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Preparation and evaluation of novel self-assembled apo-transferrin-stearic acid nanoparticles in cancer targeting
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | Beom-Jin Lee | - |
| dc.contributor.author | AMIN HARDIK HASTIBHAI | - |
| dc.date.accessioned | 2018-11-08T08:28:56Z | - |
| dc.date.available | 2018-11-08T08:28:56Z | - |
| dc.date.issued | 2017-08 | - |
| dc.identifier.other | 26037 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/14200 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :약학,2017. 8 | - |
| dc.description.tableofcontents | ChapterⅠ.Preparation and characterization of self-assembled apotransferrin- stearic acid nanoparticle and their evaluation in cancer targeting. 1 Abstract 2 1. Introduction 4 2. Materials and methods. 8 2.1. Materials 8 2.2. Preparation of apo-transferrin-stearic acid (TfS) conjugates. 8 2.3. Characterization of TfS conjugates. 9 2.4. Preparation of NPs. 10 2.5. Characterization of NPs 10 2.6. Cell culture and maintenance. 12 2.7. In-vitro cytotoxicity. 12 2.8. Cell targeting and uptake 13 2.9. Route of endocytosis 14 2.10. Statistics. 14 3. Results and discussion. 16 3.1. Characterization of TfS conjugates 16 3.2. Characterization of NPs 18 3.3. Drug release study 21 3.4. Cell toxicity by MTT assay 23 3.5. Cell targeting and uptake of NPs 26 3.6. Uptake of NP in cancer cell lines 31 3.7. Quantitative uptake of NP in cells using FACS flow cytometry 33 3.8. Route of cellular uptake 35 4. Conclusions 39 ChapterⅡ.Paclitaxel loaded apo-transferrin-stearic acid nanoparticle in targeting and killing breast cancer cells 40 Abstract 41 1. Introduction 42 2. Materials and methods 45 2.1. Materials 45 2.2. Preparation of TfS +PAC and TfS + C6 45 2.3. Drug loading efficiency (LE) and encapsulation efficiency (EE) 45 2.4. Characterization of NPs 46 2.5. Cell culture and maintenance 47 2.6. In-vitro cytotoxicity 48 2.7. Cellular uptake by confocal microscopy 48 2.8. Cellular uptake by flow cytometry 49 2.9. Cell apoptosis assay 50 2.10. Route of endocytosis 50 2.11. Statistics 51 3. Results and discussion 52 3.1. Characterization of NPs 52 3.2. Drug release study. 55 3.3. Cell toxicity by MTT assay 57 3.4. Confocal microscopy study 59 3.5. Uptake of NP in cancer cell line through flow cytometry 61 3.6. Apoptosis assay 63 3.7. Route of cellular uptake 66 4. Conclusions 68 Supplementary Information 69 1. Confirmation of the structure of TfS conjugates from FTIR and MALDI-TOF spectra 69 1.1 FTIR Spectra 69 1.2 MALDI-TOF Spectra 75 2. Degree of conjugation of stearic acid to apo-Tf 77 References 79 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | Preparation and evaluation of novel self-assembled apo-transferrin-stearic acid nanoparticles in cancer targeting | - |
| dc.title.alternative | Preparation and evaluation of novel self-assembled apo-transferrin-stearic acid nanoparticles in cancer targeting | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | AMIN HARDIK HASTIBHAI | - |
| dc.contributor.department | 일반대학원 약학 | - |
| dc.date.awarded | 2017. 8 | - |
| dc.description.degree | Doctoral | - |
| dc.identifier.localId | 872315 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000026037 | - |
| dc.subject.keyword | transferrin | - |
| dc.subject.keyword | nanoparticle | - |
| dc.subject.keyword | self-assembly | - |
| dc.subject.keyword | paclitaxel | - |
| dc.subject.keyword | doxorubicin | - |
| dc.description.alternativeAbstract | The major hurdle in cancer treatment is to precisely target drugs at cancer site. Transferrin-Transferrin receptor interaction is widely exploited to target cancer cells due to over-expression of transferrin receptor by many cancer cells. In this study, novel apo-transferrin-stearic acid (TfS) conjugates were prepared and characterized by FTIR, MALDI-TOF mass spectroscopy and TNBS assay to confirm their structure. The prepared TfS conjugates showed self-assembly in water and formed nanoparticle which were evaluated by SEM, TEM, particle size and zeta potential analysis. Doxorubicin HCl was adsorbed on surface of NP by incubation method and prepared doxorubicin containing TfS NP (NP+DOX) were analyzed for their cell targeting and cell killing properties in transferrin receptor over-expressing cell lines A549 and HCT116 cells by MTT assay, confocal microscopy and fluorescent assisted cell sorting flow cytometry. The data showed that NP+DOX exhibited improved targeting and cell killing effects compared to doxorubicin. The cell killing efficiency of NP+DOX was similar, to that of Doxil with improved cell uptake as seen from confocal microscopy. The prepared drug delivery system has potential to target transferrin receptor over-expressing cancer cells and prevents possible off-target effects. In this study, paclitaxel was encapsulated in self assembled apo-transferrin-stearic acid nanoparticles and the prepared nanoparticles were evaluated for their transferrin receptor targeting and cell killing effects. The nanoparticles exhibited spherical smooth surface in SEM and TEM studies. The particle size of paclitaxel loaded nanoparticle was 326.97 ± 2.03 nm with loading and encapsulation efficiency being 7.94 ± 1.60 and 71.10±4.12. Paclitaxel loaded nanoparticle showed 7 fold reduction in LC50 in breast carcinoma cells MCF-7 compared to that of free paclitaxel, accounting for increased cytotoxicity due to effective targeting of cells. Confocal study confirmed this observation as uptake of nanoparticle was inhibited in presence of transferrin receptor blocking treatment. Flow cytometry data also confirmed above observations depicting time dependent uptake of nanoparticles and reduction in uptake in case of receptor blocking. These data suggest for advantage of developed drug delivery platform in cancer cell targeting and subsequently cell killing effect. | - |
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- Graduate School of Ajou University > Department of Pharmacy > 4. Theses(Ph.D)
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