Tail Vein Administration Research Articles

A Nanocapsule System Combats Aging by Inhibiting Age-Related Angiogenesis Deficiency and Glucolipid Metabolism Disorders.

Insufficient angiogenic stimulation and dysregulated glycolipid metabolism in senescent vascular endothelial cells (VECs) constitute crucial features of vascular aging. Concomitantly, the generation of excess senescence-associated secretory phenotype (SASP) and active immune-inflammatory responses propagates within injured vessels, tissues, and organs. Until now, targeted therapies that efficiently rectify phenotypic abnormalities in senescent VECs have still been lacking. Here, we constructed a Pd/hCeO2-BMS309403@platelet membrane (PCBP) nanoheterostructured capsule system loaded with fatty acid-binding protein 4 (FABP4) inhibitors and modified with platelet membranes and investigated its therapeutic role in aged mice. PCBP showed significant maintenance in aged organs and demonstrated excellent biocompatibility. Through cyclic tail vein administration, PCBP extended the lifespan and steadily ameliorated abnormal phenotypes in aged mice, including SASP production, immune and inflammatory status, and age-related metabolic disorders. In senescent ECs, PCBP mediated the activation of vascular endothelial growth factor (VEGF) signaling and glycolysis and inhibition of FABP4 by inducing the synthesis of hypoxia-inducible factor-1α, thereby reawakening neovascularization and restoring glycolipid metabolic homeostasis. In conclusion, the PCBP nanocapsule system provides a promising avenue for interventions against aging-induced dysfunction.

Degradation of the α-Carboxyl Terminus 11 Peptide: In Vivo and Ex Vivo Impacts of Time, Temperature, Inhibitors, and Gender in Rat.

In previous research, a synthetic α-carboxyl terminus 1 (αCT1) peptide derived from connexin 43 (Cx43) and its variant (αCT11) showed beneficial effects in an ex vivo ischemia-reperfusion (I/R) heart injury model in mouse. In an in vivo mouse model of cryo-induced ventricular injury, αCT1 released from adhesive cardiac patches reduced Cx43 remodeling and arrhythmias, as well as maintained cardiac conduction. Whether intravenous injection of αCT1 or αCT11 produces similar outcomes has not been investigated. Given the possibility of peptide degradation in plasma, this study utilized in vivo I/R cardiac injury and ex vivo blood plasma models to examine factors that may limit the therapeutic potential of peptide therapeutics in vivo. Following tail vein administration of αCT11 (100 μM) in blood, no effect on I/R infarct size was observed in adult rat hearts on day 1 (D1) and day 28 (D28) after injury (p > 0.05). There was also no difference in the echocardiographic ejection fraction (EF%) between the control and the αCT11 groups (p > 0.05). Surprisingly, αCT11 in blood plasma collected from these rats was undetectable within ∼10 min after tail vein injection. To investigate factors that may modulate αCT11 degradation in blood, αCT11 was directly added to blood plasma isolated from normal rats without I/R and peptide levels were measured under different experimental conditions. Consistent with in vivo observations, significant αCT11 degradation occurred in plasma within 10 min at 22 and 37 °C and was nearly undetectable by 30 min. These responses were reduced by the addition of protease/phosphatase (PTase/PPTase) inhibitors to the isolated plasma. Interestingly, no significant differences in αCT11 degradation in plasma were noted between male and female rats. We conclude that fast degradation of αCT11 is likely the reason that no beneficial effects were observed in the in vivo I/R model and inhibition or shielding from PTase/PPTase activity may be a strategy that will assist with the viability of peptide therapeutics.

Comparative pharmacokinetic study of Anisodamine Hydrobromide tablets and injection in septic acute lung injury rats.

Aim: We aimed to establish a sensitive LC-MS/MS method to analyze the pharmacokinetics of Ani HBr tablets and injection.Methods: Around 10mmNH4Ac containing 0.1% formic acid and acetonitrile were used as the mobile phase. Acute lung injury in septic and normal rats, respectively, were administered Ani HBr tablets at doses of 12.5, 25 and 50mg/kg and injection at doses of 4, 8 and 16mg/kg, followed by extraction of the drugs from plasma using ethyl acetate for subsequent analysis.Results & conclusion: The method met the requirements for biological analysis. Ani HBr tablets absorbed slowly in rats with disease, tail vein administration was a more promising approach for treating septic acute lung injury.

Ameliorative effect of anisodamine (654-1/654-2) against myocardial dysfunction induced by septic shock via the NF-κB/NLRP-3 or the PI3K-AKT/NF-κB pathway

BackgroundSeptic shock, an extremely dangerous condition that causes impairment of organ function, always largely contributes to mortality in intensive care units. The impact of septic shock-induced organ damage on morbidity and mortality is substantially influenced by myocardial dysfunction. However, it remains unclear whether and in what manner anisodamine (654-1/654-2) ameliorates myocardial dysfunction caused by septic shock. PurposeThis study is the pioneering investigation and validation about the protective efficacy of anisodamine (654-1/654-2) against LPS-induced myocardial dysfunction in septic shock rats. It also aims to explore the differences in the underlying molecular mechanisms of both drugs. MethodsA septic shock model was established in SD rats by after tail vein administration of LPS. 64 rats were distributed into eight groups, such as LPS group, control group, LPS+654-1 group (1.25, 2.5, and 5 mg/kg), and LPS+654-2 group (1.25, 2.5, and 5 mg/kg). The hemodynamics, echocardiography, immunohistochemical analysis, TEM, TUNEL assay, and H&E staining were utilized to assess the septic shock model and myocardial function. Lactic acid, inflammatory markers (IL-1β, IL-6, and TNF-α), endothelial injure markers (SDC-1, HS and TM) and myocardial injury markers (CK, c-TNT, and NT-pro BNP) were assessed using ELISA or biochemical kits. Additionally, the mechanisms of 654-1/654-2 were analyzed using RNA-seq and bioinformatics, and validated using western blotting and RT-PCR. ResultsAdministration of 654-1/654-2 significantly restored hemodynamics and improved myocardial and endothelial glycocalyx injury in septic shock rats. Furthermore, 654-1/654-2 dose-dependently reduced plasma levels of lactic acid, inflammatory cytokines, and markers of endothelial and myocardial injury. Analyses using RNA-seq, WB and RT-PCR techniques indicated that 654-1/654-2 could mitigate myocardial and endothelial injury by inhibiting the NF-κB and NLRP-3 pathways, and activating the PI3K-AKT pathway. ConclusionsThese findings demonstrated that 654-1/654-2 could alleviate myocardial damage in septic shock rats. Specifically, 654-1 inhibited the NF-κB/NLRP-3 pathway, whereas 654-2 promoted the PI3K-AKT pathway and inhibited the NF-κB pathway, effectively mitigating the inflammatory response and cell apoptosis.

Production and synthesis of a novel 191Pt-labeled platinum complex and evaluation of its biodistribution in healthy mice

We previously reported that our sugar-conjugated platinum complex (cis-dichloro [(2-fluoro-α-d-glucopylanosidyl) propane-1,3-diamine] platinum: FGC-Pt) has low toxicity and tumor growth inhibitory effect comparable to that of cisplatin. We focused on radioactive Pt isotopes in order to analyze the kinetics of FGC-Pt using gamma-ray imaging techniques, assuming that FGC-Pt could be used for chemotherapy in the future. Therefore, in this study, we aimed to develop a non-invasive method to analyze the biodistribution of FGC-Pt using 191Pt-labeled FGC-Pt ([191Pt]FGC-Pt).191Pt was produced via the (n,2n) reaction induced by accelerator neutrons. [191Pt]FGC-Pt was prepared using two different methods. In the first method, [191Pt]FGC-Pt (method A) was obtained through the accelerator neutron irradiation of FGC-Pt. In the second method, [191Pt]FGC-Pt (method B) was synthesized using [191Pt]K2PtCl4, which was obtained by the accelerator neutron irradiation of K2PtCl4. Highly purified [191Pt]FGC-Pt was obtained using the latter method, which suggests that the synthetic method using a 191Pt-labeled platinum reagent is suitable for the radioactivation of platinum complexes.We also aimed to investigate whether a significant correlation existed between the biodistribution of FGC-Pt and [191Pt]FGC-Pt in healthy mice 24 h after tail vein administration. FGC-Pt and [191Pt]FGC-Pt were similarly distributed in healthy mice, with a higher accumulation in the liver and kidney 24 h post injection. In addition, a significant correlation (p < 0.05, r = 0.92) between the 191Pt radioactivity concentration (%ID/g (gamma counter)) and platinum concentration (%ID/g (ICP-MS)) was observed in 13 organs. These results suggest that 191Pt-labeled compounds, synthesized using radioactive platinum reagents, can be used to confirm the biodistribution of platinum compounds. Our study on the biodistribution of [191Pt]FGC-Pt is expected to contribute to the development of novel platinum-based drugs in the future.

A Golgi‐Targeted Platinum Complex Plays a Dual Role in Autophagy Regulation for Highly Efficient Cancer Therapy

AbstractRegulating autophagy to control the homeostatic recycling process of cancer cells is a promising anticancer strategy. Golgi apparatus is a substrate of autophagy but the Golgi‐autophagy (Golgiphagy) mediated antitumor pathway is rarely reported. Herein, we have developed a novel Golgi‐targeted platinum (II) complex Pt3, which is ca. 20 times more cytotoxic to lung carcinoma than cisplatin and can completely eliminate tumors after intratumoral administration in vivo. Its nano‐encapsulated system for tail vein administration also features a good anti‐tumor effect. Mechanism studies indicate that Pt3 induces substantial Golgi stress, indicated by the fragmentation of Golgi structure, down‐regulation of Golgi proteins (GM130, GRASP65/55), loss of Golgi‐dependent transport and glycosylation. This triggers Golgiphagy but blocks the subsequent fusion of autophagosomes with lysosomes, that is a dual role in autophagy regulation, resulting in loss of proteostasis and apoptotic cell death. As far as we know, Pt3 is the first Golgi‐targeted Pt complex that can trigger Golgi stress‐mediated dual‐regulation of autophagic flux and autophagy‐apoptosis crosstalk for highly efficient cancer therapy.

A Golgi-Targeted Platinum Complex Plays a Dual Role in Autophagy Regulation for Highly Efficient Cancer Therapy.

Regulating autophagy to control the homeostatic recycling process of cancer cells is a promising anticancer strategy. Golgi apparatus is a substrate of autophagy but the Golgi-autophagy (Golgiphagy) mediated antitumor pathway is rarely reported. Herein, we have developed a novel Golgi-targeted platinum (II) complex Pt3, which is ca. 20 times more cytotoxic to lung carcinoma than cisplatin and can completely eliminate tumors after intratumoral administration in vivo. Its nano-encapsulated system for tail vein administration also features a good anti-tumor effect. Mechanism studies indicate that Pt3 induces substantial Golgi stress, indicated by the fragmentation of Golgi structure, down-regulation of Golgi proteins (GM130, GRASP65/55), loss of Golgi-dependent transport and glycosylation. This triggers Golgiphagy but blocks the subsequent fusion of autophagosomes with lysosomes, that is a dual role in autophagy regulation, resulting in loss of proteostasis and apoptotic cell death. As far as we know, Pt3 is the first Golgi-targeted Pt complex that can trigger Golgi stress-mediated dual-regulation of autophagic flux and autophagy-apoptosis crosstalk for highly efficient cancer therapy.

Abstract 2713: Modulation of the blood-tumor barrier by 6-bromoindirubin acetoxime increases intra-tumoral drug accumulation and extends survival in murine glioblastoma models

Abstract Introduction: Glioblastoma (GBM) is the most frequent primary malignant adult brain tumor. Median survival is ~15 months after diagnosis. Therapies to effectively manage glioblastoma are hindered due to the presence of the blood-tumor barrier (BTB). Indirubins are 3,2’-bisindole compounds of the Indigo naturalis used in traditional Chinese medicine to treat proliferative diseases. Herein, we demonstrate that systemic delivery of 6-bromoindirubin-3’-acetoxime (BIA), an indirubin derivative, increases cisplatin delivery to GBM tumors through alteration of the BTB integrity and synergy of BIA with platinum therapeutics to potentiate anti-tumor killing. Methods: Pre-clinical studies involved G9-pCDH glioblastoma cell line implantation in naïve nude mice. Cisplatin uptake 24 hours after BIA tail-vein administration into healthy brain and GBM murine tumors was assessed via inductively-coupled plasma mass spectrometry (ICP-MS). DNA-damage was assessed by γH2AX immunofluorescence. BIA combination with cisplatin in vitro studies were performed via luminescent viability assays. Treatment of human cerebrovascular endothelial cells (HCMEC/D3) with BIA for BBB studies in vitro involved trans-endothelial electrical resistance (TEER) measurement, transcriptome analysis, VE-cadherin western blotting and immunofluorescence. Results: Combined addition of cisplatin and BIA showed enhanced anti-GBM killing effects than cisplatin alone in G9-pCDH (IC50 cisplatin=1, BIA+cisplatin=0.2), G34-pCDH (IC50 cisplatin=3, BIA+cisplatin=1.7) and G30-LRP (IC50 cisplatin=1, BIA+cisplatin=0.3) lines. Efficacy studies using the G9-pCDH xenograft murine model showed extended survival when using cisplatin in combination with BIA (median 27d, p=0.0016) when compared to cisplatin (18.5d), BIA (21.5d) and control (18d). Increased intra-tumoral accumulation of cisplatin was observed when used in combination with BIA (mean 100 pg/mg, p=0.012) rather than cisplatin alone (5 pg/mg). No increased platinum uptake was seen in healthy brain tissue under any conditions. γH2AX foci in tumor were more frequent in the BIA and cisplatin combination arm (p=0.0086) than each single agent and control groups. Evaluation of HCMEC/D3 cells treated with BIA showed a decrease in TEER levels (p&amp;lt;0.001) and transcriptomic dysregulation of members of the WNT pathway and downregulation of VE-cadherin (~4-fold). BIA exerted cytoskeleton changes in HCMEC/D3 cells and decreased VE-cadherin expression at the protein level and its distribution on cellular surface, suggesting loss of adherens junctions as a potential mechanism for BIA-induced permeability of the BTB. Conclusion: Overall, our data support the applicability of BIA for modulating the BTB permeability and increasing intra-tumoral drug accumulation for improving GBM therapeutic efficacy. Citation Format: Jorge L. Jimenez Macias. Modulation of the blood-tumor barrier by 6-bromoindirubin acetoxime increases intra-tumoral drug accumulation and extends survival in murine glioblastoma models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2713.

Cerium (IV) Oxide Nanoparticles Enhance Hepatotoxic and Nephrotoxic Effects of Paraquat, Cisplatin, or Acetaminophen in Mice

Nanomaterials are central to nanotechnology and have many useful properties. Cerium(IV) oxide nanoparticles (nCeO) have catalytic and antioxidant activity, and hold promise in industrial and medical products. However, the potential biological hazards of nCeO have not been addressed. We therefore investigated the in vivo effects and drug interactions of nCeO. Tail vein administration of nCeO did not induce liver and kidney injury in mice but induced liver or kidney damage when co-administered with paraquat, cisplatin or acetaminophen. Therefore, our findings indicate that nCeO are potentially hepatotoxic and nephrotoxic due to drug interactions.

Modulation of Extravascular Binding of Recombinant Factor IX Impacts Hemostatic Efficacy in Mouse Models

Introduction: Hemophilia B (HB) is a X-linked bleeding disorder characterized by the deficiency in coagulation factor IX (FIX). Replacement treatment with recombinant human FIX (rFIX) is a safe approach to prevent bleeding in HB patients. There is an emerging concept that FIX resides extravascularly, by potentially binding to collagen IV in the subendothelial basement membrane, where it contributes to hemostasis. Our work evaluates hemostatic efficacy of extravascular FIX using animal models of tail clip bleeding and ferric chloride (FeCl3)-induced thrombus formation. Methods: The pharmacokinetic (PK) profile of wildtype rFIX (rFIXWT) and rFIX mutants which are stated to have an enhanced (rFIXK5R) or reduced (rFIXK5A) binding to components of the extravascular space (EVS) was evaluated in C57BL/6J HB mice (B6.129P2-F9<tm1Dw1s>). Blood and liver samples were collected at 0.08, 24, and 72 hours after tail vein administration of rFIX proteins (n=3-5 per timepoint). Plasma activity levels of rFIX were determined by a one stage clotting assay (OSCA). The content of FIX in the liver sections was analyzed using immunofluorescence staining. The direct binding of rFIX to collagen IV was evaluated for the first-time using surface plasma resonance (SPR). The in vivo efficacy of these proteins in HB mice was compared in tail clip bleeding and FeCl3-induced thrombus formation models. Results: PK studies in HB mice showed that the total exposure was higher for the rFIXK5A mutant which exhibited a dose normalized (dn) AUC0-last of 0.442±0.025 IU/mL*h/IU, followed by lower dnAUC0-last of rFIXWT (0.069±0.023 IU/mL*h/IU) and rFIXK5R (0.056±0.031 IU/mL*h/IU). The predicted dose normalized maximum concentrations (dnCmax) at 5 minutes post administration were comparable between rFIXWT (0.013±0.003 IU/mL/IU), rFIXK5R (0.013±0.001 IU/mL/IU) and slightly higher for rFIXK5A (0.018±0.001 IU/mL/IU). At 5 minutes following intravenous injection, all three rFIX proteins were detected in liver sections with similar mean fluorescence intensities (MFI). However, at 24 hours following intravenous injection, rFIXK5R had higher MFI compared to rFIXK5A and rFIXWT groups (rFIXK5R vs. rFIXWT p<0.001). rFIXK5R was detectable in the liver tissue up to 72 hours even in the absence of circulating FIX antigen levels (Figure 1). Though rFIXK5A was detected in the plasma until 336 hours, the presence in liver tissue was observed only at 5 minutes. Human FIX and murine collagen IV co-staining in liver sections of rFIX treated HB mice showed an overlap of FIX staining mostly with collagen IV-stained regions. However, there were certain liver vascular beds stained specifically for collagen IV with no detectable FIX in that region. To further investigate this, direct binding of FIX to collagen IV was evaluated in SPR and no binding was observed between collagen IV and rFIX wildtype or rFIX mutant proteins. In the tail clip bleeding model, rFIXK5R displayed significant hemostatic protection against bleeding incidence for up to 72 hours post intravenous administration, whereas for rFIXK5A this was only observed at the 15 minutes time point. In the mesenteric artery thrombus model, the time to occlusion of the artery was comparable in both rFIXK5R and rFIXWT treated groups. In contrast, rFIXK5A lacked the ability to form occlusive clots and was comparable to vehicle group. In line with this observation, rFIXWT and rFIXK5R treated HB mice developed significantly larger clots. In contrast, rFIXK5A mutant with decreased binding to components of the EVS exhibited smaller clots compared to rFIXK5R and rFIXWT. Conclusion: The results of the PK study confirmed a longer exposure of rFIXK5A in plasma compared to rFIXK5R, as expected from decreased binding affinity of rFIXK5A to components of the EVS. The histological evaluations of the harvested liver tissues confirmed a longer accumulation of rFIXK5R, which is expected to have increased binding affinity to components of the EVS. The efficacy studies revealed that the ability of FIX to bind to components of the EVS has an impact on the hemostatic response in HB mouse models. Using two different in vivo models of hemostasis, we demonstrate that mutated rFIX protein with enhanced binding to components of the EVS (rFIXK5R) confers prolonged hemostatic efficacy while the duration of efficacy of rFIXK5A was relevantly shorter after intravenous administration of the recombinant proteins. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Adeno-associated virus-based caveolin-1 delivery via different routes for the prevention of cholesterol gallstone formation

BackgroundHepatic caveolin-1 (CAV1) is reduced in cholesterol gallstone disease (CGD). Mice with CAV1 deficiency were prone to develop CGD. However, it remains unknown whether restored hepatic CAV1 expression prevents the development of CGD.MethodsC57BL/6 mice were injected with adeno-associated virus 2/8 (AAV2/8) vectors carrying the CAV1 gene (AAV2/8CAV1) via intravenous (i.v.) or intraperitoneal (i.p.) route and then subjected to a lithogenic diet (LD) for 8 weeks. Uninjected mice were used as controls. The functional consequences of rescuing CAV1 expression by either i.v. or i.p. AAV2/8CAV1 treatment for CGD prevention and its subsequent molecular mechanisms were examined.ResultsCAV1 expression was reduced in the liver and gallbladder of LD-fed CGD mice. We discovered that AAV2/8CAV1 i.p. delivery results in higher transduction efficiency in the gallbladder than tail vein administration. Although either i.v. or i.p. injection of AAV2/8CAV1 improved liver lipid metabolic abnormalities in CGD mice but did not affect LD feeding-induced bile cholesterol supersaturation. In comparison with i.v. administration route, i.p. administration of AAV2/8CAV1 obviously increased CAV1 protein levels in the gallbladder of LD-fed mice, and i.p. delivery of AAV2/8CAV1 partially improved gallbladder cholecystokinin receptor (CCKAR) responsiveness and impeded bile cholesterol nucleation via the activation of adenosine monophosphate-activated protein kinase (AMPK) signaling, which induced a reduction in gallbladder mucin-1 (MUC1) and MUC5ac expression and gallbladder cholesterol accumulation.ConclusionCGD prevention by i.p. AAV2/8CAV1 injection in LD-fed mice was associated with the improvement of gallbladder stasis, which again supported the notion that supersaturated bile is required but not sufficient for the formation of cholesterol gallstones. Additionally, AAV treatment via the local i.p. injection offers particular advantages over the systemic i.v. route for much more effective gallbladder gene delivery, which will be an excellent tool for conducting preclinical functional studies on the maintenance of normal gallbladder function to prevent CGD.

The use of melittin to enhance transgene expression mediated by recombinant adeno-associated virus serotype 2 vectors both in vitro and in vivo

ObjectiveMelittin, a cell-penetrating peptide, improves the efficiency of many non-viral gene delivery vectors, yet its application in viral vectors has not been well studied. The non-pathogenic recombinant adeno-associated virus (rAAV) vector is an ideal in vivo gene delivery vector. However, its full potential will only be achieved after improvement of its transduction efficiency. To improve the transduction efficiency of rAAV2 vectors, we attempted to develop a melittin-based rAAV2 vector delivery strategy. MethodsThe melittin peptide was inserted into the rAAV2 capsid either in the loop VIII of all viral proteins (VPs) or at the N terminus of VP2. Various rAAV2-gfp or -fluc vectors were subjected to quantitative real-time polymerase chain reaction and Western blot assays to determine their titers and integrity of capsid proteins, respectively. Alternatively, the vectors based on wild-type capsid were pre-incubated with melittin, followed by transduction of cultured cells or tail vein administration of the mixture to C57BL/6 and BALB/c nude mice. In vivo bioluminescence imaging was performed to evaluate the transgene expression. ResultsrAAV2 vectors with melittin peptide inserted in the loop VIII of VPs had low transduction efficiency, probably due to dramatically reduced ability to bind to the target cells. Fusing the melittin peptide at the N-terminus of VP2 produced vectors without the VP2 subunit. Interestingly, among the commonly used rAAV vectors, pre-incubation of rAAV2 and rAAV6 vectors with melittin significantly enhanced their transduction efficiency in HEK293 and Huh7 cells in vitro. Melittin also had the ability to increase the rAAV2-mediated transgene expression in mouse liver in vivo. Mechanistically, melittin did not change the vector-receptor interaction. Moreover, cell counting kit-8 assays of cultured cells and serum transaminase levels indicated melittin had little cytotoxicity. ConclusionPre-incubation with melittin, but not insertion of melittin into the rAAV2 capsid, significantly enhanced rAAV2-mediated transgene expression. Although further in vivo evaluations are required, this research not only expands the pharmacological potential of melittin, but also provides a new strategy to improve gene therapy mediated by rAAV vectors.

Biomimetic hybrid-cell membrane nanoparticles loaded with panaxytriol for breast cancer combinational therapy

Breast cancer is the most prevalent cancer with an unfavorable prognosis in women, which seriously threatened women’s health worldwide. In our previous study, a phytoconstituent, panaxytriol (PA), isolated from a Tujia ethnomedicine Panax japonicus, showed excellent inhibitory effects on APE1 activity, an enzyme tightly related with the development of breast cancer. Considering the tumor cell-killing ability and poor bioavailability of PA in vivo, Prussian blue nanoparticles with dopamine surface modification were applied to enhance its therapeutic efficacy against breast cancer by combining with photodynamic agent of indocyanine green. The hybrid-cell membrane (microphage and erythrocyte membranes) camouflage of the nano-delivery system significantly improved the solubility and targeting ability to tumor regions with increased immune evasion. This Prussian blue nanoparticle-based biomimetic formulation, introduced via tail vein administration, was validated as an efficient PA delivery vehicle for inhibiting the growth of breast cancer xenografts in nude mice. Molecular experiments revealed that the antitumor activity was mediated through cell proliferation inhibition and apoptosis induction. Our results prove that comprehensive therapy combining PA with nanomaterials may provide an effective strategy for treating breast cancer in vivo.

Pharmacokinetics and Pharmacodynamics of Fungal Defensin NZX Against Staphylococcus aureus-Induced Mouse Peritonitis Model.

Staphylococcus aureus (S. aureus) is one of the most common pathogenic bacteria responsible for causing a life-threatening peritonitis disease. NZX, as a variant of fungal defensin plectasin, displayed potent antibacterial activity against S. aureus. In this study, the antibacterial and resistance characteristics, pharmacokinetics, and pharmacodynamics of NZX against the S. aureus E48 and S. aureus E48-induced mouse peritonitis model were studied, respectively. NZX exhibited a more rapid killing activity to S. aureus (minimal inhibitory concentration, 1 μg/ml) compared with linezolid, ampicillin and daptomycin, and serial passaging of S. aureus E48 for 30 days at 1/2 × MIC, NZX had a lower risk of resistance compared with ampicillin and daptomycin. Also, it displayed a high biocompatibility and tolerance to physiological salt, serum environment, and phagolysosome proteinase environment, except for acid environment in phagolysosome. The murine serum protein-binding rate of NZX was 89.25% measured by ultrafiltration method. Based on the free NZX concentration in serum after tail vein administration, the main pharmacokinetic parameters for T1/2, Cmax, Vd, MRT, and AUC ranged from 0.32 to 0.45 h, 2.85 to 20.55 μg/ml, 1469.10 to 2073.90 ml/kg, 0.32 to 0.56 h, and 1.11 to 8.89 μg.h/ml, respectively. Additionally, the in vivo pharmacodynamics against S. aureus demonstrated that NZX administrated two times by tail vein at 20 mg/kg could rescue all infected mice in the lethal mouse peritonitis model. And NZX treatment (20 mg/kg) significantly reduced CFU counts in the liver, lung, and spleen, especially for intracellular bacteria in the peritoneal fluid, which were similar or superior to those of daptomycin. In vivo efficacies of NZX against total bacteria and intracellular bacteria were significantly correlated with three PK/PD indices of ƒAUC/MIC, ƒCmax/MIC, and ƒT% > MIC analyzed by a sigmoid maximum-effect model. These results showed that NZX may be a potential candidate for treating peritonitis disease caused by intracellular S. aureus.

Cardioprotective Effects of HO-1-Loaded Collagen-Targeted Phase Change Nanoparticles on Cardiomyocytes Following Acute Myocardial Infarction

Acute myocardial infarction (MI) is a major cause of death worldwide. This study utilized collagen-targeted phase change material (PCM) nanoparticles (NPs) to co-encapsulate HO-1 and explored the efficacy of composite PCM NPs on cardiomyocyte progression and development of MI. In this study, we enrolled 32 acute MI patients and 32 healthy participants, and ELISA assay was used to assess the content of creatine kinase isoenzyme (CK-MB). Mice with MI received tail vein administration of HO-1-loaded PCM NPs, followed by RT-qPCR detection of expressions of hypoxia-reoxygenation related genes (SpA, SpB, SpC, Occludin, KGF, and CK18). Patients with acute MI had a higher level of CK-MB. Treatment with HO-1-loaded collagen-targeted PCM NPs decreased expressions of SpA, SpB, SpC, Occludin, KGF, and CK18, facilitating repair of damaged tissues. Of note, NPs loaded with siRNA HO-1 up-regulated the expression of these genes. Collagen-targeted PCM NPs carrying HO-1 effectively promoted the repair of myocardial cells and relieved MI through down-regulation of hypoxia-reoxygenation related genes, which may enhance prevention and treatment for acute MI.

Exosomes derived from human umbilical cord mesenchymal stem cells alleviate acetaminophen-induced acute liver failure through activating ERK and IGF-1R/PI3K/AKT signaling pathway

This study aimed to investigate the therapeutic potential of human umbilical cord mesenchymal stem cells derived exosomes (hUCMSC-Exo) in acute liver failure (ALF) in mice as well as its underlying mechanism. We found that a single tail vein administration of hucMSC-Exo effectively enhanced the survival rate, inhibited apoptosis in hepatocytes, and improved liver function in APAP-induced mouse model of ALF. Furthermore, the deletion of glutathione (GSH) and superoxide dismutase (SOD), generation of malondialdehyde (MDA), and the over production of cytochrome P450 E1 (CYP2E1) and 4-hydroxynonenal (4-HNE) caused by APAP were also inhibited by hucMSC-Exo, indicating that hucMSC-Exo inhibited APAP-induced apoptosis of hepatocytes by reducing oxidative stress. Moreover, hucMSC-Exo significantly down-regulated the levels of inflammatory cytokines IL-6, IL-1β, and TNF-α in APAP-treated livers. Western blot showed that hucMSC-Exo significantly promoted the activation of ERK1/2 and IGF-1R/PI3K/AKT signaling pathways in APAP-injured LO2 cells, resulting in the inhibition of apoptosis of LO2 cells. Importantly, PI3K inhibitor LY294002 and ERK1/2 inhibitor PD98059 could reverse the function of hucMSC-Exo on APAP-injured LO2 cells in some extent. Our results suggest that hucMSC-Exo offer antioxidant hepatoprotection against APAP in vitro and in vivo by inhibitiing oxidative stress-induced apoptosis via upregulation of ERK1/2 and PI3K/AKT signaling pathways.

Preparation, Biocompatibility and Antitumor Activity of Nanodelivery System Targeting Breast Cancer Base on a Silica Nanoparticle.

BackgroundBreast cancer (BC) is the most common type of cancer among women worldwide, and about 30% of males will have recurrent disease.MethodsIn order to treat recurrent BC, we designed a type of silica nanodelivery system loaded with epirubicin and curcumin (composite nanoparticles, CNPs). To promote CNPs clinical application, the stability, the blood, immune and cell compatibility, skin stimulation experiments, anti-tumor activity in vivo and in vitro were studied.ResultsIn our study, the CNPs had a particle size of 73.9 nm and a uniform size and morphology; moreover, they maintained physical and chemical stability in the blood protein environment. Additionally, results showed that nanoparticles had good blood and immune compatibility, and they did not affect intracellular superoxide dismutase (SOD) and intracellular catalase (CAT). Skin stimulation experiments showed that CNPs did not cause any obvious irritative damage to the intact skin of rabbits. In the cytotoxicity study, CNPs showed strongest antitumor activity. The results of cell cycle and apoptosis studies showed that CNPs could mainly induce apoptosis of S and G2/M phase cells. In vivo, CNPs showed strongest aggregation in the tumor after 6 h of tail vein administration, and a large amount of CNPs continued to accumulate in the blood after 12 h of administration, indicating that CNPs had long circulation ability. The in vivo antitumor activities showed that CNPs had the strongest antitumor activity and tumor targeting ability, and hematoxylin-eosin staining of internal organs showed no obvious difference between treatment groups and negative control.ConclusionCNPs have an ideal biosafety and therapeutic effect for recurrent BC, and they have potential clinical application value.

Superparamagnetic α-Fe2O3/Fe3O4 Heterogeneous Nanoparticles with Enhanced Biocompatibility.

A novel type of magnetic α-Fe2O3/Fe3O4 heterogeneous nanoparticles was prepared via a facile solution combustion process with ferric nitrate and urea as raw materials, and they were characterized by XRD, SEM, TEM, and VSM techniques. The effects of the calcination temperature, the calcination time, the ratio of ferric nitrate and urea, and the heating rate on the relative content of Fe3O4 in the heterogeneous nanoparticles were investigated. The toxicity of α-Fe2O3/Fe3O4 heterogeneous nanoparticles to human hepatocytes L-02, the blood routine, and the histopathological section observation of mice were explored. The results showed that the ratio of ferric nitrate and urea was a key factor to affect the relative content of Fe3O4 in the heterogeneous nanoparticles. The calcination temperature and the calcination time had similar influences, and the corresponding calcination temperature and the calcination time were selected according to their own needs. The CCK8 results initially revealed that α-Fe2O3/Fe3O4 heterogeneous nanoparticles had no effect on cell viability when the concentration of the heterogeneous nanoparticles was less than 100 ng/mL, which suggested their excellent biocompatibility. At the same time, the tail vein administration concentration of 0.9 mg/kg had good biological safety.

MSCs derived from amniotic fluid and umbilical cord require different administration schemes and exert different curative effects on different tissues in rats with CLP-induced sepsis

BackgroundMesenchymal stem cells (MSCs) are derived from multiple tissues, including amniotic fluid (AF-MSCs) and the umbilical cord (UC-MSCs). Although the therapeutic effect of MSCs on sepsis is already known, researchers have not determined whether the cells from different sources require different therapeutic schedules or exert different curative effects. We assessed the biofunction of the administration of AF-MSCs and UC-MSCs in rats with caecal ligation and puncture (CLP)-induced sepsis.MethodsCLP was used to establish a disease model of sepsis in rats, and intravenous tail vein administration of AF-MSCs and UC-MSCs was performed to treat sepsis at 6 h after CLP. Two phases of animal experiments were implemented using MSCs harvested in saline with or without filtration. The curative effect was measured by determining the survival rate. Further effects were assessed by measuring proinflammatory cytokine levels, the plasma coagulation index, tissue histology and the pathology of the lung, liver and kidney.ResultsWe generated rats with medium-grade sepsis with a 30–40% survival rate to study the curative effects of AF-MSCs and UC-MSCs. MSCs reversed CLP-induced changes in proinflammatory cytokine levels and coagulation activation. MSCs ameliorated CLP-induced histological and pathological changes in the lung, liver and kidney. AF-MSCs and UC-MSCs functioned differently in different tissues; UC-MSCs performed well in reducing the upregulation of inflammatory cytokine levels in the lungs and inhibiting the inflammatory cell infiltration into the liver capsule, while AF-MSCs performed well in inhibiting cell death in the kidneys and reducing the plasma blood urea nitrogen (BUN) level, an indicator of renal function.ConclusionsOur studies suggest the safety and efficacy of AF-MSCs and UC-MSCs in the treatment of CLP-induced sepsis in rats and show that the cells potentially exert different curative effects on the main sepsis-affected tissues.

A pilot study of neonatal GALT gene replacement using AAV9 dramatically lowers galactose metabolites in blood, liver, and brain and minimizes cataracts in GALT-null rat pups.

Classic galactosemia (CG) is a rare metabolic disorder that results from profound deficiency of galactose-1-P uridylyltransferase (GALT). Despite early detection by newborn screening and rapid and lifelong dietary restriction of galactose, which is the current standard of care, most patients grow to experience a broad constellation of long-term complications. The mechanisms underlying these complications remain unclear and likely differ by tissue. Here we conducted a pilot study testing the safety and efficacy of GALT gene replacement using our recently-described GALT-null rat model for CG. Specifically, we administered AAV9.CMV.HA-hGALT to seven GALT-null rat pups via tail vein injection on day 3 of life; eight GALT-null pups injected with PBS served as the negative control, and four GALT+ heterozygous pups injected with PBS served as the positive control. All pups were returned to their nursing mothers, weighed daily, and euthanized for tissue collection 2 weeks later. Among the AAV9-injected pups in this study, we achieved GALT levels in liver ranging from 64% to 595% wild-type, and in brain ranging from 3% to 42% wild-type. In liver, brain, and blood samples from these animals we also saw a striking drop in galactose, galactitol, and gal-1P. Finally, all treated GALT-null pups showed dramatic improvement in cataracts relative to their mock-treated counterparts. Combined, these results demonstrate that GALT restoration in both liver and brain of GALT-null rats by neonatal tail vein administration using AAV9 is not only attainable but effective.