Preparation

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Preparation
Of Mouse
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Of Mouse
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Bone for
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Nanoinden
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tation
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Testing
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• Client
• Robert Blank, MD, PhD
• UW Medical School, Endocrinology
• Advisor
• J. Aura Gimm, PhD
• UW Engineering, Biomedical Engineering

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• Authors
• Kathleen Agard
• Sean Bruggink
• Ryan Fischer
• Chris Klundt
• Kristin La Fortune
• Kyle Quinn
• Megan Toth
• Rebecca Vanderpool

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Abstract

• Dr. Blank is interested in the Youngs Modulus
  of the outer cortical layer of the mouse tibia
  bone. Nanoindentation testing is the most
  accurate test in determining stiffness. In order
  to perform the nanoindentation test, the bone
  must be cut and sanded into flat, smooth
  sections. The goal of this project is to create
  both a cost and time effective procedure for
  preparing the mouse bone for nanoindentation
  testing.

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Problem Statement

• A nanoindentation must be made on the cortical
  surface of a mouse tibia bone in order to find
  the Youngs Modulus of the material. The
  surface of the sample must be flattened and
  smoothed. The sample will be analyzed using a
  Hysitron nanoindentation machine. The goal of
  the project is to create both a cost and time
  effective procedure for preparing mouse tibia
  bone for nanoindentation testing.

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Motivations

• Finding an accurate Youngs Modulus of cortical
  mouse bone
• Relating bone stiffness and bone dimension
• Evaluating FTIR Spectroscopy and trace element
  spectromicroscopy

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Background
Load Displacement Curve

• Youngs Modulus
• Ystress/strains/e
• Unloading curve describes Youngs Modulus
• Loading curve describes hardness

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Background

• Designed specimen holding device for MTS Bionix
  Machine
• Inaccuracy of MTS Bionix Machine
• Vibrations of machine exceed minimum constraints
• Decided to use Berkovich indentation tip
• Found available nanoindentation device on campus
  for client

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Design Criteria

• Outer cortical layer of bone must be accessible
• Surface variations must be less than 3 µm
• Specimen must have a diameter of less than 1 cm
  and thickness of 1-2 mm
• Must be attached to ferrous disc

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Final Design Selection

• Broke problem into 3 parts
• Embedding
• Cutting
• Smoothing
• Combined best alternatives from each section to
  make an optimal process.

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Embedding Alternatives

• PMMA
• Extremely exothermic
• Meticulous method
• Duco Cement
• 80 evaporation
• Liquid Nails
• Opaque

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Cutting Instrument Alternatives

• Diamond Saw
• Must rent time
• Smoothes bone as it cuts
• Must eyeball cut
• Surgical Saw
• Must purchase (gt1500)
• Too abrasive
• Must eyeball cut

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Specimen Holding Alternatives

• Two block holding device
• Undesired lateral movement
• Limited number of cuts

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Smoothing Alternatives

• Lathe table
• Difficult to hold specimen orthogonal to table
• Travel time involved to MSE building
• Sanding/polishing by hand
• Time consuming
• Inconsistent surface quality

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Final Design

• Embed the bone in an epoxy mold
• Sand bone with disc sander and specimen holder
• Polish bone with diamond cloth
• Cut sections off with rotary microtome
• Attach to ferrous disc
• Nanoindention testing

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Rotary Microtome

• Available on campus
• Effectively cuts thin specimens
• Sliding Microtome also available
• Easy to use

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Specimen Holder
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Specimen Holder
92
SLIDING PIECE
58
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TEST TUBE
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BASE
25
25
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118
19
29
13
13
3
19
118
7
BASE
121
132
All Dimensions are in mm.
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Estimated Costs

• Initial cost of process 190
• Includes Delta Disc Sander, Diamond Cloth, and
  Specimen Holder
• Cost per Bone 11.93
• Includes Labor, Silicon Carbide Paper, Test
  Tubes, and Epoxy

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Future Work

• Explore the option of working with Vicki
  Kalscheur
• Test our final design process for variations of
  less than 3 µm on the bone surface via
  nanoindenation
• Purchase Delta Disc Sander and Berkovich
  indentation tip

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References

• Allied High Tech Products Inc.
• Blank, Rob M.D. Personal communication. 20
  Sept. 2002.
• Bright Instruments Inc. 18 Nov. 2002. lt
  www.brightinstruments.comgt.
• Devcon. Duco Cement. 12 Dec. 2002
  lthttp//www.devcon.com/techinfo/207.pdfgt.
• Devcon. 5-Minute Epoxy. 12 Dec. 2002
  lthttp//www.devcon.com/techinfo/175.pdfgt.
• Escarcega, Tony. Personal communication. 20
  Sept. 2002.
• Hysitron Inc. 8 Oct. 2002. lthttp//www.hysitron.
  com/webpage2/HomePage.htmgt.
• John, Research and Development Engineer at MTS.
  Telephone interview. 15 Oct. 2002.
• Kalscheur, Vicki L. Personal Interview. 5
  Dec. 2002.
• Kishen A, et al. Experimental studies on the
  nature of property gradients in the human
  dentine. Actuators and sensors Strategic
  Research Program, School of Mechanical and
  Productional Engineering, Nanyang Technological
  University, Singapore. 2000.
• Lakes, Rod. Personal Interview. 25 Nov. 2002.
• Lamb. ltwww.ralamb.netgt. 2002.
• Liquid Nails. Small Projects Repairs Adhesive
  (LN-600). 12 Dec. 2002 lthttp//www.liquidnails.com
  /adhesives/ln-600.htmlgt.
• Liquid Nails. Small Projects Repairs Adhesive,
  Clear (LN-275). 12 Dec. 2002 lthttp//www.liquidnai
  ls.com/adhesives/ln-275.htmlgt.
• Mehta, Drew N. Sales Manager at Vibratome
  Company. Telephone interview. 19 Nov. 2002.
• Miller, David R. Engineer at Unique Instruments.
  Email. 3 Dec. 2002.
• Shaw, Gordan. Personal Communication. 21 Nov.
  2002.
• Stone, Donald. Personal Interview. 8 Nov. 2002.
  
• Tangram Technology. Polymethyl Methacrylate -
  PMMA (Acrylic). 12 Dec. 2002 lthttp//www.tangram.c
  o.uk/TI-Polymer-PMMA.htmlgt.

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MTS Machine