Team Members: Sagah Ahmed, Natalie Anzures, Zachary Bosley, Brendan Bui, Ariana Feizi, Sudabeh Jawahery, Courtney Koenig, Katherine Lakomy, Megan Lin, Poorna Natarajan, Eisha Nathan, Hiba Sayed, Eduardo Solano

Mentor: Dr. John Fisher

E.L.E.C.T.R.O.D.E.

Evaluating Linear-Radial Electrode Conformations for Tissue Repair and Optimizing a Device for Experimentation

Introduction

The Problem

ϟ  Prevalence of Diabetes

ϟ Formation of Diabetic Ulcers

ϟ Issues with Current Treatments

Groups of Interest

ϟ Individuals with Diabetes

ϟ Medical Researchers

ϟ Healthcare Professionals

Wound Healing and Diabetic Ulcersϟ Endogenous wound healing

ϟ Angiogenesis and inflammation

ϟ Diabetic ulcers and inflammation

Electrical Stimulation and Wound Healingϟ Body’s response to electrical stimulation

ϟ Cell migrationϟ Proliferation

ϟ Electrical stimulation and angiogenesis

Deprived cell migration Elect

ricField

VEGF, bFGF

Limited angiogenesis

Specific Aims

Specific Aim 1: To examine the effects of an optimized linearly applied electric field on cell proliferation, VEGF expression, bFGF expression and cell migration rates

Specific Aim 2: To examine the effects of an optimized radially applied electric field on cell proliferation, VEGF expression, bFGF expression and cell migration rates

Specific Aim 3: To examine the effects of an optimized (either radially or linearly) applied electric field on cell proliferation, VEGF expression, bFGF expression and cell migration rates in vivo

Specific Aims 1 and 2: In Vitro Experiments Rationale

ϟ Verifies that electrical stimulation can promote angiogenesis

ϟ Provides preliminary data to build in vivo device

Rat aortic endothelial cells will be grown in Dulbecco’s Modified Eagle Medium with 10% Fetal Bovine Serum

Specific Aim 1: In Vitro Experiment

Linear Electric Field Apparatus

ϟ Coat electrodes with high-purity titanium

ϟ Induces constant electric field at all points

ϟ Connected to signal generatorExperimental set-up of a

linearly applied electric field. The anode and cathode are 1.5-in apart.

Specific Aim 2: In Vitro Experiment

Radial Electric Field Apparatus

ϟ Outer electrode made from polyethylene

ϟ Inner electrode made with commercial titanium rod

ϟ Non-constant electric field

ϟ Analyze effects on angiogenic factors in different areas

Experimental set-up of a radially applied electric field. The outer hoop is positively charged, whereas the inner cylindrical electrode is negatively charged.

Specific Aims 1 and 2: In Vitro Experiments

Experimental Design

ϟ Pulsed monophasic current at (ideally) 0.1 V, 15 µA and 50 Hz

ϟ Applied for 30 minute periods, once per day for 2 days

ϟ Five control cultures will not receive any electrical stimulation

Pulsed monophasic current.

ϟ Cell Proliferationϟ Counting using hemocytometer

ϟ Measure growth factor expression (VEGF, bFGF)ϟ ELISAϟ qPCRϟ Immunohistochemistryϟ Western Blotting

ϟ Cell Migrationϟ Microscopy

Specific Aims 1 and 2: In Vitro Experiments Cell Analysis

Specific Aims 1 and 2: In Vitro Experiments

Success Criterion

ϟ Significant increase in cell migrationϟ Increased migration is associated with

increased healing rates

ϟ Significant increase in cell proliferationϟ Increased proliferation is associated with

increased healing rates

ϟ Increase in angiogenic growth factors and expressionϟ Increased growth factors are associated with

increased reparative cell activity

Specific Aim 3: In Vivo Experiments

Diabetic Rat Model

ϟ Induce wounds in diabetic rats

ϟ Use best electrode setup determined from in vitro trials to treat wounds

ϟ Set-up will be tested on 4-8 week old Sprague-Dawley rats

Prototype Design

ϟ Model off of in vivo device

ϟ Build cheap and small prototype for a more marketable device

Electrodes

Battery/Power Cable

Projected Timeline

Projected Budget

Categories Expense (dollars)

Cell Culture 800

Electrode Materials 25

Circuitry 180

In vivo Model 9420

Housing and Care for Rats 200

Cell Assay Supplies 2000

TOTAL 12,625

Questions?