Biomedical
Applications of
Nanotechnology
What is Nanotechnology?
The creation of useful, functional materials,
devices, and systems through:
1. Controlling and ma...
Potential market for nanotechnology ?
$1 trillion business within the next 10 to 15 years.
Nanotechnology in Medicine
• Biological imaging for medical diagnostics.
• Advanced drug delivery systems.
• Biosensors f...
Targeted Drug Delivery
J Leukoc Biol 2005;78:585
Cardiovascular Obesity /
Disease Diabetes
Risk Factors Inflammatio...
Development of
In Vivo and In Vitro
Inflammatory Disease
Models
In Vivo Brain Inflammation Models
• Environmental Toxicants-Mediated
Brain Inflammation.
• Bacterial Infection-Mediat...
Environmental Toxicants & Brain Inflammation
Isolation of brain regions
C57/BL6 Mouse
...
20 *
Control
(TNF-α mRNA/GAPDH mRNA)
...
25
(TNF-α mRNA/GAPDH mRNA) Control
CPF, 20 mg/kg *
...
6
Control *
CPF, 4 h
(IL...
7
Control
(IL-6 mRNA/GAPDH mRNA)
CPF, 20 mg/kg
...
6 *
Control
(MCP-1 mRNA/GAPDH mRNA)
...
5
(MCP-1 mRNA/GAPDH mRNA) Control *
CPF, 20 mg/kg ...
3.0 *
(E-selectin mRNA/GAPDH mRNA)
Control
...
Bacterial Infection & Brain Inflammation
Isolation of brain regions
C57/BL6 Mouse
Det...
40 30
* Control
(TNF-α mRNA/GA...
35 12
(IL-6 mRNA/GAPDH mRNA)
30
...
25 6
* Control *
(IL...
120 20
Control
(MCP-1 mRNA/...
15 Control
10
(ICAM-1 mRNA/GAPDH mRNA...
20 15
(E-selectin mRNA/GAPDH mRNA)
Control
...
In Vitro Brain Inflammation Models
• Brain
Microvascular
Endothelial Cells
(BMEC)
• Astrocytes
• Microglia
Pro-inflammatory Stimuli-Mediated
Inflammation in Brain Cells
Pro-inflammatory Stimuli
[100 ng/ml of LPS]
...
Brain Microvascular Endothelial Cells (bEnd.3)
120 ...
Brain Microvascular Endothelial Cells (bEnd.3)
20 ...
Brain Microvascular Endothelial Cells (bEnd.3)
30 * ...
Relative Fold Induction
(TNF-α mRNA/GAPDH mRNA)
0
10
2...
Relative Fold Induction
(IL-6 mRNA/GAPDH mRNA)
0
1
...
Relative Fold Induction
(TNF-α mRNA/GAPDH mRNA)
0
1
...
Relative Fold Induction
(MCP-1 mRNA/GAPDH mRNA)
0
10
20
...
In Vitro Ischemia/Reperfusion Model
A Novel Design of Double-
Layer Parallel-Plate Flow
Chamber & Its Biomedical
...
In Vitro Flow Chamber Systems
Cone-Plate Orbital Shaker
Artificial Capillary Parallel-Plate
Parallel-plate flow chambers (PPFC) have been
most commonly used for its simplicity of concept
...
Conventional PPFCs have shown weaknesses and
problems in several aspects of its design
To eliminate these problems, we designed and
developed a new double-layer PPFC
• Accepts up to four glass slides ...
The system becomes much simpler with the new chamber.
The new double-layer PPFC consists of separate
layers of different materials and thicknesses
A...
The new double-layer PPFC consists of separate
layers of different materials and thicknesses
S...
The new double-layer PPFC consists of separate
layers of different materials and thicknesses
G...
The new double-layer PPFC consists of separate
layers of different materials and thicknesses
M...
The new double-layer PPFC consists of separate
layers of different materials and thicknesses
To set ...
A flow loop system provides a constant
hydrostatic pressure to the PPFC
Upper
...
• The streamlines near the lateral walls were not disturbed
ensuring that the lateral wall effects are negligible.
• The...
RBE4 HMEC-1
Static
Flow
1.4
1.2 Static
...
In Vitro Ischemia/Reperfusion Model
7
Normal Flow
...
In Vitro Ischemia/Reperfusion Model
5
Normal Flow ...
In Vitro Ischemia/Reperfusion Model
3
(ICAM-1 mRNA/β-Actin mRNA)
...
In Vitro Ischemia/Reperfusion Model
6
(VCAM-1 mRNA/β-Actin mRNA)
...
In Vitro Ischemia/Reperfusion Model
5
(E-selectin mRNA/β-Actin mRNA)
...
of 52

Nanotechnology Bio

Published on: Mar 3, 2016
Published in: Technology      Business      
Source: www.slideshare.net


Transcripts - Nanotechnology Bio

  • 1. Biomedical Applications of Nanotechnology
  • 2. What is Nanotechnology? The creation of useful, functional materials, devices, and systems through: 1. Controlling and manipulating matter on the nanometer-length scale (1-100 nm), and 2. Exploiting novel phenomena and properties (physical, chemical, biological, mechanical, electrical) at the nanoscale. “Going Small for Big Advances” “Going Small for Big Advances”
  • 3. Potential market for nanotechnology ? $1 trillion business within the next 10 to 15 years.
  • 4. Nanotechnology in Medicine • Biological imaging for medical diagnostics. • Advanced drug delivery systems. • Biosensors for airborne chemicals or other toxins. • Regenerative medicine: More durable, rejection-resistant artificial tissues and organs. NANOMEDICINE NANOMEDICINE
  • 5. Targeted Drug Delivery J Leukoc Biol 2005;78:585
  • 6. Cardiovascular Obesity / Disease Diabetes Risk Factors Inflammation Nanotechnology Tumor Neurological Angiogenesis Disorder / Metastasis
  • 7. Development of In Vivo and In Vitro Inflammatory Disease Models
  • 8. In Vivo Brain Inflammation Models • Environmental Toxicants-Mediated Brain Inflammation. • Bacterial Infection-Mediated Brain Inflammation.
  • 9. Environmental Toxicants & Brain Inflammation Isolation of brain regions C57/BL6 Mouse Determination of pro-inflammatory mediators - Cytokines: TNF-α, IL-1β, IL-6 - Chemokine: MCP-1 Chlorpyrifos - Adhesion molecules: E-selectin, ICAM-1, VCAM-1 [OP Pesticide]
  • 10. 20 * Control (TNF-α mRNA/GAPDH mRNA) CPF, 8 h CPF, 16 h Relative Fold Induction 15 CPF, 24 h * 10 * 5 * * * * * * * 0 HIP COR STR CER
  • 11. 25 (TNF-α mRNA/GAPDH mRNA) Control CPF, 20 mg/kg * CPF, 70 mg/kg 20 Relative Fold Induction CPF, 140 mg/kg 15 10 * * * 5 * * * 0 HIP COR STR CER
  • 12. 6 Control * CPF, 4 h (IL-6 mRNA/GAPDH mRNA) 5 CPF, 8 h Relative Fold Induction CPF, 24 h 4 * * 3 * * * 2 * * 1 0 HIP COR STR CER
  • 13. 7 Control (IL-6 mRNA/GAPDH mRNA) CPF, 20 mg/kg * 6 CPF, 70 mg/kg Relative Fold Induction CPF, 140 mg/kg 5 4 * * 3 * * 2 1 0 HIP COR STR CER
  • 14. 6 * Control (MCP-1 mRNA/GAPDH mRNA) CPF, 8 h 5 CPF, 16 h Relative Fold Induction CPF, 24 h 4 * * 3 * * * 2 1 0 HIP STR CER
  • 15. 5 (MCP-1 mRNA/GAPDH mRNA) Control * CPF, 20 mg/kg * CPF, 70 mg/kg 4 Relative Fold Induction CPF, 140 mg/kg 3 * * 2 1 0 HIP STR CER
  • 16. 3.0 * (E-selectin mRNA/GAPDH mRNA) Control * CPF, 70 mg/kg 2.5 Relative Fold Induction * 2.0 1.5 1.0 0.5 0.0 HIP COR STR CER
  • 17. Bacterial Infection & Brain Inflammation Isolation of brain regions C57/BL6 Mouse Determination of pro-inflammatory mediators - Cytokines: TNF-α, IL-1β, IL-6 - Chemokine: MCP-1 Lipopolysaccharide - Adhesion molecules: E-selectin, ICAM-1, VCAM-1 [LPS]
  • 18. 40 30 * Control (TNF-α mRNA/GAPDH mRNA) 35 LPS * 25 Relative Fold Induction * 30 * * 20 25 * 20 15 15 10 10 5 5 0 0 HIP COR STR CER HIP COR STR CER 4 h Exposure 24 h Exposure
  • 19. 35 12 (IL-6 mRNA/GAPDH mRNA) 30 Control * * LPS Relative Fold Induction 25 9 20 * 6 15 * 10 * 3 * 5 0 0 HIP COR STR CER HIP COR STR CER 4 h Exposure 24 h Exposure
  • 20. 25 6 * Control * (IL-1β mRNA/GAPDH mRNA) LPS 5 20 Relative Fold Induction 4 * 15 3 * * 10 * 2 * 5 1 0 0 HIP COR STR CER HIP COR STR CER 4 h Exposure 24 h Exposure
  • 21. 120 20 Control (MCP-1 mRNA/GAPDH mRNA) LPS * * 100 Relative Fold Induction * 15 80 * * 60 10 40 * * 5 20 0 0 HIP COR STR CER HIP COR STR CER 4 h Exposure 24 h Exposure
  • 22. 15 Control 10 (ICAM-1 mRNA/GAPDH mRNA) * LPS * * Relative Fold Induction 12 * 8 9 6 * * 6 * 4 * 3 2 0 0 HIP COR STR CER HIP COR STR CER 4 h Exposure 24 h Exposure
  • 23. 20 15 (E-selectin mRNA/GAPDH mRNA) Control LPS * * Relative Fold Induction 12 15 * * 9 * 10 * * 6 5 * 3 0 0 HIP COR STR CER HIP COR STR CER 4 h Exposure 24 h Exposure
  • 24. In Vitro Brain Inflammation Models • Brain Microvascular Endothelial Cells (BMEC) • Astrocytes • Microglia
  • 25. Pro-inflammatory Stimuli-Mediated Inflammation in Brain Cells Pro-inflammatory Stimuli [100 ng/ml of LPS] Culture Media Brain Cells Determination of pro-inflammatory mediators - Cytokines: TNF-α, IL-1β, IL-6 - Chemokine: MCP-1 - Adhesion molecules: E-selectin, ICAM-1
  • 26. Brain Microvascular Endothelial Cells (bEnd.3) 120 10 * (TNF-α mRNA/GAPDH mRNA) (IL-1β mRNA/GAPDH mRNA) 100 * 8 Relative Fold Induction Relative Fold Induction 80 6 60 * 4 40 * 2 20 * * 0 0 Control 1 2 4 8 Control 1 2 4 8 Exposure Time (hours) Exposure Time (hours)
  • 27. Brain Microvascular Endothelial Cells (bEnd.3) 20 100 * (MCP-1 mRNA/GAPDH mRNA) * (IL-6 mRNA/GAPDH mRNA) 80 Relative Fold Induction Relative Fold Induction 15 * 60 * 10 * 40 * 5 20 * * 0 0 Control 1 2 4 8 Control 1 2 4 8 Exposure Time (hours) Exposure Time (hours)
  • 28. Brain Microvascular Endothelial Cells (bEnd.3) 30 * 12 (E-selectin mRNA/GAPDH mRNA) (ICAM-1 mRNA/GAPDH mRNA) * 25 Relative Fold Induction Relative Fold Induction * 9 20 * 15 6 * 10 3 5 * 0 0 Control 1 2 4 8 Control 1 2 4 8 Exposure Time (hours) Exposure Time (hours)
  • 29. Relative Fold Induction (TNF-α mRNA/GAPDH mRNA) 0 10 20 30 40 50 Control * LPS Relative Fold Induction 0 (IL-1β mRNA/GAPDH mRNA) 1 2 800 900 1000 1100 Microglia (BV-2) Control * LPS
  • 30. Relative Fold Induction (IL-6 mRNA/GAPDH mRNA) 0 1 2 6500 7000 7500 8000 Control * LPS Relative Fold Induction 0 (MCP-1 mRNA/GAPDH mRNA) 10 20 30 40 50 60 70 80 90 Microglia (BV-2) Control * LPS
  • 31. Relative Fold Induction (TNF-α mRNA/GAPDH mRNA) 0 1 2 330 360 390 420 Control * LPS Relative Fold Induction (IL-6 mRNA/GAPDH mRNA) 0 5 10 15 20 25 30 35 40 45 Astrocytes (C6) Control * LPS
  • 32. Relative Fold Induction (MCP-1 mRNA/GAPDH mRNA) 0 10 20 30 40 50 60 70 80 Control * Astrocytes (C6) LPS
  • 33. In Vitro Ischemia/Reperfusion Model A Novel Design of Double- Layer Parallel-Plate Flow Chamber & Its Biomedical Application
  • 34. In Vitro Flow Chamber Systems Cone-Plate Orbital Shaker Artificial Capillary Parallel-Plate
  • 35. Parallel-plate flow chambers (PPFC) have been most commonly used for its simplicity of concept x y l h w z Flow enters the parallel plates at the origin and exits where x equals the length of the chamber, l
  • 36. Conventional PPFCs have shown weaknesses and problems in several aspects of its design
  • 37. To eliminate these problems, we designed and developed a new double-layer PPFC • Accepts up to four glass slides facing each other so that the flow within the channel is exclusively formed by endothelial cells. • Provides a total of 96 cm2 cell monolayer per chamber. • Placing glass slides in series shortens the duration of procedure. • The multilayer design only requires 2D cutting, which is easier and faster to manufacture and modify.
  • 38. The system becomes much simpler with the new chamber.
  • 39. The new double-layer PPFC consists of separate layers of different materials and thicknesses Acrylic sheets of 0.08 inch thickness Acrylic sheets of 0.5 inch thickness Each acrylic layer was cut by Laser Computer Aided Modeling and Manufacture (LaserCAMM) machine. The system is a computerized laser cutter that uses a laser beam to cut sheet materials into intricate patterns with a high degree of accuracy.
  • 40. The new double-layer PPFC consists of separate layers of different materials and thicknesses Silicone gaskets of 0.03 inch thickness Silicone gasket of 0.01 inch thickness Silicone gaskets of 0.03 inch thickness serve as a firm grip for glass slides. The silicone gasket in the middle constitutes the channel height, h, and the width, w.
  • 41. The new double-layer PPFC consists of separate layers of different materials and thicknesses Glass slides to fill up space Glass slides which will have cells seeded Up to four glass slides can be entered in a chamber. Glass slides or endothelial monolayers are placed between the gasket in the middle. Placing endothelial monolayers on both sides of channel minimizes pressure loss while having a larger effective area.
  • 42. The new double-layer PPFC consists of separate layers of different materials and thicknesses Media enters through the inlets. Fills up a small reservoir formed in the gasket. Spreads evenly across width through a thin slit. Flows across the endothelial monolayer. Escapes the chamber through the thin slit, the small reservoir, and the outlets.
  • 43. The new double-layer PPFC consists of separate layers of different materials and thicknesses To set up the chamber bubble-free, the layers are installed in the order from the bottom to the top layers where the flow channel, reservoirs are filled up with media by means of syringe as each layer is piled up.
  • 44. A flow loop system provides a constant hydrostatic pressure to the PPFC Upper Reservoir Flow Flow Meter Peristaltic Pump Lower PPFC Reservoir
  • 45. • The streamlines near the lateral walls were not disturbed ensuring that the lateral wall effects are negligible. • The chamber clearly applies a uniform magnitude of shear stress throughout the entire surface where endothelial cell monolayer will be placed.
  • 46. RBE4 HMEC-1 Static Flow
  • 47. 1.4 1.2 Static 1.2 Flow (Relative Fold Induction) (Relative Fold Induction) IL-6 Gene Expression Gene Expression 0.9 1.0 0.8 0.6 * * 0.6 * 0.4 0.3 * 0.2 0.0 0.0 Static Flow ICAM-1 VCAM-1 E-selectin
  • 48. In Vitro Ischemia/Reperfusion Model 7 Normal Flow (IL-6 mRNA/β-Actin mRNA) Relative Fold Induction 6 Ischemia/Reperfusion * 5 4 3 2 1 0 0.5 1 6 12 24 (hours) Ischemia Reperfusion
  • 49. In Vitro Ischemia/Reperfusion Model 5 Normal Flow * (MCP-1 mRNA/β-Actin mRNA) Ischemia/Reperfusion 4 * Relative Fold Induction 3 2 1 0 0.5 1 6 12 24 (hours) Ischemia Reperfusion
  • 50. In Vitro Ischemia/Reperfusion Model 3 (ICAM-1 mRNA/β-Actin mRNA) Normal Flow Ischemia/Reperfusion * Relative Fold Induction 2 1 0 0.5 1 6 12 24 (hours) Ischemia Reperfusion
  • 51. In Vitro Ischemia/Reperfusion Model 6 (VCAM-1 mRNA/β-Actin mRNA) Normal Flow Relative Fold Induction 5 Ischemia/Reperfusion * 4 3 * 2 1 0 0.5 1 6 12 24 (hours) Ischemia Reperfusion
  • 52. In Vitro Ischemia/Reperfusion Model 5 (E-selectin mRNA/β-Actin mRNA) Normal Flow Ischemia/Reperfusion * 4 * Relative Fold Induction 3 2 1 0 0.5 1 6 12 24 (hours) Ischemia Reperfusion

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