Jessica Lopez, Yuriy Bazylev, Henry Kwong, and Jamie Adkins

Mentor: Dr. Vilupanur A. Ravi, Department of Chemical and Materials Engineering, Cal Poly Pomona,
3801 West Temple Ave., Pomona, CA, 91768, jelopez@csupomona.edu, ysbazylev@csupomona.edu,
hkwong@csupomona.edu, jladkins@csupomona.edu

Mechanical Properties of Spider Silk

Abstract:

Spider silk is a natural material that has unique mechanical properties. These include high

mechanical strength and toughness, and strength-to-weight ratios five times that of steel fibers.

The structure of spider silk is of particular interest at the molecular level due to its effects on the

properties of the material, particularly its mechanical behavior. Research at Cal Poly Pomona

over the past few years has focused on evaluating the mechanical properties of fibers from

different spider species with a view to elucidating structure-property interrelationships that

would form the basis for new material designs. Our focus in the current project has been on the

determination of the mechanical properties of spider silk produced by the Araneus gemma,

known as the orb weaver, and the Nephila clavipes, known as the giant golden orb weaver.

These two types of spider hang-line silk were tested in uniaxial tension using a mechanical test

frame - the Instron 3342. A 5 N load cell was used to obtain the load-extension data for the

analysis. The properties of these silks were compared to the silk from the Lactrodectus

geometricus, known as the brown widow spider, which was tested by the previous teams. The

effect of engineering strain rate on the ultimate tensile strength, extensibility, etc., of these silks

will be discussed. The behavior of spider silk will be discussed as a viscoelastic polymer. Ideas

for future research, e.g., silk-reinforced agar composites, cellular growth along silk strands,

viscoelastic characterization using a Dynamic Mechanical Analyzer (DMA), and further

characterization of the hang-line silk at different strain rates using the Instron tensile tester will

also be discussed.

1
Narrative:

The motivation for this project is derived from the fascinating properties of spider silk that

include its high specific strength and structure. Spider silk is a biodegradable material that is

composed of very specific protein sequences that can potentially lead to biomedical applications

such as regenerative nerve growth in patients who have sustained nerve damage due to injury or

genetic predisposition [Allmeling, et al]. Other possible applications may include fluid body

armor that is able to conform to the human body and biodegradable thread that can be used for

surgical sutures. The silk formation occurs within the spider’s spinneret glands, which hold a

liquid protein that crystallizes upon ejection from the spinnerets (Figure 1).

Figure 1: Spider Silk Formation (Photograph by Dennis Kunkel)

Each spinneret gland shown in green releases the liquid crystal protein-shown in purple. The
liquid strands cling to each other, forming a larger strand which then solidifies upon ejection
from the abdomen.
The specific mechanism of liquid-to-solid protein phase conversion is unknown; however, it is

theorized that the highly pressurized conditions the liquid protein is subjected to upon ejection

from the spinnerets causes them to crystallize and form the solid strands that we are able to

detect [Vollrath]. The hang-line silk itself is composed of many individual silk filaments which

2
can be categorized into two key strands that compose the single visible strand. Those two strands

are the major and the minor ampullate, which are produced by the Major Ampullate Gland and

the Minor Ampullate Gland, respectively. The major ampullate acts as the main anchor which

supports the majority of the spider’s weight as it traverses its surroundings, while the minor

ampullate acts as the safety hook in the event that the major ampullate fractures. The

misconception about silk is that it is a woven fiber. In actuality, the two main silk strands are

ejected from the spider’s abdomen parallel to each other and are cemented together at the

Piriforme gland, located at the end of the spider’s abdomen.

The majority of the hang-line silk is composed of glycine and alanine rich protein regions, giving

the silk its unique properties. The glycine regions compose on average about 42% of the silk in

its protein structure, which greatly contribute to the silks’ high extensibility [Spider Silk &

Venom]. The alanine rich regions compose about 25% of the silk and its hydrogen bonded

structure gives the silk its incredible specific strength. These polyalanine regions are highly

crystalline structures due to the numerous hydrogen bonds which create beta-pleated sheets.

Though hydrogen bonds are weaker than ionic, covalent, and metallic bonds in materials, the

multitude of these bonds create a strong and dense structure. When the applied load reaches the

yield strength, these chains begin to slip past one another and as they do, they attempt to reform

the broken bonds and are thus able to re-support the applied load [Brehm, et al].

To properly characterize the properties of spider hang-line silk, we must have fresh silk samples

because of the rapid degradation of these fibers upon exposure to air. The spiders and frames

were kept in a room where the humidity and temperature were held constant. Their diet consisted

of crickets and bees. A limit of one spider per frame was used due to the cannibalistic nature of

the spiders. Specimens were collected on campus and silk diameter measurements were taken

3
immediately upon collection using the Leica 4000-Dm optical microscope. After spiders were

collected, they were cared for utilizing an open air frame measuring 20” x 12” x 48” constructed

from tree branches and a cardboard box. To obtain silk diameter measurements, several readings

were taken from each visible strand under a 100X objective lens. The diameters of individual

strands that comprise a fiber were measured in at least 5 different longitudinal locations. The

average measurements for the individual strands were then summed to obtain the total diameter

of the silk fiber. These measurements showed that the L. geometricus had a silk with diameters

ranging between 2 and 5 µm. The A. gemma silk diameters ranged from 5 to 8 µm, and the N.

clavipes silk ranged from 10 to 17 µm in diameter. Tensile tests were then conducted using an

Instron 3342 equipped with a 5 N load cell.

The silk samples were subjected to a constant engineering strain rate tensile test. As the silk

sample is extended, a continuously increasing load is applied until the sample fractures

completely. During the test, load and extension are continuously recorded. Using this

information, stress and strain are calculated by the software. Stress is the amount of load applied

to the sample, P, over the original cross sectional area of the spider silk, A0.

P
Tensile Stress σ=
A0

Strain is the change in length, ΔL, over the original test length of the sample.

ΔL
Tensile Strain ε=
L

Following completion of the test, a tensile stress (MPa) versus tensile strain (mm/mm) graph was

obtained. An example of a stress-strain curve for A. gemma is shown in Figure 2 for an

4
engineering strain rate of 0.39 %/s. The engineering strain rate is calculated using the crosshead

speed, v(t), and the gage length of the specimen, l0 as shown in the equation below.

v (t )
Strain Rate ε =
L

Figure 2: Stress vs Strain Graph for the A. Gemma Spider

(Graph generated by Instron 3342. Each separate colored curve represents a single strand
sample, with each related modulus shown by the linear blue lines.)

There are several regions of interest for the stress strain curve. First, there is a linear, elastic

modulus portion occurring at the first 5% of strain. This elastic portion is due to the silk’s

stiffness as the hydrogen bonds are being loaded. The second portion is observed by the decrease

in modulus. The stiffness of the silk is reduced because the hydrogen bonds of the silk are

beginning to break due to the load and reform. Lastly, a single silk strand fractures at

approximately 400 MPa and the remaining silk strands attempt to carry the applied load. As the

specimen continues to be stretched, other silk strands continue to fracture until total silk fiber

fracture occurs. All silk samples exemplified these regions regardless of species.

5
In order to compare the data that we obtained from the three spider species, we averaged the data

at the ultimate tensile stress and produced several graphs. Comparing tensile strength to strain

rate, it was expected that the tensile strength would increase with increasing strain rate.

However, the N. clavipes data displays an increasing trend initially, followed by a more complex

behavior with eventual decrease in strength as the strain rate continues to increase (Figure 3).

This contrasts with the behavior of the other silks which show an initial decrease in strength

followed by a more typical monotonic increase with increasing strain rate. The latter follows the

strain hardening behavior associated with typical polymers [Ravi].

Tensile  Strength  vs  Strain  Rate  

1400  
Gemma  
1200  
Geometricus  
Tensile  Strength  (MPa)  

1000  
Nephilia  
800  

600  

400  

200  

0  
0   5   10   15   20   25   30   35   40   45  
Strain  Rate  (%/s)  

Figure 3: Tensile Strength vs. Strain Rate

Maximum tensile strain was also compared to strain rate in order to study their relationship

(Figure 4). For the N. clavipes, the maximum tensile strain initially decreases followed by an

increase and a subsequent decrease. On the other hand, both the L. Geometricus and A. Gemma

displayed a different behavior in regards to the same parameter. Figures 3 and 4 are

6
complementary, i.e., increases in strength are associated with decreases in ductility. Our results

for N. Clavipes compare favorably with literature values [M. Elices et al].

0.3  
Max  Tensile  Strain  vs  Strain  Rate  
Max  Tensile  Strain    (mm/mm)  

0.25  

0.2  

0.15  

0.1  
Gemma  
0.05   Geometricus  
Nephilia  
0  
0   5   10   15   20   25   30   35   40   45  
Strain  Rate  (%/s)  

Figure 4: Maximum Tensile Strain vs. Strain Rate

In conclusion, the mechanical properties of the A. Gemma and the N. Clavipes spider silk were

determined, e.g., tensile strength, the tensile strain, etc. Comparisons were made between the

properties of the two types of silk above and L. Geometricus which was tested by the previous

team. We noticed all the silk samples that were tested followed a similar trend with the first 5%

of the tensile strain having a high modulus and then reaching a maximum tensile strain where the

different silk strands begin to break and the other strands take on the load showing a staggered

failure.

In the future, we plan to continue tensile testing with larger sample group sizes and at different

strain rates. We then plan to further characterize mechanical properties using the Dynamic

Mechanical Analyzer. Understanding the mechanical behavior will allow us to develop

biological composites based on agar matrices reinforced by a silk weave. We will then perform

7
degradation tests of the silk fibers in various biologically relevant media, such as saline, blood,

etc. The data collected from this experiment will allow us to approximate the period of time silk

will retain its useful properties in physiological conditions. To determine practical biomedical

applications of the silk, we will also experiment with controlled celled growth along the strand of

silk to determine any possible antimicrobial properties and biocompatibility. The combination of

the observed mechanical properties, biodegradability, microbial properties, and composite design

has the potential to lead to the development of a more readily available aid for generating nerve

re-growth in afflicted patients.

References:

Allmeling, C, A Jokuszies, K Reimers, S Kall, C. Y Choi, G Brandes, C Kasper, T Scheper, M

Guggenheim, and P. M Vogt. Spider Silk Fibres in Artificial Nerve Constructs Promote
Peripheral Nerve Regeneration. Cell Proliferation, 41.3 (2008): 408.

Brehm, Denise. "Nanotechnology Today." : Protein's Strength Lies in H-bond Cooperation VIDEO. N.p.,
26 Feb. 2008. Web. 16 Mar. 2013. <http://nanotechnologytoday.blogspot.com/2008/02/proteins-strength-
lies-in-h-bond.html>.

M. Elices, J. Pérez-Rigueiro, G. Plaza, G.Guinea. Finding Inspiration in Argiope Trifasciata

Spider Silk Fibers. JOM. 2005.

"Spider Silk & Venom." Spider Silk. University of Bristol, n.d. Web. Last accessed on 3/15/2013.
<http://www.chm.bris.ac.uk/motm/spider/page3.htm>.

Vollrath, Fritz. Strength and Structure of Spiders’ Silk. Reviews in Molecular Biotechnology.
74th ed. 2000: 67-83.

V. Ravi, Personal Communication.

8
9