The document compares the regenerative properties of scaffold material derived from pig adipose tissue and bladder extracellular matrix (ECM). It first reviews that ECM plays an important signaling role in tissue development and healing. Studies show pig ECM is a viable biomaterial for tissue engineering due to its structural proteins. The document proposes an experiment to compare the wound healing effects of scaffold material prepared from pig adipose tissue versus bladder ECM on human subjects. It hypothesizes that bladder ECM will have superior regenerative results. The independent variables are the adipose and bladder materials, while the dependent variable is the healing outcomes in humans.

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Effects of regenerative properties based on different tissue E.C.M scaffold material.
Juan A. Barrera
University of Houston Downtown
CSP 110N
Dr.Parker
May 9, 2013

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Table of Contents
I. Abstract……………………………………………………………………..…………2
II. Introduction………………………………………………….……………..…….……3
III. Literature Review………………………………….….…………………..……...……4
A. Functions and Properties of ECM………………………...………..………….4
B. Healing Power and Use of ECM………………………………………………5
C. The use of pig ECM…………………………...………………………………5
IV. Problem Statement…………………………………………………………………….7
V. Hypothesis……………………………………………………………………….….…7
VI. Variables…………………………………………………………………...……….…7
VII. Methods…………………………………………………………………………..……7
VIII. References………………………………………………………………………….….9

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Abstract
The human body’s ability to regenerate and heal damaged body parts is limited. In many
cases the only option for the repair of trauma is tissue transplantation or injectable tissue fillers.
These procedures sometimes leave long term drawbacks that affect the survivability of the
transplanted tissue. Luckily in recent years there has been ongoing research on the effects of
mammalian Extracellular Matrix (ECM) prepared as scaffold material. The ECM of various
tissues and organs are appropriate substrates for the support of,”… tissue specific cell phenotype
and function” due in fact to its composition. The ECM consists of secreted products from its
resident cell population which is affected by signaling of the ECM. The resident cell population
and ECM share an ongoing mutual exchange. Pig adipose and bladder tissue offer an abundant
source of ECM and is the main source of scaffold material. Porcine tissue is used because it is
an easy, “off the shelf” material to obtain and work with (Brown, B., Freund, J., Han, L., Rubin,
J., Reing, J., Jeffries, E., & Badylak, S. 2011, p 411).
In this proposal there will be a comparison between pig’s adipose and bladder tissue on
healing/regenerative properties of humans. The independent variables are both the adipose and
bladder tissue. The dependent variables are the results of the regenerative properties on humans.

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Introduction
The ability of humans beings to be able to repair, regrow, and regenerate damaged body
parts has long been a topic of science fiction. The fact is the human body is limited in its
regenerative abilities. It is impossible for a human being to lose an appendage and regrow it or be
able to replace a malfunctioning organ. In recent years research on the Extra Cellular Matrix has
revealed surprising facts. The Extracellular Matrix (ECM) is the adhesion that keeps all cells
together. Every different type of cell whether its muscle cells, skin cells, or brain cells are all
fixed in a template known as the ECM. The ECM can be extracted from animals and
manipulated into a powdered, sheet or gel form for topical use. When the ECM is placed in a
wound it can signal the body to start regrowth.
When an amputation occurs,” …local responses at the site of the wound play important
roles in the initiation of regenerative processes.” (King, R., & Newmark, P., 2012, p 554). The
E.C.M is able to trigger responses from the surrounding cells to initiate growth. In most
regenerating organisms, the process of regeneration requires for the creation of new cells. When
a salamander loses a limb a series of events unfold that include the formation of a clump of
cells known as the blastema at the site of the injury, and this regenerates the missing body parts.
(Sánchez Alvarado, A., 2009).The ECM when applied to a wound acts very much like the clump
of cells from a salamander. The ECM applied to the wound creates a thin layer that signals the
surrounding cells to fill in the wound. Over time the thin layers completely fill the wound back to
the way it was before injury.

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Literature Review
Function and Properties of ECM
The extracellular matrix (ECM) has long been believed to merely be the glue that held
cells together. In recent years research has revealed the ECM has a greater role in the lives of
cells. It turns out that by collaborating with a cell via integrins, the ECM can control a cell’s
behavior (Campbell, N. A., & Reece, J. B., 2005). Furthermore the ECM has the ability to
support tissue development and remodeling. (Thiriet, Marc, 2011).The ECM is made up of
carbohydrate-containing molecules and glycoproteins secreted by cells. The most abundant
glycoprotein in the ECM is collagen, which forms strong fibers through the external part of the
cell. These fibers are attached to fibronectin and integrins that are built into the plasma
membrane of a cell.
Figure 1. Extracellular Matrix(ECM) of an animal cell (Campbell, N. A., & Reece, J. B. 2005)

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Healing power and use of scaffold ECM
All of these properties of the ECM make it a very good candidate for the healing process.
When a small cut is presented in tissue, blood flows to the wound signaling the surrounding cells
to begin the healing process. When a very deep cut is presented it is impossible for blood to fill
in the cut. There cannot be any healing or filling in of tissue where there is none. The ECM with
its tissue formation properties allows for the formation of a thin layer of scaffold material where
the surrounding cells can come in and start healing. ECM has been used, “… for the repair of
absent or damaged human organs and has been established in preclinical and in human clinical
applications” (Ringel, R., Kahane, J.,Hillsamer, P., Lee, A., & Badylak, S.,2006, p 196) . ECM
tissue engineering procedures have become an accepted, viable, and “…greater approach for
tissue restoration and reconstruction Biological matrix material as an inductive scaffold
material has already been used in more than 300,000 human patients treated in the past 5 years.
(Ringel, R., Kahane, J.,Hillsamer, P., Lee, A., & Badylak, S.,2006). This approach of the use of
ECM for tissue repair is much more convenient both economically and functionally. This
technique as compared to whole organ transplants, insertion of mechanical devises, and tissue
rejection problems is a much more viable and practical option.
The use of pig ECM
One of the main reasons why ECM scaffold material from pigs is used because it
includes the practical and structural proteins that are part of the natural mammalian ECM (Liu,
L., Li, D., Wang, Y., Xu, H., Ge, L., & Liang, Z. 2011). Pigs offer and abundant source of tissue
that would otherwise be thrown away. Most of the material that is used to extract ECM

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comes from the pigs skin, small intestine and urinary bladder which are waste in the agriculture
industry. “Porcine adipose tissue is easily obtained in large quantities from commonly discarded
food waste” (Choi, Y., Choi, J., Kim, B., Kim, J., Yoon, H., & Cho, Y. 2012, p 866). All this
waste tissue is being put to good use in research. In a recent study conducted by Hanyang
University in Korea revealed that porcine adipose tissue “supported the adhesion and growth of
human cells in vitro”. They also concluded the adipose tissue could be useful as an alternative
biomaterial for tissue engineering. (Choi, Y., Choi, J., Kim, B., Kim, J., Yoon, H., & Cho, Y.
(2012). Another study conducted in 2006 applied the use of pig urinary bladder ECM for the
repair of the larynx in dogs. The results concluded the ECM repair resulted macroscopic and
microscopic reconstruction of the laryngeal tissue (Ringel, R., Kahane, J.,Hillsamer, P., Lee, A.,
& Badylak, S.,2006). Other studies have gone on to study the morphological, bioactive and
biomechanical properties of these diverse pig ECM scaffold materials (Liang, R., Fisher, M.,
Yang, G., Hall, C., & Woo, S. (2011).It is important to understand that different ECM material
from different tissue types have their own unique and distinct properties. Therefore it is of
massive importance to know what ECM from what specific tissue has the best healing or
regenerative effect. ECM scaffolds prepared from different tissue have been proven to show their
own distinct biological properties. Some of these properties include, “three-dimensional
organization, water absorption ability, matrix degradation, mechanical properties, antimicrobial
activity, and cell attachment ability, etc,” (Liu, L., Li, D., Wang, Y., Xu, H., Ge, L., & Liang, Z.
(2011, p 222).

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Problem Statement
How would adipose ECM and bladder ECM extracted and prepared as scaffold material
from pigs differ in healing/regenerative results on humans?
Hypothesis
A proposed hypothesis for this experiment would be that the urinary bladder ECM
scaffold material will have superior results as opposed to the adipose tissue. One of reasons for
this particular hypothesis is the fact that there have been more experiments conducted with pigs
urinary bladder as opposed to adipose tissue.
Variables
The independent variables are both the pig adipose and bladder materials. These are the
materials that are being manipulated to see a difference. The dependent variables are the results
of the use of the independent variables. The dependent variables are the healing/regenerative
results on human subjects. The control is the wound being treated. It is essential to keep the
control constant to obtain reliable results.
Methods
The proposed method for this experiment calls for the obtainment of both pig bladder and
adipose tissue. These tissues can easily be obtained from a local butcher shop or meat market.
Once the tissues are obtained they must be frozen at -80.0 degrees Celsius. The next step is to
decellurized the tissues through any of the 3 methods described in the Brown experiment
(Brown, B., Freund, J., Han, L., Rubin, J., Reing, J., Jeffries, E., & ... Badylak, S. (2011). In this
experiment, all 3 methods yielded positive results that represented effective methods for use in
tissue engineering (Brown, B., Freund, J., Han, L., Rubin, J., Reing, J., Jeffries, E., & ... Badylak,

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S. 2011). Once both the bladder and adipose ECM has been processed they are ready for topical
use. It is important to understand that the only way to obtain accurate results is to have similar
experiments with both the bladder and adipose tissue. It is therefore necessary to obtain human
subjects with similar injuries in size and scale for comparison in results. The ECM scaffold
material prepared should be placed on the on the injury and covered with bandage material.
After several weeks of reapplying ECM material, results should be obtained and recorded.

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References
Brown, B., Freund, J., Han, L., Rubin, J., Reing, J., Jeffries, E., & ... Badylak, S. (2011).
Comparison of three methods for the derivation of a biologic scaffold composed of
adipose tissue extracellular matrix. Tissue Engineering. Part C, Methods, 17(4), 411-421.
doi:10.1089/ten.TEC.2010.0342
Campbell, N. A., & Reece, J. B. (2005). Biology. San Francisco, Calif: Pearson, Benjamin
Cummings.
Choi, Y., Choi, J., Kim, B., Kim, J., Yoon, H., & Cho, Y. (2012). Decellularized extracellular
matrix derived from porcine adipose tissue as a xenogeneic biomaterial for tissue
engineering. Tissue Engineering. Part C, Methods, 18(11), 866-876.
doi:10.1089/ten.TEC.2012.0009
King, R., & Newmark, P. (2012). The cell biology of regeneration. The Journal Of Cell
Biology, 196(5), 553-562. doi:10.1083/jcb.201105099
Liang, R., Fisher, M., Yang, G., Hall, C., & Woo, S. (2011). Alpha1,3-galactosyltransferase
knockout does not alter the properties of porcine extracellular matrix bioscaffolds. Acta
Biomaterialia, 7(4), 1719-1727. doi:10.1016/j.actbio.2011.01.001
Liu, L., Li, D., Wang, Y., Xu, H., Ge, L., & Liang, Z. (2011). Evaluation of the biocompatibility
and mechanical properties of xenogeneic (porcine) extracellular matrix (ECM) scaffold
for pelvic reconstruction. International Urogynecology Journal, 22(2), 221-227.
doi:10.1007/s00192-010-1288-9
Ringel, R., Kahane, J., Hillsamer, P., Lee, A., & Badylak, S. (2006). The application of tissue
engineering procedures to repair the larynx.Journal Of Speech, Language, And Hearing
Research: JSLHR, 49(1), 194-208.

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Sánchez Alvarado, A. (2009). Developmental biology: A cellular view of
regeneration. Nature, 460(7251), 39-40. doi:10.1038/460039a
Thiriet, Marc (2011). Cell and Tissue Organization in the Circulatory and Ventitilatory Systems