RESUMO
This study investigated the efficacy of using chitosan/alginate nanoparticles loaded with recombinant human bone morphogenetic-2 (rhBMP-2) and SMAD4 encoding plasmid to enhance the chondrogenesis of human bone marrow mesenchymal stem cells (hBM-MSCs) seeded on an extracellular matrix (ECM). The research treatments included the stem cells treated with the biological cocktail (BC), negative control (NC), hBM-MSCs with chondrogenic medium (MCM), hBM-MSCs with naked rhBMP-2 and chondrogenic medium (NB/C), and hBM-MSCs with naked rhBMP-2 and chondrogenic medium plus SMAD4 encoding plasmid transfected with polyethyleneimine (PEI) (NB/C/S/P). The cartilage differentiation was performed with real-time quantitative PCR analysis and alizarin blue staining. The data indicated that the biological cocktail (BC) exhibited significantly higher expression of cartilage-related genes compared to significant differences with MCM and negative control (NC) on chondrogenesis. In the (NB/C/S/P), the expression levels of SOX9 and COLX were lower than those in the BC group. The expression pattern of the ACAN gene was similar to COL2A1 changes suggesting that it holds promising potential for cartilage regeneration.
Assuntos
Alginatos , Proteína Morfogenética Óssea 2 , Cartilagem Articular , Quitosana , Condrogênese , Matriz Extracelular , Células-Tronco Mesenquimais , Nanopartículas , Regeneração , Transdução de Sinais , Proteína Smad4 , Alicerces Teciduais , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Quitosana/química , Quitosana/farmacologia , Alginatos/química , Alginatos/farmacologia , Humanos , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Cartilagem Articular/citologia , Proteína Morfogenética Óssea 2/metabolismo , Proteína Morfogenética Óssea 2/genética , Nanopartículas/química , Condrogênese/efeitos dos fármacos , Alicerces Teciduais/química , Proteína Smad4/metabolismo , Proteína Smad4/genética , Transdução de Sinais/efeitos dos fármacos , Matriz Extracelular/metabolismo , Regeneração/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Fatores de Transcrição SOX9/metabolismo , Fatores de Transcrição SOX9/genética , Proteínas Recombinantes/farmacologia , Fator de Crescimento Transformador betaRESUMO
BACKGROUND: Natural products comprise a large section of pharmaceutical agents in the field of cancer therapy. In the present study, the organic extracts and fractions of various parts of Ornithogalum bungei were investigated for in vitro cytotoxic properties on three human cancer cell lines, hepatocellular carcinoma (HepG2), prostate cancer (PC3), and leukemia (K562) cells. METHODS: The present experimental study was conducted at Tehran University of Medical Sciences (Tehran, Iran) during 2017-2019. Separately extracted plant materials, including bulbs, stems, and flowers of O. bungei were assessed by the tetrazolium dye-based colorimetric assay (MTT). The selected extracts were submitted to fractionation using vacuum liquid chromatography and after MTT assay, the half maximal inhibitory concentration (IC50 (value for each fraction was determined. The data were analyzed using One-way ANOVA followed by Tukey's post hoc test. P<0.05 was considered statistically significant. RESULTS: The cytotoxicity of the bulb's methanol extract and the dichloromethane extract of aerial parts increased in a concentration-dependent manner. Additionally, cell viability decreased in a dose-dependent manner. In the HepG2 cell line, the best IC50 values of fractions from DCM extracts of aerial parts were determined to be 19.8±10.2 µg/mL after 24 hours of exposure and 19.39±6.4 µg/mL following 48 hours of exposure. In the PC3 cell line, after 48 hours of exposure, the IC50 values of fractions were unaccountable, while the percentage of inhibition for A6 to A11 in 24 hours of exposure was more than 40 µg/mL. CONCLUSION: O. bungei growing in Iran showed significant potentials as a cytotoxic agent with selective effects on different cancer cell lines.