Modification of Mesenchymal Stem/Stromal Cell-Derived Small Extracellular Vesicles by Calcitonin Gene Related Peptide (CGRP) Antagonist: Potential Implications for Inflammation and Pain Reversal.

During the progression of knee osteoarthritis (OA), the synovium and infrapatellar fat pad (IFP) can serve as source for Substance P (SP) and calcitonin gene-related peptide (CGRP), two important pain-transmitting, immune, and inflammation modulating neuropeptides. Our previous studies showed that infrapatellar fat pad-derived mesenchymal stem/stromal cells (MSC) acquire a potent immunomodulatory phenotype and actively degrade Substance P via CD10 both in vitro and in vivo. On this basis, our hypothesis is that CD10-bound IFP-MSC sEVs can be engineered to target CGRP while retaining their anti-inflammatory phenotype. Herein, human IFP-MSC cultures were transduced with an adeno-associated virus (AAV) vector carrying a GFP-labelled gene for a CGRP antagonist peptide (aCGRP). The GFP positive aCGRP IFP-MSC were isolated and their sEVs' miRNA and protein cargos were assessed using multiplex methods. Our results showed that purified aCGRP IFP-MSC cultures yielded sEVs with cargo of 147 distinct MSC-related miRNAs. Reactome analysis of miRNAs detected in these sEVs revealed strong involvement in the regulation of target genes involved in pathways that control pain, inflammation and cartilage homeostasis. Protein array of the sEVs cargo demonstrated high presence of key immunomodulatory and reparative proteins. Stimulated macrophages exposed to aCGRP IFP-MSC sEVs demonstrated a switch towards an alternate M2 status. Also, stimulated cortical neurons exposed to aCGRP IFP-MSC sEVs modulate their molecular pain signaling profile. Collectively, our data suggest that yielded sEVs can putatively target CGRP in vivo, while containing potent anti-inflammatory and analgesic cargo, suggesting the promise for novel sEVs-based therapeutic approaches to diseases such as OA.

Measuring Rates of Visual Field Progression in Linear Versus Nonlinear Scales: Implications for Understanding the Relationship Between Baseline Damage and Target Rates of Glaucoma Progression.

PURPOSE: The purpose of this study is to test the hypothesis that the relationship between baseline visual field damage and the rate of progression depends upon the use of logarithmic (dB) versus linear (1/Lambert) scale. METHODS: A total of 60 eyes (60 patients) with treated, established glaucoma and at least 5 reliable 24-2 visual fields were included. Baseline visual field mean deviation (MD) in dB was transformed to 1/Lambert using standard equation. Mixed effects linear regression was used to calculate the slopes (MD rates of progression over time) with linear and nonlinear scales. We tested the relationship between baseline MD and MD slopes for each scale of measure. RESULTS: In dB scale, worse baseline visual field loss was associated with faster MD slopes (P=0.037), whereas the opposite effect was seen in 1/Lambert (P=0.001). For a similar rate of progression in dB/y, eyes with mild visual field damage lost more linear sensitivity over a given period of time than those with more severe baseline damage. CONCLUSIONS: There is a significant relationship between baseline visual field severity and rates of MD progression, although the direction of this association depends on the scale sensitivity is measured. The definition of fast versus slow visual field progression should be revised and take into account that sensitivity in linear scales show a better correlation with structural loss than when conventionally measured in nonlinear scale.