Abstract:The objective of this study was to investigate the use of cationized Pleurotus eryngii polysaccharide (CPEPS) as a nonviral gene delivery vehicle to transfer plasmid DNA encoding transforming growth factor beta-1 (pTGF-β1) into mesenchymal stem cells (MSCs) in vitro. Crude P. eryngii polysaccharide was purified, and then cationized by grafting spermine onto the backbone of the polysaccharide. Agarose gel electrophoresis, transmission electron microscopy, and a Nano Sense Zetasizer (Malvern Instruments, Malvern, UK) were used to characterize the CPEPS-pTGF-β1 nanoparticles. The findings of cytotoxicity analysis showed that when the nanoparticles were formulated with a CPEPS/pTGF-β1 weight ratio $ 10:1, a greater gel retardation effect was observed during agarose gel electrophoresis. The CPEPS-pTGF-β1 nanoparticles with a weight ratio of 20:1, respectively, possessed an average particle size of 80.8 nm in diameter and a zeta potential of +17.4 ± 0.1 mV. Significantly, these CPEPS-pTGF-β1 nanoparticles showed lower cytotoxicity and higher transfection efficiency than both polyethylenimine (25 kDa) (P = 0.006, Student's t-test) and Lipofectamine TM 2000(P = 0.002, Student's t-test). Additionally, the messenger RNA expression level of TGF-β1 in MSCs transfected with CPEPS-pTGF-β1 nanoparticles was significantly higher than that of free plasmid DNA-transfected MSCs and slightly elevated compared with that of Lipofectamine 2000-transfected MSCs. Flow cytometry analysis demonstrated that 92.38% of MSCs were arrested in the G1 phase after being transfected with CPEPS-pTGF-β1 nanoparticles, indicating a tendency toward differentiation. In summary, the findings of this study suggest that the CPEPS-pTGF-β1 nanoparticles prepared in this work exhibited excellent transfection efficiency and low toxicity. Therefore, they could be developed into a promising nonviral vector for gene delivery in vitro.
Introduction: An en-masse retraction with mini implant (MI) anchorage may be associated with unwanted intrusion/extrusion and uncontrolled tipping of anterior teeth. An optimum combination of MIs and hooks heights is required for proper treatment results. Materials and Methods: Maxillary finite element models were constructed from a cone beam CT scan of a patient’s orofacial region. The initial tooth displacement at 200g force with 0.019 × 0.025-in stainless steel working archwires engaged in 0.022 brackets slot was assessed. The three-dimensional displacement was examined at various MI and AAH heights. Results: The lower MI position caused extrusion of the central incisors, but the teeth were intruded at higher (6- and 8-mm) MI heights. While the shorter (2- and 4-mm) hooks extruded the central incisors, the higher (6- and 8-mm) intruded the teeth. The higher MI and hooks reduced the palatal tipping of central incisors. The distobucal cusp of the first molar was intruded, while the mesiobucal cusp was extruded in all models: Nonetheless, the shorter hooks and low MI had small molar tipping effects. Conclusions: The higher MIs caused intrusion and less palatal tipping of the central incisors crowns. The increase in hook height resulted into extrusion and reduction in palatal tipping of the central incisors crowns.
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