18 research outputs found
TNF-α expression.
<p>TNF- α in 50 µl of serum was estimated using Rat TNF-alpha platinum ELISA kit (Bender MedSystems) as per the procedure supplied with the kit. LPS was used as positive control. The immunogenicity of lactoferrin and apotransferrin is evident by the increased expression of TNF-α, a proinflammatory cytokine when treated with soluble forms of both proteins. The results showed maximum expression of TNF-α in LPS treatment (positive control). But when administered with Lactdoxonano (LD nano) and Apodoxonano (AD nano),TNF-α level is minimal as that of untreated control suggesting negligible inflammation caused by the nanoformulations.</p
Efficacy against hepatocellular carcinoma.
<p>Panel -A shows changes of rat body weight during treatments. The body weights of rats were decreased drastically when treated with Doxo and increased when given saline. During the treatment with nanoformulations , a slight increase in body weight was observed. Panel-B shows tumour nodules on the surface of liver. Tumour nodules are reduced significantly when treated with Apodoxonano and Lactodoxonano, when compared with Doxo and untreated rat (control). Panel C: After treatment of hepatocellular carcinoma, animals were sacrificed to collect the organs for their subsequent estimation of Doxo levels. Here the highest levels of drug were found only in the liver of rats treated with nanoformulations due to its specificity when compared to Doxo.</p
Molecular marker analysis.
<p>Analysis using molecular markers showed that <b>t</b>umour reduction was better during Lactodoxonano and Apodoxonano treatment when compared to Doxo, which is indicated by the over expression of the major tumoursuppressor genes namely p53 and p21. On the other hand, reduced vasculature due to decreased levels of VEGFR during nanoformulation treatment also reinforces its efficacy.</p
Treatment of hepatocellular carcinoma.
<p>Hepatocellular carcinoma was induced by adding 100 mg/L of DENA in drinking water for eight weeks. After one week of gap from the DENA treatment, animals received 2 µg/g of drug equivalent of Apodoxonano, Lactodoxonano and Doxo through <i>intra venous</i> administration on 1, 7, 14, 21 and 28 day. Panel Ashows untreated liver with maximum number of nodules which were reduced to some extent when treated with doxo (panel B). HCC was significantly reduced when treated with Lactodoxonano (Panel C) and Apodoxonano (Panel D) , which is indicated by the reduction in the number of nodules. It is further analyzed by H & E analysis of liver nodules along with treatments.</p
Safety analysis in healthy rats.
<p>Lactodoxonano, Apodoxonano and Doxo (800 µg of drug equivalent) were administered intravenously (<i>i.v.</i>). Safety analysis was carried out using biochemical kits. Heart toxicity was measured in terms of Glutathione peroxidase and LDH. Both the enzymes showed minimal expression when treated with nanoformulations as that of the control but gets increased during Doxo treatment. Kidney toxicity was evaluated by the levels of Creatinine, Blood Urea Nitrogen, whereas the Liver toxicity was estimated in terms of SGOT and SGPT levels. The level of these 4 markers also followed the same pattern as that of Glutathione peroxidase and LDH i.e., they showed minimal levels as that of control when treated with nanoformulations opposite to Doxo.</p
Pharmacokinetics profile of Lactodoxonano, Apodoxonano and Doxo.
<p>Values in the parenthesis indicates the concentration of doxorubicin in micrograms per ml of blood/per gram of tissue;</p>*<p>value of t<sub>1/2β.</sub></p><p><b>Pharmacokinetic parameters</b>.</p><p><b>AUC:</b> The integral of the concentration-time curve (after a single dose or in steady state).</p><p><b>AUMC:</b> Partial area under the moment curve between t start and t end.</p><p><b>t ½:</b> The time required for the concentration of the drug to reach half of its original value. In serum, t<sub>1/2α</sub> is referred duirngdistribution phase and t<sub>1/2β</sub> during elimination phases.</p><p><b>C<sub>max</sub> :</b>The peak plasma concentration of a drug after IV administration.</p><p><b>T<sub>max</sub> :</b>Time to reach C<sub>max.</sub></p
Characterization of lactoferrin nanoparticle size by SEM, TEM and AFM.
<p>Characterization of lactoferrin nanoparticle size by SEM, TEM and AFM.</p
Intravenous administration of Lactodoxonano, Apodoxonano and Doxo.
<p>Lactodoxonano, Apodoxonano and oxo (800 µg of drug equivalent) were administered intravenously (<i>i.v.</i>) and after indicated time points animals were sacrificed. Brain, Liver, Heart, Kidney, Spleen, Lungs, Bone marrow and Blood were collected. Proteins were precipitated in 30% AgNO<sub>3</sub> and drug was extracted in methanol. Drug was separated and estimated using absorption at 480 nm.</p
Improved Safety, Bioavailability and Pharmacokinetics of Zidovudine through Lactoferrin Nanoparticles during Oral Administration in Rats
<div><p>Zidovudine (AZT) is one of the most referred antiretroviral drug. In spite of its higher bioavailability (50–75%) the most important reason of its cessation are bone marrow suppression, anemia, neutropenia and various organs related toxicities. This study aims at the improvement of oral delivery of AZT through its encapsulation in lactoferrin nanoparticles (AZT-lactonano). The nanoparticles (NPs) are of 50–60 nm in size and exhibit 67% encapsulation of the AZT. They are stable in simulated gastric and intestinal fluids. Anti-HIV-1 activity of AZT remains unaltered in nanoformulation in acute infection. The bioavailability and tissue distribution of AZT is higher in blood followed by liver and kidney. AZT-lactonano causes the improvement of pharmacokinetic profile as compared to soluble AZT; a more than 4 fold increase in AUC and AUMC in male and female rats. The serum C<sub>max</sub> for AZT-lactonano was increased by 30%. Similarly there was nearly 2-fold increase in T<sub>max</sub> and t<sub>1/2</sub>. Our in vitro study confirms that, the endosomal pH is ideal for drug release from NPs and shows constant release from up to 96h. Bone marrow micronucleus assay show that nanoformulation exhibits approximately 2fold lower toxicity than soluble form. Histopathological and biochemical analysis further confirms that less or no significant organ toxicities when nanoparticles were used. AZT-lactonano has shown its higher efficacy, low organs related toxicities, improved pharmacokinetics parameter while keeping the antiviral activity intact. Thus, the nanoformulation are safe for the target specific drug delivery.</p></div