troubleshooting a bending device with tissue and non-tissue verification laura gump 1, chad eckert...

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Troubleshooting a Bending Device with Tissue and Non-Tissue Verification Laura Gump 1 , Chad Eckert 2 , Michael Sacks 2 1 Bioengineering & Bioinformatics Summer Institute, Dept. of Computational Biology, University of Pittsburgh, 15260 2 Department of Bioengineering, The University of Pittsburgh, 15219 Introduction The Bending Device Methods Results Future Implications Acknowledgements References Flexure testing is needed in order to properly determine the mechanical properties of various tissues in the body which deform by bending in vivo. Such a device works by tracking changes in curvature resulting from applied loads as described by the Euler-Bernoulli equation. However, before any reliable measurements for tissue can be ascertained using such a device, the exact methodology for its use must be determined. The validation of the accuracy of the results was done by testing a variety of reference and tissue samples. The effective modulus of elasticity (E) for each sample aligned with published data, indicating a reliable device. The Bending Device (BD) [Fig.1] is comprised of three main parts, aside from the computer program. The program is a custom program in LabView that tracks fiducial markers on the sample to compute moment and curvature. The other parts are: 1. Bending Beam-A beam of known stiffness is used to determine the force being placed on the sample. 2. Transverse Bar-This connects the sample to the Bending Beam 3. Tank-holds the sample in place while still allowing it to move. Also holds PBS or other fluid used to keep sample pliable and to mimic in vivo conditions. 4. Track-Allows the tank to actuate up and down to perform tests, as well as left and right to adjust the camera view. This device will be used in future studies in the lab to test micromechanical properties of both natural human tissue as well as tissue engineered specimens. The tests will further the pursuit of a perfectly engineered replacement for various tissues, specifically heart valve leaflets. One of the more valuable facets of this device is its ability to track the hysteresis of a tissue, something that has been done only rarely before but will help establish elasticity of various tissues. Some of the first samples that will be tested on this BD will be tissue engineered heart valve leaflets that were implanted into and then explanted from sheep at different time stamps. The national BBSI program (http://bbsi.eeicom.com) is a joint initiative of the NIH-NIBIB and NSF-EEC, and the BBSI @ Pitt is supported by the National Science Foundation under Grant EEC-0234002. Engelmayr Jr. GC, Sacks MS Prediction of extracellular matrix stiffness in engineered heart valve tissues based on nonwoven scaffolds. Biomechan Model Mechanobiol 2008; 7(4):309-21 Engelmayr Jr, GC and Sacks, MS. A structural model for the flexural mechanics of nonwoven tissue engineering scaffolds. J Biomech Eng. 2006 Aug; 128(4): 610-22 Li WC, et all Biomechanical properties of ascending aorta and pulmonary trunk in pigs and humans Xenotransplantation 2008; 15:384-89 Merryman WD, Huang SH-Y, et all The effects of cullular contraction on aortic valve leaflet flexural stiffness J. Biomechanics 2006; 39:88-96 Resolution: The resolution for the camera was determined by measuring the number of pixels/mm in triplicate and averaging them. Calibration: Bending Beams with varying stiffnesses are calibrated by adding known weights to the Transverse Bar and measuring the displacement in triplicate. The average of the slopes of best fit lines is taken as the calibration constant. Loading Curves: The BD allows the user to test both with and against the natural curvature of the sample. LabView measures the curvature and moment by tracking markers on the sample. Excel was used to analyze the moment vs. curvature graphs to find E for each marker. Equations used included the second moment of area (I=t 3 *w/12) and the Bernoulli-Euler equation (M=κEI). Unloading/Hysteresis Curve: The goal of this portion of the test is to test the hysteresis E of the samples in the same manner as above. I would also like to thank: Chad Eckert Dr. Michael Sacks Ms. Eunice Yi The Bioengineering Department at the University of Pittsburgh Figure 1: The Bending Device 1 4 2 3 Figure 2: A Bend Down test using rubber Figure 3: A Bend Up test using rubber Resolutions 0.03333 mm/pixel Sample nearer Camera .034091 mm/pixel Sample Alone .033708 mm/pixel Sample in PBS Figure 4: Resolution values Calibrations 0.804 ± 0.017 g/mm Stiff Beam 0.076 ± 0.005 g/mm 10.0 Beam 0.067 ± 0.002 g/mm 8.0 Beam 0.037 ± 0.003 g/mm 4.0 Beam Figure 5: Calibration constants for beams used Loading Curves -Tests gives more accurate results if the samples have initial curvature -The more intensely red the tissue markers, the more accurate the test Sample Accepted E Experimenta l E Porcine Aorta 357 kPa ±118 kPa 280 kPa ±97kPa Tissue Scaffold 206 kPa ± 16 kPa 218 kPa ± 18 kPa Rubber Sample 2300 kPa ± 100 kPa 2220 kPa ± 160 kPa Figure 7: Accepted and Experimental values of E for tested samples Figure 6: Typical combined Bend Up and Bend Down graph for porcine aorta -0.04 -0.02 0 0.02 0.04 0.06 -30 -20 -10 0 10 20 30 40 50 Porcine Aorta Marker 1 Marker 2 Marker 3 Marker 4 Marker 5 Curvature Moment (N*mm) Unloading/Hysteresis Curves -Attempted to gather information with Bend Up tests, but the weight of the Bending Bar prevents the sample from returning to its original shape. -Bending Down tests allow the sample to regain original shape -The average value of E for loading was 2219kPa; for unloading, it was 2051kPa -0.02 0 0.02 0.04 0.06 -7 -6 -5 -4 -3 -2 -1 0 1 f(x) = − 111.378186081462 x − 0.543254068507563 R² = 0.974583333612687 f(x) = − 103.258466786439 x + 0.106365738929905 R² = 0.962061018441974 Rubber Marker 2 Loading Unloading Loading Linear (Loading) Unloading Linear (Unloading) Curvature Moment (N*mm) Figure 8: Example of loading and unloading curves for rubber

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Page 1: Troubleshooting a Bending Device with Tissue and Non-Tissue Verification Laura Gump 1, Chad Eckert 2, Michael Sacks 2 1 Bioengineering & Bioinformatics

Troubleshooting a Bending Device with Tissue and Non-Tissue Verification

Laura Gump1, Chad Eckert2, Michael Sacks2 1Bioengineering & Bioinformatics Summer Institute, Dept. of Computational Biology, University of Pittsburgh, 15260

2Department of Bioengineering, The University of Pittsburgh, 15219

Introduction

The Bending Device

Methods

Results

Future Implications

Acknowledgements

References

Flexure testing is needed in order to properly determine the mechanical properties of various tissues in the body which deform by bending in vivo. Such a device works by tracking changes in curvature resulting from applied loads as described by the Euler-Bernoulli equation. However, before any reliable measurements for tissue can be ascertained using such a device, the exact methodology for its use must be determined. The validation of the accuracy of the results was done by testing a variety of reference and tissue samples. The effective modulus of elasticity (E) for each sample aligned with published data, indicating a reliable device.

The Bending Device (BD) [Fig.1] is comprised of three main parts, aside from the computer program. The program is a custom program in LabView that tracks fiducial markers on the sample to compute moment and curvature. The other parts are:

1. Bending Beam-A beam of known stiffness is used to determine the force being placed on the sample.

2. Transverse Bar-This connects the sample to the Bending Beam

3. Tank-holds the sample in place while still allowing it to move. Also holds PBS or other fluid used to keep sample pliable and to mimic in vivo conditions.

4. Track-Allows the tank to actuate up and down to perform tests, as well as left and right to adjust the camera view.

This device will be used in future studies in the lab to test micromechanical properties of both natural human tissue as well as tissue engineered specimens. The tests will further the pursuit of a perfectly engineered replacement for various tissues, specifically heart valve leaflets. One of the more valuable facets of this device is its ability to track the hysteresis of a tissue, something that has been done only rarely before but will help establish elasticity of various tissues. Some of the first samples that will be tested on this BD will be tissue engineered heart valve leaflets that were implanted into and then explanted from sheep at different time stamps.

The national BBSI program (http://bbsi.eeicom.com) is a joint initiative of the NIH-NIBIB and NSF-EEC, and the BBSI @ Pitt is supported by the National Science Foundation under Grant EEC-0234002.

Engelmayr Jr. GC, Sacks MS Prediction of extracellular matrix stiffness in engineered heart valve tissues based on nonwoven scaffolds. Biomechan Model Mechanobiol 2008; 7(4):309-21

Engelmayr Jr, GC and Sacks, MS. A structural model for the flexural mechanics of nonwoven tissue engineering scaffolds. J Biomech Eng. 2006 Aug; 128(4): 610-22

Li WC, et all Biomechanical properties of ascending aorta and pulmonary trunk in pigs and humans Xenotransplantation 2008; 15:384-89

Merryman WD, Huang SH-Y, et all The effects of cullular contraction on aortic valve leaflet flexural stiffness J. Biomechanics 2006; 39:88-96

Resolution: The resolution for the camera was determined by measuring the number of pixels/mm in triplicate and averaging them.

Calibration: Bending Beams with varying stiffnesses are calibrated by adding known weights to the Transverse Bar and measuring the displacement in triplicate. The average of the slopes of best fit lines is taken as the calibration constant.

Loading Curves: The BD allows the user to test both with and against the natural curvature of the sample. LabView measures the curvature and moment by tracking markers on the sample. Excel was used to analyze the moment vs. curvature graphs to find E for each marker. Equations used included the second moment of area (I=t3*w/12) and the Bernoulli-Euler equation (M=κEI).

Unloading/Hysteresis Curve: The goal of this portion of the test is to test the hysteresis E of the samples in the same manner as above.

I would also like to thank:

• Chad Eckert

• Dr. Michael Sacks

• Ms. Eunice Yi

• The Bioengineering Department at the University of Pittsburgh

Figure 1: The Bending Device

14

2

3

Figure 2: A Bend Down test using rubber

Figure 3: A Bend Up test using rubber

Resolutions

0.03333 mm/pixel

Sample nearer Camera

.034091 mm/pixel

Sample Alone

.033708 mm/pixel

Sample in PBS

Figure 4: Resolution values

Calibrations

0.804 ± 0.017 g/mmStiff Beam

0.076 ± 0.005 g/mm10.0 Beam

0.067 ± 0.002 g/mm8.0 Beam

0.037 ± 0.003 g/mm 4.0 Beam

Figure 5: Calibration constants for beams used

Loading Curves-Tests gives more accurate results if the samples have initial curvature

-The more intensely red the tissue markers, the more accurate the test

SampleAccepted

EExperimental

EPorcine

Aorta357 kPa

±118 kPa 280 kPa ±97kPa 

Tissue Scaffold

206 kPa ±16 kPa 

218 kPa ±18 kPa  

Rubber Sample

2300 kPa ± 100 kPa 

2220 kPa ± 160 kPa 

Figure 7: Accepted and Experimental values of E for tested samples

Figure 6: Typical combined Bend Up and Bend Down graph for porcine aorta

-0.04-0.03-0.02-0.01 0 0.01 0.02 0.03 0.04 0.05 0.06

-30

-20

-10

0

10

20

30

40

50

Porcine Aorta

Marker 1Marker 2Marker 3Marker 4Marker 5

Curvature

Mom

ent

(N*m

m)

Unloading/Hysteresis Curves

-Attempted to gather information with Bend Up tests, but the weight of the Bending Bar prevents the sample from returning to its

original shape.

-Bending Down tests allow the sample to regain original shape

-The average value of E for loading was 2219kPa; for unloading, it was 2051kPa

-0.01 0 0.01 0.02 0.03 0.04 0.05 0.06

-7

-6

-5

-4

-3

-2

-1

0

1

f(x) = − 111.378186081462 x − 0.543254068507562R² = 0.974583333612687

f(x) = − 103.258466786439 x + 0.106365738929905R² = 0.962061018441975

Rubber Marker 2

LoadingUnloadingLoadingLinear (Loading)UnloadingLinear (Unloading)

Curvature

Mom

ent

(N*m

m)

Figure 8: Example of loading and unloading curves for rubber