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OBJECTIVE: We aimed to develop an electroactive antibacterial material for the treatment of skin wound diseases. METHOD: To this aim, we modified chitosan (CS), a biocompatible polymer, by coupling it with graphene (rGO) and an antimicrobial polypeptide DOPA-PonG1. The material's effect on skin injury healing was studied in combination with external electrical stimulation (EEM). The structure, surface composition, and hydrophilicity of the modified CS materials were evaluated using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and contact angle measurements. We studied NIH3T3 cells cultured with modified materials and subjected to EEM to assess viability, adhesion, and tissue repair-related gene expression. RESULTS: SEM data demonstrated that rGO was distributed uniformly on the surface of the CS material, increasing surface roughness, and antimicrobial peptides had minimal impact on surface morphology. FTIR confirmed the uniform distribution of rGO and antibacterial peptides on the material surface. Both rGO and DOPA-PonG1 enhanced the hydrophilicity of CS materials, with rGO also improving tensile strength. The dual modification of CS with rGO and DOPA-PonG1 synergistically increased antibacterial efficacy. Cellular events and gene expression relevant to tissue repair process were enhanced by these modifications. Furthermore, EEM accelerated epidermal regeneration more than the material alone. In a rat skin wound model, DOPA-PonG1@CS/rGO dressing combined with electrical stimulation exhibited accelerated healing of skin defect. CONCLUSION: Overall, our results demonstrate that CS materials modified with rGO and DOPA-PonG1 have increased hydrophilicity, antibacterial characteristics, and tissue regeneration capacities. This modified material in conjunction with EEM hold promise for the clinical management for dermal wounds.
Assuntos
Antibacterianos , Quitosana , Camundongos , Ratos , Animais , Células NIH 3T3 , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Quitosana/farmacologia , Quitosana/química , Estimulação Elétrica , Bandagens , Di-HidroxifenilalaninaRESUMO
Regeneration and functional recovery of nerves after peripheral nerve injury is the key to peripheral nerve repair. One of the putative therapeutic strategies is to use anti-adhesion polymer films, made of polymeric biomaterials. Recently, a novel biodegradable poly (DL-lactic acid) (PDLLA) film has been prepared using a method of phase transformation with biodegradable polylactic acid polymer as the substrate. This novel, anti-adhesion film has a porous structure, which provides better mechanical properties, better flexibility, more complete diffusion through the polymer of tissue biologic factors like growth factors, and more controllable degradation compared to traditional non-porous films. Little is known, however, about the in vitro and in vivo biocompatibility and cytotoxicity of this type of PDLLA film. Therefore, our aim was to evaluate the biocompatibility and cytotoxicity of this novel PDLLA film using various experimental methods, including a skin irritation test, MTT analysis, and the mouse bone marrow cell micronucleus test, as well as hematology or clinical chemistry measurements in rats after receiving sciatic nerve transection and anastomosis with wrapping of the anastomosis with DLLA films. We demonstrated that exposure to PDLLA film extracts did not generate apparent erythema or edema in rabbit skin and had no effect on the proliferation of Vero cells. Additionally, treatment with PDLLA film extracts did not alter the incidence of micronucleated polychromatic erythrocytes as compared with saline Treated group. Furthermore, implantation of PDLLA film did not alter liver or renal function as measured by serum levels of ALT, AST, TP, A/G, Cr, and BUN, and pathologic examinations showed that implantation of PDLLA film did not cause pathologic changes to the rat liver, kidney, pancreas, or spleen. Taken together, these results suggest that PDLLA films have excellent biocompatibility and no obvious toxicity in vivo, and may be used to prevent nerve adhesion, thereby promoting nerve regeneration.
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Omentin is a novel adipokine, which is expressed in and released from omental adipose tissue. In the present study, the effect of omentin on neural stem cells (NSCs) was investigated. NSCs are a subtype of stem cell in the nervous system, which are able to selfrenew and generate neurons and glia for repairing neural lesions. Mouse NSCs were isolated and cultured in vitro. Treatment with recombinant omentin for 3 and 5 days significantly increased the size of NSC neurospheres (P<0.01) and enhanced NSC cell viability in normal conditions. In addition, omentin protected against the decrease in cell viability induced by the proinflammatory cytokine tumor necrosis factorα. In the NSCs, incubation of omentin for 2, 4, 6, 8 and 16 h enhanced the phosphorylation of Akt at the Thr308 site and of AS160 at the Ser318 site, peaking 6 h after treatment. Additionally, treatment with LY294002 (10 µM), a specific inhibitor of phosphatidylinositol 3kinase/Akt signaling, eliminated the omentininduced increase in neurosphere size and cell viability. Overall, the present study provided the first evidence, to the best of our knowledge, that omentin promotes the growth and survival of NSCs in vitro through activation of the Akt signaling pathway. These results may contribute to the understanding of the role of omentin in the nervous system.
Assuntos
Citocinas/farmacologia , Lectinas/farmacologia , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Recombinantes/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Proteínas Ligadas por GPI/farmacologia , Humanos , Camundongos , Estresse FisiológicoRESUMO
Endothelial progenitor cells (EPCs), a group of bone marrow-derived pro-angiogenic cells, contribute to vascular repair after damage. EPC dysfunction exists in diabetes and results in poor wound healing in diabetic patients with trauma or surgery. The aim of the present study was to determine the effect of quercetin, a natural flavonoid on high glucoseinduced damage in EPCs. Treatment with high glucose (40 mM) decreased cell viability and migration, and increased oxidant stress, as was evidenced by the elevated levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase in bone marrow-derived EPCs. Moreover, high glucose reduced the levels of endothelial nitric oxide synthase (eNOS) phosphorylation, nitric oxide (NO) production and intracellular cyclic guanosine monophosphate (cGMP). Quercetin supplement protected against high glucoseinduced impairment in cell viability, migration, oxidant stress, eNOS phosphorylation, NO production and cGMP levels. Quercetin also increased Sirt1 expression in EPCs. Inhibition of Sirt1 by a chemical antagonist sirtinol abolished the protective effect of quercetin on eNOS phosphorylation, NO production and cGMP levels following high glucose stress. To the best of our knowledge, the results provide the first evidence that quercetin protects against high glucoseinduced damage by inducing Sirt1-dependent eNOS upregulation in EPCs, and suggest that quercetin is a promising therapeutic agent for diabetic patients undergoing surgery or other invasive procedures.