Shock Wave Therapy for Soft Tissue Wounds: Phase II ... · Bioelectric dressing Microelectric...

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The views expressed in this presentation are

those of the presenter, and do not reflect the

official policy of the;

- Trideum Corporation

- Department of Defense, or

- United States Government

Disclaimer

Financial Disclosure

No financial interest or investment in the technologies presented

- In 1676, observed bacteria and other microorganisms using a single-

lens microscope he designed.

- Shocking discovery at that time. Things living inside humans!!!

Replica of one of his microscopes

Anton van Leeuwenhoek

Scientific Bias Disclosure

· What’s your story? What excites you? How did you get to where you are

today?

· What research/work do you conduct?

· What STEM skills and knowledge do you actively need to be successful in

your job?

· What courses or extracurricular activities have helped you in your career?

· What guidance would you provide to students to prepare them to enter the

workforce and manage challenges they may encounter?

· Do you offer any resources to support teachers or outreach programs for

students and teachers to explore your research area or STEM field?

My Story

Mina Izadjoo, PhD

Chief Science Officer

Trideum Biosciences

Trideum Biosciences

We conduct test and evaluation of therapeutics and devices

Troubleshoot and recommend path to advance new technologies

Success Track record

2017 Innovation in North America Award

Gold Award for Excellence - Best Poster

2016 Military Health Systems

Research Symposium (MHSRS)

2016 "Best Poster Award” at the 5th

World Union of Wound Healing

Societies (WUWHS) conference in Florence, Italy

Where are we Located?

Trideum Biosciences laboratories are located in Frederick, MD near Fort Detrick

Trideum Biosciences is an associate company with the Hudson Alpha Institute of Biotechnology in Huntsville near the Trideum Corporation headquarter office.

www.trideumbiosciences.com Info@trideumbiosciences.com

Who are Our People?

VP & Chief Science Officer - Dr. Mina Izadjoo

◦ Ph.D., Microbiology and Immunology, LSU

◦ Former Director of Diagnostics & Translational

Research Center for the Henry M. Jackson Foundation for Advancement of Military Medicine

◦ Former Division Chief of Wound Biology & Translational Research, Armed Forces Institute of Pathology

◦ Associate Professorship at Uniformed Services University for Health Sciences (USUHS)

◦ President Emeritus of the Washington Academy of Sciences

Advancing medical products in support of Armed Forces

Armed Forces Institute of Pathology Walter Reed Army Medical Center

Walter Reed National Military Medical

Center

Uniformed Services University of the

Health Sciences (USU)

Humans and Microorganisms………….

Bacteria: Friends or Foes?

We are outnumbered by our bacterial tenants.

We provide them room and board and in return they help us with digestion,

vitamin production and innate immune response.

Microbial World: A Challenging Frontier

The physical diversity of microorganism is a reflection of underlying

genetic differences.

http://textbookofbacteriology.net

Bacteria could have traveled through

space on comets. If even one microbe

survived space travel to Earth, it would be

enough to start a colony on our planet. Courtesy of NASA

Air Force Proficiency Testing Program

7900 Fast Real Time PCR

ABI 7500 Fast DX

Swine Flu Pandemic

-This is a 14 year-old girl who presented with symptoms of a space occupying

lesion in the posterior fossa consistent with a brain tumor.

Diagnosis: Medulloblastoma

Helicobacter pylori bacteria in the stomach has been linked to ulcers

and stomach cancers.

SPL / BARCROFT MEDIA

Association of Wound Healing and Cancer

J Wound Care. 2014 Jun;23(6):314, 316-9.

Wound infections and healing: are they contributing factors for

carcinogenesis? Park S, M. Izadjoo.

Prostate cancer cells, after

interacting with normal wound

healing cells have become more

invasive and more able to

metastasize.

Tumor cells secrete signals that call in

wound healing cells to the tumor site. In

the process, the normal wound healing

cells make the tumor cells more

aggressive and able to metastasize.

http://www.ns.umich.edu/new/

21432

Combat Casualty in Recent Conflicts in Iraq and Afghanistan

News Story (Wednesday, August 12, 2009)

AFIP Partners With Walter Reed to Help Advance

Combat Wound Care Program

Human Skin

Skin is the interface between human body and the

environment, prevents loss of moisture, blocks

entry of pathogens and foreign material.

The skin microbiome may have beneficial or

detrimental effects on wound healing.

Skin generates microcurrents

Wound Healing: Complex and Dynamic Process

http://en.wikipedia.org/wiki/Wound_healing#mediaviewer/File:Wound

_healing_phases.png

• Normal wound healing is a well-coordinated and progressive series of

events designed to restore the barrier function and mechanical integrity of

the skin.

• Wound healing involves interactions between cells and their

microenvironment.

• Traditional Wound Care (Wound-closure Products; Tissue Adhesives,

Sealants, and Glues; Fibrin-based Sealants; Collagen-based Sealants;

Synthetic Adhesives/Glues; Anti-infective Dressings)

• Basic Wound Care (Tapes; Dry Dressings; Cleansing)

• Advanced Wound Care (Films; Foam Dressings; Collagen; Alginates;

Hydrocolloids; Hydrogels; Super Absorbers)

• Bio-Active Wound Care (Artificial Skin and Skin Substitutes)

• Therapy Devices (Negative Pressure Wound Therapy Devices; Pressure

Relief Devices; Electrical Stimulation Devices; Ultra-Violet Devices;

Oxygen and Hyperbaric Oxygen Equipment; Whirlpool Therapy Devices;

Electromagnetic Therapy Devices; Ultrasound Devices).

Wound Care Products

$15.6 billion (2014) 18.3 billion (2019)

http://www.marketsandmarkets.com/Market-Reports/wound-care-market-371.html

The use of silver as an antimicrobial for infection spans hundreds of

years. The ancient Greeks and Romans used silver to disinfect their

water and food supplies.

Silver: An age-old treatment modality in modern times (Surg Nurs. 2010 Apr-Jun;30(2):90-3)

• Silver was also used in ancient times to treat burns and wounds as a

wound dressing.

• Silver solutions were approved by FDA in the 1920s.

Procellera Technology Microcurrent generating antimicrobial wound dressing

wireless

microcell

batteries Silver

In presence of a conductive fluid (i.e. saline, hydrogel, wound exudate) batteries are activated and generate microcurrents

Zinc

Inspired by the Body

Electricity is Essential to Wound Healing

An electric potential exists across intact skin.

The body naturally creates and uses electrical energy to promote healing.

When skin is wounded, a change in electric

potential occurs. This stimulus is the earliest

guidance signal to initiate cell migration and

re-epithelialization, and is essential to wound

healing.

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

Log

(No

. of

viab

le c

ells

/ml)

Wound isolates

Initial (CFU/mL) After (CFU/mL)

Figure 1. Antibacterial properties of the wound dressing showing reduction of bacteria after 24 hours exposure against clinical wound isolates.

ground

Positive

Bioelectric dressing

Microelectric Measurement System Set Up

Microelectric potential can be

measured by moving the positive

probe on the wetted dressing with

fluids (culture medium, bacteria

cultures and saline). During

measurement, the ground probe

stayed on the white place.

Bacterial Biofilm is a major barrier to wound healing

• Complex microbial communities

• Microbial community produce and secrete a protective surrounding substance that

allows it to stick to surfaces, living or non living.

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

Log

(No

. of

viab

le c

ells

/ml)

Wound isolates

Initial (CFU/mL) After (CFU/mL)

Figure 1. Antibacterial properties of the wound dressing showing reduction of bacteria after 24 hours exposure against clinical wound isolates.

• Biofilms are everywhere and it is a tragedy to use clinical microbiology

techniques for their identification. The clinical microbiology is for floating

bacteria.

• We need other methodologies for detection and treating biofilm diseases.

• Patent: Device and Methods for Creating and Testing Biofilms (Application

# 62075343)

Planktonic vs Biofilm

Efficacy Testing of Procellera against A. baumannii biofilm

1. Add 1 mL A. baumannii biofilm and swill the culture onto a TSA plate (Fig. 1A & 2A)

2. Place Procellera directly onto the biofilm and incubate at 37°C for 24 h (Fig. 1B & 2B)

3. Check a bacterial growth (Fig. 1C & 2C)

Bacteria killed

1A

1B 1C

2A

2B 2C

Biofilm of Acinetobacter baumannii on a Poloxamer

hydrogel plate

1 day-old A. baumannii

onto a 20% Poloxamer hydrogel

plate

4 day-old A. baumannii

onto a 20% Poloxamer hydrogel

plate

1 day-old A. baumannii

onto a TSA plate

All plates were incubated at 37°C, 48 h

Procellera + incubation at 37°C, 24 h

There was no significant bacterial growth.

Bacterial cultures are mixed with 30% poloxamer hydrogels in

Mueller-Hinton broth (MHB). Mono-species biofilms were formed in

poloxamer hydrogels placed onto glass coverslips (JWC in press).

A

B

D

C

A new poloxamer biofilm model

Anti-Biofilm Efficacy using Drip-Flow Reactor (DFR)

Acinetobacter

baumannii

Pseudomonas

aeruginosa

blank Procellera Procellera

Fig. 2. Formation of A. baumannii and P. aeruginosa biofilms on the sample dressings of blank and

Procellera in colony DFR biofilm model (A). Microscopic observations of A. baumannii and P.

aeruginosa biofilms stained by crystal violet in colony DFR (B).

A B

blank

Procellera

Procellera

Patent: Device and Methods for Creating and Testing

Biofilms (Application # 62075343)

A new platform technology for standardized biofilm

formation and testing.

Wound infections are commonly polymicrobial and interspecies interactions

play a significant role in wound infections.

S. aureus is commonly isolated from wound infections and has become an

alarming problem due to emergence of several methicillin-resistant strains

(MRSA).

More than 90% of all S. aureus isolates from human infections are gold

pigmented.

Golden pigment is the manifestation of staphyloxanthin biosysnthesis, a

virulence factor to survive stressful environment.

P. aeruginosa is often isolated along with S. aureus from wound infections.

Wound Microbiome Communications

Pseudomonas aeruginosa induced pigment production and

enhanced virulence in a white phenotypic variant of

Staphylococcus aureus

We isolated two pigment variants of S. aureus and one strain of P.

aeruginosa from the same clinical sample of a patient with soft tissue

wound.

S. aureus variants produced white and yellow colonies.

We demonstrated that white variant can be triggered to produce golden

pigment by the P. aeruginosa isolate.

Effect of co-culture with different bacteria on

pigment production by the S. aureus white variant

Control W

+

E. coli

W

+

Acinetobacter

W

+

Enterobacter

W

+

P. aeruginosa

• Medical community is beginning to recognize the significance of

polymicrobial infections and many therapies are now take into

account the polymicrobial nature of wound infections.

• At present, it is impossible to define the role of each participating

bacterium in these communities and the wound outcome.

• It is critical to understand the interaction between bacterial species in

wound.

Importance of Understanding Microbiome Communications

Biofilm Therapeutics

Serum Wound Effluent Urine

Bioflux 200

Cellular adhesion at

2 hours and 6 hours

under shear flow.

Efficacy of novel antibacterial agents for

treating polymicrobial wound infections

Distinctive stages and strain variation of A. baumannii biofilm development

2-D 3-D

A. baumannii isolates from human wounds showed attachment occurred

at the moment of contact. The first report to show the effectiveness of

this organism in attaining irreversible attachment.

Feng et al JWC 2013, 22(4): 1-9

Clinical Evaluation of a Bioelectric Dressing System for

Acute Wound Management

Wound Healing Assessment

Martin Army Community Hospital

A critical project addressing joint force health protection capability.

Team Effort

Application of a Novel Cold Atmospheric

Plasma for Water Decontamination

Plasma: Fourth State of Matter

The three known states of matter are solids, liquids and gases. Plasma is the

fourth state of matter.

Plasma make 99 percent of the known universe. Plasmas are formed when

atoms are stripped of their electrons, producing ionized gas flows.

Plasma does not contain molecules. It is a gas that is composed of ions.

Technology Description

Cold Plasma

Cold Plasma Device

This pocket-size cold plasma technology is the next generation technology due to its

size, portability, precision, reduced cost and ease of application with no requirement

for any noble gas. Ideal device for field application.

Cold Plasma Application

Medical equipment sterilization

Food decontamination

Antimicrobial device

Decontaminated military gear

Protective shield around sensitive electronic

devices

Stealth technology

Nuclear/chemical detection

Biosensors/Biosignature

Medicinal/Healthcare Application is relatively

young (15 yrs)

Efficacy against Pseudomonas aeruginosa, A Bacterial Pathogen

Efficacy against Trichophyton rubrum

A Toe Fungal Pathogen

B C D A

Treatment with Cold Plasma No Treatment

Microbial Biofilms

A major medical, industry and environmental

problem

TESTING FOUNDATIONS FOR BUSINESS APPLICATIONS

A NEW STANDARD FOR ANTIMICROBIAL AND ANTI-BIOFILM

TESTING

Biofilms

• Complex microbial communities

• Microbial community produce and secrete a protective surrounding substance

that allows it to stick to surfaces, living or non living.

Next Steps……………….

Manufacture, Validate, Market

Human Microbiome………

There are 100 trillion bacterial cells living with each one of us.

This is ten times the number of our own cells.

We need to find out how microorganisms contribute to health and disease.

Human DNA is only a small fraction of the genetic

material in our bodies.

New Frontiers

Role of Women in Advancement of Military Medicine

Women in Warfare and the Military

The second-century carving

two female gladiators (Ancient

Era)

522 BC: Leutenant Artunis,

Commander of Persian Army,

daughter of Artebaz, Sepahbod of

Darius the Great.

A woman using a compass to measure

distances on a diagram (c. 1310 AD)

Women and Science

1775 - 1783

Women served traditional roles within the U.S. Army

such as cooks, laundresses, nurses and seamstresses.

During the American Revolution some women served in

combat alongside their husbands or disguised as men.

There were some courageous women took on roles as

spies.

Deborah Sampson (1760 - 1827)

She had a desire to join the Continental Army.

First enlisted as Timothy Thayer and then Robert Shurtleff (1782).

She wrote to her family that was working in “ a large but well regulated family”.

Augusta Ada Byron Lovelace (1815-1852)

Computer Scientist

The world’s first computer programmer; the

programming language “Ada” was named

in her honor.

Maria Mitchell (1818-1889)

Astronomer

In 1847, her discovery of a comet invisible

to the naked eye won her international

fame and a medal from the king of

Denmark.

Florence Bascom (1862-1945)

Geologist

She was the first women and first geologist

to be awarded a Ph.D. from Johns Hopkins

University.

Maria Sklodowska Curie (1867-1934)

Physicist and Chemist

Dr. Elizabeth Blackwell (1821-1910)

First Female to receive a medical

degree in US.

Dr. Mary Edwards Walker (1832-

1919)

First Female surgeon

First Female US Army Surgeon

(1863)

1970: Women never made up more than 6% of any medical school in

US.

Rebecca Lee Crumpler

First African-American women to

receive an MD (1864)

Eliza Ann Grier, (ex-slave) the first

African-American women licensed

to practice medicine (1898).

Clara Barton (1821 – 1912)

Founder and the First President of the American Red Cross

Floreance Sabin, the first female

faculty of Johns Hopkins School of

Medicine (1903).

“A pioneer for women in science”

Margaret Craighill, the

first woman MD to join

US military (1943)

1972: Congress passes the Higher Education Act prohibiting any sex

discrimination in educational institutional that receive federal funds.

Lt. Gen. Patricia D.

Horoho became the first

female and first nurse to

become surgeon

general of the Army

Dec. 7, 2011.

Maj. Gen Marcia M.

Anderson became the

Army's first-ever female

African-American officer to

obtain the rank of major

general in Oct. 1, 2011.

U.S. Army female soldiers in Boldak

district in Afghanistan's Kandahar

province (2013).

Deborah Sampson

(1760 - 1827)

Nancy Grace Roman, Margaret Hamilton, Sally Ride, and Mae Jemison

“ It is not the strongest of the species that

survive, nor the most intelligent, but the

one most responsive to change.”

Adapt to Change

The past is connected to the present and to the future through

knowledge and challenges faced by mankind.