Grant of the Month | March 2012

Peter Wearden

Bedside Monitor to Quantify Cardiac Shunt Flow in Newborns and Small Children

There is no current technology for routine measurement of shunt flow (Qp:Qs – ratio of pulmonary to systemic blood flow) in newborns and small children in the intensive care unit (ICU). Current methods either require placement of highly invasive catheter or depend on assumptions, leading to risky and less accurate measurement of shunt flow. Timely and accurate quantitative assessment of Qp/Qs permits successful pharmacologic, ventilator or fluid therapy or in time surgical intervention. Hence routine measurement of shunt flow is vital in the management of critically ill newborn and small children with cardiac defects.

This SBIR grant will allow us to develop mathematical models and algorithms accounting for various shunts, clinical and physiological conditions. These will then be implemented into a monitor that could be used clinically at the bedside in a non-invasive manner with patients having in situ arterial and central venous catheters. The approach is based on well-established indicator dilution principles using innocuous isotonic saline as an indicator. These factors make the proposed monitor eminently suitable for used with neonatal and pediatric ICU patients.

Main objectives of this proposal include –

• Development of mathematical models and software algorithms for accurately quantifying shunt flow
• Perform feasibility and validation testing of shunt flow measurements in animal models
• Perform feasibility and clinical comparison of shunt flow measurements in newborns and small children.

These objectives will be achieved in a concerted effort by Transonic’s R&D staff, and its collaborators at University of Pittsburgh, PA and Crouse Hospital, NY.

Transonic Systems

09/01/11 – 07/31/13

$60,000 total in Year 1: $50,000 total in Year 2

Grant of the Month | February 2012

Stephen F. Badylak

Optimization of Surgical Mesh Materials

This project involves a combination of in vitro and preclinical in vivo methods to develop and evaluate biologic surgical mesh materials.  The work involves a combination of well described benchtop assays and animal models which can evaluate in vivo biocompatibility for novel surgical mesh materials. 

CR Bard

01/01/12 – 12/31/12

$600,000

Grant of the Month | january 2012

David Hackam, MD, PhD and John March, PhD

Generation of an Artificial Intestine for the Treatment of Short Bowel Syndrome in Children

The clinical condition in which the body is unable to absorb food after significant loss of the intestine is called short bowel syndrome (SBS). While its true incidence is unknown, in the United States the condition affects over 5000 children, with an estimated 15,000 older patients requiring long-term home parenteral nutrition. SBS can be caused by loss of large portions of functioning intestine – such as occurs typically as a consequence of necrotizing enterocolitis (NEC), Crohn's disease, or as a result of a birth defect in which the intestines do not develop normally. Because food cannot be adequately absorbed by the shortened intestine, nutrients must be administered directly into the circulation through a vein. Although this approach can supply adequate calories, children who receive nutrition directly into the circulation commonly suffer from intravenous catheter infections and severe liver toxicity, with mortality around 30%. Only about one third of patients with SBS can expect to be weaned from parenteral nutrition. The majority of children with short bowel syndrome require intestinal transplantation and if toxicity from the administered nutrition is severe enough, liver transplantation, as well. While the outcome after intestinal transplantation is improving, this procedure is limited by a lack of suitable donors and complications from immunosuppressive therapy. To address the difficulty of managing short bowel syndrome in children, Hackam and March propose constructing an artificial intestine using cultured intestinal stem cells from the recipient’s intestine that can grow on a synthetic 3-dimensional bioscaffold.

The Hartwell Foundation

2012-2015

$543,571 in direct costs over three years

Grant of the Month | December 2011

Marina Kameneva

Continuous Red Blood Cell Production (Phase II)

A ready supply of safe and effective red blood cells is a critical component in the treatment of battlefield and civilian trauma. Conventional approaches to this challenge center around voluntary donation of whole blood, testing, processing, extended storage, shipping and therapeutic transfusion of blood or fractionated components. Many of the steps in this conventional approach are prone to error, are inefficient, and in some pathologies can be ineffective. We intend to transform this conventional approach by developing methods and systems to produce erythrocytes (and at a future time, other blood components) from readily available and expandable human non-embryonic progenitor cell populations in a safe, effective, robust and limited footprint in vitro manufacturing system.

DARPA

07/01/11 - 06/30/12

$240,755

Grant of the Month | November 2011

James Funderburgh

Stem Cells for Corneal Engineering

The proposed research builds upon the pioneering work from the laboratory of Dr. Wagner in developing novel degradable biomaterials for a variety of soft tissue applications.  Dr. Wagner’s group links polymer chemists, bioengineers and surgeons in this effort, and the proposed research takes advantage of this expertise.  They have synthesized, characterized and processed a variety of biodegradable polymeric biomaterials and evaluated their performance for treating tissue insufficiency in vivo.  The thermoplastic elastomeric materials proposed in this project have been synthesized and evaluated in the rat model in several different locations.  Most of this in vivo work has involved the cardiovascular system for cardiac wall and blood vessel scaffolding.  Other studies have evaluated application of the material in the abdominal wall and as subcutaneous implants for first level biocompatibility assessments.  Dr. Wagner and Dr. Funderburgh have been collaborating over the past several years to begin translation of the Wagner lab’s materials expertise to the ophthalmic area.

NIH

04/01/11 – 03/31/15

Total DC: $307,303

Grant of the Month | October 2011

Joerg Gerlach, Eva Schmelzer, Ian Nettleship

Innovative In Vivo-Like Model for Vascular Tissue Engineering

The shortage of donor organs for transplantation suggests a need to develop engineered tissue transplants. Proper in vitro vascularization, a key prerequisite for the development of functional engineered tissue constructs, would enable adequate mass exchange, gas supply, and functional mediator exchange in high-density tissue cultures. The impact of physical and mechanical factors supporting endothelial differentiation has been investigated, but not in three-dimensional (3D) co-culture models. We propose to address this gap in cellular models and technology model systems, by analyzing neo-vascularization in an organ-like environment in vitro designed to mimic human organogenesis and that can vary physical conditions, such as flow- and pressure changes in the rhythm of the heart rate.

In the fetal liver in vivo, angiogenesis occurs in hematopoietic and hepatic tissues that develop together. In our cell model for enabling vascularization in vitro, we therefore propose to investigate second trimester human fetal liver derived endothelial progenitors within fetal parenchymal cells, which contribute to hematopoietic and hepatic tissue vascularization.

In the culture technology model, we propose to apply physical forces to control vascular structure formation, shear stress, perfusion flow and pressure changes. Additionally we will investigate the effects of calcium liberating hydroxyapatite scaffolds that mimics natural bone on formation of hematopoietic vascular sinusoids in the stem cell niche.

RFP transfection labeled progenitors (hemangioblasts and angioblasts) and non-endothelial fetal liver cells will be cultured in 3D perfusion and the response to various physical-mechanical cues determined. Harvested cells will be analyzed by histology, flow cytometry, and gene expression, and compared to prenatal organ explants and postnatal organ tissues in vivo. The prior labeling of hemangioblasts will allow us to selectively distinguish between original hemangioblasts, endothelial- and non-endothelial cell types.

The bioreactor model provides four independent interwoven hollow fiber compartments, enabling 3D perfusion with low gradients by decentral mass exchange and integral oxygenation.  This has been proven to support vascularized tissue-like structure formation at high cell densities. We have already demonstrated that our 3D perfusion bioreactors support the spontaneous neo-tissue formation with neo-vascular hepatic structures and functionality in the laboratory and in clinical application for extracorporeal liver support.

The innovation of our project is the specific experimental model that mimics the mass exchange in the native organ environment, allowing the fate of labeled fetal vascular progenitors to be studied during tissue formation, depending on different physical conditions.

NIH / NHLBI

09/01/11 – 05/31/15

1st year direct costs:  $345,672
1st year indirect costs: $159,737

Total direct costs: $1,422,839

Grant of the Month | September 2011

Harvey Borovetz

Coulter Translational Research Award in Biomedical Engineering

The Coulter Translational Research Award program provides funding for Professors in established Biomedical Engineering Departments within the United States. The award seeks to support biomedical research that is translational in nature, and to encourage and assist eligible biomedical engineering investigators to establish themselves in academic careers involving translational research. The translational research projects are directed at promising technologies with the goal of progressing toward commercial development and entering clinical practice.

Wallace H. Coulter Foundation

5 Years

Grant of the Month | August 2011

Thomas Gilbert

Basic and Clinical Studies of Cystic Fibrosis: Ex Vivo Model of Cystic Fibrosis

Cystic fibrosis (CF) affects approximately 30,000 individuals in the United States, causing an accumulation of thick, sticky mucus that adversely impacts normal mucociliary clearance.  The lack of proper clearance predisposes patients to chronic pulmonary infections, injury to the conducting airways in the form of bronchiectasis and bronchiolitis obliterans, and ultimately can lead to respiratory failure.  CF is the leading diagnosis in children that require lung transplantation.  Considerable resources have been applied to the study of CF with the hope of developing a treatment or cure, but progress has not been as rapid as anyone would desire.  In the current research environment, the only way to determine if a treatment strategy has an effect on airway function is to move from in vitro studies to clinical studies in patients, which poses a very high bar to acceptance.  The primary purpose of this proposal is to develop a humanized ex vivo model of the CF airway as a means to investigate the effect of new therapies on function, including ion transport and mucociliary clearance.  A bioreactor has been developed that is capable of simulating the conditions in the trachea during respiration.  With the expertise from the P30, decellularized tracheas will be seeded with airway epithelial cells from patients with and without cystic fibrosis and the culture conditions will be optimized.  Then testing strategies will be developed to assess airway epithelial cell function without disrupting the trachea construct and the impact of various known medical treatments on the cultures will be tested.  Finally, airway tissue will be obtain from patients with and without CF, the tissue will be decellularized and seeded with airway epithelial cells  to begin to understand if the composition and structure of airways in CF contributes to the dysfunction.  The end result of this work will be a robust testing platform that will enable functional testing of the airway which will allow a deeper understanding of the pathogenesis of CF and more robust testing of new drug therapies.

NIDA Core “Center of Excellence” Grant Program

06/01/11 – 05/31/13

Year 1 DC: $75,000
Year 1 Total: $113,625

Total DC: $150,000
Total: $227,250

Grant of the Month | July 2011

Peter Wearden

Rick Koepsel, Alan Russell, Tom Gilbert

Nanotechnology Based Infection Control for Ventricular Assist Devices

Control of infection and thrombosis in total artificial heart technology has been of great concern for the last five decades. Even for totally implantable total artificial hearts, infection control is necessary because patients still need percutaneous lines for collecting post-implant hemodynamic data. During the Phase I contract with NIH/NHLBI, ND Life and University of Pittsburgh developed a highly effective anti-infection coating system based on nanotechnology that can be easily applied to the treatment of drivelines for ventricular assist devices (VADs). A novel green processing technique has been explored to immobilize silver nanoparticles on Dacron in Phase I. Irreversibly immobilized silver nanoparticles on Dacron showed significant reduction of bacterial challenges in Phase I. In the Phase II, ND Life and University of Pittsburgh will optimize the nanotechnology-based antimicrobial coating systems by a novel green processing technique through in vitro and in vivo studies. The system we propose here will provide a simple and highly effective anti-infection coating technology for VAD drivelines, with relevance to a broad range of other implantable medical devices.

NanoDynamics Life Sciences, Inc. DBA LIG Sciences  (via an NIH SBIR)

05/15/11 – 04/30/13

Grant of the Month | June 2011

Kang Kim

William Wagnerl

Non-invasive Monitoring of Tissue-Engineered Construct by US Elasticity Imaging

Non-invasively monitoring the extent of tissue scaffold degradation, cellular growth, and tissue development will greatly help tissue engineers to non-destructively evaluate candidate scaffold performance in vivo. Biodegradable polymer scaffolds are used to support cells and growing tissues until they are replaced by the body’s own extracellular matrix (ECM). Two main challenges in creating the ideal biodegradable polymer scaffold are: (1) the scaffold must have a defined shape and porous internal architecture suitable for direct tissue ingrowth but with appropriate mechanical and degradation properties and (2) the scaffold must have the right surface properties to provide favorable conditions for cells to attach, differentiate, and lay down ECM. To design scaffolds which appropriately transfer their mechanical load over time to the ingrowing tissue, temporal data are required that verify the mechanical viability of the remodeling construct. Current analysis methods are destructive, requiring animal euthanasia and explanting the construct for histological and direct mechanical characterization. In addition, different samples are prepared and measured at varying times, but high growth deviation between specimens makes analysis difficult. Ideally, tissue engineers need a system that can noninvasively monitor growth in the same specimen over time. Other imaging methods, such as magnetic resonance imaging (MRI) and computed tomography (CT), provide internal scaffold structural information, but they are limited to providing only morphological information. Ultrasound easticity imaging (UEI) based on phase-sensitive speckle tracking can characterize the mechanical, structural, and functional change of the implanted engineered tissues at very high resolution and sensitivity. Local UEI offers the potential to radically improve the biomaterial scaffold design and engineered tissue growth techniques. The long term goal of this research program is to develop a novel noninvasive functional imaging modality in the field of tissue engineering and regenerative medicine. The objective of the current project is to evaluate UEI as noninvasive imaging tool to assess mechanical, structural, and functional characteristics of the scaffold degradation and tissue ingrowth. The specific aims are: (1) Establish the in vitro relationship between noninvasive UEI and the mechanical and structural characteristics of the biomaterial scaffold degradation. (2) Establish the in vivo relationship between noninvasive UEI and the mechanical, structural, and functional characteristics of simultaneous tissue growing and scaffold degradation. These specific aims will be evaluated using novel polyurethane-based soft tissue scaffolds with three different degradation rates. In vivo feasibility will also be demonstrated using the rat abdominal repair model.  If successful, UEI integrated into a commercial ultrasound scanner can also be rapidly translated into clinical practice since it is based upon novel processing of ultrasound data that can be obtained conveniently and non-invasively from human subjects.

National Science Foundation

04/01/11 – 03/31/13

Year 1 Direct: $30,340
Year 1 IDC: $15,625

Total Direct: $61,197
Total IDC: $31,516

Grant of the Month | May 2011

Eric Lagasse

The Role of mTOR (mammalian Target Of Rapamycin) Complex 1 and 2 in liver regeneration and ectopic liver organogenesis

Liver failure is a major cause of morbidity and mortality in the Western world. Although the liver is mitotically a quiescent organ in adult life, parenchymal regeneration can occur through the proliferation of the two major types of hepatic epithelial cells, hepatocytes and biliary epithelial cells. Akt and its downstream mTOR signal are likely to contribute to this process. Indeed, transient increase of activated forms of Akt, mTOR, TSC2, p70S6K1 and 4E-BP1 was correlated with the growth response following hepatotectomy in animal models. Accordingly, the mTOR inhibitor rapamycin was effective in retarding proliferation of hepatocytes in the same model of liver injury.

Recent findings indicate that mTOR is shared by two different complexes with specific funtions, mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2), and a careful revision of the literature suggests that the previous liver regeneration studies were limited to the analysis of only mTORC1 activity. The major difference between the two complexes is the substrate specificity factors, raptor in mTORC1 and rictor in mTORC2. Briefly, mTORC1 regulates mRNA translation and ribosome biogenesis while mTORC2 regulates actin cytoskeleton. Importantly, according to recent investigations, Akt, apart from being an upstream activator of mTORC1, is also a downstream effector of mTORC2, being this latter responsible for its full activation. Importantly, mTORC2-mediated Akt activation is emerging as a critical inducer of cell growth and motility.

In the mouse model of human tyrosinemia type I (the FAH-/- mouse) -a form of acute liver failure where spontaneous liver regeneration is impaired- intraperitonelly-injected hepatocytes migrated and repopulated the lymph nodes, rescuing the animal from lethal hepatic failure. mTORC2 pathway could contribute to ectopic liver organogenesis inducing transplanted cells to migrate to the extrahepatic sites and propagate.

In the above-mentioned study, hepatocytes behave as metastatic cell being able to migrate to the lymph nodes. Therefore, it is likely that a complex network of chemokines and their receptors contributes to the observed ectopic liver organogenesis. Importantly, chemokine receptor/ligand interactions have been shown to activate mTORC1 in some cancer types. Thus, an intricate regulation of the mTOR system could allow hepatocytes to migrate, enter the lymph nodes, be retained and proliferate.

RiMED Foundation

03/09/11 – 1/9/12

Grant of the Month | April 2011

Alan Russell

Richard Koepsel

Miniature Biofuel Cell from Gold Microfiber Electrodes

Evolving research on implantable sensors, drug-delivery systems and other power consuming implantable devices like pacemakers and insulin pumps requires the matching development of power sources that can be used together with the implantable device.  A great deal of research performed on miniature, lightweight, long-lived batteries resulted in the development of the small lithium ion battery [1]. The common battery is an energy source that contains reacting chemicals securely encased in an impermeable cell, and provides only the electrical leads to connect to devices.  Current lithium iodide pace maker batteries have an open circuit voltage of 2.8 V, weigh about 13 g and have a relatively large volume of 5-8 ml. [2]  Cardiac pacemaker battery design poses a number of special challenges including: the development of biocompatible materials; prevention of corrosion;  preventing leakage of the contents; ensuring high reliability; and accurate determination of  the end of the battery life.  Many of these concerns and many of the inherent risks involved in battery replacement could be alleviated with a longer lasting biomimetic power source.

Another example is the insulin pump where most modern models are external and operate on common batteries that need to be replaced every 3-4 weeks.  Many of the limitations of batteries can be eliminated by replacement with a fuel cell.  A fuel cell has reactants that are fed from external reservoirs to the power cells.  A biofuel cell based on enzymatic redox reactions which uses reactants that are always present in vivo holds great potential in applications and devices that would benefit from implantable power sources.  A biofuel cell can be made smaller than batteries that require containment, would be biocompatible and environmentally friendly, could potentially run indefinitely, and can produced be cost-effectively.

The biofuel cell consists of an anode containing an oxidizing enzyme, typically glucose oxidase (GOX), and a cathode containing a reducing enzyme, usually either laccase or bilirubin oxidase (BOD). The fuel for the power generation is glucose which is oxidized by GOX to gluconolactone and hydrogen peroxide resulting in a 2 electron transfer to the electrode. The electrons flow from the anode to the cathode and are utilized by the reducing enzyme, laccase or BOD, to reduce dissolved oxygen to water.

The literature provides many examples of biofuel cells with different configurations of enzyme immobilization and electron shuttling mechanisms between enzyme and electrode [3-7]. The reported configurations suffer from several limitations: (a) Limited supply of oxygen to the cathode thus the power output is lower than the theoretical value [3]; (b) The electron transfer between the enzyme and the electrode is facilitated by the use of an electron mediator, either soluble or immobilized on the enzyme or on surrounding supporting polymer and these mediators are most often highly toxic species such as osmium complexes [4]; and (c) The size of the electrode is most often large (several centimeters in both diameter and length) and while reduction of the electrode size is possible it is most often accompanied with lower amounts of immobilized enzyme [5] and correspondingly lower power. Thus, the overall performance of biofuel cells is far from optimal, and inhibits the application of the fuel cells in implantable devices.

National Science Foundation

05/01/11 – 04/30/13

Grant of the Month | March 2011

Partha Roy

Marina Kameneva

Drag-reducing polymers to curb breast cancer metastasis

Adhesion of circulating tumor cells to microvascular endothelial cells is key for extravasation of tumor cells and therefore an important step for tumor metastasis. There is growing evidence that systemic inflammation facilitates adhesion of circulating tumor cells to endothelial cells hence promoting metastasis and progression of cancer. It has been hypothesized that leukocytes enhance attachment of tumor cells to endothelial cells by creating formation of a tripartite linkage between these three different cell types. Presence of leukocytes in the tumor microenvironment also leads to local release of cytokines that further promotes junctional disruption of endothelial cells and extravasation of tumor cells. Current strategies to inhibit extravasation which involve molecular targeting of either a single adhesion receptor on tumor cells or a specific signaling pathway are therapeutically inefficient because of involvement of multiple adhesion receptors and signaling pathways in the extravasation process. In complete contrast to these currently envisioned strategies, we proposed a conceptually novel paradigm that hemodynamic perturbation that inhibits attachment of inflammatory cells to endothelial cells is an efficient way to impair tumor cell attachment to endothelium thereby reducing extravasation and metastasis. Systemic administration of so called drag reducing polymers (DRP - long-chain viscoelastic polymers that are non-toxic and blood-soluble) at nanomolar concentrations was shown to reduce/eliminate the near-wall cell-free layer naturally existing in microvessels (Fåhraeus effect) and to increase blood flow in microcirculation. DRP-induced occupation of the near-wall space by red blood cells and increasing of near-wall shear rates may inhibit leukocyte rolling and attachment to blood vessel wall which can drastically reduce inflammatory responses (demonstrated in animals implanted with biodegradable scaffolds) and transendothelial migration of tumor cells. We therefore propose a working postulate that “systemic administration of DRP is a novel interventional approach to reduce extravasation and metastasis of tumor cells” and this hypothesis will be tested by

• Determining whether presence of DRP impairs the ability of circulating breast cancer cells to attach and transmigrate through endothelial monolayer in vitro (Aim 1), and
• Determining whether systemic administration of DRP reduces spontaneous metastasis of breast cancer cells in vivo (Aim 2).

Department of Defense, USAMRMC (Concept Award)

2/1/2011 - 2/29/2012 (Research Ends 1/31/2012)

Grant of the Month | February 2011

Harvey Borovetz

Pumps for Kids, Infants and Neonates (PumpKIN) Preclinical Program: The PediaFlow™ Pediatric VAD

For the past five years our consortium, consisting of the University of Pittsburgh and the Children’s Hospital of Pittsburgh, Carnegie Mellon University, LaunchPoint Technologies and WorldHeart Inc., has undertaken an ambitious program to develop a pediatric blood pump, motivated by the critical need to treat infants and toddlers with congenital and acquired heart diseases. We have relied on first principles to develop, de-novo, a miniature blood pump specifically intended for this population. The first phase of this program has produced the PediaFlow™ PF3, believed to be the world’s smallest magnetically levitated (maglev) blood pump.
With a flow rate range between 0.3 -1.5 LPM and a footprint approximating a AA cell battery, the clinical PediaFlow VAD will provide circulatory support for neonates, infants, and children less than 20 kg who experience cardiac failure and circulatory collapse due to congenital and acquired cardiovascular disease. Our consortium is uniquely poised to carry this forward to clinical use, fulfilling the needs of the PumpKIN program. The individual and collective strengths of our individual organizations and our unique and close collaborations over decades have resulted in innovative implantable blood pumps introduced to clinical use and trials following regulatory approvals.

National Heart, Lung and Blood Institute, NIH

4 years

Grant of the Month | January 2011

William Wagner

Biodegradable, Thermoresponsive Hydrogels to Treat Ischemic Cardiomyopathy

Cardiac failure incurs a major economic and social burden on the United States populace, while also providing a distinct technical challenge since options for treating this condition remain highly limited. In ischemic cardiomyopathy ventricular wall thinning is coupled with dilation of the ventricular cavity. This remodeling process is associated with elevated ventricular wall stress that positively drives the thinning and dilation process towards end-stage heart failure. In the proposed work we will create novel designs for injectable biomaterials to bulk the thinning, post-infarct cardiac wall, reducing elevated wall stress, and potentially improving cardiac remodeling outcomes. The design objectives include synthesizing materials with tensile properties suitable for reducing wall stresses, degradation properties that maintain the hydrogel in the infarcted wall for a period of months during the remodeling process, and drug delivery properties that allow the controlled release of multiple growth factors that may stimulate beneficial cardiac remodeling. We will evaluate 3 distinct hydrogel designs with increasing complexity, utilizing both rat and porcine models of ischemic cardiomyopathy and a minimally invasive robotic technology (the HeartLander device) designed to effectively deliver the targeted hydrogel injections. The project specific aims are to:

1) Evaluate the functional and histopathologic effect of injecting the thermoresponsive hydrogel, poly(NIPAAm-co-acrylic acid-co-2-hydroxyethyl methacrylate-poly(trimethylene carbonate) into the central and border regions of a myocardial infarct in a porcine model for ischemic cardiomyopathy using a modified HeartLander minimally invasive robotic system.

2) Evaluate the functional and histopathologic effect of injecting the thermoresponsive hydrogel, poly(NIPAAm-co-HEMA-co-polylactide-methacrylate) into the central and border regions of a myocardial infarct in rat and then porcine (with HeartLander) models for ischemic cardiomyopathy. Hydrogel design optimization will be based on in vitro and rat in vivo results.

3) Develop and characterize the thermoresponsive hydrogel poly(NIPAAm-co-N-hydroxysuccinimide-methacrylate-co-HEMA-co-MAPLA) and the ability of this gel to deliver bFGF in vitro, as well as the ability to load this gel with microspheres containing IGF-1 for more extended controlled release.

4) Evaluate the functional and histopathologic effect of injecting growth factor loaded hydrogel from Aim #3, into infarcted myocardium in rat and porcine (with HeartLander) models for ischemic cardiomyopathy.

National Heart, Lung, and Blood Institute, National Institutes of Health

01/01/11 – 11/30/15

Grant of the Month | December 2010

Steven Little

The objective of this project is to obtain preclinical data for new treatments for periodontitis that restore immunological homeostasis.           

Wallace H. Coulter Foundation

9/1/2011 – 8/31/2013

$150,000

Grant of the Month | November 2010

ARM-IV Postdoctoral Program  (Four Positions)

1. “Rational Synthesis of Triggerably-Dissolvable Materials for Minimally Invasive Removal of WoundCAP Delivery Devices”
Mentors:  Steven Little, Ph.D. and William Wagner, Ph.D.
Objective: The ultimate objective is to develop a robust, hollow fiber-based system (WoundCAP) to deliver regenerative growth factors to a wound site while including the means for minimally invasive removal/dissolution of the delivery system.  We hypothesize that the resulting hollow fibers wound cap will have robust mechanical properties to maintain stable structures, but will dissolve rapidly upon application of a trigger, either a temperature change or enzyme solution injection.
2.  “Composite Scaffolds for Bone and Soft Tissue Attachments Targeted to Limb and Digit as Well as Craniofacial”
Mentors: Charles Sfeir, DDS, PhD  and Alejandro Almarza, PhD (Basic Science Mentors); Bernard J. Costello, MD (Clinical Mentor)
Objective:  The overall objective of this project is to engineer a regeneration therapy for the bone/tendon complex to provide both hard and soft tissue integration after injury.  The study will use hybrid scaffolds to provide the appropriate microenvironment for both bones and tendon cells.  Three dimensional manufacturing techniques will be used to print the biphasic scaffold where bone marrow derived stem cells will be seeded to form the hard/soft tissue constructs. These constructs will be implanted in-vivo to determine their efficacy to regenerate the bone/tendon interface.
3. “Matrix Mediated Enhancement of Musculotendinous Tissue Regeneration”
Mentor: Stephen F. Badylak, DVM, PhD, MD
Objective: To determine the mechanism(s) by which multipotential stem/progenitor cells are recruited to the site of constructive remodeling/regeneration when biologic scaffolds are used as a therapeutic intervention for traumatic musculotendinous injury, and to characterize the “genetic signature” of the mammalian multipotential cells at the regenerative site and compare this genetic profile to cells that compose a true “blastema” found in regenerating species such as the newt.
4. “Scaffolds for Vascularized Bone Regeneration”
Mentor: Yadong Wang, Ph.D.
Objective:Bone regeneration is very important in limb and digit regenerative medicine because bone formation is a basic step in limb pattern regeneration.  Our long-term goal is to restore function in damaged bone tissues via holistic bone tissue engineering that includes closely-related tissues such as blood vessels and nerve. We propose to achieve this by using scaffolds containing multi-regenerative factors. The objective of this proposal is co-regeneration of bone and blood vessels.

PTEI/DOD

10/01/10 – 09/15/12

$800,768

Grant of the Month | October 2010

Robert Squires

A Multi-Center Group to Study Acute Liver Failure in Children

Our goal is to improve short- and long-term outcomes for pediatric acute liver failure (PALF) through a better understanding of patient phenotypes, reassessment of risk classifications, and associating early events to outcome at one year.  We will integrate two research efforts (Vodovotz-3UO1DK-072146-05S1 and Roberts-1R21DK084201-01) currently collaborating with the PALF Study Group (NIH/NIDDK UO1 DK072146-05) which are (1) modeling PALF as a complex biological system using physiological and inflammatory biomarkers and (2) developing models to represent the liver transplant (LT) decisions in PALF.  To examine our hypotheses that clinical, biochemical, genomic, proteomic, metabolomic, immunologic, and cytokine analyses in PALF can be used to accurately define phenotypes that respond favorably to directed therapy (e.g., immunomodulation) as well as predict disease progression, including potential for spontaneous recovery or risk of death, all of which will provide a platform on which computer/informatics-based (e.g., in silico) studies can inform the design and conduct of clinical trials, and evaluate the impact of therapeutic decisions, including LT;  we propose these Aims: Aim 1: To comprehensively characterize PALF phenotypes utilizing traditional clinical, biochemical, diagnostic, and management profiles supplemented by immune, inflammatory and liver regeneration markers to identify factors that explain variations in outcomes for PALF phenotypes. Outcomes include survival, LT, neurocognitive function, health-related quality of life (HRQOL), depression and post-traumatic stress disorder (PTSD) 6 months and 1 year after enrollment.  Aim 2: To model the dynamics of PALF within and between distinct phenotypes using serially collected clinical, physiological, and biomarker data.  Statistical modeling techniques will be augmented with models used to represent complex biological systems to more accurately reflect the dynamic nature of PALF.  The data and models will be utilized to create a computer-based or “in silico” analog of PALF to simulate interventional studies and to assess treatment, including LT decision processes and to estimate the impact of improved decision-making on organ allocation.

NIH ARRA

09/15/10 – 08/31/15

Year 1:  $24,907
Year 2:  $300,000
Year 3:  $250,000
Year 4:  $250,000
Year 5:  $250,000

Grant of the Month | September 2010

Thomas Gilbert

Cardiac Remodeling with Organ Specific Extracellular Matrix Scaffolds

Improved materials for cardiac reconstruction of congenital defects and heart failure are needed. Current surgical approaches for cardiac reconstruction utilize synthetic materials that slow the progression of disease, but do not provide any contractile function and do not have the ability to grow with the patient. Recently, porcine urinary bladder matrix (UBM) has been used to repair myocardial tissue. The remodeled UBM contributed to regional function in both canine and porcine models, but did not fully restore myocardial tissue. Cardiac extracellular matrix (C-ECM) may promote faster reconstruction of functional tissue by providing a scaffold with a composition and architecture similar to the tissue that it is intended to replace. The proposed study will determine the morphologic and functional differences in cardiac remodeling after repair with C-ECM, UBM, and Dacron patches. Furthermore, the study will include analysis of the recruitment and fate of bone marrow derived progenitor cells at the site of remodeling.

NIH

06/01/10 – 05/31/11

$71,929 (Year 2), $144,674 (Total)

Grant of the Month | August 2010

Kacey Marra

3D Culture of Adipose Tissue for Screening Obesity-related Drugs

We have developed a novel, 3D bioreactor technology that permits the long-term culture of adipocytes, which is not possible using traditional 2D cell culture methods. In this study, we will utilize our technology to rapidly and effectively screen the effects of drugs on human adipose tissue function. We will examine the function of adipocytes in both obese and non-obese patients. One of the parameters we will study is cytokine/adipokine expression. With the bioreactor technology, we are able to rapidly and easily analyze daily expression of cytokines in the media. New drugs may target cytokine expression of adipocytes. It has been shown that involved cytokine behavior in obesity includes the increased expression of, but not limited to: MCP-1 (monocyte chemotactic protein-1, which can recruit macrophages to adipose tissue), TNF-α (tumor necrosis factor-α, a pro-inflammatory mediator secreted by macrophages), and IL-8 (interleukin-8, a pro-inflammatory cytokine secreted by macrophages). Also of interest is the expression of anti-inflammatory cytokines, such as IL-10 (interleukin-10). While these mediators have been examined using human and murine adipose tissue in 2D in vitro culture, improved experimental systems are necessary to allow the development of high throughput assays for drug discovery. Therefore, the specific aims of this proposal are to: 1) Isolate and characterize human adipose-derived stem cells from both male and female patients, age 40-60 years, (non-obese, vs. obese patients); 2) Develop a novel, multi-compartment, hollow fiber 3D perfusion bioreactor technology for ASC culture in 3D bioreactor; 3) Utilize the 3D perfusion bioreactor system as a tool to study the effects of drug therapies on adipose function. In summary, cell-cell contact in a 3D culture system mimicking natural adipose tissue represents an improvement over current petri-dish technologies aimed at developing high throughput assays for drug discovery.

The National Institutes of Diabetes and Digestive and Kidney Diseases

09/01/2010 – 08/31/2013

$993,776

Grant of the Month | July 2010

Stephen Badylak

Biologic Scaffold Development

This collaborative research and development effort involves the characterization of various forms of extracellular matrix, especially porcine dermal derived extracellular matrix, for development and use as a biologic scaffold for pelvic floor reconstruction and general surgical use.  The effort characterizes and identifies novel biomaterials for general surgical applications; particularly pelvic floor reconstruction and hernia repair. We will apply principles of regenerative medicine to principles of general surgery.

C. R. Bard

05/01/2010 – 04/30/2011

$250,000

Grant of the Month | June 2010

Eric Lagasse

Cancer Stem Cells from HBV-Associated Hepatocellular Carcinoma

This research shall be conducted in the laboratories of Drs. Steve Strom and Eric Lagasse with collaborative work packages performed at Vertex. This collaboration shall focus on identification and characterization of HBV-HCC cancer stem cells. The collaboration shall include, but shall not be limited to, the following 4 stages. The objective of Stage I shall be to ascertain whether HBV-HCC primary human tumor tissue contains a cancerous stem cell population. The objective of Stage II shall be to characterize these HBV-HCC cancer stem cells in more extensive in vitro assays (e.g., expansion of the cancer stem cells in bioreactors) and in vivo assays (e.g., tumorigenicity, passage into naïve animals). The objective of Stage III shall be to compare/contrast these HBV-HCC cancer stem cells with candidate cancer stem cells from other forms of HCC. The objective of Stage IV shall be to complete comparative studies with HCC cancer stem cells from liver metastases in remote sites (e.g., lung) or circulating in blood.

Vertex Pharmaceuticals, Inc.

05/01/10 – 04/30/12

$655,629

Grant of the Month | May 2010

Alan J. Russell, Stephen F. Badylak, J. Peter Rubin, Bernard J. Costello, Prashant N. Kumta, Charles Sfeir, Paul Kemp

Limb Salvage and Regenerative Medicine Initiative

This program will advance technologies that return wounded personnel to active duty, restore their limb, muscular, and skin form or function, and assist them in reclaiming independence, dignity, and self-confidence in the tasks of daily living. The program will fund rapid research, development and validation of innovative technologies to improve the clinical outcome of burn and blast injured personnel. Technology refers to integrated systems based on combinations of hardware, software, pharmaceuticals, biologics, and surgical methods. This initiative will advance medical technologies from their existing levels of maturation, through FDA trials and approval, to significantly improve upon current standard treatments for use by the Department of Defense, Veteran’s Administration, public health, and commercial health systems.

Program Areas:

1)  Muscle Tissue Regeneration with the Use of Biologic Scaffolds for Patients Suffering from Massive Loss of Skeletal Muscle Tissue

2)  Bone Scaffolding for Craniofacial Regeneration

3)  Research and Implementation of Clinical Trials of Allogenic Human Dermal Fibroblasts for Remodeling Scar contractors using ICX-RHY Technology

DOD

04/01/10-3/31/12

$12,183,808

Grant of the Month | April 2010

Stephen Badylak and Michael Sacks

Mechanobiology and Regenerative Medicine

Regenerative medicine approaches for the reconstitution of missing or injured tissues and organs involves the use of scaffolds, cells, and bioactive molecules.  The use of biologic scaffolds seeded with cells is a common approach and several applications have been successfully translated to clinical medicine including lower urinary tract, gastrointestinal tract, musculotendinous, and dermal skin regeneration.  The principles that guide tissue remodeling and regeneration are only partially understood but the influence of biomechanical loading upon the remodeling process is accepted as an important variable.  However, there is an almost complete absence of systematic, quantitative studies to determine the effect of this controllable factor upon tissue remodeling, especially tissues with a smooth muscle wall component.

This study is a quantitative, hypothesis driven study that determines the effects of mechanical loading upon smooth muscle phenotype in vitro and in vivo and the related changes to the architecture of the scaffold upon which they are seeded.  A biologic scaffold derived from the extracellular matrix (ECM) of a porcine urinary bladder will be seeded with smooth muscle cells derived from different sources: the vascular wall, urinary bladder, and esophagus.  The influence of those organ specific mechanical loading regimens upon the remodeling process and the ability to modulate the remodeling process by changing the mechanical loading pattern will be investigated.  Two specific aims are described in which: 1) ECM seeded with the three different types of smooth muscle will be subjected to carefully selected mechanical loading regimens and the effect upon cell phenotype and matrix organization will be quantitatively evaluated and 2) two smooth muscle cells types will be evaluated upon ECM used within an organ culture model (rat bladder wall) to evaluate the effect of cellular and ECM remodeling when adjacent normal tissue cells are present.

NIH (2nd year funding)

05/15/09 – 14/30/11

$723,556 (total for 2 years)

Grant of the Month | March 2010

Marina Kameneva

Multi-scale model of thrombosis in artificial circulation

The objective of the project is to advance the accuracy and utility of a predictive model for thrombosis in blood-wetted cardiovascular devices. The research is built upon a combination of a previous model developed by the PI and colleagues for shear-mediated thrombosis and recent progress in modeling cellular-scale hemodynamics. Further incorporation of a model for synergy of platelet agonists is intended to yield a comprehensive design tool that is practical for design optimization of cardiovascular devices.  

CMU via R01 from the NIH

02/01/09 – 01/31/14 (2nd year funding received)

$829,618

Grant of the Month | February 2010

Harvey Borovetz

Pumps for Kids, Infants and Neonates (PumpKIN) Preclinical Program: The PediaFlow™ Pediatric VAD

For the past five years our consortium, consisting of the University of Pittsburgh and the Children’s Hospital of Pittsburgh, Carnegie Mellon University, LaunchPoint Technologies and WorldHeart Inc., has undertaken an ambitious program to develop a pediatric blood pump, motivated by the critical need to treat infants and toddlers with congenital and acquired heart diseases. We have relied on first principles to develop, de-novo, a miniature blood pump specifically intended for this population. The first phase of this program has produced the PediaFlow™ PF3, believed to be the world’s smallest magnetically levitated (maglev) blood pump with the following outstanding features:

• Unparalleled biocompatibility, due to our maglev technology, streamlined single-flowpath design, and computer-optimization design process.
• Exceptionally small, due to our supercritical (above resonance frequency) rotordynamic technology.
• Valveless turbodynamic design with one moving part to minimize size.
• Optimized computationally using first principles of bioengineering and physics.
• Lightweight controller and battery designed for reliability and failsafe operation.
• Human factors design specifically for pediatric operation.
• Cannulation designed to facilitate removal for bridge to recovery.

With a flow rate range between 0.3 -1.5 LPM and a footprint approximating a AA cell battery, the clinical PediaFlow VAD will provide circulatory support for neonates, infants, and children less than 20 kg who experience cardiac failure and circulatory collapse due to congenital and acquired cardiovascular disease. Our consortium is uniquely poised to carry this forward to clinical use, fulfilling the needs of the PumpKIN program. The individual and collective strengths of our individual organizations and our unique and close collaborations over decades have resulted in innovative implantable blood pumps introduced to clinical use and trials following regulatory approvals.

The overall objectives of our response to the PumpKIN RFP are to finalize device development and conduct pre-clinical qualification testing necessary to apply for an IDE for a US clinical trial within 2.5 years. Specifically:

I. Finalize the current PediaFlow (PF3) pediatric VAD, leading to the clinical PediaFlow VAD design;

II. Conduct all necessary pre-clinical in-vitro and in-vivo testing with the clinical PediaFlow VAD design;

III. Submit an application and obtain IDE approval for the PediaFlow VAD;

IV. Collaborate with the PumpKIN Data Coordinating Center (DCC) and the other PumpKIN pre-clinical contractors to develop the clinical protocol and monitoring procedures which will be used in the PumpKIN program clinical study; and

V. Provide regulatory, manufacturing, training, and technical support for the PediaFlow VAD while the clinical study is underway.

National Heart, Lung and Blood Institute, NIH

4 years

$5,630,969

Grant of the Month | January 2010

William J. Federspiel

Anaerobically stored red blood cells with extended shelf-life

As part of the overall project, the University of Pittsburgh, McGowan Institute of Regenerative Medicine under the leadership of Dr. William Federspiel, will develop the conceptual design for the Oxygen Depletion Device (ODD) for NHSi’s Hemanext Anaerobic Storage Platform (HASP) red blood storage system.

The long-term objective of the overall project is to develop a novel blood storage system that will extend the shelf life of additive system red cell units, and at the same time, deliver red cells of higher efficacy and lower toxicity for transfusion therapy. In this system, red cells are stored in a modified additive solution under oxygen-depleted condition (anaerobic storage).  The final product will have extended shelf life (9 weeks or more) as well as higher efficacy and lower toxicity compared to same-aged blood stored by conventional methods (more viable cells, higher oxygen delivery capacity immediately after transfusion and more deformable cells for better capillary perfusion).

The new storage system is design to be readily accommodated by the current blood banking operation without incurring major alteration in procedures or equipments.  In the Phase I of this project, an experimental procedure was used to demonstrated that anaerobic storage yields; i) a significantly higher post-transfusion recovery compared to the conventional method after 6 weeks of storage; and ii) a comparable 9-week recovery compared to 6-week storage under conventional conditions.

NIH STTR

7/1/09 – 6/30/10

$139,583 (University Sub Contract; Prime to New Health Sciences)

Grant of the Month | December 2009

Stephen F. Badylak, DVM, PhD, MD

Control of the Microenvironmental Niche to Promote Epimorphic Regeneration in Amputated Digits

a. Background: The proposed studies address one of the most pressing needs of the wounded soldier population: the replacement of lost digits and limbs.  These studies challenge a fundamental tenet of the mammalian response to tissue injury; specifically, the concept that regeneration of complex tissues following severe injury is not possible in adult mammals.  We hypothesize that such regeneration is indeed possible and, to support this hypothesis, we have successfully recruited endogenous stem cells to the injury site thru the use of chemotactic matricryptic peptides derived from extracellular matrix (ECM). We call this cell mass a “multipotential cell cluster” and it is similar, but not identical, to the true blastema of regenerating species such as the newt.  We now seek support to identify the genetic signature, the appropriate microenvironmental niche, and inductive molecular signals required to stimulate functional tissue formation from the multipotential cell cluster. 

Concept:  We can effectively recruit abundant endogenous multipotential cells to the site of digit amputation in our well established mouse model.  Proposed work would include isolation of individual cells that have been dissociated from the MCC and characterization of the transcriptome.  The number of cells will be very low and will require cell sorting technology that our team has recently developed.  Characterization of the transcriptome will be conducted by the J. Thomson/R. Stewart group at Madison, Wisconsin. Realization of the regenerative potential of the multipotential cell cluster will require control of the microenvironmental niche through development of a “biodome”.  This work will be conducted by the team of Susan Braunhut in Boston, Massachusetts.  The critical components of the proposed work include: 1) the mouse model and creation of the MCC, 2) MCC isolation and cell sorting, 3) characterization of the genomic profile and transcriptome, and 4) development of a biodome. This multidisciplinary project can only be accomplished through the combined efforts of multiple investigators and multiple novel technologies.  The problem is challenging but the potential payoff is huge.

b. Objective/Hypothesis: To induce functional tissue formation in adult mammals by controlling the microenvironmental niche into which multipotential stem cells are recruited.

c. Specific Aims:  1) To identify the genetic signature that characterizes constructive tissue remodeling as opposed to scar tissue formation.  2) To produce a prototype biodome that can locally manipulate and control factors such as hydration state, pH, oxygen tension, electrical potential, and nutrient composition at the site of a multipotential cell cluster in a mouse model of digit amputation.  3) To form bone and functional contractile tissue at the site of digit amputation in a mouse model.  4) To develop and document the formation of a nerve and vascular plexus into the regenerated digit tissue. 

d. Study Design: This two year project will utilize the mid-second Phalanx digit amputation model in C57Bl/6 mice.  The work will be a progressive investigation of variables in the biodome that effect functional tissue reconstruction.  In year one, in addition to developing the prototype biodome, we will evaluate oxygen tension and nutrient composition of the biodome fluid as variables that effect the cell transcriptome and functional tissue reconstruction.  During year two, we will utilize the second version of the biodome device in which electrical potential, pH, and replacement of the biodome fluid can be investigated in systematic fashion.  In addition, in year two, the analytic transcriptome comparison of the multipotential cell cluster (MCC) that develops in the C57Bl/6 mice will be compared against a carrier treated mouse control group and the transcriptome data that has been developed in separately funded work using the red spotted newt.  This comparative transcriptome analysis will provide invaluable information regarding the efficacy of controlling cell phenotype and functional tissue reconstruction.

Continued iterative versions of the biodome will be utilized over the two years of this work.  As opposed to what would be possible in one year, which is development of a single prototype, the second year will provide the opportunity to add controllable variables and develop a more user friendly version of the prototype.  In addition, the shared number of biodomes developed and the size of the animal studies will increase dramatically thus adding to the study output. 

ARM 3

09/30/09 – 09/29/11

DC: $374,903 (for 2 years)
IDC: $52,226

Grant of the Month | November 2009

Eric Lagasse

Organogenesis of Ectopic Tissue in Lymph Node

Our proposal addresses some of the solutions to the development of complex 3-
dimensional tissue models and a new paradigm by using lymph node as in vivo

National Institutes of Health

09/25/09 – 08/31/14

Year 1
DC = $560,526
IDC = $237,171
TC = $797,697

Total
DC = $2,533,749
IDC = $1,235,381
TC = $3,787,133

Grant of the Month | October 2009

J. Peter Rubin

Biomedical Translational Initiative: Structural Fat Grafting for Craniofacial Trauma

Facial trauma injuries, especially those sustained in military combat, are characterized by destruction of bone and soft tissue anatomy. While the bony skeleton can often be reconstructed, the overlying soft tissue is difficult to restore. Importantly, it is the structure of the soft tissue that imparts the normal human form, and adequate reconstruction of soft tissue defects allows trauma victims to reintegrate into society. Current procedures for soft tissue reconstruction of the face primarily involve tissue flap reconstruction procedures. Synthetic (e.g silicone) implants for soft tissue trauma of the face have no practical role and are fraught with complications and poor results. Tissue flap operations are extensive, often including microvascular surgery, and do not precisely correct the deformities. Autologous fat grafting, performed through a minimally invasive means, has the potential to correct deformities with much greater precision and lower morbidity.

While autologous adipose tissue grafting may provide a minimally invasive means of accurately restoring facial soft tissue structure after trauma, graft resorption is a significant limitation. In this study, we capitalize upon two enabling technologies to make fat grafting effective for the wounded soldier and validate outcomes: 1) Specialized instrumentation for fat tissue harvest (Lipokit, Medi-Khan USA, Inc , California) that concentrates adipose stem cells, and 2) Specialized instrumentation for fat tissue injection (Coleman Cannula System, Mentor Medical, California) that allow for precise placement of appropriately sized aliquots of fat within the injured tissues and adjust to irregularly shaped and scarred tissue beds. Both of these devices are approved by the FDA.

We hypothesize that results from fat grafting for facial trauma, made possible by these two enabling technologies, will show good restoration of tissue volume and craniofacial form. Additionally, we hypothesize that the results will be durable and patient quality of life improved. Validation will be accompanied by a plan to make this technology broadly available to physicians treating combat injuries.

The specific aims of the study are:

1) Treat disfiguring craniofacial injuries in 20 soldiers with fat grafting to improve form with a high level of precision. Facial appearance and persistence of treatment effect will be assessed using aesthetic grading scales, state of the art 3D photography, and high resolution CT scanning with 3D reconstruction. Patients will be followed for 9 months after treatment to define long term outcomes.

2) Assess biologic properties of the cells within the fat graft and correlate with clinical outcomes. This will include adipose stem cell yield per volume of fat tissue, cell proliferation, capacity for adipogenic differentiation, lipolysis, and cell sub-population analysis by multiparameter flow cytometry. Results of these assays will be correlated with graft volume retention to search for predictors of good clinical outcome that are related to variation on adipose biology between subjects.
 

Department of Defense

10/01/09 to 3/31/2010

$1,618,653 ($1,141,964 direct / $ 476,689 indirect)

Grant of the Month | September 2009

Joseph Ahearn

Molecular and Functional Characterization of the Lupus Platelet

The platelet plays critical roles in control of abnormal bleeding, in formation of pathologic thromboses, and in molecular and cellular mechanisms of inflammation, wound healing, and the immune response.  Recently it has been found that platelets can also participate in atherothrombosis and inflammation through the formation and release of membrane-bound vesicles called microparticles.  These platelet-derived microparticles (PMP) readily circulate in the vasculature and exhibit procoagulant activity.  Platelets and PMP are likely to contribute to myriad manifestations of lupus, although they appear to be relatively understudied in this regard. We have recently discovered that platelets bearing complement activation product C4d (PC4d) are highly specific for lupus versus other inflammatory diseases (98%) and versus healthy subjects (100%).  PC4d-positivity in patients with lupus is associated with neuropsychiatric and thrombotic manifestations of the disease. Through imaging and flow cytometric studies we have since discovered that PC4d associates not only with intact platelets, but also with membrane-bound PMP. This study has two central hypotheses:  first, that C4d deposition on platelets and PMP has functional consequences that contribute to thrombotic manifestations in lupus, and second, that platelets  bearing C4d hold molecular clues to a pathway responsible for the PC4d phenotype and to pathways that lead to the functional consequences of PC4d deposition. This application will specifically focus on determining differences, both functional and molecular, between PC4d-positive and PC4d-negative platelets in patients with SLE.  This will be accomplished through three specific aims.  The first aim is to investigate functional differences between PC4d-positive and PC4d-negative platelets with regard to thrombogenesis and atherogenesis.  The second aim is to investigate the functional effect of PC4d-positive versus PC4d-negative PMP on immune cells and other cellular components of the vasculature.  The third aim is to identify specific protein differences in C4d-positive platelets compared to C4d-negative platelets in patients with SLE and in healthy controls.  Successful completion of the proposed studies should advance our understanding of the functional role of platelets, PMP, and complement in the thrombotic manifestations of lupus.  In addition, identification of a lupus platelet proteomic signature would elucidate important molecular and cellular mechanisms of thrombotic and other complications of lupus and would identify potential therapeutic targets.

Department of Defense

7/1/09 – 6/30/12

$183,101

Grant of the Month | August 2009

W. P. Andrew Lee MD

Jörg Gerlach,MD, PhD and Jack Patzer, PhD

Long-term immunosuppression-free survival of a combined composite tissue allograft (CTA) and autologous skin in a swine model

Wide spread use of Composite Tissue Allografts such as a hand or face transplant are currently limited by the high dose and long-term immunosuppressive treatment required to prevent graft rejection. This is predominantly related to the high immunogenicity of the skin.

Pittsburgh Tissue Engineering Initiative via DOD

04/01/09 – 09/30/10

$450,450

Grant of the Month | July 2009

Stephen Badylak and Michael Sacks

Thomas Gilbert

Mechanobiology and Regenerative Medicine

Regenerative medicine approaches for the reconstitution of missing or injured tissues and organs involves the use of scaffolds, cells, and bioactive molecules.  The use of biologic scaffolds seeded with cells is a common approach and several applications have been successfully translated to clinical medicine including lower urinary tract, gastrointestinal tract, musculotendinous, and dermal skin regeneration.  The principles that guide tissue remodeling and regeneration are only partially understood but the influence of biomechanical loading upon the remodeling process is accepted as an important variable.  However, there is an almost complete absence of systematic, quantitative studies to determine the effect of this controllable factor upon tissue remodeling, especially tissues with a smooth muscle wall component. 

The present proposal seeks support to conduct a quantitative, hypothesis driven study that determines the effects of mechanical loading upon smooth muscle phenotype in vitro and in vivo and the related changes to the architecture of the scaffold upon which they are seeded.  A biologic scaffold derived from the extracellular matrix (ECM) of a porcine urinary bladder will be seeded with smooth muscle cells derived from different sources: the vascular wall, urinary bladder, and esophagus.  The influence of those organ specific mechanical loading regimens upon the remodeling process and the ability to modulate the remodeling process by changing the mechanical loading pattern will be investigated.  Two specific aims are described in which: 1) ECM seeded with the three different types of smooth muscle will be subjected to carefully selected mechanical loading regimens and the effect upon cell phenotype and matrix organization will be quantitatively evaluated and 2) two smooth muscle cells types will be evaluated upon ECM used within an organ culture model (rat bladder wall) to evaluate the effect of cellular and ECM remodeling when adjacent normal tissue cells are present.

NIH

05/15/09 – 14/30/11

$361,332 (Year 1), $723,556 (total for 2 years)

Grant of the Month | June 2009

PI

Steven Little, PhD

Title

Temporal Delivery of Growth Factors for Wound Healing Using Porous Hollow Fibers

Description

Our objective is to optimize wound healing through temporal delivery of growth factors using porous hollow fibers extending into a wound site.  As an extension to the wound-cap technology (artificial capillary bed delivery system), these fibers can be made from materials that dissolve in the presence of a chemical or temperature-based trigger following the wound healing process.  Because angiogenesis is, in many cases, one of the first steps towards wound healing, we propose to demonstrate enablement of this technology by mimicking the natural sequence of stimuli that directs angiogenesis.  Our hypothesis is that sequential delivery of appropriate angiogenesis-promoting factors from our externally-regulated delivery system, as opposed to simultaneous delivery of multiple factors, will result in more mature and integrated neo-vasculature.

Source

PTEI via DOD

Term

April 1, 2009 – September 30, 2010

$91,667

Grant of the Month | May 2009

PI

Thomas Gilbert, PhD

Title

Cardiac Remodeling with Organ Specific Extracellular Matrix Scaffolds

Description

Improved materials for cardiac reconstruction of congenital defects and heart failure are needed. Current surgical approaches for cardiac reconstruction utilize synthetic materials that slow the progression of disease, but do not provide any contractile function and do not have the ability to grow with the patient. Recently, porcine urinary bladder matrix (UBM) has been used to repair myocardial tissue. The remodeled UBM contributed to regional function in both canine and porcine models, but did not fully restore myocardial tissue. Cardiac extracellular matrix (C-ECM) may promote faster reconstruction of functional tissue by providing a scaffold with a composition and architecture similar to the tissue that it is intended to replace. The proposed study will determine the morphologic and functional differences in cardiac remodeling after repair with C-ECM, UBM, and Dacron patches. Furthermore, the study will include analysis of the recruitment and fate of bone marrow derived progenitor cells at the site of remodeling.  The study will be conducted in collaboration with Drs. Badylak, Wagner, and Tobita, and members of their respective laboratories.

Source

NIH-NIBIB – RO3

Term

June 1, 2009—May 31, 2011

$145,400

Grant of the Month | April 2009

PI

Johnny Huard

Title

Tissue Engineered Skeletal Muscle (TESM) from Muscle Progenitor Cells: A Model for Studying Insulin Resistance and Muscle Metabolism

Description

The overall specific aims are as follows:

• To optimize a protocol for the isolation and differentiation of human MPC
• To develop a regimen and bioreactor for development of a tissue engineered skeletal muscle model (TESM) from MPC that will provide a means to study nutrient metabolism and insulin function
• To utilize human MPC and the TESM system to analyze pathways involved with glucose homeostasis and fatty acid uptake in healthy and insulin resistant phenotypes

Source

Pfizer

Term

March 1, 2009 – February 28, 2011

$475,000

Grant of the Month | March 2009

Steven Belle and Kyong-Mi Chang

Robert Carithers, Adrian Di Bisceglie, Michael Fried, Marc Ghany, Steven Han, E. Jenny Heathcote, W. Ray Kim, Daryl Lau, William Lee, Anna Lok, Mitchell Shiffman, KathleenSchwarz, and Norah Terrault

Multi-Center Clinical Trials of Novel Therapies and Diagnostics for Patients with Chronic Hepatitis B

In establishing the Hepatitis B Research Network, the NIDDK wishes to consider the potential application of diagnostics and therapeutics for patients of all ages with chronic hepatitis B. The overall goal of the Hepatitis B Network will be to perform clinical, epidemiological and therapeutic research in patients with chronic hepatitis B using a standardized and coordinated approach to the evaluation and therapy of chronic hepatitis B and to provide sufficient numbers of patients for the research. This will be done by development of a database on chronic hepatitis B patients including clinical information as well as liver, serum and DNA samples.

National Institute of Diabetes and Digestive and Kidney Diseases

2009 - 2016

$11 million

Grant of the Month | February 2009

Steven F. Badylak, DVM, PhD, MD

Regenerative Medicine Approach to the Treatment of Abdominal Compartment Syndrome in a Dog Model

This study involves a combination of preclinical animal work and the treatment of clinical patients at Fort Sam Houston in San Antonio, Texas at the Institute for Surgical Research.  The work is based upon the bioinductive properties of an extracellular matrix (ECM) scaffold derived from porcine urinary bladder.  This biologic scaffold contains a bimodal surface architecture which is supportive of epithelial cell growth on one surface and integration into an exposed wound on the opposite surface.  The preclinical animal studies show the ability of the material to integrate into the full thickness wound bed.  Seven patients have been treated and results show that the preclinical studies were predictive of the excellent biointegration that occurred in these patients.  Complete epithelialization with no contracture was observed when the material was used to treat the donor sites of patients requiring split thickness skin grafts.  This pilot safety study sets the stage for subsequent clinical applications and primary full thickness wounds in patients.

PTEI (STRaC)

9/1/08 – 11/30/08

$95,000 add-on

Grant of the Month | January 2009

Rick Koepsel

Sharon Marx and Gabriel Amitai

Temperature responsive modification of microfiber tissue perfusion devices.

Tissue perfusion devices based on hollow fibers have been developed for use in bioreactors and for wound healing.  In both cases the perfusion devices are in direct contact with cells and tissues.  The hollow fiber tubing that makes up the devices is fabricated from hydrophilic materials, which resist the direct attachment of cells and proteins in the short term but over time cells will attach to the devices.  With cells and tissue attached to the device, removal of the device can disrupt the structure of the tissue that was the intended when the device was implanted.  This project will extend the development of tissue perfusion devices by providing smart polymer coatings which, when activated, will facilitate the removal of the device.

Smart polymers are materials that exhibit a response to a physical stimulus.  For this project we will concentrate on the temperature responsive polymer poly (N-isopropylacrylamide) (pNIPAAm).  As with many temperature responsive materials, pNIPAAm in aqueous solution changes structure at a temperature called the lower critical solution temperature (LCST).  In the case of pNIPAAm the LCST is about 320C, below the LCST the polymer is soluble in aqueous solution while above the LCST a change in the polymer structure causes the polymer to be come hydrophobic and form micelles in solution.  When pNIPAAm is immobilized on a surface it still responds to temperature.  A surface coated with pNIPAAm is hydrophobic below the LCST but becomes hydrophilic above the LCST.  Cells grown on tissue culture dishes coated with pNIPAAm will attach and proliferate normally while the dishes are kept at 370C but when the temperature is lowered below the LCST the cells are sloughed off of the surface.  If the cells have grown to a confluent monolayer, they will come off the surface as a cohesive sheet showing that the underlying protein layer to which the cells are attached is also sloughed from the surface. Applying a layer of pNIPAAm to the hollow fiber perfusion tubing should therefore allow a temperature change to eliminate the adhesions between the cells or tissues and the perfusion device allowing the device to be removed with considerably less trauma to the insertion site.

PTEI/DOD

11/1/08 – 10/31/09