RESUMEN
Melanoma has gained considerable attention due to its high mortality and morbidity rate worldwide. The currently available treatment options are associated with several limitations such as nonspecificity, drug resistance, easy clearance, low efficacy, toxicity-related issues, etc. To this end, nanotechnology has garnered significant attention for the treatment of melanoma. In the present manuscript, we have demonstrated the in vitro and in vivo anticancer activity of silver nitroprusside nanoparticles (abbreviated as AgNNPs) against melanoma. The AgNNPs exhibit cytotoxicity against B16F10 cells, which has been investigated by several in vitro experiments including [methyl 3H]-thymidine incorporation assay, cell cycle and apoptosis analysis by flow cytometry, and ROS generation through DCFDA, DHE, and DAF2A reagents. Further, the internalization of nanoparticles was determined by ICPOES analysis, while their colocalization was analyzed by confocal microscopy. Additionally, JC-1 staining is performed to examine mitochondrial membrane potential (MMP). Cytoskeleton integrity was observed by phalloidin staining. Expression of different markers (Ki-67, cytochrome c, and E-cadherin) was checked using an immunofluorescence assay. The in vivo therapeutic efficacy of AgNNPs has been validated in the melanoma model established by inoculating B16F10 cells into the dorsal right abdomen of C57BL/6J mice. The intraperitoneal administration of AgNNPs reduced melanoma growth and increased the survivability of tumor-bearing mice. The in vivo immunofluorescence studies (Ki-67, CD31, and E-cadherin) and TUNEL assay support the inhibitory and apoptotic nature of AgNNPs toward melanoma, respectively. Furthermore, the various signaling pathways and molecular mechanisms involved in anticancer activity are evaluated by Western blot analysis. These findings altogether demonstrate the promising anticancer potential of AgNNPs toward melanoma.
Asunto(s)
Antineoplásicos , Apoptosis , Proliferación Celular , Ensayos de Selección de Medicamentos Antitumorales , Ratones Endogámicos C57BL , Nitroprusiato , Plata , Animales , Ratones , Antineoplásicos/farmacología , Antineoplásicos/química , Nitroprusiato/farmacología , Nitroprusiato/química , Apoptosis/efectos de los fármacos , Plata/química , Plata/farmacología , Proliferación Celular/efectos de los fármacos , Tamaño de la Partícula , Ensayo de Materiales , Melanoma/tratamiento farmacológico , Melanoma/patología , Nanopartículas del Metal/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Nanopartículas/química , Supervivencia Celular/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Línea Celular Tumoral , Especies Reactivas de Oxígeno/metabolismo , Melanoma Experimental/tratamiento farmacológico , Melanoma Experimental/patología , Melanoma Experimental/metabolismoRESUMEN
Melanoma is the most invasive and lethal form of skin cancer that arises from the malignant transformation of specialized pigment-producing cell melanocytes. Nanomedicine represents an important prospect to mitigate the difficulties and provide significant benefits to cure melanoma. In the present study, we investigated in vitro and in vivo therapeutic efficacies of copper nitroprusside analogue nanoparticles (abbreviated as CuNPANP) towards melanoma. Initially, in vitro anti-cancer activities of CuNPANP towards melanoma cells (B16F10) were evaluated by several experiments such as [methyl-3H]-thymidine incorporation assay, cell cycle and apoptosis assays using FACS analysis, ROS generation using DCFDA, DHE and DAF2A reagents, internalization of nanoparticles through ICP-OES analysis, co-localization of the nanoparticles using confocal microscopy, JC-1 staining to investigate the mitochondrial membrane potential (MMP) and immunofluorescence studies to analyze the expressions of cytochrome-c, Ki-67, E-cadherin as well as phalloidin staining to analyze the cytoskeletal integrity. Further, the in vivo therapeutic effectiveness of the nanoparticles was established towards malignant melanoma by inoculating B16F10 cells in the dorsal right abdomen of C57BL/6J mice. The intraperitoneal administration of CuNPANP inhibited tumor growth and increased the survivability of melanoma mice. The in vivo immunofluorescence studies (Ki-67, CD-31, and E-cadherin) and TUNEL assay further support the anti-cancer and apoptosis-inducing potential of CuNPANP, respectively. Finally, various signaling pathways and molecular mechanisms involved in anti-cancer activities were further evaluated by Western blot analysis. The results altogether indicated the potential use of copper-based nanomedicines for the treatment of malignant melanoma.
Asunto(s)
Apoptosis , Cobre , Melanoma Experimental , Ratones Endogámicos C57BL , Nitroprusiato , Animales , Ratones , Línea Celular Tumoral , Melanoma Experimental/tratamiento farmacológico , Melanoma Experimental/patología , Melanoma Experimental/metabolismo , Apoptosis/efectos de los fármacos , Cobre/química , Cobre/farmacología , Nitroprusiato/farmacología , Nitroprusiato/química , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Melanoma/tratamiento farmacológico , Melanoma/patología , Melanoma/metabolismo , Antineoplásicos/química , Antineoplásicos/farmacología , Nanopartículas/química , Neoplasias Cutáneas/tratamiento farmacológico , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/metabolismo , Proliferación Celular/efectos de los fármacos , Nanopartículas del Metal/química , Nanopartículas del Metal/uso terapéuticoRESUMEN
Critical limb ischemia (CLI) refers to a severe condition resulting from gradual obstruction in the supply of blood, oxygen, and nutrients to the limbs. The most promising clinical solution to CLI is therapeutic angiogenesis. This study explored the potency of pro-angiogenic terbium hydroxide nanorods (THNR) for treatment of CLI, with a major focus on their impact on ischemia-induced maladaptive alterations in endothelial cells as well as on vascularization in ischemic limbs. This study demonstrated that, in hypoxia-exposed endothelial cells, THNR improve survival and promote proliferation, migration, restoration of nitric oxide production, and regulation of vascular permeability. Based on molecular studies, these attributes of THNR can be traced to the stimulation of PI3K/AKT/eNOS signaling pathways. Besides, Wnt/GSK-3ß/ß-catenin signaling pathways may also play a role in the therapeutic actions of THNR. Furthermore, in the murine model of CLI, THNR administration can integrally re-establish blood perfusion with concomitant reduction of muscle damage and inflammation. Additionally, improvement of locomotor activities and motor coordination in ischemic limbs in THNR treated mice is also evident. Overall, the study demonstrates that THNR have the potential to be developed as an efficacious and cost-effective alternative clinical therapy for CLI, using a nanomedicine approach.
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Isquemia , Nanotubos , Animales , Nanotubos/química , Ratones , Isquemia/tratamiento farmacológico , Isquemia/patología , Isquemia/metabolismo , Humanos , Hidróxidos/química , Hidróxidos/farmacología , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Masculino , Ensayo de Materiales , Proliferación Celular/efectos de los fármacos , Tamaño de la Partícula , Ratones Endogámicos C57BL , Células Endoteliales de la Vena Umbilical Humana , Miembro Posterior/patologíaRESUMEN
Triple negative breast cancer (TNBC) is an aggressive form of tumor, more prevalent in younger women resulting in poor survival rate (2nd in cancer deaths) because of its asymptomatic existence. The most popular and convenient approach for the treatment of TNBC is chemotherapy which is associated with several limitations. Considering the importance of nanotechnology in health care system, in the present manuscript, we have designed and developed a simple, efficient, cost effective, and ecofriendly method for the synthesis of copper nitroprusside analogue nanoparticles (Cu[Fe(CN)5NO] which is abbreviated as CuNPANP that may be the potential anti-cancer nanomedicine for the treatment of TNBC. Copper (present in CuNPANP) is used because of its affordability, nutritional value and various biomedical applications. The CuNPANP are thoroughly characterized using several analytical techniques. The in vitro cell viability (in normal cells) and the ex vivo hemolysis assay reveal the biocompatible nature of CuNPANP. The anti-cancer activity of the CuNPANP is established in TNBC cells (MDA-MB-231 and 4T1) through several in vitro assays along with plausible mechanisms. The intraperitoneal administration of CuNPANP in orthotopic breast tumor model by transplanting 4T1 cells into the mammary fat pad of BALB/c mouse significantly inhibits the growth of breast carcinoma as well as increases the survival time of tumor-bearing mice. These results altogether potentiate the anti-cancer efficacy of CuNPANP as a smart therapeutic nanomedicine for treating TNBC in near future after bio-safety evaluation in large animals.
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Cobre , Especies Reactivas de Oxígeno , Neoplasias de la Mama Triple Negativas , Animales , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Neoplasias de la Mama Triple Negativas/patología , Femenino , Ratones , Cobre/química , Cobre/farmacología , Cobre/administración & dosificación , Humanos , Línea Celular Tumoral , Especies Reactivas de Oxígeno/metabolismo , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Antineoplásicos/química , Antineoplásicos/administración & dosificación , Ratones Endogámicos BALB C , Nanopartículas del Metal/uso terapéutico , Nanopartículas del Metal/química , Supervivencia Celular/efectos de los fármacos , Modelos Animales de EnfermedadRESUMEN
Sodium nitroprusside (SNP), U.S approved drug has been used in clinical emergency as a hypertensive drug for more than a decade. It is well established for its various biomedical applications such as angiogenesis, wound healing, neurological disorders including anti-microbial applications etc. Apart from that, SNP have been considered as excellent biomedical materials for its use as anti-cancer agent because of its behavior as NO-donor. Recent reports suggest that incorporation of metals in SNP/encapsulation of SNP in metal nanoparticles (metal nitroprusside analogues) shows better therapeutic anti-cancer activity. Although there are numerous reports available regarding the biological applications of SNP and metal-based SNP analogue nanoparticles, unfortunately there is not a single comprehensive review which highlights the anti-cancer activity of SNP and its derivative metal analogues in detail along with the future perspective. To this end, the present review article focuses the recent development of anti-cancer activity of SNP and metal-based SNP analogues, their plausible mechanism of action, current status. Furthermore, the future perspectives and challenges of these biomedical materials are also discussed. Overall, this review article represents a new perspective in the area of cancer nanomedicine that will attract a wider spectrum of scientific community.
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Fármacos Cardiovasculares , Neoplasias , Nitroprusiato/metabolismo , Nitroprusiato/farmacología , Nitroprusiato/uso terapéutico , Metales , Neoplasias/tratamiento farmacológicoRESUMEN
Recently, we have demonstrated casein manganese oxide nanoparticles (CMnNP) that exhibit pro-angiogenic property established through different in vitro and in vivo experiments. The CMnNP was explored for therapeutic angiogenesis for treatment of wounds and recovery of hindlimb ischemia in pre-clinical mouse prototypical. It is well known that to translate any therapeutic nanoparticle for future clinical applications, their biosafety evaluation in small and large animals is essential. Herein, in the current study, the biosafety and bioavailability of the CMnNP have been explored by a systematic toxicity profiling study in C57BL/6J mice model. Initially, the in vitro cytotoxic effects of CMnNP were validated in RAW 264.7 cells. Later, the CMnNP was administered intraperitoneally with different doses (50, 300, and 2000 mg/kg b.wt./day) at different time points of exposure (acute: 2 weeks, sub-chronic: 4 weeks as well as chronic exposure: 8 and 20 weeks) with reference to the maximum tolerable dose (MTD) of CMnNP as per the OECD guidelines. The blood hematological and serum biochemical parameters of CMnNP treatment groups indicate negligible changes similar to untreated group. The histopathological examination of CMnNP-treated vital organs (lung, spleen, liver, brain, kidney, and heart) illustrates no major changes even at higher doses. Further, the biodistribution and excretion study depicts normal clearance of CMnNP. Additionally, the serum cytokine levels were normal in the therapeutic dose of CMnNP. The results altogether indicate that the non-toxic nature of CMnNP makes them useful as future therapeutic angiogenic agent for the treatment of various diseases where angiogenesis plays an important role.
Asunto(s)
Caseínas , Compuestos de Manganeso , Nanopartículas , Óxidos , Ratones , Animales , Caseínas/toxicidad , Distribución Tisular , Ratones Endogámicos C57BL , Nanopartículas/toxicidadRESUMEN
Our earlier reports established that zinc oxide nanoflowers (ZONF) show significant pro-angiogenic properties, where reactive oxygen species, nitric oxide and MAPK-AKT-eNOS cell signaling axis play an essential task. Considering the significance of angiogenesis in healthcare, our research group has recently demonstrated the in vivo therapeutic application of ZONF (10 mg/kg b.w.) for treating peripheral artery disease. Moreover, based on the angio-neural crosstalk between vascular and neuronal systems, we have further demonstrated the neuritogenic and neuroprotective characteristics of pro-angiogenic nanoflowers (10 mg/kg b.w.) for the treatment of cerebral ischemia. However, it is crucial for a therapeutic material to be non-toxic for its practical clinical applications and therefore assessment of its in vivo toxicity and adverse effect is highly important. Herein, for the first time, we investigate a detailed nanotoxicology of therapeutically active ZONF in Swiss albino mice to evaluate their safety profile and comprehend their aspects for future clinical applications. The maximum tolerated dose (MTD) of ZONF was found to be 512.5 mg/kg b.w. which was employed for acute exposure (2 weeks), showing slight toxicity. However, sub-chronic (4 weeks) and long term chronic (8-12 weeks) studies of nanoflowers exhibited their non-toxic nature particularly at lower therapeutic doses (1-10 mg/kg b.w.). Additionally, in depth genotoxicity study revealed that lower therapeutic dose of ZONF (10 mg/kg b.w.) did not exhibit significant toxicity even in genetic level. Overall, the present nanotoxicology of ZONF suggests their high biocompatible nature at therapeutic dose, offering the basis of their future clinical applications in ischemic and other vascular diseases.
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Óxido de Zinc , Ratones , Animales , Óxido de Zinc/toxicidad , Especies Reactivas de OxígenoRESUMEN
Myocardial hypoxia reperfusion (H/R) injury is the paradoxical exacerbation of myocardial damage, caused by the sudden restoration of blood flow to hypoxia affected myocardium. It is a critical contributor of acute myocardial infarction, which can lead to cardiac failure. Despite the current pharmacological advancements, clinical translation of cardioprotective therapies have proven challenging. As a result, researchers are looking for alternative approaches to counter the disease. In this regard, nanotechnology, with its versatile applications in biology and medicine, can confer broad prospects for treatment of myocardial H/R injury. Herein, we attempted to explore whether a well-established pro-angiogenic nanoparticle, terbium hydroxide nanorods (THNR) can ameliorate myocardial H/R injury. For this study, in vitro H/R-injury model was established in rat cardiomyocytes (H9c2 cells). Our investigations demonstrated that THNR enhance cardiomyocyte survival against H/R-induced cell death. This pro-survival effect of THNR is associated with reduction of oxidative stress, lipid peroxidation, calcium overload, restoration of cytoskeletal integrity and mitochondrial membrane potential as well as augmentation of cellular anti-oxidant enzymes such as glutathione-s-transferase (GST) and superoxide dismutase (SOD) to counter H/R injury. Molecular analysis revealed that the above observations are traceable to the predominant activation of PI3K-AKT-mTOR and ERK-MEK signalling pathways by THNR. Concurrently, THNR also exhibit apoptosis inhibitory effects mainly by suppression of pro-apoptotic proteins like Cytochrome C, Caspase 3, Bax and p53 with simultaneous restoration of anti-apoptotic protein, Bcl-2 and Survivin. Thus, considering the above attributes, we firmly believe that THNR have the potential to be developed as an alternative approach for amelioration of H/R injury in cardiomyocytes.
Asunto(s)
Daño por Reperfusión Miocárdica , Nanotubos , Animales , Ratas , Miocitos Cardíacos/metabolismo , Terbio/metabolismo , Terbio/farmacología , Terbio/uso terapéutico , Fosfatidilinositol 3-Quinasas/metabolismo , Línea Celular , Hipoxia/metabolismo , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/metabolismoRESUMEN
The advancement of nanotechnology has led to the experimental development of cancer therapeutics, which may overcome the shortcomings of commercially available drugs and facilitate improved clinical outcomes. Recently, several metal nanoparticles, especially silver, have been evaluated by scientists globally as useful chemotherapeutic agents due to their multi-functionality and well-recognized biological activity. Herein, we developed silver nitroprusside nanoparticles (abbreviated as AgNNPs) with slight modifications in the reaction conditions and demonstrated their application for breast cancer therapy using in vitro assays and in vivo experiments in a mouse model. Initially, the modified AgNNPs were thoroughly characterized using several analytical techniques. AgNNPs were found to be biocompatible according to in vitro experiments in normal cell lines (HEK-293 and EA.hy926), which was further validated by a hemolysis assay (ex vivo experiment) using mouse red blood cells. In contrast, the cell viability assay using the MTT reagent showed the cytotoxic nature of the AgNNPs against several cancer cell lines (MDA-MB-231, 4T1, B16F10, and PANC-1). Their detailed anticancer activity was investigated using 4T1 (mouse specific) and MDA-MB-231 (human specific) cells through various in vitro assays. The nanoparticles inhibited the formation of blood vessels in the chick embryo model, highlighting their anti-angiogenic behavior. Furthermore, the administration of AgNNPs significantly inhibited orthotopic breast tumor growth (4T1; BALB/c mice) and increased the survivability of the tumor-bearing mice. Also, we demonstrated the plausible molecular mechanisms for the anti-cancer activity of AgNNPs through various in vitro assays and in vivo experiments. Overall, the results support that AgNNPs can be used as an alternative generalized nanomedicine for the treatment of breast and other cancers after proper biosafety evaluation in near future.
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Antineoplásicos , Neoplasias de la Mama , Nanopartículas del Metal , Embrión de Pollo , Humanos , Animales , Ratones , Femenino , Neoplasias de la Mama/patología , Nitroprusiato/farmacología , Nitroprusiato/uso terapéutico , Plata/farmacología , Línea Celular Tumoral , Células HEK293 , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Nanopartículas del Metal/uso terapéutico , Apoptosis , Ratones Endogámicos BALB CRESUMEN
Tweetable abstract Nanotechnology and its role in hair growth by enhancing drug delivery for the management of alopecia #nanotechnology #hair #drug_delivery.
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Nanomedicina , Nanopartículas , Humanos , Nanotecnología , Cabello , Alopecia/tratamiento farmacológico , Sistemas de Liberación de MedicamentosRESUMEN
Nanotechnology has immensely advanced the field of cancer diagnostics and treatment by introducing potential delivery vehicles as carriers for drugs or therapeutic agents. In due course, mesoporous silica nanoparticles (MSNs) have emerged as excellent vehicles for delivering drugs, biomolecules, and biomaterials, attributed to their solid framework and porosity providing a higher surface area for decorating with various functional ligands. Recently, the metal tin (Sn) has gained huge importance in cancer research owing to its excellent cytotoxicity and ability to kill cancer cells. In the present work, we synthesized MSNs, conjugated them with organotin compounds, and characterized them using various physicochemical techniques. Subsequently, the biological evaluation of MSN (S1), MSN-MP (S2) and tin-conjugated MSNs (S3: MSN-MP-SnPh3) (MP = 3-mercaptopropyltriethoxysilane) revealed that these nanoconjugates induced cytotoxicity, necrosis, and apoptosis in MCF-7 cells. Moreover, these nanoconjugates exhibited anti-angiogenic properties as demonstrated in the chick embryo model. The increase of reactive oxygen species (ROS) was found as a one of the plausible mechanisms underlying cancer cell cytotoxicity induced by these nanoconjugates, encouraging their application for the treatment of cancer. The tin-conjugated MSNs demonstrated less toxicity to normal cells compared to cancer cells. Furthermore, the genotoxicity studies revealed the clastogenic and aneugenic effects of these nanoconjugates in CHO cells mostly at high concentrations. These interesting observations are behind the idea of developing tin-conjugated MSNs as prospective candidates for anticancer therapy.
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Antineoplásicos , Dióxido de Silicio , Estaño , Animales , Embrión de Pollo , Cricetinae , Humanos , Antineoplásicos/farmacología , Supervivencia Celular , Cricetulus , Portadores de Fármacos/química , Nanoconjugados , Dióxido de Silicio/química , Estaño/farmacologíaRESUMEN
In the current communication, a simple, environmentally compatible, non-toxic green chemistry process is used for the development of silver nanoparticles (AgZE) by the reaction between silver nitrate (AgNO3) and the ethanolic leaf extract of Zinnia elegans (ZE). The optimization of AgZE is carried out using a series of experiments. Various physico-chemical techniques are utilized to characterize the nanomaterials. The cell viability assay of AgZE in normal cells (CHO, HEK-293T, EA.hy926, and H9c2) shows their biocompatible nature, which is supported by hemolytic assay using mouse RBC. Interestingly, the nanoparticles exhibited cytotoxicity towards different cancer cell lines (U-87, MCF-7, HeLa, PANC-1 and B16F10). The detailed anticancer activity of AgZE on human glioblastoma cell line (U-87) is exhibited through various in vitro assays. In vivo the AgZE illustrates anticancer activity by inhibiting blood vessel formation through CAM assay. Furthermore, the AgZE nanoparticles when intraperitoneally injected in C57BL6/J mice (with and without tumor) exhibit fluorescence properties in the NIR region (excitation: 710 nm, emission: 820 nm) evidenced by bioimaging studies. The AgZE biodistribution through ICPOES analysis illustrates the presence of silver in different vital organs. Considering all the results, AgZE could be useful as a potential cancer therapeutic agent, as well as an NIR based non-invasive imaging tool in near future.
RESUMEN
Nanotechnology is the most promising technology to evolve in the last decade. Recent research has shown that transition metal nanoparticles especially manganese (Mn)-based nanoparticles have great potential for various biomedical applications due to their unique fundamental properties. Therefore, globally, scientists are concentrating on the development of various new manganese-based nanoparticles (size and shape dependent) due to their indispensable utilities. Although numerous reports are available regarding the use of manganese nanoparticles, there is no comprehensive review highlighting the recent development of manganese-based nanomaterials and their potential applications in the area of biomedical sciences. The present review article provides an overall survey on the recent advancement of manganese nanomaterials in biomedical nanotechnology and other fields. Further, the future perspectives and challenges are also discussed to explore the wider application of manganese nanoparticles in the near future. Overall, this review presents a fundamental understanding and the role of manganese in various fields, which will attract a wider spectrum of the scientific community.
Asunto(s)
Nanopartículas del Metal , Nanopartículas , Nanoestructuras , Iones , Manganeso , Nanopartículas del Metal/toxicidad , NanotecnologíaRESUMEN
Critical limb ischemia (CLI) is a severe type of peripheral artery disease (PAD) which occurs due to an inadequate supply of blood to the limb extremities. Patients with CLI often suffer from extreme cramping pain, impaired wound healing, immobility, cardiovascular complications, amputation of the affected limb and even death. The conventional therapy for treating CLI includes surgical revascularization as well as restoration of angiogenesis using growth factor therapy. However, surgical revascularization is only suitable for a small percentage of CLI patients and is associated with a high perioperative mortality rate. The use of growth factors is also limited in terms of their poor therapeutic angiogenic potential, as observed in earlier clinical studies which could be attributed to their poor bio-availability and non-specificity issues. Therefore, to overcome the aforesaid disadvantages of conventional strategies there is an urgent need for the advancement of new alternative therapeutic biomaterials to treat CLI. In the past few decades, various research groups, including ours, have been involved in developing different pro-angiogenic nanomaterials. Among these, zinc oxide nanoflowers (ZONFs), established in our laboratory, are considered one of the more potent nanoparticles for inducing therapeutic angiogenesis. In our earlier studies we showed that ZONFs promote angiogenesis by inducing the formation of reactive oxygen species and nitric oxide (NO) as well as activating Akt/MAPK/eNOS cell signaling pathways in endothelial cells. Recently, we have also reported the therapeutic potential of ZONFs to treat cerebral ischemia through their neuritogenic and neuroprotective properties, exploiting angio-neural cross-talk. Considering the excellent pro-angiogenic properties of ZONFs and the importance of revascularization for the treatment of CLI, in the present study we comprehensively explore the therapeutic potential of ZONFs in a rat hind limb ischemia model (established by ligating the hind limb femoral artery), an animal model that mimics CLI in humans. The behavioral studies, laser Doppler perfusion imaging, histopathology and immunofluorescence as well as estimation of serum NO level showed that the administration of ZONFs could ameliorate ischemia in rats at a faster rate by promoting therapeutic angiogenesis to the ischemic sites. Altogether, the present study offers an alternative nanomedicine approach employing ZONFs for the treatment of PADs.