Experiment 6: INSTRON Tensile Testing: Fracture Properties of Chicken Skin and

Sutured Chicken Skin

It was stated in the manual for Experiment 2 that surgical suturing techniques are formulated to
optimize a number of different outcomes, including strength, speed, and gap-width of a wound.
For Experiment 2, different stitching techniques were used and loaded to determine which stitch
would display the least deformation, however, it was noted that this is not an accurate judge of
which is the best stitch. Ideally, a suture will have similar, if not stronger, material properties
of unbroken skin under a load. Considering displacements under a load can give only one-
dimensional insight about the sutures responses to loading, neglecting their failure
characteristics and material properties such as Youngs modulus. Taking the suturing techniques
of Experiment 2 in combination with the material property analysis of Experiment 3, the
appropriateness and fitness of a suture as a skin surrogate can be tested. In Experiment 3, chicken
skin was tested at a loading rate of 25 mm/min to determine its material and failure properties;
the failure strain of whole (un-sutured) chicken skin samples averaged 0.6940.169 mm/mm,
and failure displacement averaged 17.634.28mm (Appendix1). Similar protocol can be used to
determine the failure strain and displacement of sutured chicken skin along with other material
properties such as elastic modulus. These data can be compared through appropriate t-tests
(unpaired two-tailed to test if sutured samples have different a failure displacement, elastic
modulus, and/or failure strain as unbroken skins) in order to support or nullify the hypothesis.

Hypothesis/Objectives:

It is hypothesized that unbroken (whole) chicken skin samples will have similar material
properties as sutured skin samples. Thus, these properties (failure displacements, failure strains,
and elastic modulus) as obtained by the INSTRON 4444 and LabView Software, will show no
significant difference when compared with a t-test. Beyond simply testing this hypothesis, aims
of this experiment also include appropriate allocation of supplies and correct interpretation and
use of INSTRON data. The force-deformation plot given by the LabView software in
combination with the appropriate measurements will be used to create a stress-strain plot in order
to determine Youngs moduli and failure properties. The limited number of samples and amount
of time requires experimenters to properly plan ahead in order to gather all the correct
information. Experimenters will become familiar with real time data acquisition and the
challenges associated with it. Meanwhile, hands-on experience will be gained with the
difficulties of controlled preparation and testing of biological samples and use of the INSTRON
4444. Also, like in Experiment 2, this lab will yield practice with suturing techniques but with
biological samples and real suturing materials.
Equipment:

1. Major Equipment:
INSTRON 4444 table-top mechanical testing machine with static load cell pulling
attachment, Serial No. UK740. Needed to apply uniform uniaxial loading to samples
while recording sample deformation over time.
LabView Software to record and display force-deformation plot for analysis of material
and failure properties.

2. Lab Equipment:
Weight set for calibrating INSTRON 4444.
Calipers and Rulers for preparing samples and gathering sample dimensions used when
creating stress-strain plot.
Scalpels and scissors for sample cutting.
Marker/pen

3. Supplies:
Chicken skins (5 skins each group=100 skins total) this should be enough to yield 10-15
samples of adequate size (~1x 3) allowing for two sample groups with n~5.
3 Boxes: NYLON SUTURES, 900788 5-0 Nylon Blk Mono 18" DS-12 Sharpoint #AC-
0529D Bx/36. Suture-needle equipment for performing sutures. Each group gets 5.

4. Newly Purchased Equipment:

Other than the chicken skins and suturing materials listed above, nothing new needs to be
purchased.
Proposed Methods and Analysis:
Set up and Calibration of INSTRON 4444
Specify speed of the crosshead and the direction of movement before starting a run.
Verify load transducer settings in the INSTRON according to the procedures laid out in
step 2 of the Experiment 3 lab manual.
Set up INSTRON for sample testing as dictated in step 4 of Experiment 3 lab manual.
Specimen Harvest and Preparation:
Remove skins from 5 chicken legs provided.
Cut skins to desired dimensions as determined through observation of the INSTRON
clamps and based on experience with suturing in Experiment 2 (a recommended
dimension is 1x 3 which should allow for adequate clamping as well as suturing
length).
Separate into two groups with n=5: one to remain whole and the other to be cut in half
and then sutured back together.
Prepare the sutured group (n=5) by cutting the samples in half and repairing them with
five stitches of a pre-selected type (e.g. Running Locked, Interrupted, etc.).
INSTRON Specimen Tensile Testing:
Define a no-load position and loading protocol to be kept the same for all specimens.
Be sure to note each specimens dimensions (i.e. height, width, and thickness).
Subject each sample to loading in the INSTRON according the steps 2 and 3 of the
Experiment 3 lab manual.
Repeat testing for all specimens saving the force-displacement plots for each.
Interpreting the Data:
Use Force-displacement plots to determine the average failure displacements of each
group.
With the information recorded on specimen dimensions, convert force displacement data
to stress-strain data and re-plot with Matlab or Excel software (Appendix 1).
From the stress-strain data, determine the average elastic modulus of the groups as well
as average failure strain (Appendix 1).
Compare these data with an unpaired two-tailed t-test (=0.05) to test the hypothesis.
Analysis of Expected Results:
The INSTRON/LabView software setup will display force-displacement graphs for each
sample that will resemble those gathered in Experiment 3. Converting these plots to stress-strain
plots will yield plots that are also similar (at least for the whole chicken skin samples) to those
derived in Experiment 3.
If the posed hypothesis is correct, statistical analysis of the data sets for both groups will
show no significant difference between their failure displacements, failure strains, or elastic
moduli. On the other hand, if the two groups are significantly different, then a two-tailed t-test
will show low p-values, implying that the material properties of sutured skin are not the same as
those of whole skin.
Students may alternatively choose to test failure energy. Analysis of force-displacement
curves with Matlab software to estimate the area under the curve (up to the point of failure) will
yield failure energy (see Appendix 1). This too can be compared via a two-tailed unpaired t-test
amongst the two groups to judge the appropriateness of the suture as a surrogate.
Potential Pitfalls:
Combining Experiments 2 and 3 will provide new information on the strength of sutures
as skin surrogates when used to repair wounds. Unfortunately, the combination also yields a
combination of the potential problems of both experiments. Using the experience gained in lab
this semester, several of these potential pitfalls as well as potential remedies are presented below.

Though ideally the samples within groups would display similar properties, it was
observed in Experiment 3 that the results were highly variable. This high variance was
attributed to possible inconsistencies in sample composition/dimensions as well as
sample preparation. For example, some specimens may have been damaged or stretched
more than others when being cut or sutured. A possible solution or way to avoid such an
issue is to select samples more carefully with attention to sample uniformity and
gentleness of preparation.
Another problem encountered in Experiment 3 was the relatively small sample size. With
an n of 5, the data were highly variable and difficult to interpret. Larger sample sizes
would provide better insight into the material properties of both groups, but this is limited
to the resources provided.
Much like in Experiment 2, a problem may arise from inconsistencies or inadequacies in
suturing technique. When making and testing samples in Experiment 2, it was observed
that occasionally stitches would come undone, distributing the load amongst the other
stitches, or would have uneven lengths of thread, resulting in inconsistent load
distribution. A possible remedy for this is to have one person perform all sutures
(selecting the stitch he/she is most confident doing) with a controlled amount of thread
and measured stitch widths.
In Experiment 3, it was observed that the clamps used to secure the specimens sometimes
applied a force that was unaccounted for causing the samples to rupture at or around the
clamps and not in the center. The best way to handle such a situation is to note which
samples display such results, if it occurs at all, and keep it in mind as a cause of
variability or inconsistency of data.
Budget:
The only materials that need to be purchased for this experiment are the chicken samples and the
suturing materials.
100 chicken samples (enough for 20 groups to have 5 skins each) ~ $120
3 boxes of nylon suturing thread with needle: NYLON SUTURES, 900788 5-0 Nylon Blk
Mono 18" DS-12 Sharpoint #AC-0529D Bx/36. Three boxes will provide 108 sutures
allowing each group to perform 5 sutures with a few left over in case. 3 @ $120.36 =
$361.081
Total expected expenditure: $481.08

1
Prices taken from STAT Pharmaceuticals online medical suppliers.
(http://www.fluvaccine.com/Virtual/VirtualPages/Item_Page_MERCHANT_6.asp?
ItemCode=3L1)
Appendix 1:
Loading Rate 25 mm/min
Trial Failure Failure Failure Strain Stiffness
Displacement Force (mm/mm) (N/mm)
(mm) (N)
1 13.39 8.75 0.527 1.23
2 23.79 10.83 0.937 0.51
3 21.52 23.56 0.847 3.01
4 17.50 24.20 0.689 2.36
5 16.28 17.37 0.641 2.00
6 13.29 25.84 0.523 3.38
Average 17.63 18.43 0.694 2.08
4.28 7.31 0.169 1.08
Table 1: Failure and material properties of chicken skin under a load rate of 25 mm/min from Experiment 3.
Discussion of determining failure points, stiffness, and Youngs Modulus
Matlab software was employed in creating the stress-strain plots and to determine the elastic
modulus of the samples. Figure 1 is an example force-displacement plot.
Using Matlab software, the linear portion of each curve was isolated and a linear fit was
performed to find the slope of the line. R2 values were analyzed to make sure that the line was a good fit
for the graph. The same limits and procedure were used for the elastic modulus since the linear portions
of the force-displacement and stress-strain are the same (only with different proportions).

Figure 2. A sample force-displacement plot to demonstrate how failure properties, stiffness, and elastic modulus
were determined. Failure energy can also be determined from such a plot (image taken from Agres et. al.) 2.
Stress = Force/Cross-sectional Area
Strain = change in length/original length
Youngs (elastic) Modulus = stress/strain
Sample calculation of stress from force and strain from displacement:
Original Width=25.40 mm, Original Length=25.40 mm, and Original Thickness=6.35 mm
Failure Displacement/Original Length=Failure Strain
41.20/25.40=1.62
Failure Force/Cross Sectional Area=Failure Stress
9.11/(25.40 x 6.35)=0.056 N/mm2

2
Agres et. al., Failure Energy: Suture Performance under Uniaxial Tension, 4/28/06