An Introduction to 3D Bioprinting - TeachEngineering

An Introduction to 3D Bioprinting Lesson goals Introduce engineering problem (Bill) Define and analyze different types of 3D bioprinters Define the basics of tissue engineering Identify current applications and limitations of 3D bioprinting Start figuring out how to help Bill! Why do we care about 3D bioprinting? Watch this video Image 1

Bills Injuries Image 2 Missing skin on the left arm Image 3 Ripped rectus femoral muscle Severely broken femoral shaft

We need your help! Image 4 What is 3D bioprinting? Image 5 Image 6 Image 7 We need to learn about the different types of bioprinters! Will be used in the activity

inkjet Inkjet Bioprinting laser Image 8 B Image 9 extrusion

Image 10 Types of bioprinters: Inkjet Image 8 Inkjet Bioprinting Analogy: inkjet printer Limitations Low

viscosity Bio-ink must solidify Cell densities Best application = quickly creating skin grafts Types of bioprinters: Inkjet Watch this video Types of bioprinters: Laser Assisted Analogy: placing polka dots on a dress to create a pattern

Limitations Low printing speed Cannot print multiple layers easily Wasteful

Best application = placing cells precisely into solid structures Image 9 Types of bioprinters: Laser Assisted Watch this video Types of bioprinters: Extrusion Will be used in the activity Analogy:

squeezing ketchup out of a bottle Limitations Lower Low resolution Slow Best cellular viability

print speed application = creating large 3D structures Image 10 Types of bioprinters: Extrusion Watch this video Parts of an extrusion bioprinter Image 11 Reservoir 2

Reservoir 1 Printing stage Image 11 + Control system Types of bioprinters: Summary inkjet Inkjet Bioprinting laser

extrusion Basics of tissue engineering design: 5 Steps 1. Identify function being replaced 2. Determine cell types 3. Determine biomaterial types 4. Determine construction method 5. Construction! Determine cell types Choose

cell type for its function! Constraints: Strength of cells Rejection and immune responses Determine biomaterial types Natural biomaterials:

Collagen Elastin Synthetic Polys biomaterials: Cell survival during printing Image 13 Image 14 Image 12

O2 oxygen nutrients temperature Applications of 3D bioprinting Image 15 Current

Image 16 Tissue mimics for drug testing and screening Non-transplantable tissues and vessels aortic heart valve Image 18 Near

Image 19 Transplantable tissues Image 21 Far future (~20+ years) blood vessels future (~15 years) Image 20

Image 17 cartilage Organs kidney heart skin

Applications Watch this video Image 22 Limitations Vascularization Immune rejection Image 23 Image 24

Biocompatibility Lesson Goals: Summary Introduce engineering problem (Bill) Define and analyze different types of 3D bioprinters Define the basics of tissue engineering Identify current applications and limitations of 3D bioprinting Start figuring out how to help Bill! Activity Instructions 1. Review your assignments (~5 min) 2. Learn to use your mock bioprinter (~2 min) 3. Engineering sketch of your plan > get approval (~10 min)

4. Get biomaterials and print! (~20 min) 5. Present your design and limitations (~2 min for each group) Mock 3D Bioprinter Instructions for Use Show this video Engineering Sketch Diagram with labels Measurements and scale Axes, for reference Image 25

Image 1 Bills Injuries Image 2 Missing skin on the left arm Image 3 Ripped rectus femoral muscle Severely broken

femoral shaft Image Source/Rights Image 1: Skin anatomy diagram | File name: skin1.jpg Source/Rights: 2013 Anatomy Box, Creative Commons Attribution Share License http://www.anatomybox.com/chapters/skin/ Caption: An example of a skin tissue. Bill has injured skin. Image 2: Muscle anatomy diagram | File name: muscle.jpg Source/Rights: WikiJournal of Medicine Gallery of Blausen Medical, 2014. CC Licensure 3.0. http://teachmeanatomy.info/the-basics/ultrastructure/histology-muscle/ Caption: An example of a muscle. Bill has a ripped femoral muscle. Image 3: Anatomy of a long bone | Image file name: bone.jpg Source/Rights: 2016 Carl Fredrick, Wikimedia Commons CC BY-SA 4.0 https://commons.wikimedia.org/wiki/File:603_Anatomy_of_a_Long_Bone.jpg Caption: An example of a bone and underlying tissue. Bill has a broken femoral shaft.

Image 4: Uncle Sam saying I want you to learn about 3D bioprinting. | Image file name: unclesam.jpg Source/Rights: 2011 Library of Congress (public domain) https://www.loc.gov/exhibits/treasures/trm015.html AND https://www.loc.gov/item/93509735/ Caption: Some U.S. scientific research funding is going towards 3D bioprinting research. Image 5: A photograph shows a 3D bioprinter with 4 extrusion heads | Image file name: regenhu.jpg Source/Rights: 2016 RegenHu. All rights reserved. Used with Permission. http://www.aniwaa.com/product/3d-printers/regenhu-3ddiscovery/ Image 6: A photograph shows a 3D bioprinted tissue being taken out of growing media. | Image file name: tissue.jpg Source/Rights: 2016 Anderson, Ojada, Nguyen. Governors School of Architecture. All rights reserved. Used with permission. https ://govschoolagriculture.com/tag/3d-bioprinting/ Caption: A 3D bioprinted tissue that is kept in media to allow cells to grow. Image 7: A photograph shows of 3D bioprinted structure in the shape of an ear. | Image file name: ear.jpg Source/Rights: Wake Forest Institute for Regenerative Medicine. Non-published. Used with permission. http://www.wakehealth.edu/WFIRM/ Caption: 3D bioprinters can build structures that are in the shape of an ear. Image 8: Diagram shows a model of an inkjet bioprinter. | Image file name: inkjet.jpg Source/Rights: 2016 Arslan-Yildiz, Assal, Chen, Guven, Inci, Demirci. Used with permission. http://iopscience.iop.org/article/10.1088/1758-5090/8/1/014103

Caption: Inkjet bioprinters disperse cells over a surface covering much area quickly. Image Source/Rights Image 9: A diagram of a laser-assisted 3D bioprinter. | Image file name: laser.jpg Source/Rights: 2016 Arslan-Yildiz, Assal, Chen, Guven, Inci, Demirci. Used with permission. http://iopscience.iop.org/article/10.1088/1758-5090/8/1/014103 Caption: A laser-assisted bioprinter use lasers to force cells onto specific locations of the printing surface. Image 10: A diagram of an extrusion 3D bioprinter. | Image file name: extrusion.jpg Source/Rights: 2016 Arslan-Yildiz, Assal, Chen, Guven, Inci, Demirci. Used with permission. http://iopscience.iop.org/article/10.1088/1758-5090/8/1/014103 Caption: Extrusion bioprinters extrude cells within a filament. The filament/cell mixture forms the structure we see. Image 11: A photograph of a 3D bioprinter with 2 extrusion heads printing onto a cell culture plate called a 96-well plate. | Image file name: organovo.jpg Source/Rights: 2016 Organovo. Used with Permission http://www.csmonitor.com/Technology/2015/0519/3D-printing-human-skin-The-end-of-animal-testing Caption: Extrusion 3D bioprinters have multiple heads to print from. They can also print on different surface. A computer system controls the rate of extrusion, the location,

and which material is extruded. Image 12: A graphic of a cell having oxygen delivered to it. | Image file name: oxygen.jpg Source/Rights: 2017 Nick Asby (author), UVa Department of Biomedical Engineering. Caption: Cells need proper oxygen concentrations to survive. Image 13: A picture of bottle of cell media | Image file name: media.jpg Source/Rights: 2011 Lillly_M, Max Planck Institute for Molecular Cell Biology and Genetics in Dresden, Wikimedia Commons CC BY-SA 3.0 https ://commons.wikimedia.org/wiki/File:Glasgow_MEM_cell_culture_medium.jpg Caption: Cells are kept in cell media while they grow. Media provides nutrients, proper pH, water, and other compounds need to make cells grow. Image 14: A graphic of a thermometer | Image file name: temperature.jpg Source/Rights: 2004 Microsoft Corporation, One Microsoft Way, Redmond, WA 98052-6399 USA. All rights reserved Caption: Cells need to be kept at proper temperatures to replicate and survive. Image 15: A picture of 3D bioprinter printing into a cell culture container. The one in this picture is called a 96-well plate. | Image file name: drugTest.jpg Source/Rights: 2016 Organovo. Used with permission. http://www.csmonitor.com/Technology/2015/0519/3D-printing-human-skin-The-end-of-animal-testing Caption: 3D bioprinters are utilized by companies and researchers to create testing tissue for pharmaceutical research. Image 16: A still image taken from an animation/video of beating bioprinted aortic valve. | Image file name: valve.gif

Source/Rights: 1989 Valveguru, Wikimedia Commons CC BY-SA 3.0 https://commons.wikimedia.org/wiki/File:Aortic_valve.gif Caption: Researchers use 3D bioprinters to create valves. These are used for research purposes and cannot be placed in humans yet. Original caption: This is still image pulled from a video clip of a living, beating pig heartan aortic valvethat was bioprinted in a lab. The heart was arrested, connected to the perfusion system and restarted. The working fluid was oxygenated balanced saline solution. Image Source/Rights Image 17: A picture of cells laid down to create a blood vessel. | Image file name: vessel.jpg Source/Rights: 2004 Microsoft Corporation, One Microsoft Way, Redmond, WA 98052-6399 USA. All rights reserved Caption: Scientists have methods of creating blood vessels with 3D bioprinting methods. Image 18: A picture of bioprinted cartilage in the shape of an ear. | Image file name: cartilage.jpg Source/Rights: Wake Forest Institute for Regenerative Medicine. Non-published. Used with permission. http://www.wakehealth.edu/WFIRM/ Caption: Scientists can bioprint cartilage in the shape of an ear for research purposes. However, it is not safe to use these ears on humans.

Image 19: A picture of bioprinted skin being held. | Image file name: skin2.jpg Source/Rights: Wake Forest Institute for Regenerative Medicine. Non-published. Used with permission. http://www.wakehealth.edu/WFIRM/ Caption: Scientists can print human skin for drug testing purposes. The skin does not have the exact structure of human skin, but it is a close replicate. Image 20: A photo of a kidney being printed. | Image file name: kidney.jpg Source/Rights: Wake Forest Institute for Regenerative Medicine. All rights reserved. Nonpublished. Used with permission. https://govschoolagriculture.com/tag/3d-bioprinting/ Image 21: A picture of a human heart in vitro (outside the body) | Image file name: heart.jpg Source/Rights: 2007 alexanderpiavas134, Wikimedia Commons (public domain) https://commons.wikimedia.org/wiki/File:Humhrt2.jpg Caption: Scientists are working towards bioprinting hearts. Image 22: A picture of a complex blood vessel system. | Image file name: vasculature.jpg Source/Rights: 2004 Microsoft Corporation, One Microsoft Way, Redmond, WA 98052-6399 USA. All rights reserved Caption: Incorporating blood vessel structure into tissues and organs requires complicated computer algorithms. Image 23: A picture of a body rejecting pathogens. | Image file name: immune.jpg Source/Rights: 2004 Microsoft Corporation, One Microsoft Way, Redmond, WA 98052-6399 USA. All rights reserved Caption: Scientists are working to reduce of immune rejection when implanting a bioprinted tissue or organ into the body. Image 24: A picture of pancreas attached to the gall bladder and bile duct. | Image file name: pancreas.jpg

Source/Rights: 2004 Microsoft Corporation, One Microsoft Way, Redmond, WA 98052-6399 USA. All rights reserved Caption: Biocompatible bioprinted organs have the functionality, longevity, and mechanical properties that the original organ possessed. Image 25: A multi-view engineering drawing | Image file name: engineeringSketch.png Source/Rights: 1938 Frank R. Leslie, Historic American Engineering Record, National Park Service; Record MO-1105, Library of Congress, Prints & Photographs Division, MO1105 (public domain) https://commons.wikimedia.org/w/index.php?curid=3715658 Caption: Engineering sketches include measurements, scales, dimensions and multiple views of the same design at different angles.

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