differentiation of DPSCs on PCL/PLLA/HA nanofibers treated with SIM, moreover the alizarin red assay ensured significant (p<0.05) higher mineralization of this group. Finally, real time PCR confirmed the positive regulation (P<0.05) of the expression of osteo/odontogenesis markers BMP2, Osteocalcin, DSPP and RUNX2 genes in PLLA-PCL-HA (0.1)-SIM group. Conclusion: As a result, addition of simvastatin with incorporation of hydroxyapatite in PCL-PLLA scaffolds might increase the expression of osteogenesis markers in the DPSCs, with a possible increase in cell differentiation and bone formation.
Highlights
In recent decades the medical sciences have been revolutionized due to the introduction of treatment modalities with the application of stem cells and the synthesis of various tissues and organs
The MTT assay showed that PCL/Poly lactic acide (PLLA)/HA scaffolds seeded with DPSCs has significant (p
Fabrication of PCL/PLLA/HA nanofiber Nanofibers were prepared from PCL and PLLA that both of them are well known biocompatible and biodegradable polymers enable the successful cultivation of human DPSCs (hDPSCs) and osteogenic cells
Summary
In recent decades the medical sciences have been revolutionized due to the introduction of treatment modalities with the application of stem cells and the synthesis of various tissues and organs. Stem cells are progenitor cells that can be differentiated into cells with specific functions under some physiologic or laboratory conditions These cells have indefinite potential for proliferation, can remain undifferentiated and preserve their capacity to differentiate into every cell type in the body.[1] Stem cells have been isolated from the human primary tooth pulps and periodontal ligaments in various studies to date.[2,3] Human dental pulp is one of the most available sources for stem cells. The effects of different materials have been evaluated on the proliferation and differentiation of these cells, and the role of different materials in the induction or inhibition of these cells has been studied
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