Pharmacology 4T12: 32nd Annual Undergraduate Thesis Defense Day March 19, 2022 by Stephanie C.

HONOURS BIOLOGY & PHARMACOLOGY Co-op Program Presents:

Pharmacology 4T12: 32nd Annual Undergraduate Thesis Defense Day

March 19, 2022

Welcome to the 2022 4T12 defence day! Message from Prof. Jan Huizinga, Director of our BioPharm program. Today is a highlight of our program where everyone can see how far you have come in acquiring research skills, progression in your critical thinking and your actual contributions to the ever-expanding knowledge of physiology and pharmacology and the ever-expanding solutions to the many clinical problems that still face us. One of the highlights for me is reading the evaluation of your thesis work by your supervisors, related to your determination, your desire to acquire skills and your motivation to be excellent. It is why our program should live forever. I am most grateful to all the faculty members mentioned in this booklet who supervised our students in their laboratories! This supervision is an essential and very exciting part of our BioPharm program for our students! My most profound appreciation goes to the Faculty and Students who have contributed and will be contributing to the 2022 examination process of our 4T12 course!

Jan Huizinga Jan D. Huizinga Program Director

Jan D. Huizinga PhD Professor, Faculty of Health Sciences. Examiner for today’s defence presentations, Director of the BioPharm program. Dr. Huizinga is an expert on the basic science of Gastrointestinal Motility and has a CIHR and NSERCfunded laboratory within the Farncombe Institute for basic and clinical studies to understand the pathophysiology of motility disorders. Jihong Chen MD PhD Associate Professor, Faculty of Health Sciences. Gastroenterologist and Researcher at the Faculty of Health Sciences. Examiner for today’s defence presentations Dr. Chen has a clinic focussing on severe gastrointestinal motility disorders and is developing new techniques for diagnosis and treatment related to patients having dysfunction of the autonomic nervous system. She is a principal investigator in CIHR funded research and clinical trials. Luke Janssen PhD Professor Emeritus, McMaster University. Department of Medicine, Respirology. Dr. Janssen directs the 3B06 course in our program and has evaluated written theses for the present 4T12 course. Dr. Janssen is an expert on airway smooth muscle and lung function. Derek Lobb PhD Professor, Faculty of Health Sciences. Department of obstetrics and gynecology. Dr. Lobb teaches 3A06 in our program and has evaluated written theses for the present 4T12 course. Dr. Lobb is an expert in reproductive endocrinology. Lijun Liu BSc Ph.D. student in the Medical Sciences and the Farncombe Institute. Lijun will be an examiner and organizer for the defences. Lijun did her undergraduate at the University of Toronto and is now working in the Chen-Huizinga laboratory to understand the pathophysiology of patients with severe refractory colon motor disorders, develop techniques to measure autonomic dysfunction, and evaluate treatment involving neuromodulation of the autonomic nervous system. Amer Hussain BSc MSc - Ph.D. student in the Medical Sciences and the Farncombe Institute. Amer will be an examiner and organizer for the defences. Amer did his undergraduate in our Honours Biology-Pharmacology Coop Program and remembers well his participation in 4T12, what was then called 4F09. He is now working in the Chen-Huizinga laboratory to evaluate colon motor functioning in healthy subjects and patients using ultrasound imaging. He combines this with the development of techniques to assess autonomic dysfunction.

Stephanie Cusick BA Administration, organization and booklet design

PHARMAC 4T12 THESIS PRESENTATIONS MARCH 19, 2022 TIME 8.30

8.55

STUDENT

TITLE

SUPERVISOR

Jeniszka Gill

Evaluating plectin-targeted phototheranostic agent forpancreatic ductal adenocarcinoma

Dr. Gang Zheng

Tenzin Gyaltsen

9.20

Kruti Bhakta

9.45

Sabrina Dam

10.10

Amro ElRafie

10.45

Roshan Ahmad

11.10

Carina Yan

11.35

Rachel Tu

12.00

Mackenzie Thrope

Confirming CD125 Expression on ILC2s: Effects of TSLP on CD125 Upregulation and Dexamethasone

Dr. Roma Sehmi

Exploring the role of the Health and Wellness Center professional health care team in the provision of mental health support to uniDr. Lisa Dolovich versity students to develop effective medication management practices in the new academic pharmacy SMAC Debio 1143 and avelumab combination immunotherapy elicDr. Jonathan Bramson its an increase in the proportion of activated T cells in solid tumour patients Induction of leukemic regenerDr. Allison Boyd & Dr. Mick ating cells in acute Bhatia myeloid leukemia relapse 10.30 -10.45. BREAK Mixed-Method Data Collection for Development of an Electronic Application to Promote Home-Based SelfCare in Older Heart Failure (HF) Patients

Generating rat islet cell reaggregates and characterizing their function when encapsulated in biocompatible hydrogel formulations Development of caSTAT5 and muIL-15 engineered CAR T cells to enhance adoptive cell therapy Investigating the efficacy of a novel vaccine delivery method targeting the oral mucosal membrane

Dr. Catherine Demers

Dr. Harald Stöver

Dr. Jonathan Bramson

Dr. Mark Larché

12.25

1250

14.10

14.35

15.00

15.25

15.50

Tasfiah Tasnim

Rachel Snelgrove

Identification of Rare Genetic Variants in a European Family with Co-Presentation of Postural Tachycardia Syndrome, Hypermobile Ehlers-Danlos Syndrome and Mast Cell Activation Syndrome Through Whole Exome Sequencing

Interactions of chronic metformin exposure with the gut microbiome in adult zebrafish Dr. Karen Kidd & Dr. Oana Birceanu

13.10 -14.10 BREAK. LUNCH WILL BE PROVIDED Effects of Upadacitinib for the Treatment of Moderate-to-Severe Active Rheumatoid Arthritis: A Systematic Nimah Siddiqui Review and Meta-analysis of Randomized Controlled Trials

Tyler Seto

Dr. Guillaume Pare and Dr. Matthew Lanktree

Biopanning a T7Select phage display α1-AT M358R library hypervariable at positions P7 -P3 of the reactive center loop with human kallikrein

Dr. Hammad Qazi

Dr. William P. Sheffield

Srustie Patel

The effects of high dietary salt intake and nifedipine treatment on renal function and pathology

Dr. Jeffrey Dickhout

Louise Limoges

Effects of cannabidiol on mitochondria and endoplasmic reticula of human trophoblasts

Dr. Sandeep Raha

Yejin Kang

The role of nonsense mediated decay in Caenorhabditis elegans models of neurodegeneration.

Dr. Lesley MacNeil

16.10- 16.25 BREAK

16.25

16.50

15.15

15.40

Papiha Joharapurikar

Data-Driven Predictive Analytics Framework for Analysis of Prognostic Factors and Health Outcomes in Pediatric Intestinal Failure Patients

Dr. Nikhil Pai

Zarin Hossain

Characterization of UVA biophoton emission and survival of α-irradiated HCT116+/+ cells via radium exposure

Dr. Carmel E. Mothersill

Heidi Ho

In vitro biofilm model to assess dosing regimens in phage therapy for inflammatory bowel disease

Dr. Zeinab Hosseinidoust

Effects of Plasma Rich in Growth Factor Versus Hyaluronic Acid for The Treatment of Knee Osteoarthritis: A Mackenzie Haggstrom Systematic Review and Metaanalysis of Randomized Controlled Trials

Dr. Hammad Qazi

Evaluating Plectin-Targeted Phototheranostic Agent for Pancreatic Ductal Adenocarcinoma Jeniszka Gill Supervisor: Dr. Gang Zheng Background: Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers with less than a 10% 5-year survival rate. Due to a lack of PDAC specific symptoms and diagnostic modalities, most patients are diagnosed at late stages of this cancer. The complexity of the organs surrounding the pancreas also make eligibility for surgery bleak. Thus, there is a dire clinical need for additional diagnostic methods, particularly non-invasive methods that can detect earlier PDAC stages, while being suitable for the vast majority of patients with unresectable tumours. Aim: This project investigated a novel phototheranostic agent (Pyro-PTP), comprised of a pyropheophorbide photosensitizer and a PDAC specific plectin-targeting peptide. The targeting peptide allows for drug accumulation at the tumour site, and when paired with the conjugated photosensitizer, this site can be located via fluorescence imaging and treated with photodynamic therapy (PDT). Methods: Pyro-PTP toxicity and photodynamic activity were evaluated in two PDAC cell lines. Pyro-PTP’s behaviour was compared with its long-circulating form Pyro-pep-PTP, which contains a pharmacokinetic-modulating peptide linker. Cellular uptake of these agents was compared between PDAC cell lines and a healthy pancreatic cell line using confocal microscopy. Biodistribution of Pyro-PTP was evaluated in a subcutaneous PDAC mouse model with in-vivo and ex-vivo fluorescence imaging and with homogenized tissue fluorescence. Furthermore, the behaviour of Pyro-PTP and Pyro-pep-PTP biodistribution was compared in an orthotopic mouse model. Results: PDT on a subcutaneous model was shown to be effective. Within the subcutaneous model a 6h and 24h drug accumulation period did not show specific accumulation within the tumour. Through fluorescence imaging it was observed that Pyropep-PTP accumulates in the orthotopic pancreatic tumour to a higher extent than surrounding healthy pancreatic tissue and other benign tissue. Conclusion: With further modifications these principles can be utilized for fluorescence imaging and PDT treatment in PDAC patients. Overall, Pyro-PTP requires further validation in vitro and in vivo to evaluate its targeting capabilities but it shows promise as a PDAC-targeting photodynamic therapy agent.

Confirming CD125 Expression on ILC2s: Effects of TSLP on CD125 Upregulation and Dexamethasone Extension Study Tenzin Gyaltsen Supervisor: Dr. Roma Sehmi

Abstract: ILC2s belong to a family of lineage-negative lymphocytes that are primarily tissue-resident, found at mucosal interfaces in various body areas. Basal ILC2 levels maintain integrity of tissue, but once increased in number and overactivated, ILC2s contribute to the recruitment and activation of inflammatory cells that lead to tissue damage; this is achieved primarily through the production of type 2 cytokines following activation. The IL-5 and IL-5R axis plays a central role in eosinophilic asthma, but its presence on ILC2s has been contested. To confirm the expression of CD125 on ILC2s, as well as identify possible up and down-regulatory factors for this expression, overnight coculture and flow cytometry experiments were conducted on ILC2s isolated from peripheral blood. CD125 was observed on ILC2s, with a dose-dependent trend of TSLP on CD125 upregulation seen, although not reaching significance. A trend in dexamethasone attenuation of CD125 upregulation was also seen, though again not reaching significance. Power calculations for the aforementioned trends were conducted to identify ideal sample sizes for future analysis. Identification of CD125-high patient characteristics may provide clues into patient phenotypes most responsive to anti-IL-5 benralizumab in the treatment of eosinophilic asthma.

Exploring the role of the Health and Wellness Center professional health care team in the provision of mental health support to university students to develop effective medication management practices in the new academic pharmacy

Kruti Bhakta Supervisor: Dr. Lisa Dolovich Background: The global age-of-onset distributions for mental health illnesses shows that 70% of cases are reported in patients that are 24 years of age or younger. In Ontario, the transitional youth group experience many challenges in accessing mental health medication support. Local pharmacies are well positioned to fill this gap and meet current health care trends. The Discovery Pharmacy at the University of Toronto aims to provide medication management services for students with mental health needs, in partnership with the Health and Wellness Center. Objectives: The objective of this study was to understand the role of the Health and Wellness Center health care professionals, explore how the Discovery Pharmacy operations can be integrated with the Health and Wellness Center to support mental health services, and identify strategies that can be implemented in the pharmacy to approach psychotropic medication management for students. Methods: This study took an exploratory approach based in the philosophies of interpretivism. Qualitative data was collected by conducting one-on-one interviews with members of the Health and Wellness Center – 3 participants were recruited. The data was analyzed using NVivo 12 qualitative content analysis software and organized into categories and themes. Results: Descriptive content analysis of the interviews produced many themes for further investigation. The participants provided an in-depth review of their role, offered suggestions for how to integrate the academic pharmacy into the circle of primary care, outlined strategies to target mental health medication management for students, and noted potential barriers that need to be overcome. Discussion: This thesis provided an outlook for how the pharmacy team can implement mental health medication management services in the Discovery Pharmacy. Several models of primary care are available to provide a framework of interdisciplinary health care. The main findings from this study can help develop an investigation that targets students to understand their perspective as a patient that faces the challenges of mental health illness.

SMAC Debio 1143 and avelumab combination immunotherapy elicits an increase in the proportion of activated T cells in solid tumour patients Sabrina Dam Supervisors: Ying Wu, Jamie McNicol, and Jonathan Bramson ABSTRACT Despite the development of the healthcare system in the past several decades, 50% of Canadians will be diagnosed with cancer in their lifetime and this number is projected to rise in the next 20 years with the growing and aging population. A relatively new concept in cancer therapy is to use the immune system to combat tumour cells by enhancing the innate anti-tumour immune response, specifically the T cell response. Activation of T cells is modulated by co-stimulatory and co-inhibitory immune checkpoints. Unfortunately, these checkpoints are also targeted by tumour cells, such as the programmed death-1 ligand (PD-L1) and its receptor. The immune checkpoint inhibitor (ICI) avelumab binds to PD-L1 on tumour cells, preventing tumour-induced T cell exhaustion. Inhibitor of apoptosis proteins (IAPs) are also exploited by tumour cells as they promote cell survival. Debio 1143 is a second mitochondrial-derived activator of caspase (SMAC) mimetic compound that inhibits specific IAPs to make tumour cells more susceptible to T cell killing. Thus, the NCT03270176 study aims to investigate the combination of avelumab and SMAC Debio 1143 in their ability to engage T cells and sensitize tumour cells to T cell killing. Polychromatic flow cytometry was employed for immunophenotyping T cells in the peripheral blood and dimension reduction algorithms were utilized to cluster the data. The treatment showed an increase in peripheral activated T cells, however further research is required to explore the potential of this drug combination in cancer immunotherapy.

Induction Of Leukemic Regenerating Cells in Acute Myeloid Leukemia Relapse Amro ElRafie Supervisors: Dr. Allison Boyd & Dr. Mick Bhatia Background: Acute myeloid leukemia (AML) is a debilitating disease that results from the proliferation and clonal expansion of aberrant hematopoietic cells, commonly referred to as leukemic stem cells (LSCs). Among acute leukemias that affect adults, AML is the most common with a five-year overall survival rate of 28.7%. To date, AML therapy that leads to disease-free survival mainly consists of chemotherapy regimens, sometimes followed by an allogeneic hematopoietic cell transplant. Unfortunately, around 60% of high-risk AML pa-

tients will relapse despite progress in targeted therapy accompanying chemotherapy. Previous work has demonstrated a novel transient leukemic cell population, leukemicregenerating cells (LRCs), that arise post-chemotherapy (Ara-C) and can only be detected in vivo. These LRCs are thought to be a specific progenitor population, harboring an aberrant self-renewal and hematopoietic differentiation capacity. LRCs were shown to be molecularly distinct from therapy naïve LSC progenitors and possibly susceptible to therapeutic targeting. While Ara-C chemotherapy could only stimulate LRC emergence in vivo, serum collected from mice treated with Ara-C was shown to increase progenitor activity and enrich for LRC markers in primary AML cells via in vitro liquid culture. Progenitor activity was quantitatively measured by colony-forming unit (CFU) assays, in which cells are grown in specialized media that allows for the formation of distinct colonies arising from individual progenitors. Hypothesis: Given that progenitors are thought to drive regrowth of AML to produce relapse, signals in the serum must be involved in stimulating the outgrowth of leukemic progenitors. If injury signals in the microenvironment trigger the aggressive outgrowth of rare residual leukemic cells, drugs could interrupt the interaction and prevent the sequence of events that lead to AML relapse when used in conjunction with chemotherapy. Methods: Here, serum-borne elements arising post-chemotherapy in AML patients were identified and analyzed to determine candidates for screening (via the CFU assay) and investigation of biological relevance to LRC induction and AML relapse. Additionally, progress towards optimizing and validating a high-throughput workflow for protein screening laid the groundwork for investigating potential LRC factors.

Results: AML progenitors display heterogeneity in their response to serum-borne elements spiked in CFU culture. Optimization of a high-throughput workflow is stagnated by the capacity of AML progenitors to form CFUs in a new format. IL-6 (0.80nM-4.00nM) and TNF-α (7.72nM-77.2nM) growth factors were established as positive and negative controls respectively for the high throughput workflow, given their consistent effects across multiple AML patient samples. Conclusion: We have partially developed a high-throughput CFU assay workflow that can be used to screen biologics. Additionally, we have demonstrated that the CFU stimulating potential of serum-borne elements can be multifactorial.

Mixed-Method Data Collection for Development of an Electronic Application to Promote Home-Based Self-Care in Older Heart Failure (HF) Patients

Roshan Ahmad Supervisor: Catherine Demers (MD)

ABSTRACT Background: Heart failure (HF) is a chronic illness that greatly impacts older patients and is one of the leading conditions which results in rehospitalizations. HF poses as a huge financial burden on the health-care system due to an increasingly aging population. Certain key home-based self-care strategies can help manage heart failure symptoms and severity. These strategies include daily weight monitoring, diuretic adjustments and treatment adherence in addition to managing any comorbidities. Furthermore, they can be challenging for older patients with HF due to the presence of mild cognitive impairments, varying economic status, limited health knowledge about their disease and poor treatment adherence. Therefore, there is an incredible need for the introduction of mobile health (mHealth) applications designed specifically for HF. The Health Smart Scale (HHS) is an electronic tool that promotes home-based self-care in older patients with HF. The app includes audio reminders for daily weight monitoring, graphical weight trend data, a Standardized Diuretic Adjustment Tool (SDDST) and informs patients when they should contact their physician. Many mHealth applications exist, however there is a gap in evidence supporting their usability and acceptability in the end-user patient population. The purpose of this project is to assess the usability of the HSS, through mixed-method data collection. Objectives and Methods: The objective of this study was to conduct usability testing to identify any usability or functionality issues, and determine if the participants are satisfied with the design features of HSS. Feedback received from participants will aid in improving features in future iterations and catering the HSS better to the end-users. Mixed-method data collection was used including questionnaires using Likert-like items and open-ended feedback questions. The tool was delivered to the participant (N=1) at their home, and the participant completed ten questionnaires at baseline and up to 30 days after using the HSS.

Results: At baseline the participant had marginal health literacy (13), higher subjective numeracy (18), inadequate eHealth literacy (16) and scored in the highest level of the patient activation measure (90.70). The participant’s baseline to 30-day SelfCare of Heart Failure Index scores increased for self-management and monitoring, decreased for self-maintenance and remained the same for self-confidence, however statistical significance was not assessed. The HSS received a good system usability score (SUS=77.5). Three specific themes were identified regarding the usability of the HSS which include 1) Visual/Display design, 2) Functional design, and 3) End-user considerations. They all highlighted specific issues and areas of improvement for the HSS.

Conclusions: Due to a lack of power and statistical analyses, there is a lack of significance and generalizability in whether the HSS can actually help improve self-management outcomes. However, the HSS obtained a good SUS score which suggests that the tool has the potential to be a promising intervention in helping older patients self-manage their HF. It also received positive feedback on most features, but further iterations are needed to address the uncovered issues.

Generating rat islet cell re-aggregates and characterizing their function when encapsulated in biocompatible hydrogel formulations

Carina Yan Supervisor: Dr. Harald Stöver Abstract

Type 1 diabetes mellitus (T1DM) is a chronic autoimmune disorder characterized by the destruction of pancreatic β-cells. With no cure, T1DM treatment is associated with daily challenges and compliance issues. Transplantation of insulin-producing cells into T1DM patients may allow for long-term glycemic correction. Microencapsulation of these therapeutic cells is a method of delivery that can overcome the need for immunosuppression by protecting the cells from the host immune system, as well as chemical and mechanical stresses. Isolating viable and high functioning islets from donors is a challenge as the vasculature necessary for nutrient transport is destroyed during isolation. Since the transport of nutrients is consequently dependent on passive diffusion, the generation of smaller islet cell re-aggregates, less than 150 µm in size, have been found in literature to perform better than native islets in terms of survival and insulin secretion. The following research aims to investigate the function of encapsulated and non-encapsulated islet cell re-aggregates generated from rat pancreatic islets. Islet cell re-aggregates of approximately 90 µm in size, containing 250 cells each, were encapsulated in two proprietary polymer formulations: Polymers A and B. Research includes the development and optimization of a protocol for generating islet cell re-aggregates and characterizing their function through glucose-stimulated insulin secretion tests and immunofluorescence and autofluorescence imaging. Preliminary results suggest that islet cell re-aggregates may exhibit better insulin secretion than native islets. Both native islets and islet cell re-aggregates were functional when encapsulated in Polymer A but non-functional when encapsulated in Polymer B. However, more experimental replications are required to statistically determine whether the function and survival of encapsulated and non-encapsulated islet cell re-aggregates are significantly improved from that of native islets.

Abbreviations: DM, diabetes mellitus; GSIS, glucose-stimulated insulin secretion; ID, islet dissociation; IEQ, islet equivalent; IR, islet cell re-aggregation; MFI, mean fluorescent intensity; SI, stimulation index; T1DM, type 1 diabetes mellitus.

Development of caSTAT5 and muIL-15 engineered CAR T cells to enhance adoptive cell therapy Rachel Tu Supervisor: Jonathan Bramson Background: Cancer immunotherapy has progressed massively over the past few decades with the development of ACTs, involving the use of transplanted tumor infiltrating lymphocytes or genetically modified T cells to treat patients. However, ACTs come with many limitations such as low persistence in vivo, T cell exhaustion, and the requirement for a lymphodepletion regimen prior to T cell reinfusion. IL-15 and STAT5 have both been implicated in cell growth and survival pathways. These prosurvival signals can be directly provided to adoptively transferred cells through genetic engineering and have the potential to enhance the proliferative function, persistence, and antitumor efficacy of T cells. Aim and hypothesis: The aim of this project is to investigate the effects of caSTAT5 and murine IL-15 on engineered T cells. We hypothesize that the addition of a caSTAT5 or IL-15 transgene to CAR engineered murine T cells will lead to enhanced proliferative function and survival of the T cells in in vitro tumor models. Methods: Dual expression γ-retrovirus vectors were generated through molecular cloning and used to transduce murine T cells from bulk splenocytes. T cells were monitored in culture over 7 days and supplemented with IL-7 and IL-15 cytokines. Phenotype and proliferation assays were performed to evaluate expression and functional outcomes. Western blot analysis and an ELISA were performed to detect phospho-STAT5 and soluble murine IL-15 expression from engineered T cells. Results: T cells engineered with caSTAT5 or muIL-15 transgenes showed high CAR expression and proliferated upon co-culture with tumor cells. Phenotypic analysis of caSTAT5 and muIL-15 CAR T cells showed a shift in T cell population towards an effector memory phenotype. caSTAT5 CAR T cells showed the largest increase in phospho-STAT5. Soluble IL-15 expression from muIL-15 CAR T cells showed no increase compared to controls. Conclusion: The addition of a caSTAT5 or IL-15 transgene to CAR engineered murine T cells improves CAR functionality and proliferative potential of T cells. Future research should be done to investigate proliferation and survival of these engineered T cells in in vivo tumor models.

Investigating the efficacy of a novel vaccine delivery method targeting the oral mucosal membrane Mackenzie Thorpe Supervisor: Dr. Mark Larché Introduction The COVID-19 pandemic is caused by SARS-CoV-2, a positive stranded RNA corona virus. Currently, many of the candidate vaccines for COVID-19 aim to induce neutralizing antibodies against the viral spike protein, these antibodies inhibit uptake through the human ACE2 receptor and block infection. Respiratory viruses such as SARS-CoV-2 primarily invade host defenses through the mucosal surfaces within the upper respiratory tract, meaning that an ideal vaccine should induce protective immunity at mucosal sites, where the immune system first encounters the pathogen. Mucosal immunization makes vaccine delivery more accessible than traditional administration routes, improves vaccine availability, and is very suitable for mass immunization during pandemic situations. This project aims to optimize an oral vaccine delivery system using buccally administered oral thin films (OTFs) made from food-grade biodegradable polymers. Methods To determine if buccally-administered oral thin films loaded with the model antigen ovalbumin (OVA) and mucosal adjuvants are tolerated and will produce an antibody response in a murine model, OTFs loaded with OVA with and without poly-I:C or CpG were administered buccally on days 1, 14 and 29 before euthanization on day 30. Serum was collected and an ELISA to measure ovalbumin-specific serum IgG titers was completed. To determine if buccally-administered oral thin films loaded with SARS-CoV-2 spike protein antigen and mucosal adjuvants will produce an antibody response in a murine model, OTFs loaded with whole or the receptor binding domain (RBD) of SARS-CoV-2 spike protein were administered buccally on days 0-4, 22-26 and 42-46 before euthanization on day 60. Serum was collected on days 0, 20, 40 and 60 and ELISAs to measure SARS-CoV-2 Spike RBD-Specific Serum IgG titers were completed. Results OTFs containing OVA alone produced higher IgG titers than blank vaccines when applied buccally. On Day 60, OTFs containing a high dose of ML spike produced a higher IgG titer compared to the blank vaccines when applied buccally, however, all treatment groups produced lower IgG titers compared to the positive control. Conclusion This novel vaccine delivery method targeting the oral mucosal membrane displays some promise for future development of a needle free SARS-CoV-2 vaccine with the buccally administered high dose of RBD of SARS-CoV-2 spike protein presented a significantly higher IgG titer compared to the blank vaccine group (Figure 9D). However, repetitive exposure to the same antigen at the oral mucosa may have led to oral immunotolerance in some treatment groups in both experiments. A larger study investigating IgA titers as well as IgG titers to explore the site-specific immunity is required to get a larger picture of the immunity induced by the OTFs. Additionally, improving the delivery method to include nanoparticles will assist in ensure full doses of antigen and adjuvants are delivered across the buccal membrane inducing the desired immune responses.

Background: Whole exome sequencing (WES) involves the identification of genetic variants in the coding regions of an individual’s DNA. In the context of medical diagnosis and clinical research, WES has proven to be a viable method of identifying the underlying genetic basis of a patient’s disease. This investigation was focused on a proband with co-presentation of postural tachycardia syndrome (POTS), hypermobile Ehlers-Danlos syndrome (hEDS), and mast cell activation syndrome (MCAS) and her family members who are exhibiting a similar clinical presentation.

Hypothesis: It was hypothesized that there may be a genetic component to this family’s presentation, and therefore this investigation involved conducting WES to determine if a rare genetic mutation was associated with their disease.

Methods: The methodology behind this investigation involved: 1) clinical assessment and recruitment of patients to the study; 2) collection of saliva samples to collect from patients in a remote manner; 3) conduction of WES on the patients DNA samples; 4) variant filtration, prioritization, and analysis of WES results.

Results: After conducting WES in the proband and 4 other family members, no variants were found in candidate genes for POTS or MCAS. Two variants in hEDS candidate genes were observed in two patients, COL1A1 and COL5A3, however, their relevance was determined to be uncertain. Three variants in incidental genes were found in two individuals in PTEN, RYR1 and APC, all of which were deemed variants of uncertain significance through use of the ACMG criteria for rare variant interpretation. Conclusion: This study served to demonstrate the utility of genetic sequencing in a clinical context to aid in the investigation of the mechanism behind patients disease and allow for the better understanding of patients disease etiology

Interactions of chronic metformin exposure with the gut microbiome in adult zebrafish

Rachel Snelgrove Supervisors: Karen Kidd, & Oana Birceanu

Abstract Metformin (Met) is a common medication used to treat type-2 diabetes (T2D) and is one of the most used drugs worldwide by mass. Although its effects are well-studied in humans, effects on fish and other wildlife remain elusive. After human excretion, Met enters the environment through municipal wastewater relatively unchanged. Given the extent to which fish are exposed to Met through this wastewater, it is important to improve our understanding of how Met exposure may affect fish, particularly through sublethal effects such as body condition or microbiome. In this study, adult Tupfel Longfin zebrafish were exposed to 0 (control) and two environmentally relevant Met concentrations; 4 and 40 µg/L of Met for 30 days, in controlled laboratory conditions, with an equal distribution of males and females within each concentration. Gut content was collected at day 0 (just prior to the addition of Met to the exposure tanks) and at day 14 and 30 of exposure. Microbial composition, abundance, Shannon (alpha) diversity, and beta diversity were analyzed and expressed relative to length, weight, and condition factor. Sex differences were observed for virtually all measurements taken, with females being generally larger and exhibiting lower gut microbiome diversity than males. Males exposed to 40 µg/L of Met were heavier than those exposed to 4 µg/L of Met, females exposed to 4 µg/L Met had higher condition factor than controls on day 14, and Shannon diversity increased within control fish between days 14-30. Beyond this, no treatment effects were observed for length, weight, condition factor, or Shannon diversity within either sex. Treatment groups showed differing beta diversity between treatment groups in males on days 0 and 14 and in females on day 30. Since treatment effects on beta diversity were observed before the addition of Met, these findings should be further observed in future studies to more definitively parse out any effects Met may have on zebrafish gut microbiome beta diversity. In terms of gut microbiome composition, Fusobacteriota was the most abundant phylum in all groups (~60-80%), with Firmicutes being the second most abundant (~10-30%) and Proteobacteria being present as well in most groups (5-25%). Actinobacteriota was also present in a few groups, namely the 4 µg/L Met and 40 µg/L Met groups of males on day 0 (~25%). Results from this work do not show clear trends of Met affecting zebrafish health or the gut microbiome of adult zebrafish. However, sex differences were observed in virtually all measures, which does contribute to our understanding of zebrafish physiology, particularly in the measures of length, weight, condition factor, Shannon diversity, beta diversity, and gut microbiome composition at the phylum and family levels. As a follow-up to this study, zebrafish muscle and liver will be analyzed for Met and Met metabolite concentrations. Future directions after these studies could include further research on pathways involved in Met absorption, distribution, metabolism, and excretion in fish, as well as performing similar studies in juvenile and/or wild zebrafish to see if effects are different based on these demographics.

Effects of Upadacitinib for treatment of Moderate-to-Severe Active Rheumatoid Arthritis: A Systematic Review and Meta-analysis of Randomized Controlled Trials Nimah Siddiqui Supervisor: Dr. Hammad Qazi

ABSTRACT Introduction: Rheumatoid arthritis (RA) is a chronic autoimmune disorder of the diarthrodial joints. Despite standard treatment, many patients progress to disability and suffer significant morbidity over time. Therefore, there is a need to explore newer and more targeted treatment options such as Janus Kinase Inhibitors (Jakinibs). As a result, Upadicitinib (UA) developed by AbbVie is the third Jakinib for moderate-severe RA. UA has a more specific selectivity of JAK inhibition for the JAK1 pathway. The assumption that JAK2, JAK3 and Tyk2 inhibition might be more frequently associated with unwanted actions made JAK1 an attractive target in terms of selective inhibition.

Aim: The main objective of this study is to assess the efficacy of 15 mg and 30 mg Upadacitinib in the treatment of pain and severity symptoms in moderate-to-severe active rheumatoid arthritis patients. Safety was also assessed in this review as a secondary objective.

Methods: A systematic review and meta-analysis was conducted in accordance with Cochrane guidelines. All articles were evaluated at the title, abstract and full text-phase using the selection criteria; Randomized controlled trials were selected where one daily administration of 15 and 30 mg of Upadicitnib was compared to standard therapy or placebo, in individuals over the age of 18 fulfilling the ACR/EULAR diagnosis criteria. Statistical analysis was performed using RevMan 5.3.

Results: It was demonstrated that the UA 15 mg and UA 30 mg significantly improved ACR20, ACR50 and ACR70 responses from baseline to week 12 than the comparator (p < 0.00001). Additionally, the 15 mg and 30 mg UA groups were significantly more effective than the comparator in achieving clinical remission (DAS28-CRP score ≤ 2.6), low disease activity (DAS28-CRP score ≤ 2.6) and change in baseline DAS28-CRP response at Week 12 (p < 0.00001). A dose-specific comparison was conducted for the primary outcomes between 15 mg and 30 mg of UA but the results were not significant. All secondary outcomes (HAQ-DI, SF-36 PCS, Duration of Morning Stiffness FACIT-F) displayed significant improvement than the comparator (p < 0.00001), except for the UA 30 mg for FACIT-F (p = 0.13)

Conclusion: The results of this study indicate that UA 15 mg and 30 mg are more efficacious compared to placebo or standard treatment for reducing pain and severity in RA with no significant risk for adverse events. Clinicians should consider the results and take into account the cost-effectiveness of this drug for efficient evidence-based decision-making. Future directions should include conducting head-to-head trials comparing UA 15 mg and UA 30 mg, UA monotherapy trials and trials with larger sample sizes and longer follow-up periods to assess any long-term adverse effects.

Biopanning a T7Select phage display α1-AT M358R library hypervariable at positions P7-P3 of the reactive center loop with human kallikrein

Tyler Seto Supervisor: Dr. William P. Sheffield

Background: Serine protease inhibitors (serpins) are a superfamily of protease inhibitors that regulate key cellular processes such as coagulation, fibrinolysis, inflammation, and immunity (Law et al., 2006). Alpha-1 antitrypsin (α1-AT) is the most abundant serpin found in human plasma, where its primary function is to inhibit neutrophil elastase (Schmaier, 2019). In 1978, researchers identified a naturally occurring α1-AT mutant, α1-AT M358R, altered at the P1 position of the crucial reactive center loop (RCL) (Lewis et al., 1978). This novel mutation changed α1-AT from inhibiting neutrophil elastase to inhibiting multiple coagulation proteases, such as fXIa and thrombin, as well as kallikrein-kinin system proteins, such as kallikrein (Scott and Sheffield, 2020). Kallikrein has recently emerged as a target for drug development because of its role in the production of bradykinin (Kearney et al., 2021). The loss of control of kallikrein and bradykinin production has been linked to life-threatening hereditary angioedema (Zeerleder and Levi, 2016).

Hypothesis: We hypothesize that variants of α1-AT M358R exist in the sequence space defined by all residues in the reactive centre loop (RCL) that are specific for kallikrein.

Methods: Previously, the Sheffield laboratory has employed phage display and bacterial lysate screening to identify potential α1-AT variants hypervariable at different residues of the RCL (P13-P5’) that could inhibit thrombin and fXIa (Bhakta et al., 2021) (Scott et al., 2014). Here, we attempted to identify variants of α1-AT M358R hypervariable at positions P7-P3 and P2-P1 of the RCL that are specific for kallikrein using phage display and bacterial lysate screening.

Results: For the P7-P3 variants, four rounds of selection with kallikrein led to substantial enrichment of ten candidates, with QLIPS being the most enriched and most abundant, by both methods. For the P2-P1 candidates, we identified three candidates, RR, GR, and FR, from the bacterial lysate screen. These candidates had a stronger affinity for kallikrein than fXIa in the screen. We acknowledged that both the selected P7-P3 and P2-P1 candidates showed a decrease in proximity between the catalytic triad (His57, Asp102, Ser195) and the P1 Arg358 residue in comparison to α1-AT M358R. Additionally, we recognized that the QLIPS candidate was able to form a stable hydrogen bond complex with kallikrein’s Glu217 residue.

Conclusion: The decrease in proximity between the nucleophilic serine and the P1 Arg358 residue may explain why these candidates were selected for in screening. Additionally, the fact that a hydrogen bond was formed between the P3 serine of QLIPS and kallikrein’s Glu217 residue may provide evidence as to why QLIPS was the most enriched candidate. The next steps will be to purify and express the most abundant candidates from the screens in E. coli and assess their kinetics of inhibition to fXIa and kallikrein and compare them to α1-AT M358R.

The effects of high dietary salt intake and nifedipine treatment on renal function and pathology Srustie Patel Supervisor: Dr. Jeffrey Dickhout; McMaster University Background: Chronic kidney disease (CKD) is a global health concern and is characterized by the gradual loss of kidney function. CKD is strongly linked with hypertension and both conditions are known to exacerbate one another. In humans, a high dietary salt intake has been associated with an increase in blood pressure (BP). Nifedipine is an L-type calcium channel blocker and is a drug of interest that has been shown to reduce BP. Uromodulin (UMOD) has been recognized as a protein of interest; as increases in urinary UMOD levels have been associated with increased risk of CKD. Aim and Hypothesis: This thesis aims to assess how high dietary salt intake and nifedipine treatment given to SHR and WKY rats correlate with BP, proteinuria, 24-hr UMOD excretion, percent protein cast area in both cortex and medulla, and serum creatinine levels; and how this, in turn, correlates with the progression of CKD. It was hypothesized that consuming a high salt diet would lead to an increase in BP, proteinuria, 24-hr UMOD excretion, percent protein cast area in both cortex and medulla, and serum creatinine levels. Additionally, it is hypothesized that the consumption of a high salt diet combined with nifedipine treatment would improve renal function and pathology in both WKY and SHR rats. Methods: Older (49-55 weeks of age) male WKY and SHR rats were given one of three treatments: (1) normal salt diet (NS; 0.4% NaCl), (2) high salt diet (HS; 8% NaCl) or (3) high salt plus nifedipine diet (HSN; 8% NaCl plus 25mg/kg/day nifedipine) for 8 weeks. The BP was measured at baseline and after each week of treatment using radiotelemetry. The rat’s 24-hr urine sample and blood sample were collected at endpoint and the samples were used for western blotting & Bradford protein assay and creatinine assay respectively. After 8 weeks the rats were sacrificed, and their kidneys were harvested for periodic acidSchiff (PAS) staining and analysis of protein casts. Results: It was found that 8 weeks of HS feeding did not result in significant changes in BP in both WKY and SHR rats when compared to the respective NS fed groups. However, 8 weeks of HS feeding did significantly increase proteinuria, 24-hr UMOD excretion, percent protein cast area in cortex and medulla, and serum creatinine levels in SHR-HS rats when compared to SHRNS rats. Additionally, it was found that an HSN diet fed to SHR rats significantly decreased proteinuria, 24-hr UMOD excretion, percent protein cast area in cortex and medulla, and serum creatinine levels when compared to SHR-HS rats. Conclusion: In conclusion, 8 weeks of HS feeding in older SHR rats led to a decline in renal function and pathology, however, a HS diet combined with nifedipine lowered the SHR rat’s blood pressure and improved renal function and pathology.

Effects of cannabidiol on mitochondria and endoplasmic reticula of human trophoblasts

Louise Limoges Supervisor: Dr. Sandeep Raha

Background: Considering our limited knowledge of its pharmacodynamic properties, the rise in popularity of cannabidiol (CBD) products is alarming, especially for vulnerable populations such as pregnant people. CBD is the major non-psychoactive cannabinoid found in the cannabis plant and it has been shown to negatively impact both mitochondrial and endoplasmic reticulum (ER) function in a variety of in vitro models. In the placenta, mitochondrial and/or ER functional deficits have been associated with fetal growth restriction, pre-eclampsia, pre-term birth, and placental insufficiency.

Objectives: This study set out to identify the effects of 1, 5 and 10 M CBD on the expression of mitochondrial dynamics and ER stress gene markers in human trophoblasts. Additionally, a pilot study was conducted to optimize the use of a fluorescent mitochondrial dye and live-cell fluorescence microscopy for the visualization of morphology and quantification of mitochondrial length.

Methods: Both undifferentiated (cytotrophoblasts, CTBs) and differentiated (syncytiotrophoblasts, STBs) BeWo b30 cells were used in this study. Major results were generated by extracting RNA and conducting quantitative reverse transcription polymerase chain reaction (RT-qPCR). Additionally, MitoTracker® Red FM and live-cell fluorescence microscopy were utilized for the pilot study.

Results: For STBs, there was a decrease in the expression of a key mitochondrial fusion gene (MFN1) following 48-hour treatment with 10 M CBD, when compared to vehicle control. In terms of ER stress markers, differential effects were seen for CTBs and STBs. For CTBs treated with higher CBD concentrations, there were significant increases in the gene expression of ER stress markers, including ATF4 and CANX. Conversely, STBs treated with CBD showed significant decreases in the gene expression of ER stress markers IRE1 and ATF4. Additionally, a protocol utilizing fluorescence live-cell imaging was optimized to observe ultrastructural changes to mitochondria and quantify mitochondrial fragmentation. Validation utilized a known mitochondrial complex I inhibitor, rotenone.

Conclusion: The present study found differential effects of CBD on CTBs and STBs. These are some of the first findings exploring CBD’s effects on mitochondrial structure and ER function in placental cells. Given the importance of these organelles to placental health and pregnancy success, this is an area that warrants further investigation.

The role of nonsense mediated decay in Caenorhabditis elegans models of neurodegeneration. Yejin Kang

Supervisor: Dr. Lesley MacNeil Abstract: Alzheimer’s disease (AD) is the most common form of neurodegenerative disease that its prevalence is continuously rising with ageing society. Features of AD include the progressive accumulation of amyloid-beta oligomers into plaques and the formation of abnormal hyperphosphorylation tau aggregates. Previously, for the better understanding of mechanisms of AD pathogenesis, RNA-sequencing of aggregate-prone (AP) Tau Caenorhabditis elegans was performed, and differential gene expression in neurodegeneration model of C. elegans was analyzed. It was found that pseudogenes, a target of nonsense-mediated decay (NMD), was the largest class of genes whose expression levels were increased in AP-Tau model, suggesting NMD is disrupted. To investigate whether the disruption of NMD contributes to neurodegeneration, C. elegans strain carrying smg-1 mutations, as well as a GFP reporter gene for mechanosensory neurons, was generated. smg-1 mutations were also introduced into AP-tau model of C. elegans to determine whether neurodegeneration is accelerated when NMD is disrupted. Neurodegeneration levels were scored using a GFP reporter. For both strains with smg-1 and smg-1; tau, levels of neurodegeneration were higher than their respective controls without smg-1 mutations. The differences in percentage (%) neurodegeneration were both significant with p-vale less than 0.05. These results imply that disrupted NMD activity contributes to neurodegeneration in AP-Tau C. elegans model, and under normal conditions, NMD may play a role in neuroprotection.

Data-Driven Predictive Analytics Framework for Analysis of Prognostic Factors and Health Outcomes in Pediatric Intestinal Failure Patients: Developed through McMaster Children’s Hospital's Intestinal Rehabilitation Program

Papiha Joharapurkar Supervisor: Dr. Nikhil Pai

Introduction: Pediatric intestinal failure (IF) is a complex medical-surgical disorder with short- and long-term complications that can greatly affect a child’s overall wellbeing. Patients present a diverse set of etiologies, but they all struggle with intestinal adaptation to achieve enteral autonomy (EA) where they are not dependent on total parenteral nutrition (TPN) to sustain nutritional requirements. Despite improvements in clinical management, patients still suffer tremendous morbidity and an uncertain prognosis. The primary goal of intestinal rehabilitation programs is to facilitate sufficient adaptation for patients to achieve EA. Objectives: This study aims to identify predictors of EA for pediatric IF patients through machine learning (ML) and evaluate their impact on TPN duration. For patients on TPN, comparative analyses were conducted of lipid emulsion (ILE) treatments and central venous catheter (CVC) placement. Methods: Patient demographics, surgical procedures, and clinical outcomes for patients (N=17) from the McMaster Children’s Hospital Intestinal Failure Program from 1999-2021 were collected for a retrospective chart review. Predictors of EA status were obtained using a decision tree model and assessed for their impact on TPN durations with Kaplan Meier (KM) and Cox Proportional Hazards (CPH) analyses. Comparative ILE treatments were tested with the Mann-Whitney test and CVCs compared with KM, CPH, and ANOVA (P<0.05). Results: The algorithm retrieved 10 predictors of EA: CVC complications (total and frequency over 1000 catheter days), total bilirubin values, short bowel length, colon length, gestational and chronological age, time since undergoing serial transverse enteroplasty procedure (STEP) or initial resection surgeries, and weight. The model’s Area under Receiver Operator Curve and Precision-Recall Curve were 91% and 71% respectively. Sex, STEP and ileocecal valve (ICV) resection history and % remnant small bowel length were all found to be significant under KM but not CPH. Further, SMOFLipid demonstrated earlier stabilization of total bilirubin values compared to Intralipid shown at 6 months with Mantel Cox test (4.0 and 10.0 uMol/L, P<0.05). KM demonstrated greater longevity for peripherally inserted central catheter (PICC) at 847 days (95% CI: 0-2044) compared to tunnelled catheters at 268 days (95% CI: 93-443) and port CVCs at 181 days (95% CI: 144-218). ANOVA demonstrated no significant differences in complication profiles across catheters (P>0.05). Conclusions: Although no predictors were found to be significant through multivariate tests, this novel approach can be used as a framework for exploratory analyses and enhance clinical decision-making around IF management. When considering clinical management, there are preliminary indications that SMOFLipid treatments are an optimal ILE, with a potential role for IF associated liver disease (IFALD) prevention. Further, the use of PICCs for long-term CVC access may be more favorable to tunnelled catheters or ports.

Characterization of UVA biophoton emission and survival of α-irradiated HCT116+/+ cells via radium exposure

Zarin Hossain Supervisor: Dr. Carmel E. Mothersill

Background: It’s been hypothesized that biophoton production is a result of the generation of excited species followed by relaxation of these excited species to a stable state. This can occur as a result of many stressors including irradiation from radioactive materials. Cell exposure to radium (Ra-226) has not been thoroughly explored for biophoton production. Furthermore, biophoton production is often associated with oxidative stress and cell death which is concurrently also an area of exploration.

Objectives: The objective of this study was to characterize biophoton emission and additionally observe cell survival from αirradiated HCT116+/+ cells via radium exposure. We aim to expand evidence that supports increased biophoton production and decreased cell survival as a result of oxidative stress and exposure to radioactive material.

Methods: HCT116+/+ cells were standardly cultured in this study. For characterization of biophotons, cell groups were irradiated with 10, 100, 1000 and 10,000mBq/ml of Ra-226 for 24 hours. A photon counter was used to quantify counts. For characterization of cell survival, standard clonogenic assay techniques were used. Cell groups were irradiated with the same concentrations for Ra-226 and colonies were counted 7-9 days later.

Results: No significant results were seen when observing biophoton counts from Ra-226 irradiated groups compared to a background count. Furthermore, there was a significant difference between Ra-226 irradiated groups when compared to a positive control known to produce biophotons. Some significant differences were seen in the surviving fraction of directly irradiated cells when looking at the different concentrations of Ra-226. However, overall there was a correlation that could be seen where an increase in concentration of Ra-226 resulted in a decreased surviving fraction of cells.

Conclusions: The hypothesis of this study was partially supported where increased exposure and sensitivity to Ra-226 showed decreased cell survival, and assumed to have implications on biophoton production at higher concentrations although was not observed here at environmentally relevant concentrations. It was also concluded that biophoton production and oxidative stress are independent events. The significance of biophoton production as a radiation-induced bystander effect was explored as an implication of this study.

In vitro biofilm model to assess dosing regimens in phage therapy for inflammatory bowel disease

Heidi Ho Supervisor: Dr. Zeinab Hosseinidoust

Abstract The gut is home to a wide range of microbial species termed the microbiome. The microbiome makeup plays a vital role in maintaining overall health. Additionally, the gut barrier consisting of a layer of mucous and lining cells prevents the invasion of harmful bacteria. When the gut barrier integrity is compromised, there is an infiltration of harmful bacteria, which results in an imbalance of the gut microbiome and can lead to gutrelated diseases such as inflammatory bowel disease (IBD). IBD is a multifactorial disease characterized by chronic, recurring inflammation of the gut, caused by an inappropriate immune response and subsequent inflammation. The presence of adherent-invasive Escherichia coli (AIEC) in IBD patients promote gut inflammation through its increased ability to adhere and invade intestinal epithelial cells (IEC) and form biofilms. There is currently a growing interest in manipulation of the gut microbiota through phage therapy to treat chronic diseases such as IBD, since long-term use of antibiotics is impractical. However, several topics in phage therapy are currently unaddressed including the establishment of phage dosing. To perpetuate phage therapy, such questions surrounding phage dosing pharmacology needs to be better understood. To address the challenges of complex dosing in phage therapy prior to in vivo studies, an in vitro model is of fundamental importance. Thus, the objective of this thesis is to optimize an in vitro, AIEC biofilm model to better characterize phage candidates and phage therapy dosing regimens pre-clinically for IBD.

List of Abbreviations AIEC: adherent-invasive Escherichia coli; ANOVA: analysis of variance; CD: Crohn’s disease; CECAM: cell adhesion molecule; CFU: colony forming units; CV: crystal violet; DNA: deoxyribonucleic acid; GI: gastrointestinal; GI: genomic island; IBD: inflammatory bowel disease; IEC: intestinal epithelial cells; IL: interleukin; Lpf: long polar fimbriae; LPS: lipopolysaccharide; MOI: multiplicity of infection; MPS: mononuclear phagocytic system; NF: nuclear factor; PBS: phosphate buffered saline; PFU: plaque forming units; TSA: trypticase soy agar; TSB: tryptic soy broth; UC: ulcerative colitis

Effects of Plasma Rich in Growth Factor Versus Hyaluronic Acid For The Treatment of Knee Osteoarthritis: A Systematic Review and Meta-analysis of Randomized Controlled Trials

Mackenzie Haggstrom Supervisor: Dr. Hammad Qazi Honours Biology & Pharmacology Program, McMaster University, Hamilton ON

ABSTRACT Introduction: Current treatment options for osteoarthritis are targeted at reducing pain, however, osteoarthritis is characterized by degeneration of articular cartilage. Therefore, there is a need for a novel treatment option that works to rebuild articular cartilage. The main objective of this study was to determine the effects of plasma rich in growth factor versus hyaluronic acid to treat adults with knee osteoarthritis.

Methods: A systematic review and meta-analysis was done in accordance with Cochrane guidelines. All articles were evaluated at the title, abstract and full text-phase using the selection criteria. PRGF was compared to HA, in men and women over the age of 50 with a BMI less than or equal to 35 mg/kg2. Revman 5.3 software was used to perform the statistical analysis.

Results: The total WOMAC score for the PRGF group was significantly lower than the HA group at 6 months (P=0.01) and 12 months (P = 0.006). Similarly, the WOMAC pain score was significantly lower in the PRGF group compared to the HA group at 6 months (P = 0.02) and 12 months (P = 0.02). The final WOMAC sub score, physical function, found the score to be significantly lower for the PRGF group at 6 months (P = 0.02) and 12 months (P = 0.007). The Lequesne index was significantly lower in the PRGF group compared to the HA group at 6 months (P = 0.009) and 12 months (P = 0.02). The VAS pain score was significantly lower for the PRGF group compared to the HA group at 12 months (P = 0.004).

Discussion and Conclusion: The results of this study indicate that PRGF is advantageous compared to HA in the treatment of knee OA, including reduced long-term pain and increased physical function. Based on the results of this study, PRGF has no additional risk compared to PRP and HA and should be considered as a non-surgical treatment option for knee OA. In order for clinicians to use this research in a clinical setting, more RCTs need to be done comparing the safety and efficacy of PRGF versus HA. Future RCTs should be focused on optimizing the dose of PRGF as well as including longer follow-up periods due to PRGF’s latent effects.

Department of Education Services, Faculty of Health Sciences, 2021 Layout by S. Cusick