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Home > Lipid-Polymer Hybrid Nanoparticles for Potential Delivery of Chemotherapeutic Agents: Formulation and Characterization

Lipid-Polymer Hybrid Nanoparticles for Potential Delivery of Chemotherapeutic Agents: Formulation and Characterization

Thesis Info

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Author

Khan, Muhammad Muzamil

Program

PhD

Institute

The Islamia University of Bahawalpur

City

Bahawalpur

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Pharmaceutics

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/12449/1/muhammad%20muzamil%20khan%20pharmaceutics%202019%20iub%20prr.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726590154

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Nanotechnology has emerged as a hope to deliver drugs at targeted site to obtain maximum therapeutic benefits. Lipid-polymer hybrid nanoparticles have provided the platform to deliver drugs in a controlled manner with enhanced stability and biocompatibility. LPHNPs have the dual advantages of polymeric and liposomal drug delivery system and can encapsulate both hydrophilic and hydrophobic drugs. The objective of this dissertation was to develop the lipid-chitosan hybrid nanoparticles for potential delivery of chemotherapeutic agents at tumor site to achieve maximum therapeutic benefits and decrease side effects associated with chemotherapeutic agents. Lipid-chitosan hybrid nanoparticles were prepared by using chitosan as a polymer and LIPOID S75 as a lipid using modified ionic gelation method. The prepared nanoparticles have size in range of 200-300nm with PDI values less than 0.3 and surface charge showed good stability in suspension form. The transmission electron microscopy images showed spherical nanoparticles having lipoplex like structure. All prepared nanoparticles showed high entrapment efficiency (>80%) and good drug loading. The FTIR analysis confirmed the compatibility among the excipients and XRD analysis showed no sharp peaks of cisplatin in formulation and cisplatin is converted into amorphous form inside lipid-chitosan hybrid nanoparticles system. Thermal studies using differential scanning calorimetry and thermogravimetric analysis confirmed the excellent stability of prepared hybrid nanoparticles. The in vitro release showed that controlled release of drug over prolong period of time. Kinetic modeling showed that the release pattern follows super case II transport mechanism. The physicochemical evaluation confirmed the excellent stability and controlled release profile. The therapeutic efficacy of cisplatin loaded lipid-chitosan hybrid nanoparticles was evaluated by using A2780 ovarian cell lines. The results confirmed the enhanced cytotoxicity of cisplatin loaded lipid-chitosan hybrid nanoparticles as compared to cisplatin solution. Cellular interaction and cell uptake showed 8 times greater uptake as compared to control. Further in vivo pharmacokinetic studies confirmed enhanced mean residence time of LPHNPS inside the biological system. Toxicology studies confirmed the safety profile of lipid-chitosan hybrid nanoparticles. The folic acid was conjugated with chitosan for folate targeting to achieve maximum therapeutic benefits at the tumor site. The TLC analysis confirmed the purity of conjugate and absence of free folic acid while nuclear magnetic resonance spectroscopy confirmed the successful conjugation of folic acid with chitosan. The folate-chitosan conjugate was then used to prepare nanoparticle by ionic gelation method with anionic lipids. The prepared nanoparticles have particle size in range of 200nm and low polydispersity index and surface charge of greater than +20. The folate LPHNPs showed greater than 75% encapsulation with excellent drug loading. The prepared nanoparticles are spherical in shape with lipoplex like structure having folate on the outer side of nanoparticles. In vitro release profile shows sustained release of cisplatin over a period of 48 hours. The therapeutic efficacy of folate lipid-chitosan hybrid nanoparticles was evaluated on ovarian and breast cell lines. The A2780 and SKOV3 were used as ovarian cell lines and treated with folate targeted LPHNPs and untargeted LPHNPs and cisplatin solution. The results confirmed the enhanced cytotoxic effect of folate LPHNPs as compared to untargeted LPHNPs and cisplatin solution. Similar enhanced cytotoxic effect of folate LPHNPs was observed on MCF-7 breast cancer cell lines. The cell uptake studies showed two times more uptake of folate LPHNPs are compared to untargeted LPHNPs that is due to folate receptor mediated endocytosis and leads to enhanced therapeutic efficacy. The therapeutic efficacy of folate LPHNPs was further evaluated on 3D spheroids in vivo model to check the response of nanoparticles in in vivo environment. The cell viability studies on 3D spheroids confirmed the enhanced cytotoxic effect of folate LPHNPs as compared to untargeted and much more significant cytotoxic effect as compared to cisplatin solution. The fluorescence microscopy images, and flow cytometry analysis confirmed the enhanced cellular uptake of folate LPHNPs in 3D spheroids that leads to enhanced therapeutic efficacy. In conclusion, lipid-chitosan hybrid nanoparticles are suitable platform for controlled delivery of chemotherapeutic agents. Folate targeted LPHNPs with added advantage of lipid coating is suitable for active targeting with enhanced therapeutic efficacy and minimum side effects.
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سماجی رویوں کی تشکیل کا نبوی منہج اخلاق

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Fluoroquinolone Resistance in Extended-Spectrum Beta-Lactamase Esbl Producing Uropathogenic E. Coli

Escherichia coli lineage ST131 predominates across various spectra of extra-intestinal infections. Distinctive resistance profile, diverse armamentarium of virulence factors and rapid global dissemination of ST131 E. coli makes it an intriguing pathogen of significant importance. However, little is known about the prevalence and various attributes of ST131 associated with urinary tract infections in Pakistan. Evolutionary dimensions of ST131 linage and corresponding molecular markers encoding drug resistance remained largely unexplored and prevalence data are extremely scarce in South Asian countries. We scrutinized prevalence and various genetic attributes of E. coli ST131 isolates involved in urinary tract infections by processing 148 randomly selected samples during August 2012 and August 2014. Phylogenetic grouping, fumC/fimH and O-typing was done by PCR based methods. Antibiotic susceptibility assays and phenotypic detection of ESBLs were carried out according to the CLSI guidelines of year 2013 and ESBL isolates were screened for the prevalence of blaCTX-M-15. Plasmid encoded qnrA, qnrB and qnrS genes were identified by DNA amplification and sequencing. Mutations in genes gyrA, gyrB, parC and parE were identified among fluoroquinolone resistant isolates. Minimum inhibitory concentrations (MICs) for ciprofloxacin and levofloxacin were determined by using broth dilution method. Phylogenetic analysis was performed by PCR using three gene classification systems (Clermont et al., 2000). Overall, 59% of the UPEC isolates belonged to phylogenetic group B2 which was followed by group D=28%, B1=8% and A=5%. Among 18 different ST-types ST131 was the most dominant lineage as 46% of the isolates belonged to ST 131 lineage. By, CH-typing 46% of the ST 131 isolates belonged to sub-group H30. Higher numbers of identified ST 131 isolates 74% were MDR and 44% showed ESBL phenotypes while 100% of the isolates carried blaCTX-M-15. Overall, resistant patterns of ST 131 isolates confirmed least resistance against tigecycline, meropenam and amikacin. Likewise, resistance against tazocine, nitrofurantoin, sulzone and minocycline remained below threshold level. For fosfomycin, (FF) and gentamicin (CN) resistance were 13% and 28% respectively. Higher resistances was noticed against all four generations of cephalosporins, (53%-78%). Higher percentages of ST 131 isolates were resistant to co-trimoxazole (85%), amoxicillin clavulanic acid 65% and fluoroquinolones 60%. Overall, minimum inhibitory concentrations of ciprofloxacin remained above 256μg/ml in ST 131 and non ST 131 isolates. In comparison to MICs of ciprofloxacin, MICs of levofloxacin in most of the isolates varied between 8μg/ml-64μg/ml. In this study, majority of the fluoroquinolone resistant UPEC ST 131 isolates carried at least two or more non-synonymous mutations in gyrA, parC and parE genes. These isolates showed higher MICs for both ciprofloxacin and levofloxacin. Conclusively, higher percentages of the ST 131 isolates showed antibiotic resistance against trimethoprimsulphamethoxazolen, cephalosporin, fluoroquinolone, and majority of these isolates were multi-drug resistant. In addition, fluoroquinolone resistant UPECs isolates shared lineage of a clonal sub-group. An allelic combination of gyrA/parC/parE genes of sub-clone O25b-ST131 H30-R was associated with higher MICs of ciprofloxacin and levofloxacin. Overall, it is the first report from Pakistan that provides details insights about susceptibility patterns and genetic attributes of pandemic MDR ST131 strains involved in UTIs.