RESUMO
Lipid-based vesicular nanoparticles, for instance liposomes, conjugated with polyethylene glycol (PEG) have proven to be the closest to an ideal drug delivery vehicle, making way for several PEG-liposomes based nanomedicines in market. However, the synthetic nature of the nanomaterial poses a threat to stimulate immune system. Alternatively, nanovesicles derived from mammalian cells, such as RBCs, have gained interests as they may not elicit much immune response due to the presence of host specific self-recognition markers on their surface. While several reports demonstrating the superior efficacy of these naturally derived vesicles have come out in the last few years, a comparison with clinically established liposomes is still missing. Thus, we conducted an in-vitro and in-vivo comparative studies between PEG-Liposomes and nanovesicles (NVEs) derived from red blood cell (RBC) membrane with an aim to establish a biocompatible nanocarrier for efficient delivery of chemotherapeutic drugs and photothermal agents.
Assuntos
Lipossomos , Polietilenoglicóis , Animais , Sistemas de Liberação de Medicamentos , Eritrócitos , Lipídeos , MamíferosRESUMO
Bone erosion is a debilitating pathological process of osteopathic disorder like rheumatoid arthritis (RA). Current treatment strategies render low disease activity but with disease recurrence. To find an alternative, we designed this study with an aim to explore the underlying therapeutic effect of PEGylated liposomal BBR (PEG-BBR) against Wnt1/ß-catenin mediated bone erosion in adjuvant-induced arthritic (AA) rat model and fibroblast-like synoviocytes (FLS) with reference to microRNA-23a (miR-23a) activity. Our initial studies using confocal microscopy and Near-Infrared Imaging (NIR) showed successful internalization of PEG-BBR and PEG-miR-23a in vitro and in vivo respectively and was retained till 48 h. The preferential internalization of PEG-BBR into the inflamed joint region significantly reduced the gene and protein level expression of major Wnt1 signaling mediators and reduced bone erosion in rats. Moreover, PEG-BBR treatment in FLS cells attenuated the gene and protein expression levels of FZD4, LRP5, ß-catenin, and Dvl-1 through the induction of CYLD. Furthermore, inhibition of these factors resulted in reduced bone loss and increased calcium retainability by altering the RANKL/OPG axis. PEG-BBR treatment markedly inhibited the expression of LRP5 protein on par with the DKK-1 (LRP5/Wnt signaling inhibitor) and suppressed the transcriptional activation of ß-catenin inside the cells. We further witnessed that miR-23a altered the expression levels of LRP5 through RNA interference. Overall, our findings endorsed that miR-23a possesses a multifaceted therapeutic efficiency like berberine in RA pathogenesis and can be considered as a potential candidate for therapeutic targeting of Wnt1/ß-catenin signaling in RA disease condition.
Assuntos
Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Berberina/farmacologia , MicroRNAs/genética , Animais , Berberina/administração & dosagem , Feminino , Lipossomos , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Masculino , Polietilenoglicóis/química , Ratos , Ratos Wistar , Via de Sinalização Wnt/efeitos dos fármacos , Proteína Wnt1/metabolismo , beta Catenina/metabolismoRESUMO
The rate at which Plasmodium falciparum is developing resistance to clinically used antimalarial drugs is alarming. Therefore, there is a compelling need to develop an efficient drug delivery system to improve the efficacy of existing antimalarial agents and circumvent drug resistance. Here, we report the antibacterial drug doxycycline (DOXY) in liposomal formulations exhibits enhanced antiplasmodial activity against blood stage forms of P. falciparum (3D7) in culture and established Plasmodium berghei NK-65 infection in murine model. Parasite killing on blood stage forms in culture was determined by a radiolabeled [3H] hypoxanthine incorporation assay and infected erythrocytes stained with Giemsa were counted using microscopy in vivo. The 50% inhibitory concentration (IC50) of DOXY-stearylamine liposome (IC50 0.36 µM) and DOXY-SPC:Chol-liposome (IC50 0.85 µM) exhibited marked growth inhibition of parasites compared with free DOXY (IC50 14 µM), with minimal toxicity to normal erythrocytes. Administration of polyethylene glycol distearoyl phosphatidylethanolamine-methoxy-polyethylene glycol2000 (DSPE-mPEG-2000) coated liposomes loaded with DOXY at 2.5 mg/kg per day resulted in efficacious killing of blood parasites with improved survival in mice relative to the free drug in both chloroquine sensitive and resistant strains of P. berghei infection. This is the first report to demonstrate that DOXY in liposomal system has immense chemotherapeutic potential against plasmodial infections at lower dosages.
Assuntos
Antimaláricos/administração & dosagem , Doxiciclina/administração & dosagem , Portadores de Fármacos/química , Malária/tratamento farmacológico , Plasmodium berghei/patogenicidade , Plasmodium falciparum/efeitos dos fármacos , Animais , Antimaláricos/uso terapêutico , Cloroquina/administração & dosagem , Cloroquina/uso terapêutico , Modelos Animais de Doenças , Doxiciclina/uso terapêutico , Resistência a Medicamentos , Eritrócitos/efeitos dos fármacos , Eritrócitos/parasitologia , Feminino , Humanos , Concentração Inibidora 50 , Lipossomos , Malária/parasitologia , Malária/patologia , Camundongos , Fosfatidiletanolaminas/química , Plasmodium berghei/efeitos dos fármacos , Polietilenoglicóis/química , Resultado do TratamentoRESUMO
Rheumatoid arthritis is a chronic autoimmune disorder resulting in synovial inflammation. Fibroblast activation protein (FAP) is overexpressed by fibroblastlike synoviocytes in arthritic joints. Radioimmunoimaging with an anti-FAP antibody might be used to monitor the response to therapy, thus enabling tailored therapy strategies and therapeutic outcomes. The aim of this study was to assess whether a radiolabeled anti-FAP antibody could be used to monitor the efficacy of treatment with long-circulating liposomes (LCL) containing prednisolone phosphate (PLP-LCL) in a mouse model of arthritis. METHODS: Collagen-induced arthritis (CIA) was induced in male DBA/1J mice. Mice were treated with a single injection (10 mg/kg) of PLP-LCL or empty LCL as a control. SPECT and CT images were acquired 24 h after injection of 99mTc-labeled succinimidyl-hydrazinonicotinamide (99mTc-S-HYNIC)-conjugated anti-FAP antibody 28H1 at 2, 5, and 9 d after treatment. The uptake of 99mTc-S-HYNIC-28H1 in all joints was quantified and correlated with macroscopic arthritis scores. RESULTS: Treatment of CIA with PLP-LCL significantly suppressed joint swelling. At just 1 d after treatment, the macroscopic arthritis scores had decreased by 50%. Scores decreased further, to only 10% of the initial scores, at 5 and 9 d after treatment. In contrast, macroscopic arthritis scores had increased up to 600% in untreated mice at 9 d after the injection of empty LCL. 99mTc-S-HYNIC-28H1 uptake ranged from 1.5 percentage injected dose per gram in noninflamed joints to 22.6 percentage injected dose per gram in severely inflamed joints. The uptake of radiolabeled 28H1 in inflamed joints (percentage injected dose) correlated with the arthritis score (Spearman ρ, 0.77; P < 0.0001). Moreover, the uptake of 99mTc-S-HYNIC-28H1 was slightly increased at 9 d after therapy but was not seen macroscopically, indicating that SPECT/CT imaging might be more sensitive than the macroscopic arthritis scoring method. CONCLUSION: SPECT/CT imaging with 99mTc-S-HYNIC-28H1 specifically monitored the response to therapy, and tracer accumulation correlated with the severity of inflammation. In addition, SPECT/CT imaging was potentially more sensitive than the macroscopic arthritis scoring method. This study showed that SPECT/CT with 99mTc-S-HYNIC-28H1 could be used to noninvasively monitor the course of CIA in mice.
Assuntos
Anticorpos Monoclonais/imunologia , Artrite Reumatoide/diagnóstico por imagem , Artrite Reumatoide/tratamento farmacológico , Monitoramento de Medicamentos/métodos , Gelatinases/imunologia , Proteínas de Membrana/imunologia , Serina Endopeptidases/imunologia , Tomografia Computadorizada com Tomografia Computadorizada de Emissão de Fóton Único/métodos , Animais , Artrite Reumatoide/imunologia , Preparações de Ação Retardada/administração & dosagem , Endopeptidases , Glucocorticoides/administração & dosagem , Marcação por Isótopo/métodos , Lipossomos/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos DBA , Imagem Molecular/métodos , Prednisolona/administração & dosagem , Prednisolona/análogos & derivados , Compostos Radiofarmacêuticos/imunologia , Tecnécio/imunologia , Resultado do TratamentoRESUMO
Surface PEGylation on nanoparticles has greatly helped prolong their blood circulation half-lives. However, The injection of PEGylated nanoparticles into mice induced poly(ethylene glycol) (PEG)-specific IgM antibodies (anti-PEG IgMs), significantly changing PEG-liposomes' pharmacokinetics. In this study, we used various PEG-conjugates to conduct a mechanistic study of anti-PEG IgMs' binding behavior. The conventional belief has been that anti-PEG IgMs bind to PEG main chains; however, our findings reveal that anti-PEG IgMs did not bind to PEG main chains, whereas anti-PEG IgMs did bind to PEG-hydrophobic polymer blocks. The insertion of a hydrophilic polymer between each PEG chain and each hydrophobic polymer block suppressed anti-PEG IgMs' binding. We prove here that hydrophobic blocks are essential to anti-PEG IgMs' binding, and also that anti-PEG IgMs do not bind to intact PEGs without hydrophobic moiety. These results support our conclusion that anti-PEG IgMs exhibit specificity to PEG; however, the presence of a hydrophobic block at a proximity position from each PEG chain is essential for the binding. Also in the present study, we elucidate relations between anti-PEG IgMs and PEGylated nanoparticles. In one of our previous studies, anti-PEG IgMs scarcely affected the pharmacokinetics of PEG-b-poly(ß-benzyl l-aspartate) block copolymer (PEG-PBLA) micelles, whereas anti-PEG IgMs significantly decreased PEG-liposomes' blood circulation half-life. Finally, we found that the ratio of anti-PEG IgM molecules to PEG-liposome particles is critical to these pharmacokinetic changes, and that a 10-fold increase in the number of anti-PEG IgM molecules permitted them to capture the PEG-liposome particles, thus leading to the aforementioned changes.
Assuntos
Imunoglobulina M/sangue , Nanopartículas/química , Peptídeos/farmacocinética , Polietilenoglicóis/farmacocinética , Animais , Meia-Vida , Interações Hidrofóbicas e Hidrofílicas , Lipossomos , Masculino , Taxa de Depuração Metabólica , Camundongos Endogâmicos C57BL , Micelas , Peptídeos/química , Polietilenoglicóis/químicaRESUMO
Long-circulating liposomes (LCL) are often used as a drug carrier system to improve the therapeutic index of water-soluble drugs. To track these LCL in vivo, they can be radiolabelled with (111)In-oxine. For this labelling method, generally DTPA is encapsulated in the aqueous phase of LCL (DTPA-LCL). Alternatively, LCL can be labelled with (111)InCl3 after incorporation of DTPA-conjugated DSPE in the lipid bilayer (DTPA-DSPE LCL). Here, we compared the in vitro properties of DTPA-DSPE LCL with those of DTPA LCL and empty LCL. Additionally, we compared the in vivo performance of DTPA-DSPE LCL with those of DTPA LCL in mice. DTPA LCL (88 nm) and empty LCL (84 nm) were labelled with (111)In-oxine, and DTPA-DSPE LCL (83 nm) were labelled with (111)InCl3. Labelling efficiency at increasing specific activity was determined. In vitro stability of (111)In-labelled LCL was determined in human serum at 37 °C. The in vivo properties of (111)In-labelled LCL were determined in mice with a Staphylococcus aureus infection in the thigh muscle. Image acquisition, blood sampling and biodistribution studies were performed 1, 4 (blood sampling only), 24, 48 and 72 h p.i. of (111)In-labelled LCL. DTPA-DSPE LCL could be labelled efficiently at a much higher specific activity compared to DTPA LCL and empty LCL: > 90% at 15 GBq/mmol, > 90% at 150 MBq/mmol and 6065% at 150 MBq/mmol, respectively. (111)In-labelled DTPA-DSPE LCL and DTPA LCL were stable in human serum, regarding label retention, for at least 48 h at 37 °C (> 98% retention of the radiolabel). In contrast, only 68% radiolabel was retained in empty LCL after 48 h. In vivo targeting of (111)In-DTPA-DSPE LCL to the abscess was comparable to targeting of (111)In-DTPA LCL (3.5 ± 0.9%ID/g and 3.4 ± 0.9%ID/g abscess uptake respectively, 48 h p.i.). In conclusion, labelling of DTPA-DSPE LCL with (111)InCl3 represents a robust, easy and fast procedure which is preferred over the more laborious conventional labelling of DTPA-LCL with (111)In-oxine.