RESUMEN
Angiogenesis is crucial for tissue engineering, wound healing, and regenerative medicine. Nanomaterials constructed based on specific goals can be employed to activate endogenous growth factor-related signaling. In this study, based on the conventional single-stranded DNA self-assembly into tetrahedral framework nucleic acids (tFNAs), the Apt02 nucleic acid aptamer and dimethyloxallyl glycine (DMOG) small molecule are integrated into a complex via a template-based click chemistry reaction and toehold-mediated strand displacement reaction. Thus, being able to simulate the VEGF (vascular endothelial growth factor) function and stabilize HIF (hypoxia-inducible factor), a functional whole is constructed and applied to angiogenesis. Cellular studies demonstrate that the tFNAs-Apt02 complex (TAC) has a conspicuous affinity to human umbilical vein endothelial cells (HUVECs). Further incubation with DMOG yields the tFNAs-Apt02-DMOG complex (TACD), which promotes VEGF secretion, in vitro blood vessel formation, sprouting, and migration of HUVECs. Additionally, TACD enhances angiogenesis by upregulating the VEGF/VEGFR and HIF signaling pathways. Moreover, in a diabetic mouse skin defect repair process, TACD increases blood vessel formation and collagen deposition, therefore accelerating wound healing. The novel strategy simulating VEGF and stabilizing HIF promotes blood-vessel formation in vivo and in vitro and has the potential for broad applications in the vascularization field.
Asunto(s)
Células Endoteliales de la Vena Umbilical Humana , Neovascularización Fisiológica , Transducción de Señal , Factor A de Crecimiento Endotelial Vascular , Animales , Ratones , Humanos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Neovascularización Fisiológica/fisiología , Modelos Animales de Enfermedad , Ácidos Nucleicos/metabolismo , Cicatrización de Heridas/fisiología , Aptámeros de Nucleótidos/metabolismo , Aptámeros de Nucleótidos/farmacología , AngiogénesisRESUMEN
Certain miRNAs, notably miR29c, demonstrate a remarkable capacity to regulate cellular osteogenic differentiation. However, their application in tissue regeneration is hampered by their inherent instability and susceptibility to degradation. In this study, we developed a novel miR29c delivery system utilising tetrahedral framework nucleic acids (tFNAs), aiming to enhance its stability and endocytosis capability, augment the efficacy of miR29c, foster osteogenesis in bone marrow mesenchymal stem cells (BMSCs), and significantly improve the repair of critical-sized bone defects (CSBDs). We confirmed the successful synthesis and biocompatibility of sticky ends-modified tFNAs (stFNAs) and miR29c-modified stFNAs (stFNAs-miR29c) through polyacrylamide gel electrophoresis, microscopy scanning, a cell counting kit-8 assay and so on. The mechanism and osteogenesis effects of stFNAs-miR29c were explored using immunofluorescence staining, western blotting, and reserve transcription quantitative real-time polymerase chain reaction. Additionally, the impact of stFNAs-miR29c on CSBD repair was assessed via micro-CT and histological staining. The nano-carrier, stFNAs-miR29c was successfully synthesised and exhibited exemplary biocompatibility. This nano-nucleic acid material significantly upregulated osteogenic differentiation-related markers in BMSCs. After 2 months, stFNAs-miR29c demonstrated significant bone regeneration and reconstruction in CSBDs. Mechanistically, stFNAs-miR29c enhanced osteogenesis of BMSCs by upregulating the Wnt signalling pathway, contributing to improved bone tissue regeneration. The development of this novel nucleic acid nano-carrier, stFNAs-miR29c, presents a potential new avenue for guided bone regeneration and bone tissue engineering research.
Asunto(s)
Regeneración Ósea , Diferenciación Celular , Células Madre Mesenquimatosas , MicroARNs , Osteogénesis , Cráneo , MicroARNs/genética , MicroARNs/metabolismo , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/citología , Regeneración Ósea/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Animales , Cráneo/patología , Ácidos Nucleicos , Células Cultivadas , Masculino , Ratas Sprague-Dawley , Ratones , Humanos , RatasRESUMEN
Iodine oxidizes [Ru(II)(NH(3))(5)isn](2+) in mildly acidic (HClO(4)) aqueous solution at 25 degrees C according to the reaction I(2) + 2[Ru(II)(NH(3))(5)isn](2+) --> 2I(-) + 2[Ru(III)(NH(3))(5)isn](3+). The rate law is -d[Ru(II)]/dt = {2k(1)[I(2)] + 2k(2)[I(3)(-)]}[Ru(II)] with k(1) = 4.3 x 10(3) M(-)(1) s(-)(1) and k(2) = 80 M(-)(1) s(-)(1) at &mgr; = 0.10 M (NaClO(4)). An outer-sphere electron-transfer mechanism is proposed for both terms of the rate law, with the k(1) term corresponding to the formation of I(2)(-) and k(2) corresponding to the formation of I(2)(-) plus I(-). Subsequent reduction of I(2)(-) by Ru(II) to form I(-) is expected to be fast. A value of 2.7 for log(k(22)) (the I(2)/I(2)(-) self-exchange rate constant) is derived from the Marcus cross relationship.