NANOSCIENCE AND NANOTECHNOLOGY IN MEDICINE : Pre-clinical studies on targeted tumour therapy with iron oxide...
NANOTECHNOLOGYNanotechnology is the creation of USEFUL/FUNCTIONAL materials, devicesand systems (of any useful size) throu...
Unique Properties of Nanoscale Materials• Quantum size effects result in unique mechanical, electronic, photonic, and...
Source: Nanoscale Materials in Chemistry, Ed. K.J. Klabunde, Wiley, 2001
• Spherical iron nanocrystals• J. Phys. Chem. 1996, Vol. 100, p. 12142
• Adsorption is like absorption except the adsorbed material is heldnear the surface rather than inside• In bulk solids, a...
S N CF3 CH 2 CH 2 - ...
Three different cells from three different tissues of mice : Spleen, liver and fibrosarcoma tumor were incubated w...
NANOTECHNOLOGY APPLICATIONSMedicine/Health Care Energy ...
MEDICAL APPLICATIONS• Cancer Nanotechnology (i) Dignosis using Quantum Dots (ii) Tumor Targeted Delivery (iii) Imagin...
NANOTECHNOLOGY IN DRUG DELIVERY - ADVANTAGES• Prevention of drug from biological degradation• Effective Targeting• Patien...
Biosensor
• Probe molecules for a given target can be attachedto CNT tips for biosensor development• Electrochemical approach: requi...
Nanoscale electrodes create a dramatic improvement in signal detection over traditional electrodes ...
Electrochemical Detection of DNA Hybridization - by AC Voltammetry ...
300 µm 200 µm30 dies on a 4” Si wafer Potential applications: (1) La...
Target Molecule1. Chen, G.Y., Thundat, T. Wachter, E. A., Warma...
Self-Assembly of ssDNAThiolated ssDNA Au ...
Probe ssDNA Target ssDNAWu, G. et al. “Origin of nanomechanical cantilever motion generated from biomolecu...
200 PSA [BSA] = 1 mg/ml ...
DNA microarrays can be manufactured by:• Photolitography (Affymetrix, Febit, Nimblegen)• Inkjet (Agilent, Canon)• Robot s...
DNA MICROARRAYS CONSIST OF 100 - 1 MILLION DNA PROBES ATTACHED TOA SURFACE OF 1 CM BY 1 CM (CHIP).By hybridisation, they c...
FabricationFabrication via Printing DNA sequence stuck to glass substrate DNA solution pre- synthesized in the lab...
DNA Microarrays• Each probe consists of thousands of strands of identical oglionucleotides – The DNA sequences at each...
PRACTICAL APPLICATION OF DNA MICROARRAYS• DNA Microarrays are used to study gene activity (expression) – What protei...
The Process Poly-A 10% Biotin-labeled UracilCells cDNA...
Hybridization and Staining GeneChip Biotin Hybridized Array ...
DNA SequencingUsing Nanopores Goal: Very rapid gene sequencing
- Nanopore in membrane- DNA in buffer- Voltage clamp (~2nm diameter)- ...
• When there is no DNA translocation, there is a background ionic currentOpen nanopore • Whe...
After a decade of using protein pores,efforts are underway in many groups todevelop synthetic pores (such as in Si3N4) •...
nanopore chip Voltage Clamp ...
Spontaneous Blocking Events with Smaller NASA Pores +200 mV 100 pA0.5 s
C C C C TT G ...
• Tree-like polymers, branching out from a central core and subdividing into hierarchical branching units - Not more...
• Desired features of effective drug delivery - Targeted delivery, controlled release (either timed or in response t...
Future Possiblities: Oxygen Selective Pump http://www.foresight.org/Nanomedicine/
Respirocytes: A Mechanical Artifical Red Blood Cell•Bloodborne spherical 1-micron diamondoid 1000-atm pressure v...
Cancer• Cancer is one of the most common diseases in the developed world:• 1 in 4 deaths are due to cancer• 1 in 17 death...
Cancer Defined as the disturbance ofgrowth characterized by excessiveproliferation of cells without apparentrelation to t...
CancerNewspapers, magazines, radio, and television are reportingdiscoveries and breakthroughs attributing one form of canc...
Tobacco Alcohol Lack of Nutrients RISK FACTORS FOR CANCEREnvironme...
Ten Leading Sites of Cancer (1998) AAR SITE SITE AAR10.04 Lung Cervix Uteri 26.118.02 ...
900 ...
CANCER INCIDENCE & MORTALITY WORLD WIDE
Year wise total cancer prevalence in India[ICMR, 2006; ICMR, 2009].
Cancer prevalence in five metropolitan cities of India[Marimuthu, Projection of cancer incidence in five cities and cancer...
CANCER – THE SECOND LEADING CAUSE OF DEATHWorld• Every year- 10 million diagnosed 6 million dieWorldwide - Lu...
India3.4 % of all deaths – cancer7 lakh new cases detected every yearMales - mouth/oropharynx oesophagus ...
Oral cancer• 50 – 70% of all cancers diagnosed in india• Risk factors Tobacco Alcohol Precanc...
Cancer cervix• Most common cancer of women –developing countries• Causative agent - HPV• Risk Factors• Prevention and con...
Stomach CancerLung cancer• Most common cancer in the • Worlds second most world common cancer• ...
CAUSES OF CANCEREnvironmental Genetic• Tobacco Eg• Alcohol ...
Dr Suwas Darvekar
Common sites of oral cancerThe most common sites of the oral cancer is the tongue and the floor of themouth. The other com...
CANCER CONTROLPrimary Prevention Secondary Prevention • Control of tobacco & • Cancer Registration a...
Danger Signals• A lump or a hard area in breast• A change in wart or mole• A persistent change in bowel habits• A persiste...
Cancer Screening• Pre malignant lesion can be identified• Most cancers are localized in initial stages• 75% occur...
• Screening of cancer cervix – Pap Smear• Screening of breast cancer Breast self examination(BSE) Palpatio...
Oral cancer• 50 – 70% of all cancers diagnosed in india• Risk factors Tobacco Alcohol Precanc...
Cancer cervix• Most common cancer of women –developing countries• Causative agent - HPV• Risk Factors• Prevention and con...
Stomach CancerLung cancer• Most common cancer in the • Worlds second most world common cancer• ...
What causes cancer?• Cancer arises from the mutation of a normal gene.• Mutated genes that cause cancer are called oncog...
Carcinogens• Ionising radiation – X Rays, UV light• Chemicals – tar from cigarettes• Virus infection – papilloma virus can...
GROWTH OF NORMAL CELLS• Growth means size increase and proliferation• • Not all adult cells can proliferate• • Special res...
CAUSES OF CANCER• DNA Mutations– Radiation – other environmental (tobacco, alcohol,radon, asbestos, etc)– Random somatic m...
INHERITED CANCERS - a small percentage of manycancers• Breast cancer– ~3% cases between 36 and 45 years ofagehave a BR...
Types of genes which may mutate to cause cancer: • Tumour suppressor ...
cancer is a disease of the cell cycle
Tumour Growth or neoplasiaPathways that control colorectal tumorigenesis. Mutations in the APC/b-cateninpathway initiate t...
Hallmarks of CancerSix changes for cancer –found in most, if not all, cancers1. Self-sufficiency in growth signals2. Insen...
Major types of cancers• Over 200 types of cancer are known grouped into major categories Carcinomas Sarcomas Melano...
Tumour suppressor genes• The gene’s normal function is to regulate cell division. Both alleles need to be mutated or re...
oncogenes• Cellular oncogene c-onc• Viral oncogene v-onc• Proto-oncogene, activated by mutation to c-onc
Proto-oncogene activation
Types of proto-oncogene• Growth factor e.g. SIS oncogene (PDGF)
Types of proto-oncogene• Nuclear transcription factors e.g. MYC
p53• suppresses progression through the cell cycle in response to DNA damage• initiates apoptosis if the damage ...
Transformation is a multistep process
Transformation is a multistep process
The environment:Some environmental agents associated with cancer are:• Viruses• Tobacco smoke• Food• Radiation• Chemicals...
Viruses Viruses—mostly in the form of DNA viruses—have been causally linked to cancer.• human papillomaviruses—...
MODALITIES OF CANCER TREATMENT LOCAL THERAPEUTIC MODALITIES SURGERY RADIOTHERAPY SYSTEMIC FORMS OF TREATMENT CHEMOTHE...
Cost of cancer per year in U.S.
Targeted drug delivery to tumour using magnetic nanoparticles:Preclinical studies with Fe3O4 – doxorubicin nanopart...
Nanoparticles for targeting drugs to tumour- Fe2O3 nanoparticles coated with polymer –polyoxyethylene 25 propylene glycol ...
Dose-Effect Curve Desirable 100 Dose Range 80% with 60Maximal Effect 40 ...
DOXYRUBICIN. HCL – MWT 580
Doxorubicin (Adriamycin) intercalation of DNA Prevent DNA replication ...
Cancer chemotherapy - the agent exerts anticancer action through cytotoxic mechanismsCommon problems encountered: – Poor...
• Antitumor activity of water dispersible Fe3O4 nanoparticles (coated with Poly Vinyl Pyrolidone (PVP) and Poly oxy ethy...
Water dispersible Fe3O4 nanoparticlescarring doxorubicin for cancer therapyO.D.Jayakumar, R...
• XRD pattern of Fe3O4 nanoparticles coated ...
TEM images of Fe3O4-PVP- POES nanoparticles and ...
Water dispersible Fe3O4 Nanoparticles Carrying Doxorubicin for Cancer Therapy: Journal of Nanoscienceand Nanotechnology, 2...
Cytotoxic effect on DLA cells Fe3O4-PVP-POES-Doxo ...
Induction of apoptosis Apoptotic index of Fe3O4-PVP-POES- Doxorubicin was higher than that of doxorubicin or Fe3...
Effect on tumor growth • Daltons Lymphoma solid tumor on hind limbs of mice. • The treatments were started on the ...
Effect on tumor growth • The growth of the ...
Effect on cardio toxicity • Decrease in GSH - ...
Effect on cardio toxicity • Increase in the MDA ...
OXIDATIVE ENXYME THERAPY USING MAGNETIV NANOPARTICLESD-Alanine --- (DAO) ------------→ Pyruvic acid + H2O2 FTIR spectra of...
OXIDATIVE ENXYME THERAPY OF TUMOUR USING MAGNETIV NANOPARTICLESEffect of administration of ...
COMET ASSAY OF DLA CELLS TREATED WITH FE2O3-DAO AND D-ALANINE Apoptoic index in dla cells treated ...
Conclusions• Doxorubicin and the enzyme DAO can be complexed to magnetic Fe3O4 nanoparticles.• This comple...
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  • 1. NANOSCIENCE AND NANOTECHNOLOGY IN MEDICINE : Pre-clinical studies on targeted tumour therapy with iron oxide nanoparticles C.K.K.Nair Dean of Research, Pushpagiri Institute of Medical Sciences and Research Centre, Tiruvalla 689101.S ymp o s i um o n Nanoscience and Nanotechnology: Fundamentals andRecent Trends o n 1 5 F e b r u a r y 2 0 1 3
  • 2. NANOTECHNOLOGYNanotechnology is the creation of USEFUL/FUNCTIONAL materials, devicesand systems (of any useful size) through control/manipulation of matter on thenanometer length scale and exploitation of novel phenomena and propertieswhich arise because of the nanometer length scale: • PhysicalNanometer • Chemical• One billionth (10-9) of a meter • Electrical• Hydrogen atom 0.04 nm • Mechanical• Proteins ~ 1-20 nm • Optical• Feature size of computer chips 90 nm • Magnetic (in 2005) •• Diameter of human hair ~ 10 µm •
  • 3. Unique Properties of Nanoscale Materials• Quantum size effects result in unique mechanical, electronic, photonic, and magnetic properties of nanoscale materials• Chemical reactivity of nanoscale materials greatly different from more macroscopic form, e.g., gold• Vastly increased surface area per unit mass, e.g., upwards of 1000 m2 per gram• New chemical forms of common chemical elements, e.g., fullerenes, nanotubes of carbon, titanium oxide, zinc oxide, other layered compounds
  • 4. Source: Nanoscale Materials in Chemistry, Ed. K.J. Klabunde, Wiley, 2001
  • 5. • Spherical iron nanocrystals• J. Phys. Chem. 1996, Vol. 100, p. 12142
  • 6. • Adsorption is like absorption except the adsorbed material is heldnear the surface rather than inside• In bulk solids, all molecules are surrounded by and bound toneighboring atoms and the forces are in balance. Surface atoms arebound only on one side, leaving unbalanced atomic and molecularforces on the surface. These forces attract gases and molecules ⇒Van der Waals force, ⇒ physical adsorption or physisorption• At high temperatures, unbalanced surface forces may be satisfied byelectron sharing or valence bonding with gas atoms ⇒ chemicaladsorption or chemisorption - Basis for heterogeneous catalysis (key to production offertilizers, pharmaceuticals, syntheticfibers, solvents, surfactants,gasoline, other fuels, automobile catalytic converters…) - High specific surface area (area per unit mass)
  • 7. S N CF3 CH 2 CH 2 - - Cl Cl α ω CH 2 N+ N+ CH3 H H TrifluoperazineAntidepressent and antipsychotic drug. It is known to givesevere side effects such as photosensitization of skin andeye tissues TFP pK 4 and 8 TFP+ a Dimensions:TFP 25 ÅTFP+ SiO2 TFP+ SiO2 60 Å TFP+ Biophysical Chemistry, 109, 113-119, 2004
  • 8. Three different cells from three different tissues of mice : Spleen, liver and fibrosarcoma tumor were incubated with TFP and TFP/SiO2Spleen ce llstreated with TFP/SiO2 Biophysical Chemistry, 109, 113-119, 2004.
  • 9. NANOTECHNOLOGY APPLICATIONSMedicine/Health Care Energy • More Efficient And Cost•Cancer treatment Effective Technologies•Bone treatment For Energy Production•Dentistry − Solar Cells − Fuel Cells•Drug delivery − Batteries•Appetite control − Bio Fuels•Drug development•Tissue engineering/ Consumer Goods•Regenetaive medicine • Foods and beverages −Advanced packaging materials, sensors, and•Medical tools lab-on-chips for food quality testing•Diagnostic • Appliances and textiles•Imaging −Stain proof, water proof and wrinkle free textilesInformation Technology • Household and cosmeticsSmaller, faster, more − Self-cleaning and scratch free products, energy efficient and paints, and better cosmetics powerful computing and other IT-based systems
  • 10. MEDICAL APPLICATIONS• Cancer Nanotechnology (i) Dignosis using Quantum Dots (ii) Tumor Targeted Delivery (iii) Imaging (iv) Cancer Gene Therapy• DNA Vaccines for parasitic, bacterial and viral diseases• Oral and pulmonary routes for systemic delivery of proteins and peptides• Nanotechnology in Tissue Engineering
  • 11. NANOTECHNOLOGY IN DRUG DELIVERY - ADVANTAGES• Prevention of drug from biological degradation• Effective Targeting• Patient Compliance• Cost effectiveness• Product life extension
  • 12. Biosensor
  • 13. • Probe molecules for a given target can be attachedto CNT tips for biosensor development• Electrochemical approach: requires nanoelectrodedevelopment using PECVD grown vertical nanotubes• The signal can be amplified with metal ion [(P ] u ) Rby 2 3 +mediator oxidation catalyzed by Guanine. 2+ • High specificity 2+ • Direct, fast 3+ response • High sensitivity • Single molecule and cell 3+ signal e capture and detection
  • 14. Nanoscale electrodes create a dramatic improvement in signal detection over traditional electrodes Traditional Macro- or NanoelectrodeElectrode Micro- Electrode Array Insulator Nano- • Scale difference between • CNT tips are at the scale Electrode macro-/micro- electrodes and close to molecules molecules is tremendous • Dramatically reduced • Background noise on electrode background noise surface is therefore significant • Multiple electrodes results in magnified • Significant amount of target signal and desired redundance for molecules required statistical reliability. • Can be combined with other electrocatalytic mechanism for magnified signals.
  • 15. Electrochemical Detection of DNA Hybridization - by AC Voltammetry 1st #1-#2 2nd and 3rd #2-#3 1st, 2nd, and 3rd scan in AC voltammetry 1st – 2nd scan: mainly DNA signal 2nd – 3rd scan: BackgroundLower CNT Density ⇒ Lower Detection Limit J. Li, H.T. Ng, A. Cassell, W. Fan, H. Chen, J. Koehne, J. Han, M. Meyyappan, NanoLetters, 2003, Vol. 3, p. 597.
  • 16. 300 µm 200 µm30 dies on a 4” Si wafer Potential applications: (1) Lab-on-a-chip applications (2) Early cancer detection (3) Infectious disease detection (4) Environmental monitoring (5) Pathogen detection
  • 17. Target Molecule1. Chen, G.Y., Thundat, T. Wachter, E. A., Warmack, R. A., “Adsorption-induced surface stress and its effects on resonance frequency of microcantilevers,” J. Appl. Phys 77, pp. 3618-3622 (1995).2. Ratierri, R. et al., “Sensing of biological substances based on the bending of microfabricated cantilevers,” Sensors and Actuators B 61, 213-217 (1999).3. Fritz, J. et al. “Translating Biomolecular Recognition into Nanomechanics,” Science 288, 316-318 (2000).4. Wu, G. et al. “Origin of nanomechanical cantilever motion generated from biomolecular interactions,” PNAS 98(4), 1560-1564 (2001).
  • 18. Self-Assembly of ssDNAThiolated ssDNA Au - -+ -5’-HS ATCCGCATTACGTCAATC + - -+ + + -- + TAGGCGTAATGCAGTTAG-5’ (Complementary Strand) - + PB = Sodium Phosphate Buffer Wu, G. et al. “Origin of nanomechanical cantilever motion generated from biomolecular interactions,” PNAS 98(4), 1560-1564 (2001).
  • 19. Probe ssDNA Target ssDNAWu, G. et al. “Origin of nanomechanical cantilever motion generated from biomolecular interactions,” PNAS 98(4), 1560-1564 (2001).
  • 20. 200 PSA [BSA] = 1 mg/ml [fPSA] 150 60 µg/ml Analyte Deflection, h [nm] 6µg/mlRabbit Anti- 100 InjectionsHuman PSA 50 60 ng/ml DTSSP Au 6 ng/ml 0 SiNx No PSA Ab No fPSA ([fPSA] = 60 µg/ml) Glass -50 0 60 120 180 240 300 Time [min] HSA: Human Serum Albumin 80 [f P S A ] [H S A ] = 1 m g / m l 60 HP: Human Plasminogen 60 n g/m l Deflection, h [nm] 40 fPSA: free PSA I n je c t io n s 6 n g /m l 20 cPSA: complex PSA 0 Wu, G. et al., “Bioassay of Prostate Specific -20 No PSA Ab H P o n ly No PSA µ Antigen (PSA) Using Microcantilevers,” ( [f P S A ] = 6 0 g /m l) ( [ H P ] = 1 m g /m l) -40 Nature Biotechnology (Sept., 2001) 0 60 120 1 80 240 T im e [m in ]
  • 21. DNA microarrays can be manufactured by:• Photolitography (Affymetrix, Febit, Nimblegen)• Inkjet (Agilent, Canon)• Robot spotting (many providers)Nanochip− A lab on chip integrates one or more laboratory operation on a single chip− Provides fast result and easy operation− Applications: Biochemical analysis (DNA/protein/cell analysis) and bio-defense
  • 22. DNA MICROARRAYS CONSIST OF 100 - 1 MILLION DNA PROBES ATTACHED TOA SURFACE OF 1 CM BY 1 CM (CHIP).By hybridisation, they can detect DNA or RNA: If the hybridised DNA or RNA is labelled fluorescently it can be quantified by scanning of the chip.
  • 23. FabricationFabrication via Printing DNA sequence stuck to glass substrate DNA solution pre- synthesized in the labFabrication In Situ Sequence “built” Photolithographic techniques use light to release capping chemicals 365 nm light allows 20-µm resolution
  • 24. DNA Microarrays• Each probe consists of thousands of strands of identical oglionucleotides – The DNA sequences at each probe represent important genes (or parts of genes)• Printing Systems – Ex: HP, Corning Inc. – Printing systems can build lengths of DNA up to 60 nucleotides long – 1.28 x 1.28+ cm glass wafer GeneChip • Each “print head” has a ~100 µm diameter and are separated by ~100 µm. (≈ 5,000 – 20,000 probes)• Photolithographic Chips – Ex: Affymetix – 1.28 x 1.28 cm glass/silicon wafer • 24 x 24 µm probe site (≈ 500,000 probes) – Lengths of DNA up to 25 nucleotides long – Requires a new set of masks for each new array type
  • 25. PRACTICAL APPLICATION OF DNA MICROARRAYS• DNA Microarrays are used to study gene activity (expression) – What proteins are being actively produced by a group of cells? • “Which genes are being expressed?”• How? – When a cell is making a protein, it translates the genes (made of DNA) which code for the protein into RNA used in its production – The RNA present in a cell can be extracted – If a gene has been expressed in a cell • RNA will bind to “a copy of itself” on the array • RNA with no complementary site will wash off the array – The RNA can be “tagged” with a fluorescent dye to determine its presence• DNA microarrays provide a high throughput technique for quantifying the presence of specific RNA sequences
  • 26. The Process Poly-A 10% Biotin-labeled UracilCells cDNA RNA Antisense cRNA IVT AAAA L L L (In-vitro Transcription) Fragment (heat, Mg2+) Labeled fragments Hybridize Wash/stain Scan L L L
  • 27. Hybridization and Staining GeneChip Biotin Hybridized Array Labeled cRNA L L L L L + L L + L L LL L SAPE Streptavidin- phycoerythrin
  • 28. DNA SequencingUsing Nanopores Goal: Very rapid gene sequencing
  • 29. - Nanopore in membrane- DNA in buffer- Voltage clamp (~2nm diameter)- Measure currentα-hemolysin pore (very first, natural pore) Axial View Side View
  • 30. • When there is no DNA translocation, there is a background ionic currentOpen nanopore • When DNA goes through the pore, there is a drop in the background signal • The goal is to correlate the extent and duration of the drop in the signal to the individual nucleotides DNA translocation event
  • 31. After a decade of using protein pores,efforts are underway in many groups todevelop synthetic pores (such as in Si3N4) • Interaction with single nuclotides - ~20 nucleotides in αHL simultaneously • Slower translocation - 1-5 µs /nucleotide in αHL • Resistance to extreme conditions - Temperature - pH - Voltage • α - hemolysin is toxic and hard to work with
  • 32. nanopore chip Voltage Clamp Amplifier AgCl AgCl KCl KCl Data Acquisition• Voltage-clamp amplifier designed to measure pA level currents• Fast (up to 1GHz) data acquisition• Software for automatic blocking event detection and recording
  • 33. Spontaneous Blocking Events with Smaller NASA Pores +200 mV 100 pA0.5 s
  • 34. C C C C TT G A G A G G A A A GG A GPresent Future
  • 35. • Tree-like polymers, branching out from a central core and subdividing into hierarchical branching units - Not more that 15 nm in size, Mol. Wt very high - Very dense surface surrounding a relatively hollow core (vs. the linear structure in Courtesy of: http://www.uea.ac.uk/cap/wmcc/anc.htm traditional polymers)• Dendrimers consist of series of chemical shells built on a small core molecule - Surface may consist of acids or amines ⇒ means to attach functional Groups ⇒ control/modify properties - Each shell is called a generation (G0, G1, G2….) - Branch density increases with each generation - Contains cavities and channels ⇒ can be used to trap guest molecules for various applications.
  • 36. • Desired features of effective drug delivery - Targeted delivery, controlled release (either timed or in response toan external signal)• Desirable characteristics of dendrimers - Uniform size - Water Solubility - Modifiable surface functionality - Availability of internal cavity - Control of molecular weight - Control of the surface and internal structure• Number of different drugs can be encapsulated in dendrimers and injectedinto the body for delivery - Incorporating sensors would allow release of drugs where needed• Gene Therapy - Current problem is getting enough genes into enough cells to make a difference. Using viruses for this triggers immune reactions. Dendrimersprovide an alternative without triggering immune response• Cancer Therapy; Antimicrobial and Antiviral Agents
  • 37. Future Possiblities: Oxygen Selective Pump http://www.foresight.org/Nanomedicine/
  • 38. Respirocytes: A Mechanical Artifical Red Blood Cell•Bloodborne spherical 1-micron diamondoid 1000-atm pressure vessel•Active pumping powered by endogenous serum glucose•Able to deliver 236 times more oxygen to the tissues per unit volumethan natural red cells and to manage carbonic acidity http://www.foresight.org/Nanomedicine/Respirocytes.html
  • 39. Cancer• Cancer is one of the most common diseases in the developed world:• 1 in 4 deaths are due to cancer• 1 in 17 deaths are due to lung cancer• Lung cancer is the most common cancer in men• Breast cancer is the most common cancer in women• There are over 100 different forms of cancer• The division of normal cells is precisely controlled. New cells are only formed for growth or to replace dead ones.• Cancerous cells divide repeatedly out of control even though they are not needed, they crowd out other normal cells and function abnormally. They can also destroy the correct functioning of major organs.
  • 40. Cancer Defined as the disturbance ofgrowth characterized by excessiveproliferation of cells without apparentrelation to the physiological demand of the organ involved.
  • 41. CancerNewspapers, magazines, radio, and television are reportingdiscoveries and breakthroughs attributing one form of canceror another to a specific gene.Cancer of the breast, colon, prostate, and many other sites inthe body are being connected to specific genes... But themeaning of this isnt always clear.Cancer is one of the most common and severe problems of clinicalmedicine. Cancer is not a single disease but rather a name applied to agreat variety of malignant tumors that are formed by the same basicprocess of uncontrolled growth. Cell proliferation results in a mass thatinvades neighboring tissues and may metastasize to more distant sites.Some cancers, however, such as blood cancers, do not form tumors.Many aspects of cell function are controlled by a balance of positive andnegative signals received from inside and outside the cell. In normaltissues, there is a balance between cell proliferation and cell death. Intumor, this balance is lost.CANCER HAS BOTH GENETIC AND ENVIRONMENTAL CAUSES.
  • 42. Tobacco Alcohol Lack of Nutrients RISK FACTORS FOR CANCEREnvironmental Factors Sociodemographic Factors
  • 43. Ten Leading Sites of Cancer (1998) AAR SITE SITE AAR10.04 Lung Cervix Uteri 26.118.02 BOT Breast 19.375.8 Py.Fossa Esophagus 4.525.75 Esophagus Ovary 4.154.88 Larynx Myel.Leuk 3.214.15 Oth. Mouth Vagina 2.033.7 Myel.Leuk Hypopharyx 22.8 Hypopharyx Oth. Mouth/ 1.68 Brain2.74 Tonsil Lung/Other 1.51 Tongue2.59 Brain Lymp.Leuk 1.43
  • 44. 900 876.7 Age Specific Incidence Rates/lakh800 Population for all Sites 753.3 767.9700 1998600 508.1500 479.7 416.4 406.3400 372.8 365.5 318.3300 288.8 292.8 245.4 217.7200 189 178.2 149.4 102.2100 76.2 64.5 38 37 10.8 3.5 9.2 4.6 10.3 8.2 8.5 13.6 12 2220.8 5.5 0 0- 5- 20- 25- 45- 50- 70- 75- 10- 15- 30- 35- 40- 55- 60- 65- 80+ MALE FEMALE
  • 45. CANCER INCIDENCE & MORTALITY WORLD WIDE
  • 46. Year wise total cancer prevalence in India[ICMR, 2006; ICMR, 2009].
  • 47. Cancer prevalence in five metropolitan cities of India[Marimuthu, Projection of cancer incidence in five cities and cancer mortality in India. Indian J Cancer 45, 4-7.2008].
  • 48. CANCER – THE SECOND LEADING CAUSE OF DEATHWorld• Every year- 10 million diagnosed 6 million dieWorldwide - Lung Cancer (12.3 %) Breast Cancer(10.4%) Colorectal Cancer(9.4%)Death from cancer - Lung (17.8%) Stomach (10.4%) Liver (8.8 %)
  • 49. India3.4 % of all deaths – cancer7 lakh new cases detected every yearMales - mouth/oropharynx oesophagus stomach Lower respiratory tractFemales - Cervix Breast mouth/oropharynx oesophagus
  • 50. Oral cancer• 50 – 70% of all cancers diagnosed in india• Risk factors Tobacco Alcohol Precancerous lesions Cultural patternsPrevention
  • 51. Cancer cervix• Most common cancer of women –developing countries• Causative agent - HPV• Risk Factors• Prevention and controlBreast Cancer• commonest cause of death of middle aged women-developed countries (35 – 50 yrs)• Risk factors
  • 52. Stomach CancerLung cancer• Most common cancer in the • Worlds second most world common cancer• Risk factors • Risk Factors• Prevention • prevention
  • 53. CAUSES OF CANCEREnvironmental Genetic• Tobacco Eg• Alcohol • Retinoblastoma in Children• Dietary Factors • Leukemia in Mongols• Occupational exposures• Viruses• Parasites• Customs ,habits, Lifestyles• Others – sunlight, pollution, drugs
  • 54. Dr Suwas Darvekar
  • 55. Common sites of oral cancerThe most common sites of the oral cancer is the tongue and the floor of themouth. The other common sites are buccal vestibule, buccal mucosa, gingivaand rarely hard and soft palate. Cancer of bucco-pharyngeal mucosa iscommon in smokers.CANCERS OF Tongue, Cheek, Subbuccal mucosa, Buccal vestibulePRECANCEROUS LESIONSLuekoplakia Erythroplakia Sub mucous fibrosis
  • 56. CANCER CONTROLPrimary Prevention Secondary Prevention • Control of tobacco & • Cancer Registration alcohol Hospital based registries consumption Population based registries • Personal Hygiene • Early detection of cases • Radiation • Treatment • Occupational Exposures • Immunisation • Foods & drugs • Air pollution • Treatment of precancerous lesion • Legislation • Cancer Education
  • 57. Danger Signals• A lump or a hard area in breast• A change in wart or mole• A persistent change in bowel habits• A persistent cough or hoarseness• Excessive loss during menstrual periods or loss of blood outside usual dates.• Blood loss from any natural orifice• A swelling that does not get better• Unexplained loss of weight
  • 58. Cancer Screening• Pre malignant lesion can be identified• Most cancers are localized in initial stages• 75% occurs at accessible body sites Methods • Mass Screening by comprehensive cancer detection examination • Mass Screening at single sites • Selective screening - for those at special risk
  • 59. • Screening of cancer cervix – Pap Smear• Screening of breast cancer Breast self examination(BSE) Palpation Thermography MammographyScreening of lung cancer Chest radiograph Sputum Cytology
  • 60. Oral cancer• 50 – 70% of all cancers diagnosed in india• Risk factors Tobacco Alcohol Precancerous lesions Cultural patternsPrevention
  • 61. Cancer cervix• Most common cancer of women –developing countries• Causative agent - HPV• Risk Factors• Prevention and controlBreast Cancer• commonest cause of death of middle aged women-developed countries (35 – 50 yrs)• Risk factors
  • 62. Stomach CancerLung cancer• Most common cancer in the • Worlds second most world common cancer• Risk factors • Risk Factors• Prevention • prevention
  • 63. What causes cancer?• Cancer arises from the mutation of a normal gene.• Mutated genes that cause cancer are called oncogenes.• It is thought that several mutations need to occur to give rise to cancer• Cells that are old or not functioning properly normally self destruct and are replaced by new cells.• However, cancerous cells do not self destruct and continue to divide rapidly producing millions of new cancerous cells.
  • 64. Carcinogens• Ionising radiation – X Rays, UV light• Chemicals – tar from cigarettes• Virus infection – papilloma virus can be responsible for cervical cancer.• Hereditary predisposition – Some families are more susceptible to getting certain cancers. Remember you can’t inherit cancer its just that you maybe more susceptible to getting it.
  • 65. GROWTH OF NORMAL CELLS• Growth means size increase and proliferation• • Not all adult cells can proliferate• • Special reserve cells retain proliferation potenitial• – Embryonic stem cells can make any cell in the body• – Although, many stem cells are committed and have limited potential. i.e. can produce all the intestinal epithelial cells.• – Proliferation requires the cell cycle• G0,G1,S,G2, and M phase
  • 66. CAUSES OF CANCER• DNA Mutations– Radiation – other environmental (tobacco, alcohol,radon, asbestos, etc)– Random somatic mutations– Inherited germ line mutations• Genetic predisposition-– Rb, p53, APC, CDKN2A, BRCA1, BRCA2» Will discuss these later in a pathway context• Infectious agents– Viral• HPV – cervical cancer• Hepatitis – liver cancer– Vaccines have been developed and are extremelyeffective – not available– Bacterial• H. pylori – stomach cancer
  • 67. INHERITED CANCERS - a small percentage of manycancers• Breast cancer– ~3% cases between 36 and 45 years ofagehave a BRCA1 mutation– ~3% cases between 36 and 45 years ofage have a BRCA2 mutation• 1/500 people have a BRCA1 mutation–There may be other breast cancer genes– and many cancers are random
  • 68. Types of genes which may mutate to cause cancer: • Tumour suppressor genes • oncogenes • DNA repair genes • telomerase • p53
  • 69. cancer is a disease of the cell cycle
  • 70. Tumour Growth or neoplasiaPathways that control colorectal tumorigenesis. Mutations in the APC/b-cateninpathway initiate the neoplastic process, resulting in small benign tumors (adenomas).These tumors progress, becoming larger and more dangerous, as mutations in othergrowth-controlling pathway genes (such as K-Ras, B-RAF, PIK3CA, or p53)accumulate. The process is accelerated by mutations in stability genes. The top lineindicates potential clinical applications of knowledge of these pathways.
  • 71. Hallmarks of CancerSix changes for cancer –found in most, if not all, cancers1. Self-sufficiency in growth signals2. Insensitivity to growth-inhibitory signals3. Evasion of apoptosis4. Limitless replicative capacity5. Sustained angiogenesis6. Tissue invasion and metastasis
  • 72. Major types of cancers• Over 200 types of cancer are known grouped into major categories Carcinomas Sarcomas Melanomas Teratomas Leukemias and Lymphomas
  • 73. Tumour suppressor genes• The gene’s normal function is to regulate cell division. Both alleles need to be mutated or removed in order to lose the gene activity.• The first mutation may be inherited or somatic.• The second mutation will often be a gross event leading to loss of heterozygosity in the surrounding area.
  • 74. oncogenes• Cellular oncogene c-onc• Viral oncogene v-onc• Proto-oncogene, activated by mutation to c-onc
  • 75. Proto-oncogene activation
  • 76. Types of proto-oncogene• Growth factor e.g. SIS oncogene (PDGF)
  • 77. Types of proto-oncogene• Nuclear transcription factors e.g. MYC
  • 78. p53• suppresses progression through the cell cycle in response to DNA damage• initiates apoptosis if the damage to the cell is severe• acts as a tumour suppressor• is a transcription factor and once activated, it represses transcription of one set of genes (several of which are involved in stimulating cell growth) while stimulating expression of other genes involved in cell cycle control
  • 79. Transformation is a multistep process
  • 80. Transformation is a multistep process
  • 81. The environment:Some environmental agents associated with cancer are:• Viruses• Tobacco smoke• Food• Radiation• Chemicals• Pollution
  • 82. Viruses Viruses—mostly in the form of DNA viruses—have been causally linked to cancer.• human papillomaviruses—primarily types 16 and 18, which are sexually transmitted—have been linked to cervical cancer;• more than 25 other types of papillomaviruses have been linked to cancer as well• hepatitis B and C—linked to cancer of the liver• human immunodeficiency virus (HIV) —linked to Kaposis sarcoma and lymphoma• retroviruses—linked to cancers in animals other than humans
  • 83. MODALITIES OF CANCER TREATMENT LOCAL THERAPEUTIC MODALITIES SURGERY RADIOTHERAPY SYSTEMIC FORMS OF TREATMENT CHEMOTHERAPY ENDOCRINE THERAPY IMMUNOTHERAPY GENE THERAPY
  • 84. Cost of cancer per year in U.S.
  • 85. Targeted drug delivery to tumour using magnetic nanoparticles:Preclinical studies with Fe3O4 – doxorubicin nanoparticles
  • 86. Nanoparticles for targeting drugs to tumour- Fe2O3 nanoparticles coated with polymer –polyoxyethylene 25 propylene glycol stearate- Binding Drugs - Doxyrubicin , Sanazole- Effect of the complexes in vitro on EAC cellsand DLA cells- Administration to mice bearing DLA solidtumour on hind limbs for 7 days.Parameter studied: Tumour volume reduction
  • 87. Dose-Effect Curve Desirable 100 Dose Range 80% with 60Maximal Effect 40 Adverse Effects 20 0 Log Dose
  • 88. DOXYRUBICIN. HCL – MWT 580
  • 89. Doxorubicin (Adriamycin) intercalation of DNA Prevent DNA replication Common Side Effects:hair loss, mouth sores, n/v, lowered blood counts (WBCs, RBCs andplatelets),skin damage if drug leaks out of vein during infusion (necrosis)*damage to the heart muscle (cardiotoxicity from free radicals)
  • 90. Cancer chemotherapy - the agent exerts anticancer action through cytotoxic mechanismsCommon problems encountered: – Poor selectivity toward the target cancer tissue – Harm normal cells as well – Sub-therapeutic drug levels at tumor site due to varied biodistribution, fail to eradicate the tumour, stimulate overgrowth of resistant malignant cells – Possess irreversible toxic side effects. Cardio, neuro, renal toxicities• Specific targeting to tumour improves the anticancer potential of chemotherapeutic agents• Targeting by magnetic particles - a unique opportunity to treat tumors due to magnetic responsiveness.
  • 91. • Antitumor activity of water dispersible Fe3O4 nanoparticles (coated with Poly Vinyl Pyrolidone (PVP) and Poly oxy ethylene 25- propylene glycol stearate (POES)) (Fe3O4-PVP-POES) complexed with Doxorubicin (Fe3O4-PVP-POES-Doxo).• Effect of complexing Doxorubicin with Fe3O4- PVP-POES on its cardio toxic properties.
  • 92. Water dispersible Fe3O4 nanoparticlescarring doxorubicin for cancer therapyO.D.Jayakumar, R.Ganguli, A.K.Tyagi , D.K.Chandraseharan and C.K.K.NairJournal of Nanosciene and Nanotechnology, 9, 6344-6348, 2009.
  • 93. • XRD pattern of Fe3O4 nanoparticles coated with PVP-POES (a) and PVP + POES and complexed with Doxorubicin (b). • Inset shows the DC magnetization vs field of Fe3O4 nanoparticles coated with PVP measured at room temperature.Water dispersible Fe3O4 Nanoparticles Carrying Doxorubicin for Cancer Therapy: Journal of Nanoscienceand Nanotechnology, 2009, 9, 6344-6348.
  • 94. TEM images of Fe3O4-PVP- POES nanoparticles and Fe3O4-PVP-POES-Doxo complex. (a, b):Fe3O4-PVP-POES (c, d):Fe3O4-PVP-POES-DoxoWater dispersible Fe3O4 Nanoparticles Carrying Doxorubicin for Cancer Therapy: Journal of Nanoscienceand Nanotechnology, 2009, 9, 6344-6348.
  • 95. Water dispersible Fe3O4 Nanoparticles Carrying Doxorubicin for Cancer Therapy: Journal of Nanoscienceand Nanotechnology, 2009, 9, 6344-6348.
  • 96. Cytotoxic effect on DLA cells Fe3O4-PVP-POES-Doxo exhibited more toxicity towards DLA cells For Doxorubicin, the concentrations - A, B and C – corresponding to 20, 50 and 100 micrograms/ml; for the Fe3O4-PVP-POES nanoparticles the concentrations were 100, 250 and 500 micrograms/ml; and for the nanoparticle-doxorubicin complex the concentrations were 20, 50 and 100 micrograms/ml with respect to doxorubicin and 100, 250 and 500 micrograms/ml with respect to Fe 3O4-PVP-POES.Water dispersible Fe3O4 Nanoparticles Carrying Doxorubicin for Cancer Therapy: Journal of Nanoscienceand Nanotechnology, 2009, 9, 6344-6348.
  • 97. Induction of apoptosis Apoptotic index of Fe3O4-PVP-POES- Doxorubicin was higher than that of doxorubicin or Fe3O4-PVP-POES. Characteristic morphology of normal cell (A) and apoptotic cell (B) A BFor Doxorubicin, the concentrations – A and B – corresponding to 50 and 100 micrograms/ml; for the Fe 3O4-PVP-POES nanoparticles the concentrations were 250 and 500 micrograms/ml; and for the nanoparticle-doxorubicincomplex the concentrations were 50 and 100 micrograms/ml with respect to doxorubicin and 250 and 500micrograms/ml with respect to Fe O -PVP-POES.
  • 98. Effect on tumor growth • Daltons Lymphoma solid tumor on hind limbs of mice. • The treatments were started on the 13th day after tumor tranplantation. • Targeting Doxorubicin - Fe3O4 nanoparticles to tumor site - by an external magnetic field - keeping a magnet at the tumor site for 15 minutes - after the oral administration of the complex, for 7 consecutive days.Water dispersible Fe3O4 Nanoparticles Carrying Doxorubicin for Cancer Therapy: Journal of Nanoscience andNanotechnology, 2009, 9, 6344-6348.
  • 99. Effect on tumor growth • The growth of the tumor decreased in all treated animals during the period of administration. • Regression in tumor growth on administration with Doxorubicin -Fe3O4 nanoparticle complex (Fe3O4- PVP-POES-DOXO) in conjunction with magnetic treatment.Water dispersible Fe3O4 Nanoparticles Carrying Doxorubicin for Cancer Therapy: Journal of Nanoscienceand Nanotechnology, 2009, 9, 6344-6348.
  • 100. Effect on cardio toxicity • Decrease in GSH - due to the oxidative stress induced by the drug. • Magnet mediated targeting of the drug nanoparticle complex to the tumor site decreased the circulating levels of the drug complex - helped to maintain normal levels of GSH in heart tissue.n s indicate not significant and *** indicate p <0.001 when compared with untreated control.
  • 101. Effect on cardio toxicity • Increase in the MDA level - due to oxidative stress induced by the drug. • Magnet mediated targeting of the drug nanoparticle complex to the tumor site decreased the circulating levels of the drug complex - normal levels of MDA in heart tissue.n s indicate not significant, * indicate p <0.05 and *** indicate p <0.001 when compared withuntreated control.
  • 102. OXIDATIVE ENXYME THERAPY USING MAGNETIV NANOPARTICLESD-Alanine --- (DAO) ------------→ Pyruvic acid + H2O2 FTIR spectra of Fe2O3 magnetic nanoparticles FTIR spectra of Fe2O3 -DAO complex XRD pattern of Fe2O3 nanoparticles coated with PVP XRD pattern of Fe 2O3 -DAO complexD-aminoacid oxidase-Fe2O3 nanoparticle complex mediated antitumor activity in Swiss Albino mice.S.A.Divakaran, K.M.Sreekanth, K.V.Rao and C.K.K.Nair, Journal of Cancer Therapy, 2, 666-674 2011.
  • 103. OXIDATIVE ENXYME THERAPY OF TUMOUR USING MAGNETIV NANOPARTICLESEffect of administration of Fe2O3 nanoparticles coated with PVP and complexed with DAOand magnetic targeting on DLA solid tumor growth on hind limb of mice. 1 representanimals on the initial day of commencement of the experiment, 1a & 1b representsuntreated control animals, 2a & 2b represents animals treated with Fe2O3- DAO, 3a& 3brepresents animals treated with D- alanine. 4a & 4b represents animals treated with Fe 2O3-DAO and D- alanine without magnetic treatment, 5a&5b represents animals treated withFe2O3- DAO and D- alanine with magnetic treatment. The suffix ‘a’ indicates 4th day oftreatment and ‘b’ indicates15th day of treatment Journal of Cancer Therapy, 2, 666-674 2011.
  • 104. COMET ASSAY OF DLA CELLS TREATED WITH FE2O3-DAO AND D-ALANINE Apoptoic index in dla cells treated with fe2o3-dao in presence of d-alanine. Treatments Apoptoic index (%) Control 0 0.2M D-Alanine + DAO 85.2± 5.55 a (0.36U) 0.2 M D-Alanine + Fe2O3- 95.5±3.53a DAO (0.36U) 0.2M D-Alanine 6.5± 2.12b Fe2O3-DAO (0.36U) 11.0± 1.41a Apoptoic index in DLA cells treated with Representative images of DLA cells after comet assay. Fe2O3-DAO in presence of D-alanine. (‘a’ a) The untreated DLA cells. b) DLA cells treated with represents, p<0.001 compared to Fe2O3-DAO and D-alanine showing Fan-like comets respective control, ‘b’ represents, p<0.05 indicative of apoptosis c). DLA cells treated with compared to respective control) enzyme (DAO) and D-alanine showing DNA damaged and apoptotic comets and d) DLA cells treated with Fe2O3-DAO e) DLA cells treated with D-alanine.D-aminoacid oxidase-Fe2O3 nanoparticle complex mediated antitumor activity in Swiss Albino mice.S.A.Divakaran, K.M.Sreekanth, K.V.Rao and C.K.K.Nair, Journal of Cancer Therapy, 2, 666-674 2011.
  • 105. Conclusions• Doxorubicin and the enzyme DAO can be complexed to magnetic Fe3O4 nanoparticles.• This complexes can be targeted by means of an external magnetic field.• By targeting nanoparticle bound anticancer drug or administering the ezyme substrate• D-ala tumor growth can be controlled.• The targeting enhanced the efficacy of the treatment - decrease the circulatory concentrations of the drug and helps to minimize its toxic side effects.
  • 106. THANK YOU

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