Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer.

Efficient metabolism, or the means by which cells produce energy resources, is critical for proper effector function. Here, we present a protocol for examining the bioenergetics and mitochondrial fuel utilization of primary murine autoreactive immunocytes using cellular metabolism-modulating drugs. We describe steps for plate calibration, isolation of primary immunocytes, and Seahorse assay plate preparation. We then detail procedures for performing the XF Cell Mito Stress Test followed by bioenergetics calculations and statistics. For complete details on the use and execution of this protocol, please refer to Wilson et al.1.

Transcriptome analysis reveals organ-specific effects of 2-deoxyglucose treatment in healthy mice.

OBJECTIVE: Glycolytic inhibition via 2-deoxy-D-glucose (2DG) has potential therapeutic benefits for a range of diseases, including cancer, epilepsy, systemic lupus erythematosus (SLE), and rheumatoid arthritis (RA), and COVID-19, but the systemic effects of 2DG on gene function across different tissues are unclear. METHODS: This study analyzed the transcriptional profiles of nine tissues from C57BL/6J mice treated with 2DG to understand how it modulates pathways systemically. Principal component analysis (PCA), weighted gene co-network analysis (WGCNA), analysis of variance, and pathway analysis were all performed to identify modules altered by 2DG treatment. RESULTS: PCA revealed that samples clustered predominantly by tissue, suggesting that 2DG affects each tissue uniquely. Unsupervised clustering and WGCNA revealed six distinct tissue-specific modules significantly affected by 2DG, each with unique key pathways and genes. 2DG predominantly affected mitochondrial metabolism in the heart, while in the small intestine, it affected immunological pathways. CONCLUSIONS: These findings suggest that 2DG has a systemic impact that varies across organs, potentially affecting multiple pathways and functions. The study provides insights into the potential therapeutic benefits of 2DG across different diseases and highlights the importance of understanding its systemic effects for future research and clinical applications.

Dystonin modifiers of junctional epidermolysis bullosa and models of epidermolysis bullosa simplex without dystonia musculorum.

The Lamc2jeb junctional epidermolysis bullosa (EB) mouse model has been used to demonstrate that significant genetic modification of EB symptoms is possible, identifying as modifiers Col17a1 and six other quantitative trait loci, several with strong candidate genes including dystonin (Dst/Bpag1). Here, CRISPR/Cas9 was used to alter exon 23 in mouse skin specific isoform Dst-e (Ensembl GRCm38 transcript name Dst-213, transcript ID ENSMUST00000183302.5, protein size 2639AA) and validate a proposed arginine/glutamine difference at amino acid p1226 in B6 versus 129 mice as a modifier of EB. Frame shift deletions (FSD) in mouse Dst-e exon 23 (Dst-eFSD/FSD) were also identified that cause mice carrying wild-type Lamc2 to develop a phenotype similar to human EB simplex without dystonia musculorum. When combined, Dst-eFSD/FSD modifies Lamc2jeb/jeb (FSD+jeb) induced disease in unexpected ways implicating an altered balance between DST-e (BPAG1e) and a rarely reported rodless DST-eS (BPAG1eS) in epithelium as a possible mechanism. Further, FSD+jeb mice with pinnae removed are found to provide a test bed for studying internal epithelium EB disease and treatment without severe skin disease as a limiting factor while also revealing and accelerating significant nasopharynx symptoms present but not previously noted in Lamc2jeb/jeb mice.

Glucose oxidation-dependent survival of activated B cells provides a putative novel therapeutic target for lupus treatment.

Aberrant metabolic demand is observed in immune/inflammatory disorders, yet the role in pathogenesis remains unclear. Here, we discover that in lupus, activated B cells, including germinal center B (GCB) cells, have remarkably high glycolytic requirement for survival over T cell populations, as demonstrated by increased metabolic activity in lupus-activated B cells compared to immunization-induced cells. The augmented reliance on glucose oxidation makes GCB cells vulnerable to mitochondrial ROS-induced oxidative stress and apoptosis. Short-term glycolysis inhibition selectively reduces pathogenic activated B in lupus-prone mice, extending their lifespan, without affecting T follicular helper cells. Particularly, BCMA-expressing GCB cells rely heavily on glucose oxidation. Depleting BCMA-expressing activated B cells with APRIL-based CAR-T cells significantly prolongs the lifespan of mice with severe autoimmune disease. These results reveal that glycolysis-dependent activated B and GCB cells, especially those expressing BCMA, are potentially key lupus mediators, and could be targeted to improve disease outcomes.

Transcriptome Analysis Reveals Organ-Specific Effects of 2-Deoxyglucose Treatment in Healthy Mice.

OBJECTIVE: Glycolytic inhibition via 2-deoxy-D-glucose (2DG) has potential therapeutic benefits for a range of diseases, including cancer, epilepsy, systemic lupus erythematosus (SLE), and rheumatoid arthritis (RA), and COVID-19, but the systemic effects of 2DG on gene function across different tissues are unclear. METHODS: This study analyzed the transcriptional profiles of nine tissues from C57BL/6J mice treated with 2DG to understand how it modulates pathways systemically. Principal component analysis (PCA), weighted gene co-network analysis (WGCNA), analysis of variance, and pathway analysis were all performed to identify modules altered by 2DG treatment. RESULTS: PCA revealed that samples clustered predominantly by tissue, suggesting that 2DG affects each tissue uniquely. Unsupervised clustering and WGCNA revealed six distinct tissue-specific modules significantly affected by 2DG, each with unique key pathways and genes. 2DG predominantly affected mitochondrial metabolism in the heart, while in the small intestine, it affected immunological pathways. CONCLUSIONS: These findings suggest that 2DG has a systemic impact that varies across organs, potentially affecting multiple pathways and functions. The study provides insights into the potential therapeutic benefits of 2DG across different diseases and highlights the importance of understanding its systemic effects for future research and clinical applications.

Regional shear wave elastography of Achilles tendinopathy in symptomatic versus contralateral Achilles tendons.

OBJECTIVES: Ultrasound often corroborates clinical diagnosis of Achilles tendinopathy (AT). Traditional measures assess macromorphological features or use qualitative grading scales, primarily focused within the free tendon. Shear wave imaging can non-invasively quantify tendon elasticity, yet it is unknown if proximal structures are affected by tendon pathology. The purpose of the study was to determine the characteristics of both traditional sonographic measures and regional shear wave speed (SWS) between limbs in patients with AT. METHODS: Twenty patients with chronic AT were recruited. Traditional sonographic measures of tendon structure were measured. Regional SWS was collected in a resting ankle position along the entire length of the tendon bilaterally. SWS measures were extracted and interpolated across evenly distributed points corresponding to the free tendon (FT), soleus aponeurosis (SA), and gastrocnemius aponeurosis (GA). Comparisons were made between limbs in both traditional sonographic measures and regional SWS. RESULTS: Symptomatic tendons were thicker (10.2 (1.9) vs. 6.8 (1.8) mm; p < 0.001) and had more hyperemia (p = 0.001) and hypoechogenicity (p = 0.002) than the contralateral tendon. Regional SWS in the FT was lower in the symptomatic limb compared to the contralateral limb (11.53 [10.99, 12.07] vs. 10.97 [10.43, 11.51]; p = 0.03). No differences between limbs were found for the SA (p = 0.13) or GA (p = 0.99). CONCLUSIONS: Lower SWS was only observed in the FT in AT patients, indicating that alterations in tendon elasticity associated with AT were localized to the FT and did not involve the proximal passive tendon structures. KEY POINTS: • Baseline characteristics of a pilot sample of 20 subjects suffering from chronic Achilles tendinopathy showed differences in conventional sonographic measures of tendon thickness, qualitatively assessed hypoechogenicity, hyperemia, and quantitative measures of shear wave speed. • Regional shear wave speeds were lower in the free tendon but not in the proximal regions of the soleus or gastrocnemius aponeuroses in Achilles tendinopathy patients. • Using shear wave imaging to estimate tendon stiffness may prove beneficial for clinical validation studies to address important topics such as return to activity and the effectiveness of rehabilitation protocols.

Providing a Helping Hand: Metabolic Regulation of T Follicular Helper Cells and Their Association With Disease.

T follicular helper (Tfh) cells provide support to B cells upon arrival in the germinal center, and thus are critical for the generation of a robust adaptive immune response. Tfh express specific transcription factors and cellular receptors including Bcl6, CXCR5, PD-1, and ICOS, which are critical for homing and overall function. Generally, the induction of an immune response is tightly regulated. However, deviation during this process can result in harmful autoimmunity or the inability to successfully clear pathogens. Recently, it has been shown that Tfh differentiation, activation, and proliferation may be linked with the cellular metabolic state. In this review we will highlight recent discoveries in Tfh differentiation and explore how these cells contribute to functional immunity in disease, including autoimmune-related disorders, cancer, and of particular emphasis, during infection.

Use of the Independent Medical Spotter in Identifying Head Injuries in Division I Football Players.

OBJECTIVE: Sport-related concussion (SRC) is a known risk of contact sports and strategies to minimize risk have been used. We aimed to determine if an independent medical spotter (IMS) identified more SRCs than would otherwise be detected by trained sideline medical staff (SMS). DESIGN: Prospective review of SRCs during competition in the 2019 season and retrospective review of SRCs in the 2015 to 2018 seasons, which also used an IMS. SETTING: Division I football games (home and away) of a Big 10 Conference institution. PARTICIPANTS: All football team members who participated in competition. INDEPENDENT VARIABLES: Occurrence of SRC during competition and whether the IMS or SMS directly visualized the injury. MAIN OUTCOME MEASURES: The total number of SRCs that occurred during competition in the 2015 to 2019 football seasons and whom observed the SRC-SMS or IMS-or if a student athlete reported symptoms after competition. RESULTS: Over the 5-year study period, 24 SRCs occurred during competition. Of those, 19 (79.2%) were observed by SMS, 2 (8.3%) by the IMS, and 3 (12.5%) were reported to SMS after competition ended. CONCLUSIONS: Most SRCs are accurately identified by SMS, but a small number were apparent only to the IMS who seemed to add sensitivity in detecting a SRC. Instances remain in which SRC recognition and diagnosis were delayed despite trained SMS and IMS. CLINICAL RELEVANCE: An IMS allows for a small-added player protection benefit using different vantage points to identify potential SRCs during competition.

2-deoxy D-glucose treatment does not elicit a hair growth response in alopecia areata.

2-deoxy D-glucose (2DG) was tested for efficacy in treating alopecia areata using the C3H/HeJ skin graft model. 2DG has proven to be efficacious in treatment of various mouse models of autoimmunity with minimal serious side effects noted. This agent has been shown to normalize abnormally activated T-cell populations while also preventing cell surface expression of NKG2D; key factors defining alopecia areata disease progression. Daily oral ingestion of 2DG via drinking water to mice with patchy or diffuse alopecia areata for 16 weeks failed to prevent expansion of alopecia or cause regrowth of hair in treated mice. Histologically, there were no differences between treated and control groups. These results indicate that, while 2DG is effective for some autoimmune diseases, it was not efficacious for the cell-mediated autoimmune mouse disease, alopecia areata.

Enhancing the efficacy of glycolytic blockade in cancer cells via RAD51 inhibition.

Targeting the early steps of the glycolysis pathway in cancers is a well-established therapeutic strategy; however, the doses required to elicit a therapeutic effect on the cancer can be toxic to the patient. Consequently, numerous preclinical and clinical studies have combined glycolytic blockade with other therapies. However, most of these other therapies do not specifically target cancer cells, and thus adversely affect normal tissue. Here we first show that a diverse number of cancer models - spontaneous, patient-derived xenografted tumor samples, and xenografted human cancer cells - can be efficiently targeted by 2-deoxy-D-Glucose (2DG), a well-known glycolytic inhibitor. Next, we tested the cancer-cell specificity of a therapeutic compound using the MEC1 cell line, a chronic lymphocytic leukemia (CLL) cell line that expresses activation induced cytidine deaminase (AID). We show that MEC1 cells, are susceptible to 4,4'-Diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS), a specific RAD51 inhibitor. We then combine 2DG and DIDS, each at a lower dose and demonstrate that this combination is more efficacious than fludarabine, the current standard- of- care treatment for CLL. This suggests that the therapeutic blockade of glycolysis together with the therapeutic inhibition of RAD51-dependent homologous recombination can be a potentially beneficial combination for targeting AID positive cancer cells with minimal adverse effects on normal tissue. Implications: Combination therapy targeting glycolysis and specific RAD51 function shows increased efficacy as compared to standard of care treatments in leukemias.

Bicomponent ultrashort echo time T2* analysis for assessment of patients with patellar tendinopathy.

PURPOSE: To compare bicomponent ultrashort echo time (UTE) T2* parameters of patellar tendon between healthy volunteers and patients with patellar tendinopathy. MATERIALS AND METHODS: This study was performed with Institutional Review Board approval and with all subjects signing informed consent. A UTE- T2* mapping sequence was performed at 3.0T on the knees of 10 healthy volunteers and in 11 patients with patellar tendinopathy. The UTE- T2* relaxation times of the fast relaxing macromolecular bound water component ( T2*F) and the slow relaxing bulk water component ( T2*S) and the fraction of the fast relaxing macromolecular bound water component (FF ) of patellar tendon were measured in all subjects. Wilcoxon rank-sum tests were used to compare UTE- T2* parameters between healthy volunteers and patients with patellar tendinopathy. RESULTS: Mean T2*F, T2*S, and FF of the patellar tendon was 1.5 msec, 23.1 msec, and 79.5%, respectively, for healthy volunteers and 1.9 msec, 22.3 msec, and 75.5%, respectively, for patients with patellar tendinopathy. There were statistically significant differences between groups of subjects for T2*F (P = 0.01) and FF (P = 0.007) but not T2*S (P = 0.10) of the patellar tendon. CONCLUSION: Patients with patellar tendinopathy had significantly higher T2*F and significantly lower FF of patellar tendon than healthy volunteers, which suggests that bicomponent UTE- T2* parameters can detect changes in the composition and microstructure of degenerative tendon. LEVEL OF EVIDENCE: 1 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2017;46:1441-1447.

Analysis of Gene Expression in an Inbred Line of Soft-Shell Clams (Mya arenaria) Displaying Growth Heterosis: Regulation of Structural Genes and the NOD2 Pathway.

Mya arenaria is a bivalve mollusk of commercial and economic importance, currently impacted by ocean warming, acidification, and invasive species. In order to inform studies on the growth of M. arenaria, we selected and inbred a population of soft-shell clams for a fast-growth phenotype. This population displayed significantly faster growth (p < 0.0001), as measured by 35.4% greater shell size. To assess the biological basis of this growth heterosis, we characterized the complete transcriptomes of six individuals and identified differentially expressed genes by RNAseq. Pathways differentially expressed included structural gene pathways. Also differentially expressed was the nucleotide-binding oligomerization domain 2 (NOD2) receptor pathway that contributes to determination of growth, immunity, apoptosis, and proliferation. NOD2 pathway members that were upregulated included a subset of isoforms of RIPK2 (mean 3.3-fold increase in expression), ERK/MAPK14 (3.8-fold), JNK/MAPK8 (4.1-fold), and NFκB (4.08-fold). These transcriptomes will be useful resources for both the aquaculture community and researchers with an interest in mollusks and growth heterosis.

Complete mitochondrial genome of the soft-shell clam Mya arenaria.

We have sequenced and characterized the complete mitochondrial genome of the soft-shell clam, Mya arenaria, an important organism for environmental toxicology and aquaculture. Mya arenaria is located in the taxonomic order Myoida, which lacks any member with a completely annotated mitogenome. The M. arenaria mitochondrial genome is 17 947 bp in length. Like most marine bivalves, the circular mitogenome codes entirely on the heavy strand, with no introns. As with other bivalves, the gene order of the mitochondrion is highly rearranged. The mitogenome contains 12 protein-coding genes but ATP8 is missing, consistent with about half of all bivalve genera. Twenty-three tRNAs were identified. Phylogenetic analysis shows that M. arenaria is related most closely with the bivalves Sinonovacula constricta, and Moerella iridescens, of the infraclass Euheterodonta (unassigned). This, along with the close grouping of the phylogenetic trees, confirms a close tie between Myoida and Euheterodonta (unassigned).

Articular Cartilage of the Human Knee Joint: In Vivo Multicomponent T2 Analysis at 3.0 T.

PURPOSE: To compare multicomponent T2 parameters of the articular cartilage of the knee joint measured by using multicomponent driven equilibrium single-shot observation of T1 and T2 (mcDESPOT) in asymptomatic volunteers and patients with osteoarthritis. MATERIALS AND METHODS: This prospective study was performed with institutional review board approval and with written informed consent from all subjects. The mcDESPOT sequence was performed in the knee joint of 13 asymptomatic volunteers and 14 patients with osteoarthritis of the knee. Single-component T2 (T2(Single)), T2 of the fast-relaxing water component (T2F) and of the slow-relaxing water component (T2S), and the fraction of the fast-relaxing water component (F(F)) of cartilage were measured. Wilcoxon rank-sum tests and multivariate linear regression models were used to compare mcDESPOT parameters between volunteers and patients with osteoarthritis. Receiver operating characteristic analysis was used to assess diagnostic performance with mcDESPOT parameters for distinguishing morphologically normal cartilage from morphologically degenerative cartilage identified at magnetic resonance imaging in eight cartilage subsections of the knee joint. RESULTS: Higher cartilage T2(Single) (P < .001), lower cartilage F(F) (P < .001), and similar cartilage T2F (P = .079) and T2S (P = .124) values were seen in patients with osteoarthritis compared with those in asymptomatic volunteers. Differences in T2(Single) and F(F) remained significant (P < .05) after consideration of age differences between groups of subjects. Diagnostic performance was higher with F(F) than with T2(Single) for distinguishing between normal and degenerative cartilage (P < .05), with greater areas under the curve at receiver operating characteristic analysis. CONCLUSION: Patients with osteoarthritis of the knee had significantly higher cartilage T2(Single) and significantly lower cartilage F(F) than did asymptomatic volunteers, and receiver operating characteristic analysis results suggested that F(F) may allow greater diagnostic performance than that with T2(Single) for distinguishing between normal and degenerative cartilage.

Rapid in vivo multicomponent T2 mapping of human knee menisci.

PURPOSE: To compare multicomponent T2 parameters of menisci measured using Multicomponent Driven Equilibrium Single Pulse Observation of T1 and T2 (mcDESPOT) in asymptomatic volunteers and osteoarthritis (OA) patients with intact and torn menisci. MATERIALS AND METHODS: The prospective study was performed with Institutional Review Board approval and with all subjects signing written informed consent. mcDESPOT was performed on the knee joint of 12 asymptomatic volunteers and 14 patients with knee OA. Single-component T2 relaxation time (T2Single ), T2 relaxation time of the fast relaxing water component (T2F ), and the slow relaxing water component (T2S ), and fraction of the fast relaxing water component (FF ) of the medial and lateral menisci were measured. Multivariate linear regression models were used to compare mcDESPOT parameters between normal menisci in asymptomatic volunteers, intact menisci in OA patients, and torn menisci in OA patients with adjustment for differences in age between subjects. RESULTS: The mean mcDESPOT parameters for normal menisci in asymptomatic volunteers, intact menisci in OA patients, and torn menisci in OA patients were respectively 16.1 msec, 18.8 msec, and 22.7 msec for T2Single ; 9.0 msec, 10.0 msec, and 11.1 msec for T2F ; 24.4 msec, 27.7 msec, and 31.4 msec for T2S ; and 34%, 32%, 27% for FF . There were significant differences (P < 0.05) in T2Single , T2F , T2S , and FF between the three groups of menisci. CONCLUSION: The menisci of OA patients had significantly higher T2Single , T2F , and T2S and significantly lower FF than normal menisci in asymptomatic volunteers with greater changes in multicomponent T2 parameters noted in torn than intact menisci in OA patients.

Intratendinous injections of platelet-rich plasma: feasibility and effect on tendon morphology and mechanics.

BACKGROUND: Intratendinous injections may have important effects on the properties of collagen microarchitecture, morphology, and subsequent mechanical properties of the injected tendon. The purpose of this study was to examine the effects of intratendinous PRP injections; the injectant retention within tendons, the distribution of intratendinous injectant, and whether intratendinous injection or needle fenestration alters tendon morphology or mechanics. DESIGN: Controlled Laboratory Study. INTERVENTIONS: In the first part of the study, 18 lamb extensor tendons were selected to receive methylene blue-containing PRP injection (PRP/MB), methylene blue only injection (MB), or needle fenestration. The volume of retained injectant was measured and injectant distribution and tendon morphology were examined microscopically. In the second portion of the study, 18 porcine flexor tendons were divided into control, needle fenestration, or saline injection groups. Young's Modulus was then determined for each tendon under 0-4% strain. MAIN OUTCOME MEASURES: 1) Injectant volume retained; 2) Injectant distribution; 3) Post-injection/fenestration alterations in morphology, biomechanics. RESULTS: Intratendinous injectant is retained within the tendon. The difference between PRP and PRP/MB groups was not significant (p = 0.78). Intratendinous spread of the injectant solution within the tendon occurs primarily in the proximodistal direction, with very little cross-sectional penetration. Intratendinous injections resulted in microscopic morphology disruption (e.g., separation and disorganization of both the collagen bundles and cellular distribution). There were significant differences in Young's Modulus between control (Ectrl = 2415.48) and injected tendons (Einj = 1753.45) at 4% strain (p = 0.01). There were no differences in Young's Modulus between fenestrated and control tendons. CONCLUSIONS: Intratendinous PRP injections are retained within the tendon, and primarily distributes longitudinally with minimal cross-sectional spread. Intratendinous injections may alter tendon morphology and mechanics.

Evaluation of the patient with hip pain.

Hip pain is a common and disabling condition that affects patients of all ages. The differential diagnosis of hip pain is broad, presenting a diagnostic challenge. Patients often express that their hip pain is localized to one of three anatomic regions: the anterior hip and groin, the posterior hip and buttock, or the lateral hip. Anterior hip and groin pain is commonly associated with intra-articular pathology, such as osteoarthritis and hip labral tears. Posterior hip pain is associated with piriformis syndrome, sacroiliac joint dysfunction, lumbar radiculopathy, and less commonly ischiofemoral impingement and vascular claudication. Lateral hip pain occurs with greater trochanteric pain syndrome. Clinical examination tests, although helpful, are not highly sensitive or specific for most diagnoses; however, a rational approach to the hip examination can be used. Radiography should be performed if acute fracture, dislocations, or stress fractures are suspected. Initial plain radiography of the hip should include an anteroposterior view of the pelvis and frog-leg lateral view of the symptomatic hip. Magnetic resonance imaging should be performed if the history and plain radiograph results are not diagnostic. Magnetic resonance imaging is valuable for the detection of occult traumatic fractures, stress fractures, and osteonecrosis of the femoral head. Magnetic resonance arthrography is the diagnostic test of choice for labral tears.

Visualizing tendon elasticity in an ex vivo partial tear model.

Supersonic shear imaging (SSI) is evaluated as a means of visualizing changes in regional tendon elasticity caused by partial tears in a porcine model. Thirty digital flexor tendons were cut to 25% (n = 10), 50% (n = 10) and 75% (n = 10) of the tendon thickness along the deep surface. Tendon elasticity was mapped left of, centered on and right of the tear site before and after tearing from 0% to 2% strain. Shear wave speed increased at 1% (p < 0.05) and 2% (p < 0.001) strain for all regions. Deep surface shear wave speed decreased in the 25%, 50% and 75% tears (p < 0.05 and p < 0.001). Computational tendon tear models were also created to investigate regional changes in strain resulting from a tear. In the computational models, strain on the deep surface decreased progressively with increasing tear size. Visualization of tendon shear wave speed was achieved in normal and partially torn tendons, indicating the potential of SSI to add tendon shear wave speed to traditional morphologic assessment of partial tears, which may improve assessment of tendon health.

Platelet-rich plasma for the treatment of chronic plantar fasciopathy in adults: a case series.

Plantar fasciopathy (PF) is a common source of pain and disability that is often refractory to conservative management. There are no uniformly effective standard-of-care treatments for chronic recalcitrant PF. Corticosteroid injection is considered a viable treatment option when traditional therapies fail, but is limited by suboptimal long-term efficacy and potential adverse effects. Platelet-rich plasma (PRP) is an emerging injection-based treatment for various chronic degenerative soft-tissue diseases. It is postulated to promote native tissue regeneration; however, consistent scientific evidence remains lacking. A prospective case series, including 24 consecutive PF cases, was conducted to report patient-rated pain and disability following PRP injection. Foot and Ankle Ability Measure (FAAM) scores were the primary clinical outcome measure. Foot-Single Assessment Numeric Evaluation (Foot-SANE) scores, Short Form-12 Health Survey version 2 (SF-12v2) questionnaires, and PRP treatment satisfaction surveys were secondary outcome measures. Statistical analysis compared baseline and 32 weeks post-injection time points. Patients receiving PRP injection reported clinically and statistically significant improvement in all outcome measures during this interval. There were no serious adverse events associated with treatment. PRP is considered a safe therapeutic option with the ability to decrease heel pain in patients with chronic PF refractory to appropriate conservative management.