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Slide 1 :
MD.NAUSHAD ALI firstname.lastname@example.org My Mantras DUTY DEVOTION & DISCIPLINE
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SPECIALIZATION CARBON NANOTUBE GRAPHENE NANOCOMPOSITE NANOMEDICINE TECHNOLOGY TRANSFER CONSULTANCY
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Highlights EDUCATION M. Tech (Master in Technology) in Nanotechnology, AMITY University, India- SGPA 9.00/10 & CGPA 7.81/10 (1st division) 17th January, 2009 B.PHARMACY: Jamia Hamdard, New Delhi, India 2003 (57%) B.Sc. (H) Life science: Tilka Manjhi University Bhagalpur, India in 1999 (66%) Interdisciplinary technical skills covering material science, pharmaceutical sciences, electronics, chemistry, life sciences, ecology,physics & nanotechnology Experienced in the synthesis, functionalization & characterization of nanoparticles (inorganic/polymer) & carbon nanotubes Worked on Nanotoxicology (by in-vitro) studies on human cell lines for drug delivery and tissue engineering purposes Trained on TEM/SEM & AFM besides other characterization techniques
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Some involvement during master At AINT,India CVD Set up for CNT Synthesis
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ELECTRONICS LAB VAPOUR COATING UNIT ETCHING ON WAFER- MEMS/BIOMEMS
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Polymer NIPAAM nanoparticle synthesis & core shell on drug loading !! CNT AND SILANE COATING
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ULTRA MICROTONE/ TEM/Sonicator
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pNIPAAM+MCF 7 CELL Assay by MTT/PI for toxicity
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During summer ptoject At Renal Patho Lab AII India Institute of Medical Science Carbon Nanotube SYNTHESIS &BIOLOGICAL RESPONSE IHC&FACS, MTT,PI,Colorometric Assay
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MWCNT+MCF7 Cell line Control and Test Slides Dose dependent assay(MTT) CNT entry into cell
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Result & Discussion CNT synthesis, ratio of metallic catalyst, flow rate of gas,temperature and chemicals for purification must be taken into consideration. The duration of exposure may be even more predictive of damage than the nanoparticle concentration In the present study no growth of the cells were checked after 24 hours of exposure. More studies need to be done to characterize the reactive profile and toxicity of the CNT
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KNOWLEDGE ACQUIRED DURING SUMMER TRAINING CNT SYNTHESIS MICROSCOPY MCF7 CELL LINE CULTURE BIOLOGICAL RESPONSE TO CNT
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Master`s Dissertation “Engineering advanced polymeric surfaces for smart systems in Biomedicine, biology, material science and nanotechnology: A cross-disciplinary approach of Biology, Chemistry and Physics” MD.NAUSHAD ALI Enrolled as Early Stage Researcher, FP6 [BIOPOLYSURF] At Middle East Technical University Ankara Turkey METU
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TOPIC I.Nanotoxicology of Thermosensitive Smart Polymeric Nanoparticles II.Fabrication of nanopatterned polymeric intelligent surfaces by AFM & Cell Behavior BY:MD.NAUSHAD ALI Early Stage Researcher Project:BIOPOLYSURF Research Training Network MARIE CURIE ACTION FP6 Supervisor: Prof.Vasif Hasirci, BIOMAT,METU,Ankara,Turkey
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I. Nanotoxicology of Thermosensitive Smart Polymeric Nanoparticles Nanotechnology has led to the development of biodegradable self-assembled nanoparticles, which are being engineered for the development of nanomedicine. In this study, controlled nanoparticles of two thermo -sensitive polymers were synthesized through self assembly by radical polymerization. The toxicity assay on human cell lines conducted for finding the possibilities of these nanoparticles for the delivery and targeting of anticancer drugs. The target cell system was selected as the mammalian osteosarcoma cell line SaOs2 for in-vitro toxicity assay.
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Synthesis of Polymer NIPAM nanoparticle Micellar nanoparticles of polymer N-isopropyl acrylamide are hydrophilic nanoparticle, which are prepared in the aqueous media by polymerization of micelles formed spontaneously by self-assembly of amphophilic molecules above their critical micelles concentration (CMC). AB-type block copolymers consisting of a PNIPAM segment and a hydrophobic segment can form core–shell micellar structure below the LCST Experimental Set up for the polymerization
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For purification, np solution was transferred to a dialysis bag (32 mm width and of cut off 10000 KD) and then placed in distilled water with constant stirring for 24 h by changing water at every 3 hour . The image taken by Scanning Electron Microscope model JSM-6400 Electron Microscope (JEOL), confirmed the particle average size of 150 nm. SEM images of PNIPAM nanoparticle on filter paper SEM Characterization
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FTIR CHARACTERIZATION FTIR Analysis: The FTIR spectra confirm the formation of block copolymer. The Infrared transmittance spectra were measured at room temperature in a KBr pellet using a Shimadzu FTIR Infrared spectrometer in the frequency range between 750-1800 cm. As exhibited in the FT-IR spectra figure , absorbance of amide carbonyl groups in PNIPAM-NH occurs at 1641.92 cm, bending frequency of amide N–H appears at 1536.81 cm . Stretch vibration for C=O in ester in PMMA-COOH appears at 1537.78 cm. All the peaks mentioned above can be seen from the spectrum of PNIPAM-b-PMMA(green coloured) ,which supported the formation of the block copolymers. 2 -1 -1 -1 -1 -1
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wave number cm -1
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In-vitro cytotoxicity In vitro cytotoxicity of the Co-block polymer (pNIPAM-co-PMMA) and PNIPAM polymers was tested on mammalian osteosarcoma cell line SaOs2 . The cell line was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The cell line was cultured in 75 cm 2 culture flasks (Corning, USA) using RPMI media supplemented with 10% fetal bovine serum, 1% nonessential amino acids, and streptomycin (100 mg/ml) (all from Euroclone, UK). The cell culture was maintained at 37 0C, 95% relative humidity and 5% CO The cells of passage numbers 32–45 were used in the cytotoxicity tests Prior to cytotoxicity test, the cells were harvested using trypsin–ethylenediamine tetraacetic acid (EDTA)–PBS solution (0.25% trypsin–0.05mM EDTA according to the distributor’s instructions) and diluted at a density of 5x105 cells/ml in MTT assays. The cell suspension was seeded into 96-well plates (Corning, USA) at 100 ml/ well, and incubated for approximately 24 h before tests in order to reach confluency. Before the cells were seeded into 96-well plates, the plates were treated with 0.01% poly-D-lysine solution (Sigma-Aldrich,Germany). 2
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Cytotoxicity tests The cytotoxicity of the polymers was evaluated by using colorimetric cytotoxicity methods, MTT test. The test was performed with two polymers of nanometer size, three concentrations of the polymers (200 ng,600ng& 1 mg)were tested at 37 0C. The effect of incubation time for cell survival in MTT test was evaluated by performing 2 h, 24 h and 72 h.
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Standard deviation curve of cell number viability of PNIPAM
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Standard deviation curve of cell number viability of block copolymer
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Results and Discussion Amino-semitelechelic PNIPAM and carboxylic-semitelechelic PMMA were synthesized by telomerization using AET and MPA as telogens, respectively. The yield was ca. 34%. Micellar nanoparticles of polymer N-isopropyl acrylamide were prepared in the aqueous media by polymerization of micelles formed spontaneously by self-assembly of amphophilic molecules above their critical micelles concentration (CMC) .The size of the particless were 150 nm which is desirable range for the nanomedicines.The percentage yield was 37%. The cytotoxicity of these thermosensitive polymers was investigated as a function of polymer concentration and incubation time using 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The graph clearly indicating that these nanoparticles are nontoxic and thus could be used in the biological system.
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`Fabrication of nanopatterned polymeric intelligent surfaces by AFM Nanolithography` Part II
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Objective The intelligent nanostructured surface owing to its potential contributions to device miniaturization and creation of novel functional interfaces with applications ranging from electronics to biomedicine. This fabricated surface is reported to influence on the cellular adhesion, growth behavior and change the 3-dimensional structures of the adsorbed proteins . Since the invention of scanning tunneling microscopy (STM) and atomic force microscopy (AFM) these instruments have been used to fabricate surface structures ranging from ~100 nm down to atomic dimensions In this part of study ,the fabrication of Nanopattern surface of biopolymer (4%PHBV) by AFM was
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Experimental details PHBV obtained from Aldrich (batch number 01214MF). PHBV i.e. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) a biopolymer was taken as the standard materials for this study PHBV (4%) solution in chloroform was taken and heated at 35 0C for 20 minutes and at 560C for next 15 minutes. The film on petridish was prepared on pouring the solution and then drying in the hood. Dried thin film was collected from the petridish.The small pieces of film was cut 1x1 cm2 for the purpose of nanolithography These films were taken on the separate cover slips and put on the sample holder Atomic Force Microscopy (Quiscent Model of Ambious Technology,USA) was used with tapping mode AFM of SiN3tip (NSC16) . TESP (Tapping mode Etched Silicon Probe) Specifications: 20 -100 N/m Freq: 200-400kHz R tip: 3 -10nm Length: 125µm
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Lithography Characterized smooth surface of PHBV 4% film (sample B) before nanolithography and after lithography 100nm ridges After lithography
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Result and disussion The patterns formed were of desirable nanodimension range of 100 nm in breadth and 3nm in depth as seen in figures.The images were saved in the folder for the data analysis. Now these patterned films were sterilized and stored for the cell culture to see the cellular behavior Being understood the AFM handling techniques, the intelligent nanostructured surface were formed of the estimated dimension of 100 nm pattern. The patterns were made of 5-10 parallel lines. The dimension of each parallel lines were 100nm in breadth and 3 nm in depth which is the dimension of the tip of AFM. As these dimension of the patterns are even lesser than the dimension of an animal cells(in microns),during cell culture it may overlap the space and pattern could not be observable under the microscope . But cell behaves in different way and changes its morphology and motility in the nanochannel as stated above.Thus, the fluidic system will give the idea how the cells are behaving in such nanostructures. This behavior will be manipulated in miniaturization of devices..The biodegradable polymer PHBV was taken so that it can be extended for the further studies of the cell behavior.The films were separated from the cover slips, sterilized in UV radiation and stored for the further study
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CONCLUSION Nanopatterned formed were of 100nm in breadth and 3 nm in depth as clearly visible in the above AFM images. Nanolithographic technology has made nanoscale engineering a powerful tool for tissue engineering and biological applications. The nanolithography technique and engineered biomaterials are being used for tissue engineering in a variety of applications: for example, by fabricating scaffolds with control over features such as shape and pore architecture, as templates for microtissue formation, or as improved bioreactors. Thus it is possible to use the capability of AFM to sense interaction forces between atoms at the end of the probe and in the specimen to study the weak, non-covalent, usually short-range forces involved in molecular recognition reactions. It has great role to play in nanotechnology for characterization and fabrication of intelligent patterned surface. It has been proved that nanopatterning through AFM lithography is cheap, fast and reliable.
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PUBLICATIONS: as first Author 1. Growth and characterization of ZnO Nanotetrapod for the Biosensor Application- Md. N. Ali, K. Kant, S. Kaur, M. Bharti (International journal of natural and engineering sciences ,page69-70,volume 3,number 2,May 2009) 2.Toxicity assay of MWCNT (multi walled carbon nanotube)-M.N. Ali, K.Dinda,S.Mitra, L.G. Devi , C.K.Prashant(International journal of natural and engineering sciences ,page65-68,volume 3,number 2,May 2009) 3. Thermosensitive Nanoparticles of PNIPAM-co-PMMA and PNIPAM- M.Naushad Ali , M.Sengonul , N. Hasirci and V. Hasirci (4th national bioengineering congress, Turkey, 15-18 October, 2008) 4. Toxicity assay of polymeric nanoparticles for therapeutic applications - M.Naushad Ali , Nihan Ozturk , N. Hasirci and V. Hasirci (Indian Pharmaceutical Congress,New Delhi,India 12-14 December, 2008) 5. Toxicity assay of MWCNT (multi walled carbon nanotube)- Md.Naushad Ali, A.K.Dinda, Susmita Mitra ,L.Geeta Devi , C.K.Prashant (4th national bioengineering congress,Turkey 15-18 october,2008) 6. Growth and characterization of ZnO Nanotetrapod for the Biosensor Application- M. Naushad Ali, M. Kaur, Meetu Bharti and K. Kant (4th national bioengineering congress,Turkey 15-18 october,2008)
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Summary INSTRUMENTAL SKILLS acquired in relation to material science, life science, pharmaceutical science and nanoscience viz-Synthesis, characterization & functionalization of nanomaterials (nanopolymers& CNT). TEM/SEM, AFM, Vapor Coating Unit (metallization), Contact angle measurement system, Micro incubator, Confocal Microscopy, Student`s Organ Bath, Molecular biology techniques (IHC, MTT/MTS, Tissue culture, in vitro studies etc) Pharmaceutical technology, electrophoresis, UV-VIS Spectrophotometer, FTIR (ATR) Nanotoxicology became area of my thrust Great enthusiasm developed to acquire and be trained on nanotoxicology research Willing to serve the entire human kind and participate to build up healthy international harmonization strategy
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OUR GROUP MEMBERS
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Acknowledgement European Commission (Marie Curie Research Training Network for the project Biopolysurf [Project Number MRTN-CT-2004-005516] is highly acknowledged for the financial support UPDATED ON 31.08.2008
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