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Investigating the effect of laser fabricated topography on mammalian cellular behaviours

Investigating the effect of laser fabricated topography on mammalian cellular behaviours. By George Ching Tzu Goh. Supervised by: Dr. Paul French, Dr. Mark Murphy, and Dr. Martin Sharp. Group. Introduction Cell Behaviours. In the body cells make, secrete and grow within a 3-D matrix

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Investigating the effect of laser fabricated topography on mammalian cellular behaviours

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  1. Investigating the effect of laser fabricated topography on mammalian cellular behaviours By George Ching Tzu Goh Supervised by: Dr. Paul French, Dr. Mark Murphy, and Dr. Martin Sharp Group

  2. Introduction Cell Behaviours • In the body cells make, secrete and grow within a 3-D matrix • Interaction with this external matrix is crucial for development and normal tissue functioning Group

  3. Coronary Heart Attack & Surgical Procedure Percutaneous coronary intervention (PCI) Group

  4. Disease Complication-Restenosis Neointimal Hyperplasia Arterial cross-sections processed 28 days after stent placement Group

  5. Project Aims The main aim of this project are to use laser processing techniques to develop novel 3D patterned polymer substrates for improving endothelial adhesion, proliferation and migration. Whyendothelial cells so important? Endothelium act as a selective barrier and supresses intimal hyperplasia by inhibiting • Thrombus formation • Inflammation • Smooth muscle cell proliferation and migration Group

  6. Engineering works Group

  7. SPI Laser System 20W SPI 1064nm 20ns Solid State Infrared Fibre Laser System Schematic Diagram of SPI Laser Setting Laser Micromachining Lab Group

  8. Casting Procedure Biomer Technology Ltd. -Z1A1 Polyurethane 2 1 3 Z1A1 PEU+ Template Laser Microfabricated Template + Guiding Mould Cured in oven at 60˚C for 2 hours Group

  9. Cell works Group

  10. Bovine Aorta Endothelial cells (BAE-1) Adhesion assay on casted laser microfabricated and non-textured surfaces p≤0.001 BAE-1 cells adhesion assay. Numbers of cells remain attached in each well of textured and non-textured surfaces after challenged by washing were represented by final cell density recorded in individual well. Four types of laser textured Z1A1 PEU polymers were used, namely type 1A, 1B, 1C and 1D, and the control of this experiment was a non-textured PEU polymer. Group

  11. Human Coronary Artery Smooth Muscle Cells Adhesion assay on casted laser microfabricated and non-textured surfaces Phase contrast images showed that there no adhered HCASMC cells on indirect laser micro-fabricated and non-textured polyurethanes at 2 hours post-seeding. (A) Polyurethane 1A, (B) Polyurethane 1B, (C) Polyurethane 1C, (D) Polyurethane 1D and (E) Non-textured polyurethane. Group

  12. BAE-1 and HCASMC Cells Proliferation assay on casted laser microfabricated and non-textured surfaces p≤0.05 Group

  13. FAK Actin Merged Control PEU BAE-1 Cells under Static Condition Actin stained with phalloidin and vinculin complex stained with anti-mouse FITC antibody 48 Hours post seeding 1A PEU Group

  14. BAE-1 Cells Under 20 dynes/cm2 Shear Stress casted laser microfabricated and non-textured surfaces 24% 13% Group

  15. BAE-1 Cells Under 20 dynes/cm2 Shear Stress casted laser microfabricated and non-textured surfaces Actin FAK Merged NucIeus 1A PEU 48 Hours post seeding Control PEU Group

  16. Conclusion • The result clearly showed that casted laser microfabricated 1A polyurethane mediated endothelial cell adhesion and enhanced cell migration. • This could apply on stent application by micromaching microfringes and nano-coat Z1A1 polyurethane on stent surface to improve endothelial adhesion, at the same time inhibit HCASMC adhesion. Group

  17. Anyquestions? Group

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