Engineering Commons^™

Engineering An Injectable Hydrogel With Self-Assembling 3d Vasculature, Kendyl Cohn

Master's Theses

This research developed methods for culturing self-assembling capillaries in an injectable gel as a potential method for vascularizing tissue-on-a-chip models to mimic physiological drug delivery. Additionally, a mathematical model was developed as a tool for understanding nutrient delivery and comparison of potential delivery systems. Organs-on-a-chip provide novel platforms for studying biology and physiology in 3D, allow exploration of tissue engineering on a manageable scale, and serve as models for drug screening and drug-delivery testing.

Methods were first developed for co-culture of endothelial cells and fibroblasts (3T3s or HDFs) in 2D, evaluating culture time, seeding density and ratio of HUVECs and …

Using Noninvasive Calibrated Cuff Plethysmography To Observe The Effects Of Cold-Water Immersion On Arterial Compliance, Rita M. Grigorian

Biomedical Engineering: Graduate Reports and Projects

As the prevalence of cardiovascular diseases continues to exponentially grow in populations across the globe, the necessity of determining underlying factors, effective methods of diagnoses, and universally available preventive measures also grows. Early detection of endothelial dysfunction, a proven precursor of cardiovascular diseases, can be extremely impactful in encouraging preventative measures and early intervention before medical conditions become chronic. In recent years, ice plunging, a form of cryotherapy involving full body immersion in cold water, has gained popularity within circles of fitness and health practitioners, gaining the interest of people of all backgrounds. Certain parallels observed between the human physiological …

Development Of An Angiogenic Tissue-On-A-Chip Microenvironment, Eric Stuehr

Master's Theses

Preclinical testing is necessary to investigate the safety and efficacy of novel therapeutics before moving to clinical trials, yet approximately 90% of these therapies fail once tested in humans. This has led to increased interest in developing robust preclinical models that accurately mimic the complex human in vivo physiology. Microfluidic devices that can introduce dynamic conditions to 3D cell/organoid cultures, also known as tissue-on-a-chip, have emerged as physiologically relevant in vitro preclinical models that can achieve high throughput screening of therapeutics. The research presented here aimed to develop an angiogenic environment within a novel microfluidic device to stimulate formation of …

Using Noninvasive Calibrated Cuff Plethysmography To Observe The Effects Of Cold-Water Immersion On Arterial Compliance, Rita M. Grigorian

Master's Theses

As the prevalence of cardiovascular diseases continues to exponentially grow in populations across the globe, the necessity of determining underlying factors, effective methods of diagnoses, and universally available preventive measures also grows. Early detection of endothelial dysfunction, a proven precursor of cardiovascular diseases, can be extremely impactful in encouraging preventative measures and early intervention before medical conditions become chronic. In recent years, ice plunging, a form of cryotherapy involving full body immersion in cold water, has gained popularity within circles of fitness and health practitioners, gaining the interest of people of all backgrounds. Certain parallels observed between the human physiological …

Development Of A Cell-Based Regenerative Strategy To Modulate Angiogenesis And Inflammation In Ischemic Muscle, Fiona E. Serack

Electronic Thesis and Dissertation Repository

The delivery of human adipose-derived stromal cells (hASCs) to ischemic tissues represents a promising strategy to promote vascular regeneration for patients with critical limb ischemia (CLI). This thesis focused on the evaluation of hydrogels to enhance the retention and pro-angiogenic capacity of hASCs following delivery in vivo. Additionally, priming strategies to augment the paracrine function of hASCs were developed and assessed.

Recognizing the importance of endogenous macrophages in the pro-regenerative function of hASCs, delivery using a previously-developed hydrogel system, composed of peptide-functionalized methacrylated glycol chitosan (MGC-RGD) and a copolymer of poly(ethylene glycol) and poly(trimethylene carbonate) (PEG(PTMC-A)₂), was …

Angiogenic Supports For Microvascular Engineering, Zain Siddiqui

Dissertations

Ischemic tissue disease is caused by a lack of circulation / blood supply to tissue. This can be treated by introducing a number of angiogenic (pro-blood vessel forming) factors into the tissue. This work presents strategies for ischemic tissue treatment utilizing a novel proangiogenic self-assembling peptide hydrogel platform. To demonstrate the utility of this platform, its use alone as an angiogenic therapeutic (both alone as a self-assembling hydrogel and with two-component systems), and its ability to vascularize implants is explored. Due to these angiogenic scaffolds demonstrating efficacy to regenerate microvasculature, this work evaluates diseases that can be treated by the …

Establishing The Efficacy Of Non-Cellular Components Of Adipose-Derived Stromal Vascular Fraction In Promoting Angiogenesis., Daniel Benson

Electronic Theses and Dissertations

Microvascular disease is hallmarked by pathophysiological conditions such as endothelial senescence, intimal thickening which impairs vasodilation, and regression of the capillary beds causing tissue ischemia in the myocardium or in peripheral vascular networks. Adipose-derived stromal vascular fraction (SVF) has previously demonstrated the ability to revascularize tissue. Increasing evidence shows that regenerative cells elicit their therapeutic benefit by paracrine mechanisms, leaving open extracellular vesicles (EVs) as a potential crux of the cell therapy paradigm. To test this idea, three types of gelatin methacrylate hydrogels were employed: SVF gels, EV gels derived from SVF, and blank control gels, which were used in-vitro …

Bioengineered Scaffolds To Induce Alignment And Promote Axon Regeneration Following Spinal Cord Injury, Kiet Anh Tran

Theses and Dissertations

Scaffolds delivered to injured spinal cords to stimulate axon connectivity often act as a bridge to stimulate regeneration at the injured area, but current approaches lack the permissiveness, topology and mechanics to mimic host tissue properties. This dissertation focuses on bioengineering scaffolds through the means of altering topology in injectables and tuning mechanics in 3D-printed constructs as potential therapies for spinal cord injury repair. A self-assembling peptide scaffold, RADA-16I, is used due to its established permissiveness to axon growth and ability to support vascularization. Immunohistochemistry assays verify that vascularized peptide scaffolds promote axon infiltration, attenuate inflammation and reduce astrogliosis. Furthermore, …

Novel Nanoparticles To Stimulate Therapeutic Angiogenesis For The Treatment Of Peripheral Arterial Diseases, Tam Phung Nguyen

Bioengineering Dissertations

Peripheral arterial disease (PAD) is a severe impairment of arterial vessels resulting in obstruction of normal blood flow in the legs, leading to acute or chronic lower limb ischemia and subsequently high morbidity and mortality rates. Common treatments for PAD, such as medications and surgical revascularization, have several limitations. For instance, medications used to lower cholesterol, reduce high blood pressure, control blood sugar, prevent blood clots, and reduce/delay symptoms such as leg pains. Still, they cannot treat the established disease directly and often cause side-effects, including bleeding, headache, and diarrhea. Meanwhile, many elderly PAD patients cannot undergo surgical options. Therefore, …

Pulsatility Is A Predictive Marker Of Improved Cardiac Function In Patients With Liquid Matrix-Treated Left Ventricular Assist Devices, Philemon Mikail, Rinku Skaria, Marvin Slepian, Janny Garcia, Richard Smith, Zain Khalpey

The VAD Journal

Objective: Left ventricular assist devices (LVADs) are utilized as a bridge to transplant or as destination therapy for patients with end-stage heart failure. Although cardiac offloading from these devices rarely leads to complete remodeling and functional recovery, the use of mesenchymal cells to modulate heart failure has been explored in recent years due to its intrinsic regenerative properties. Current methods of evaluating cardiac function have too much variability, difficulty of access, or require too frequent follow up to create universal weaning protocols. We hypothesized that the administration of amniotic allograft liquid matrix (LM) containing amnion-derived mesenchymal stem cells (aMSCs) in …

The Influence Of Mmp14 On Angiogenesis In Chemotherapy-Treated Tumors, Abdussaboor Muhammad

Biomedical Engineering Undergraduate Honors Theses

Colorectal cancer is the third most common cancer in the world, and it is the fourth most common cause of cancer related death (1). There have been many significant advancements regarding the treatment of cancer which aim to shrink the size of tumors in patients. However, there is still more that needs to be understood about the many different factors that play a role in colorectal cancer development.

Angiogenesis is the process of forming new blood vessels from existing ones and it requires breaking down and remodeling of the extracellular matrix (ECM) in order to allow endothelial cells to migrate …

Piezoelectric Signals In Vascularized Bone Regeneration, Delfo D’Alessandro, Claudio Ricci, Mario Milazzo, Giovanna Strangis, Francesca Forli, Gabriele Buda, Mario Petrini, Stefano Berrettini, M. Jasim Uddin, Serena Danti, Paolo Parchi

Chemistry Faculty Publications and Presentations

The demand for bone substitutes is increasing in Western countries. Bone graft substitutes aim to provide reconstructive surgeons with off-the-shelf alternatives to the natural bone taken from humans or animal species. Under the tissue engineering paradigm, biomaterial scaffolds can be designed by incorporating bone stem cells to decrease the disadvantages of traditional tissue grafts. However, the effective clinical application of tissue-engineered bone is limited by insufficient neovascularization. As bone is a highly vascularized tissue, new strategies to promote both osteogenesis and vasculogenesis within the scaffolds need to be considered for a successful regeneration. It has been demonstrated that bone and …

Quantitative Flow Cytometry-Guided Protein Biomarker Characterization For Translational Proteomics And Precision Medicine, Yingye Fang

McKelvey School of Engineering Theses & Dissertations

Angiogenesis, the growth of blood vessels from pre-existing vasculature, is primarily regulated by vascular endothelial growth factor receptors (VEGFRs). Dysregulated angiogenesis is associated with cancers, obesity, and over 70 vascular diseases. Upregulated VEGFR protein expressions in diseased vasculature are promising biomarkers for predicting clinical outcomes, as indicated by non-quantitative immunohistochemical studies in patients with impaired vascularization or tumor angiogenesis. While the quantitative characterization of VEGFRs is critical in identifying biomarkers for anti-angiogenic therapies, VEGFR biomarker development presents two particular challenges: (1) The invasive tissue biopsy needed limits the amount of VEGFR data that can be collected from both normal and …

Acute Mechanical Stress In Primary Porcine Rpe Cells Induces Angiogenic Factor Expression And In Vitro Angiogenesis, Farhad Farjood, Amir Ahmadpour, Sassan Ostvar, Elizabeth Vargis

Biological Engineering Faculty Publications

Background

Choroidal neovascularization (CNV) is a major cause of blindness in patients with age-related macular degeneration. CNV is characterized by new blood vessel growth and subretinal fluid accumulation, which results in mechanical pressure on retinal pigment epithelial (RPE) cells. The overexpression of RPE-derived angiogenic factors plays an important role in inducing CNV. In this work, we investigated the effect of mechanical stress on the expression of angiogenic factors in porcine RPE cells and determined the impact of conditioned medium on in-vitro angiogenesis.

Results

The goal of this study was to determine whether low levels of acute mechanical stress during early …

Formation Of A Vascular Regenerative Microenvironment Within Implantable Human Decellularized Adipose Tissue Bioscaffolds, Christopher Leclerc

Electronic Thesis and Dissertation Repository

Cellular therapies targeted at stimulating therapeutic angiogenesis in individuals with critical limb ischemia (CLI) have been under intense investigation. Hematopoietic progenitor cells (HPC) derived from umbilical cord blood have been previously shown to support limb revascularization in animal models of CLI, despite limited cell survival at the site of ischemia. This study attempted to improve HPC survival after transplantation and prolong pro-angiogenic function using human decellularized adipose tissue (hDAT) as a novel cell delivery platform. Compared to HPC conventionally grown on tissue-cultured plastic, hDAT scaffolds were shown to promote viability and proliferation of seeded HPC, and had cell- instructive effects …

Effect Of Physical Stimuli On Angiogenic Factor Expression In Retinal Pigment Epithelial Cells, Farhad Farjood

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Age-related macular degeneration (AMD) is a major cause of blindness in adults. Abnormal growth of blood vessels in the eye during the course of AMD causes damage to the retina, resulting in irreversible blindness. The goal of this research was to determine whether physical pressure on retinal cells can contribute to the increased blood vessel formation. To replicate the tears in the cell layers, a micropatterning method was used as a means of detaching cells from each other. Two new devices were also developed to mimic slow and fast increases in mechanical pressure on cell layers of the eye. After …

The Role Of The Mechanical Environment On Cd117+ Endothelial Cell Angiogenesis, Patrick Link

Theses and Dissertations

Angiogenesis is a complex process coordinating cell migration, proliferation, and lumen formation. Changes to the microenvironment regulate angiogenesis through mechanotransduction and cytokine signals. In pulmonary hypertension, something in the process becomes abnormal, resulting in changes to the microenvironment and the formation of a glomerulus of dysfunctional capillaries, called a plexiform lesion. Endothelial cells, expressing CD117 (CD117+ EC clones) increase in the plexiform lesions of pulmonary hypertension, independent of pro-angiogenic VEGF signaling. We hypothesize that the mechanical environment and the macromolecular composition of the extracellular matrix, both, contribute to the aberrant angiogenesis. When we changed the mechanical environment, we changed the …

Nanoparticles For Gene Therapy And Protein Delivery To Induce Angiogenesis As An Alternative Treatment For Peripheral Arterial Disease, Linda Christellia Noukeu

Bioengineering Dissertations

Peripheral arterial disease (PAD) is defined as a slow, progressive disorder of the lower extremity arterial vessels characterized by chronic narrowing that often results in occlusion and is associated with loss of functional capacity. Although the PAD occurrence rate is increasing in the elderly population, outcomes with current treatment strategies are suboptimal. Hence, there is an urgent need to develop new technologies that overcome limitations of traditional modalities for PAD detection and therapy. In this research, nanoparticles have been developed and used as carriers for various therapeutic agents to treat PAD. In particular, via protein/gene delivery, we developed and formulated …

Plasma Enhanced Chemical Vapor Deposition Coating Amorphous Silica Based Implants Enhancing Angiogenesis And Mitigating Toxic Oxidative Stress In Critical Size Bone Defects, Felipe Alves Monte

Bioengineering Dissertations

The advances in bone tissue engineering search for a biomaterial that could efficiently and gradually replace the bone while the new tissue grows through the implanted structure. However, due to limitations mainly related to mechanical properties and challenges, such as the balance between tissue regeneration and material degradation, the coating of well-known inert materials still has been used in orthopedics and dental clinical practices. Bone defects are commonly related to deleterious oxidative stress induced by low oxygen levels and inflammation. Toxic oxidative stress can impair damaged tissue healing due to inhibiting new blood vessels and new bone formation. The interaction …

Quantifying The Effects Of Hydrostatic Pressure On Fibroblast Growth Factor-2 Binding By The Human Endothelium, Taylor R. Mckenty

Theses and Dissertations--Biomedical Engineering

Fluid pressures regulate endothelial cell (EC) tubulogenic activity involving fibroblast growth factor 2 (FGF-2) and its receptor, FGF receptor 2 (FGFR2). Our lab has recently shown that sustained 20 mmHg hydrostatic pressure (HP) upregulates EC sprout formation in a FGF2-dependent fashion. This upregulation of sprout formation may be due to enhanced FGF-2 / FGFR2 interactions in the presence of 20 mmHg HP. We hypothesize that exposure of ECs to 20 mmHg sustained HP enhances FGF-2 binding kinetics. We used a custom hydrostatic pressure system, immunofluorescence, and FACS to quantify FGF-2 binding by ECs in the absence or presence of a …

Three-Dimensional Endothelial Spheroid-Based Investigation Of Pressure-Sensitive Sprout Formation, Min Song

Theses and Dissertations--Biomedical Engineering

This study explored hydrostatic pressure as a mechanobiological parameter to control in vitro endothelial cell tubulogenesis in 3-D hydrogels as a model microvascular tissue engineering approach. For this purpose, the present investigation used an endothelial spheroid model, which we believe is an adaptable microvascularization strategy for many tissue engineering construct designs. We also aimed to identify the operating magnitudes and exposure times for hydrostatic pressure-sensitive sprout formation as well as verify the involvement of VEGFR-3 signaling. For this purpose, we used a custom-designed pressure system and a 3-D endothelial cell spheroid model of sprouting tubulogenesis. We report that an exposure …

Healing Of Bone Defects In A Rodent Calvarial Defect Model Using Strong Porous Bioactive Glass (13-93) Scaffolds, Yinan Lin

Masters Theses

"The main objective of this project was to evaluate the capacity of strong porous silicate (13-93) bioactive glass scaffolds prepared by a robocasting technique to regenerate bone and stimulate angiogenesis in a rat calvarial defect model. The scaffolds were created with the same grid-like microstructure but in a variety of formulations: (i) as-fabricated, (ii) pretreated in an aqueous phosphate solution to convert the glass surface to hydroxyapatite, (iii) loaded with bone morphogenetic protein-2 (BMP2) (1 µg per scaffold), and (iv) doped with copper (0.4-2.0 wt. % CuO). When compared to the as-fabricated scaffolds, the pretreated scaffolds enhanced bone regeneration at …

A New Generation Of Polymer/Ceramic Composite Biomaterials For Bone Regeneration, Mehrnaz Salarian

Electronic Thesis and Dissertation Repository

There is a substantial emerging interest for fundamental and applied research on the reinforcement of polymeric materials using nanotechnology. In the biomedical industry, development of novel bone cement composite materials with enhanced mechanical properties is of tremendous potential importance. The most universally used injectable bone cement is made of poly(methyl methacrylate) (PMMA); however, the major disadvantage of PMMA is its non-biodegradability. Polymers such as poly(propylene fumarate) (PPF) and polycaprolactone (PCL) are biodegradable, but suffer from a lack of mechanical properties. The aim of this research was to test the efficacy of these biodegradable polymers integrating nanotechnology for the development of …

Microvascular Topology And Intravascular Endothelial Cell Labeling In The Gracilis Anterior Muscle Of Balb/C Mice, Paul Heckler Ii

Biomedical Engineering

Peripheral arterial occlusive disease (PAOD) affects approximately 200 million individuals globally. The major underlying cause of PAOD is an inflammatory disease known as atherosclerosis, which results from the build-up of low-density lipoproteins (LDL) in the sub-intimal space. This initiates a complex cascade of events that lead to plaque growth. Plaque growth can then expand into the lumen of the vessel and result in occlusion and/or thrombosis. Symptoms of the disease can include claudication, ulcers, and/or gangrene, although many patients are asymptomatic. Similar to other forms of ischemic disease, risk factors for PAOD include hypertension, diabetes, and smoking. Common treatments include …

The Effects Of Hydrostatic Pressure On Early Endothelial Tubulogenic Processes, Ryan M. Underwood

Theses and Dissertations--Biomedical Engineering

The effects of mechanical forces on endothelial cell function and behavior are well documented, but have not been fully characterized. Specifically, fluid pressure has been shown to elicit physical and chemical responses known to be involved in the initiation and progression of endothelial cell-mediated vascularization. Central to the process of vascularization is the formation of tube-like structures. This process—tubulogenesis—is essential to both the physiological and pathological growth of tissues. Given the known effects of pressure on endothelial cells and its ubiquitous presence in the vasculature, we investigated pressure as a magnitude-dependent parameter for the regulation of endothelial tubulogenic activity. To …

Development Of A Protocol To Measure Gene Expression In The Mouse Tibia, Daniel Hoover

Biomedical Engineering

Numerous molecular factors active in bone tissue direct fracture repair and remodeling which can be altered by disease conditions such as Peripheral Arterial Disease (PAD) and Osteoporosis. Methods of molecular biology are commonly applied to investigate the expression and role of these molecular factors. This project presents a robust three-step protocol for examining gene expression in the mouse tibia. The protocol begins with isolating RNA from a flash frozen tibia sample. The isolated RNA is reverse transcribed into cDNA. Finally, PCR is performed to indentify expressed genes. Establishing this protocol will allow further research into the mechanisms of bone remodeling …

Three Dimensional In Vitro Model Of Head And Neck Squamous Cell Carcinoma, Anna Bulysheva

Theses and Dissertations

Head and neck squamous cell carcinomas (HNSCC) are among the leading causes of cancer related deaths throughout the world. The survival rate for this type of cancer is extremely low and has not changed significantly in recent decades. There is an imperative need to study tumor progression in a representative model in order to generate more knowledge about this disease as well as develop more effective treatment options. Multiple methods already exist for studying HNSCC and other types of cancers, including in vitro and in vivo models. Although in vivo models are more representative of the human carcinomas in terms …

Regeneration Of Electrospun Bioresorbable Vascular Grafts: A Phenomenon Associated With Vascular Graft Properties And Macrophage Phenotypes (M1/M2), Koyal Garg

Theses and Dissertations

Macrophages (MФ) and mast cells are important cell types in the context of tissue remodeling and regeneration. Mast cells participate in the early stages of wound healing and modulate the acute inflammatory responses to biomaterials. Mast cells can secrete a myriad of different cytokines by the process of degranulation; the process of regulated secretion in which preformed contents stored in their granules are rapidly released by exocytosis. Some of these cytokines such as IL-4, IL-13 and TNF-α can modulate the MФ phenotype. Macrophages (MΦ) are innate immune cells, crucial for tissue homeostasis, presentation of foreign and self-antigens following infection/injury, pathogen …

Immunohistochemical Mapping Of Hypoxia In Ischemic Mouse Hindlimb Skeletal Muscle, Emily Deckert

Biomedical Engineering

The study of blood vessel growth and remodeling is a complex endeavor. Hypoxia, the lack of oxygen in a tissue, is known to stimulate angiogenesis (the growth of new blood vessels), and have little effect on arteriogenesis (the enlargement of existing blood vessels). However, the role of hypoxia in vessel function is unknown, but may be determined using the results and methods developed in this experiment.

Supplied by the bloodstream, oxygen is required by all cells and tissues to remain healthy. If the bloodstream supplying a certain tissue with blood is disrupted, the tissue becomes ischemic, often leading to hypoxia. …

A New Pulsed Electric Field Therapy For Melanoma Disrupts The Tumor's Blood Supply And Causes Complete Remission Without Recurrence, Richard Nuccitelli, Xinhua Chen, Andrei G. Pakhomov, Wallace H. Baldwin, Saleh Sheikh, Jennifer L. Pomicter, Wei Ren, Chris Osgood, R. James Swanson, Juergen F. Kolb, Stephen J. Beebe, Karl H. Schoenbach

Bioelectrics Publications

We have discovered a new, ultrafast therapy for treating skin cancer that is extremely effective with a total electric field exposure time of only 180 mu sec. The application of 300 high-voltage (40 kV/cm), ultrashort (300 nsec) electrical pulses to murine melanomas in vivo triggers both necrosis and apoptosis, resulting in complete tumor remission within an average of 47 days in the 17 animals treated. None of these melanomas recurred during a 4-month period after the initial melanoma had disappeared. These pulses generate small, long-lasting, rectifying nanopores in the plasma membrane of exposed cells, resulting in increased membrane permeability to …