EFFECT OF

LEAD (Pb)
EXPOSURE
ON SKIN
ELASTICITY
ECOLE PRIVEE
INTERNATIONALE
TCHADO-TURQUE
CHAD-2010
 EDITOR and ADVISORY
FAYSAL YILMAZ
NTIC
Head of Biology Department
nigeria_biology@yahoo.com

ASSOCIATE EDITORS

Dayanat Mammadov
EPITT Computer Teacher

Yousouf Daoussa Deby Itno

EPITT Student
(4emeClass)
Younous Oumar Souni
EPITT Student
(4emeClass)
Haggar Bechir Ali Abdramane Haggar
EPITT Student
(5emeClass)
Idriss Ramadan Erdebou
EPITT Student
(4emeClass)
 The Effect of Lead (Pb) Exposure on Skin Elasticity

ABSTRACT
The goal of this project is to determine whether exposing chicken skin to lead metal for an extended period of time (1 week) in
DI water will change the Young’s modulus of the skin. The aim of using lead in this experiment is to be able to apply any
findings to real-world situations in which people become exposed to lead through their water pipes. If it is found that lead does
have adverse effects on chicken skin, the findings can be used as a basis for further testing with human skin so that the negative
health effects of lead can be further understood. This greater understanding could be used to generate more awareness of the
possible harms of lead exposure and to develop treatments against lead poisoning. Based on literature findings on the
composition of collagen and elastin fibers in the skin, it is hypothesized that lead exposure will make the skin more fragile and
prone to breakage than normal, resulting in a lower Young’s modulus than unexposed skin. By comparing the Young’s moduli of
the lead-exposed skin samples to the Young’s moduli of unexposed samples using a one-tailed, unpaired t-test in Excel, p<0.05
did reveal that the Young’s modulus of the exposed samples were significantly less than that of the unexposed samples.

INTRODUCTION
The goal of this project is to determine whether exposure to lead can change the elasticity of chicken
skin, as roughly measured by Young’s modulus. It is widely known that the extracellular matrix of skin (and
many other tissues) contains numerous collagen and elastin fibers. The fibers are constructed from individual
collagen or elastin molecules which are covalently cross-linked. The cross-links form between the side chains of
different lysine amino acids in the molecules and lend a higher tensile strength to the fibers and hence to the
skin.
The lysine side chains have terminal –NH3+ groups which join by forming a bond
between the N atoms (1). In the process, H atoms are given up. However, if a sufficient
number of these bonds are broken, the skin will become fragile and tear-prone (1). In order
to break the bonds, H atoms must be reintroduced through a reduction reaction. One possible
way of accomplishing this reduction might be to expose the tissue to lead, which can
undergo the reaction Pb→Pb+2 + 2e-, thereby providing electrons to initiate the reduction
reaction and break the bonds.
This project was developed to protect the public health by minimizing lead levels in
drinking water. If water is too corrosive, it can cause lead to leach out of the plumbing
materials and fixtures and enter the drinking water. Children are especially susceptible to
high levels of lead, which can cause damage to the brain, red blood cells, and kidneys.
Exposure to low levels of lead can cause low IQ, hearing impairment, reduced attention span
and poor classroom performance. High exposure to copper can cause stomach and intestinal
distress, liver and kidney damage, and complications from Wilson’s disease in genetically-
predisposed people.
High lead levels in adults have been linked to high blood pressure. Pregnant women and
their fetuses are especially vulnerable to lead exposure, which can significantly harm the
fetus, cause low birth weight and slow down normal mental and physical development.

Using lead could lend the experiment a practical objective, since exposing chicken
skin to lead in this experiment could simulate the exposure to lead that some people get from their tap water if
the pipes or solders in their homes are old (pre-1987) and lead-containing (City of N`DJAMENA/Chad Water
Department). Thus, if it is found that such exposure has the capability of changing the mechanical properties of
the skin, further experiments could be done to determine whether human skin is also at risk from such exposure.
In this project unexposed skin was used for comparison of the exposed skin.
EQUIPMENTS
Major Equipment

Handmade Instron - The Instron was used to stretch the lead-exposed skin samples. The Excel software will
record force/displacement data for each sample.

Lab Equipment

2 Large plastic containers- This was required to be large enough to soak chicken legs wrapped in lead
sheeting prior to the actual experiment. Ten legs will need to be soaked in each container.
Surgical scissors- These were used to separate the skin from the legs and cut out samples from the skin of
set dimensions.
Knife/scalpel- These were needed to help with cutting the skin off of the leg itself and to cut the skin into
sample pieces. Like the marker, they could also be used to make small markings (in the form of cuts) where
the skin should then be cut with the scissors.
Forceps- These were useful for peeling the skin off the legs and for holding the skin in place when the
samples are being cut.
Cutting board- This was used as the surface on Calipers- These were used to measure the
which the skin is stripped from the legs and cut thickness of the skin samples and the distance
into samples between the loading clamps on the Instron once
samples are loaded.

Weight set (500g, 1kg, 2kg) - These were used to Incubator- It was used to stimulate the human
verify that the load cell transducer in the body by adjusting the temperature as 37oC
Handmade-Instron is outputting correct
measurements by making measurements for
known applied loads.
Supplies

20 freshly slaughtered chicken - The skin was 20 1 ft. x 1 ft. (1 ft = 30.48 centimeters) pieces of
exposed to lead and will act as the actual test 1/32 inch (1 inch 2.54 cm) 99% pure lead sheeting-
subject in this experiment. 20 legs are needed for These are needed to act as the source of lead
each group to use 10 in an experiment. exposure for the skin.
Gloves- These were worn by anyone handling the lead
or meat to avoid contamination of their hands by
germs from the meat or by lead, which, if ingested,
can be harmful.
Paper towels wet with DI water- These was needed
to store the skin samples after they have been cut
and before they are loaded into the Instron to
prevent their drying out, which could cause
changes in their mechanical properties.
DI water- This was necessary to soak the lead-
covered legs in before the experiment.
Microscopes & Fixed Human Skin Preparets- To
be able to understand better the anatomy of human
skin, some specimens were observed unde
microscopes.
METHODS
1. One week before the lab was to be run, separate the legs from corpse and wrap each of 10 chicken legs
tightly in a 1 ft. x 1. ft. piece of lead sheet (maximize the amount of skin that is directly in contact with
the lead, especially the region of skin just above the knee). Next, submerge all of the wrapped and non-
wrapped legs in two large plastic containers independently and fill with DI water. Set the containers in
an incubator (37oC) and let it sit for a week. BE SURE TO WEAR GLOVES DURING THIS STEP
AND ALL STEPS IN WHICH THE CHICKEN SKIN OR LEAD IS HANDLED.

1a- Leg separation from corpse 1b- Lead sheet preparation

1c- Wrapping the legs 1d- Submerged two groups of legs

1e- Filling the containers with DI water 1f- Ready legs

 1g- Putting the legs in incubator 1h- One week at 37oC

2. On the day of the experiment, set up the handmade-Instron and calibrate it. Generally:
• Verify the load transducer settings using the provided set of known weights.
• Make sure the Instron is set to move up, or in other words, to stretch the samples.

2a- Main components of handmade-Instron 2b- Setting up the Instron

2c- Loading known weighs 2d- Stretching the sample

3. Remove the lead from the chicken legs and the skin from the legs. Cut the skin off so that the part above
the outside of the knee remains intact. While not working with the skins, place them in wet paper towels.
Make sure you note in what orientation you place them.
 3a- Dissection unit 3b- Getting ready for removing the skin

3c- Wetting the paper towel 3d- Cutting the skins off

3e- Placing them on wet paper towels 3f- Skin pieces are ready

4. Cut 2 samples of skin from each of the 20 specimens. The samples should be taken from just above the
outside of the knee and have dimensions of 2 cm x 4 cm (determined to be appropriate for Instron
testing). Use the calipers to make measurements. Place the samples back in the paper towels until testing
occurs.

4a- Cutting sample skins 4b- Using calipers

 4c- Prepared GI (no lead) skins on towel 4d- Prepared GII (with lead) skins on towel

5. Taking one sample, clamp it in the Instron with the 2 cm sides in the clamps. Using the calipers,
measure the distance between the top of the bottom clamp and the bottom of the top clamp. This will act
as the gage length in strain calculations and should be kept constant for all samples. Also, measure the
width of the sample in the clamps and the thickness near the center of the sample. Multiply these to find
the cross-sectional area of the sample. This was used to calculate stress. Pull the dynamometer for
different weighs and measure the distance. Record all data.

5a- Clamping the skin 5b- Measuring the thickness & width

6. Pull the dynamometer for different weighs (10-20-30 N) and measure the strain. Record all data and
repeat it for all 40 samples.

6a- Loading sample 6b- Loading weighs

 6c- Measuring the strain 6d- Recording data

ANALYSIS
1. Move the raw force (N) and displacement (mm) data for each sample into an Excel worksheet. Divide
all the force data of a sample by the cross-sectional area of that sample and then divide by 1000 to
calculate stress data in kPa. Divide the displacement data of the sample by the gage length to obtain
stress data. Repeat for all samples. (Stress=Force/Area)
2. Find the Young’s modulus (E) for each of the skin samples. This was the slope of the fit line for the
sample. (E= Stress/Strain)
3. Compare the Young’s moduli of the lead-exposed skin samples to the Young’s moduli of unexposed
samples using a one-tailed, unpaired t-test in Excel. Since the variance of the Young’s modulus of the
unexposed samples is much more than 5% of their mean, unequal variance is assumed. A p<0.05 will
reveal that the Young’s modulus of the exposed samples is significantly less than that of the unexposed
samples.

YOUNG MODULUS = STRESS/STRAIN

E= (F/A) / (∆L/L)
RESULTS
Experimental Results
STRESS                 STRAIN                                 YOUNG MODULUS              
          (F/A)  (∆L/L)  (E=STRESS/STRAIN) 

CROSS  1ST  2nd  3rd 

ST ND RD         GAGE 
SAMPLE  THICKNESS    WIDTH    SECTIONAL  1      2      3 EXTENT  EXTENT  EXTENT  MEAN  
LENGTH  (E1)  (E2)  (E3) 
  NUMBER  (mm)  (mm)  AREA  10/A 20N/A 30N/A LENGTH  LENGTH  LENGTH  (E) 
L0(mm) 
(A)(mm2)  ∆L(mm)  ∆L(mm)  ∆L(mm) 
1  0.80  20.00  16.00  0.63  1.25  1.88  28.00  0.80  1.70  3.20  21.88  20.59 16.41 19.62 
2  1.20  20.00  24.00  0.42  0.83  1.25  28.00  0.60  1.50  3.00  19.44  15.56 11.67 15.56 
3  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.75  1.56  3.10  20.74  19.94 15.05 18.58 
4  0.80  20.00  16.00  0.63  1.25  1.88  28.00  0.80  1.70  3.20  21.88  20.59 16.41 19.62 
5  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.75  1.56  3.10  20.74  19.94 15.05 18.58 
6  1.10  20.00  22.00  0.45  0.91  1.36  28.00  0.65  1.60  3.40  19.58  15.91 11.23 15.57 
7  1.20  20.00  24.00  0.42  0.83  1.25  28.00  0.60  1.50  3.00  19.44  15.56 11.67 15.56 
8  1.13  20.00  22.57  0.44  0.89  1.33  28.00  0.68  1.58  3.24  18.24  15.70 11.49 15.14 
GROUP‐ 9  1.16  20.00  23.29  0.43  0.86  1.29  28.00  0.61  1.50  2.90  19.71  16.03 12.44 16.06 
I          10  1.20  20.00  24.00  0.42  0.83  1.25  28.00  0.60  1.50  3.00  19.44  15.56 11.67 15.56 
NO  11  1.24  20.00  24.71  0.40  0.81  1.21  28.00  0.54  1.23  2.84  20.98  18.42 11.97 17.12 
LEAD  12  1.27  20.00  25.43  0.39  0.79  1.18  28.00  0.50  1.00  2.20  22.02  22.02 15.02 19.69 
13  1.31  20.00  26.14  0.38  0.77  1.15  28.00  0.20  0.80  1.95  53.55  26.78 16.48 32.27 
14  1.34  20.00  26.86  0.37  0.74  1.12  28.00  0.19  0.75  1.60  54.87  27.80 19.55 34.07 
15  0.80  20.00  16.00  0.63  1.25  1.88  28.00  0.80  1.70  3.20  21.88  20.59 16.41 19.62 
16  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.75  1.56  3.10  20.74  19.94 15.05 18.58 
17  1.00  20.00  20.00  0.50  1.00  1.50  28.00  0.70  1.20  2.90  20.00  23.33 14.48 19.27 
18  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.75  1.56  3.10  20.74  19.94 15.05 18.58 
19  1.10  20.00  22.00  0.45  0.91  1.36  28.00  0.65  1.60  3.40  19.58  15.91 11.23 15.57 
20  1.30  20.00  26.00  0.38  0.77  1.15  28.00  0.21  0.90  1.96  51.28  23.93 16.48 30.57 
 
 
           
 
  
 
 

STRESS                  STRAIN                                 YOUNG MODULUS              

          (F/A)  (∆L/L)  (E=STRESS/STRAIN) 

CROSS  1ST  2nd  3rd 

ST ND RD         GAGE 
SAMPLE  THICKNESS    WIDTH    SECTIONAL  1      2      3 EXTENT  EXTENT  EXTENT  MEAN  
LENGTH  (E1)  (E2)  (E3) 
NUMBER  (mm)  (mm)  AREA  10/A 20N/A 30N/A LENGTH  LENGTH  LENGTH  (E) 
L0(mm) 
(A)(mm2)  ∆L(mm)  ∆L(mm)  ∆L(mm) 
22  0.80  20.00  16.00  0.63  1.25  1.88  28.00  0.42  0.82  1.60  41.67  42.68 32.81 39.05 
23  1.10  20.00  22.00  0.45  0.91  1.36  28.00  0.25  0.74  1.40  50.91  34.40 27.27 37.53 
24  1.10  20.00  22.00  0.45  0.91  1.36  28.00  0.25  0.74  1.40  50.91  34.40 27.27 37.53 
25  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.35  0.78  1.55  44.44  39.89 30.11 38.15 
26  0.80  20.00  16.00  0.63  1.25  1.88  28.00  0.42  0.82  1.60  41.67  42.68 32.81 39.05 
27  1.30  20.00  26.00  0.38  0.77  1.15  28.00  0.12  0.46  0.98  89.74  46.82 32.97 56.51 
GROUP‐ 28  1.40  20.00  28.00  0.36  0.71  1.07  28.00  0.08  0.42  0.85  125.00 47.62 35.29 69.30 
II  WITH  29  1.10  20.00  22.00  0.45  0.91  1.36  28.00  0.25  0.74  1.40  50.91  34.40 27.27 37.53 
LEAD  30  1.20  20.00  24.00  0.42  0.83  1.25  28.00  0.26  0.90  1.90  44.87  25.93 18.42 29.74 
31  0.80  20.00  16.00  0.63  1.25  1.88  28.00  0.42  0.82  1.60  41.67  42.68 32.81 39.05 
32  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.35  0.78  1.55  44.44  39.89 30.11 38.15 
33  1.00  20.00  20.00  0.50  1.00  1.50  28.00  0.30  0.75  1.20  46.67  37.33 35.00 39.67 
34  1.20  20.00  24.00  0.42  0.83  1.25  28.00  0.26  0.90  1.90  44.87  25.93 18.42 29.74 
35  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.35  0.78  1.55  44.44  39.89 30.11 38.15 
36  0.90  20.00  18.00  0.56  1.11  1.67  28.00  0.35  0.78  1.55  44.44  39.89 30.11 38.15 
37  1.30  20.00  26.00  0.38  0.77  1.15  28.00  0.12  0.46  0.98  89.74  46.82 32.97 56.51 
38  1.20  20.00  24.00  0.42  0.83  1.25  28.00  0.26  0.90  1.90  44.87  25.93 18.42 29.74 
39  1.00  20.00  20.00  0.50  1.00  1.50  28.00  0.30  0.75  1.20  46.67  37.33 35.00 39.67 
40  0.80  20.00  16.00  0.63  1.25  1.88  28.00  0.42  0.82  1.60  41.67  42.68 32.81 39.05 

Table: Results of experiment and data analyzing

Statistical t-Test Results

The results of unpaired t-test

t= 7.91

Standard deviation= 8.08

Degrees of freedom = 38

Group A: Number of items= 20

29.7 29.7 29.7 29.7 37.5 37.5 37.5 38.1 38.1 38.1 38.1 38.1 39.0 39.0 39.0 39.0 39.7 56.5 56.5 69.3

Mean = 40.0

95% confidence interval for Mean: 36.33 thru 43.65

Standard Deviation = 9.92

Hi = 69.3 Low = 29.7

Median = 38.1

Average Absolute Deviation from Median = 5.43

Group B: Number of items= 20

15.1 15.6 15.6 15.6 15.6 15.6 16.1 17.1 18.6 18.6 18.6 18.6 19.3 19.6 19.6 19.6 19.7 30.6 32.3 34.1

Mean = 19.8

95% confidence interval for Mean: 16.12 thru 23.43 Degrees Probability, p
of
Freedom 0.1 0.05 0.01 0.001
Standard Deviation = 5.69
28 1.70 2.05 2.76 3.67
Hi = 34.1 Low = 15.1
29 1.70 2.05 2.76 3.66

Median = 18.6 30 1.70 2.04 2.75 3.65

40 1.68 2.02 2.70 3.55

Average Absolute Deviation from Median = 3.42
60 1.67 2.00 2.66 3.46
120 1.66 1.98 2.62 3.37
infinity 1.65 1.96 2.58 3.29
CONCLUTION
Our t value (7.91) was bigger than the table value (2.03). That proves lead exposed chicken skins had
lost their flexibility by showing low Young module values. It was known the effects of lead on internal organs,
but any scientific research about skin was not done before. We hope this project would help to understand the
effects of lead exposure better and to aware people who still use lead pipe in their installation.

References

(1) Alberts, Bruce, et al. (2002). Molecular Biology of the Cell. (4th ed., pp. 1099,1102). New
York: Garland Science.
(2) Zumdahl, Steven S. (2005). Chemical Principles. (5th ed., pp. 468).
(3) Boston: Houghton Mifflin Company.
(4) Bains, A., Baumann, B., Connie, C., & Wang, E. (2007). Tensile Testing: Elastic Properties.
(5) Meeting the Lead Standards. City of Philadelphia Water Department. Retrieved April 23,
2007 from http://www.phila.gov/water/water_quality2.html#bk_leadstandard.