Enhanced immunosuppressive capability of mesenchymal stem cell-derived small extracellular vesicles with high expression of CD73 in experimental autoimmune uveitis.

BACKGROUND: Autoimmune uveitis is an inflammatory disease triggered by an aberrant immune response. Mesenchymal stem cell-derived small extracellular vesicles (MSC-sEVs) are emerging as potential therapeutic agents for this condition. CD73, an ectoenzyme present on MSC-sEVs, is involved in mitigating inflammation by converting extracellular adenosine monophosphate into adenosine. We hypothesize that the inhibitory effect of MSC-sEVs on experimental autoimmune uveitis (EAU) could be partially attributed to the surface expression of CD73. METHODS: To investigate novel therapeutic approaches for autoimmune uveitis, we performed lentiviral transduction to overexpress CD73 on the surface of MSC-sEVs, yielding CD73-enriched MSC-sEVs (sEVs-CD73). Mice with interphotoreceptor retinoid-binding protein (IRBP)-induced EAU were grouped randomly and treated with 50 µg MSC-sEVs, vector infected MSC-sEVs, sEVs-CD73 or PBS via single tail vein injection. We evaluated the clinical and histological features of the induced mice and analyzed the proportion and functional capabilities of T helper cells. Furthermore, T-cells were co-cultured with various MSC-sEVs in vitro, and we quantified the resulting inflammatory response to assess the potential therapeutic benefits of sEVs-CD73. RESULTS: Compared to MSC-sEVs, sEVs-CD73 significantly alleviates EAU, leading to reduced inflammation and diminished tissue damage. Treatment with sEVs-CD73 results in a decreased proportion of Th1 cells in the spleen, draining lymph nodes, and eyes, accompanied by an increased proportion of regulatory T-cells (Treg cells). In vitro assays further reveal that sEVs-CD73 inhibits T-cell proliferation, suppresses Th1 cells differentiation, and enhances Treg cells proportion. CONCLUSION: Over-expression of CD73 on MSC-sEVs enhances their immunosuppressive effects in EAU, indicating that sEVs-CD73 has the potential as an efficient immunotherapeutic agent for autoimmune uveitis.

Effectiveness of toric intraocular lens implantation for correcting irregular corneal astigmatism in cataract eyes.

A retrospective cohort study was conducted to observe the correction effect of Toric intraocular lens (IOL) implantation in cataract eyes with specific types of irregular corneal astigmatism. Thirty-four eyes with either the "asymmetric bow-tie" pattern (Type I) or the "angled bow-tie" pattern (Type II) were included. Corneal topography was assessed using Pentacam HR, and changes in preoperative corneal astigmatism, visual acuity, manifest refraction, and objective visual quality were measured and compared. The average uncorrected distance visual acuity improved significantly from 0.86 ± 0.40 logMAR to 0.22 ± 0.15 logMAR (P < 0.001). Preoperative corneal astigmatism of 2.05 ± 0.90 D was corrected to a postoperative residual astigmatism of 0.78 ± 0.57 D (P < 0.001), with 32% of eyes within 0.50 D. The residual astigmatism prediction errors in Type I and Type II cases were (0.97 ± 0.68 D) and (0.66 ± 0.37 D), respectively (P = 0.100). The mean spherical equivalent prediction error in Type II cases (0.07 ± 0.36 D) was significantly smaller than that in Type I cases (- 0.29 ± 0.52 D) (P = 0.030). This study concludes that Toric IOL implantation effectively corrects specific types of irregular corneal astigmatism in cataract surgery. Eyes with the "angled bow-tie" pattern show higher accuracy in refractive predictions compared to eyes with the "asymmetric bow-tie" pattern.

Optical coherence tomography biomarkers as outcome predictors to guide dexamethasone implant use in patients with iERM: a randomized controlled trial.

BACKGROUND: We aimed to investigate the anatomical features of optical coherence tomography (OCT) and vitreous cytokine levels as predictors of outcomes of combined phacovitrectomy with intravitreal dexamethasone (DEX) implants for idiopathic epiretinal membrane (iERM) treatment. METHODS: A prospective, single-masked, randomized, controlled clinical trial included 48 eyes. They were randomly assigned in a 1:1 ratio to undergo the DEX group (combined phacovitrectomy with ERM peeling and Ozurdex implantation) and control group (phacovitrectomy only). Best-corrected visual acuity (BCVA) and central macular thickness (CMT) were assessed at 1 d, 1 week, 1 month, and 3 months. The structural features of OCT before surgery were analysed for stratified analysis. Baseline soluble CD14 (sCD14) and sCD163 levels in the vitreous fluid were measured using ELISA. RESULTS: BCVA and CMT were not significantly different in the DEX and control groups. Eyes with hyperreflective foci (HRF) at baseline achieved better BCVA (Ptime*group=0.746; Pgroup=0.043, Wald χ²=7.869) and lower CMT (Ptime*group = 0.079; Pgroup = 0.001, Wald χ²=6.774) responses to DEX during follow-up. In all patients, the mean vitreous level of sCD163 in eyes with HRF was significantly higher than that in eyes without HRF (P = 0.036, Z=-2.093) at baseline. In the DEX group, higher sCD163 predicted greater reduction in CMT from baseline to 1 month (r = 0.470, P = 0.049). CONCLUSIONS: We found that intraoperative DEX implantation did not have beneficial effects on BCVA and CMT over a 3-month period in all patients with iERM, implying that the use of DEX for all iERM is not recommended. In contrast, for those with HRF on OCT responded better to DEX implants at the 3-month follow-up and thier vitreous fluid expressed higher levels of sCD163 at baseline. These data support the hypothesis that DEX implants may be particularly effective in treating cases where ERM is secondary to inflammation. TRIAL REGISTRATION: The trail has been registered at Chinese Clinical Trail Registry( https://www.chictr.org.cn ) on 2021/03/12 (ChiCTR2100044228). And all patients in the article were enrolled after registration.

The Anatomic and Functional Outcomes of Ozurdex-Aided Vitrectomy in Proliferative Diabetic Retinopathy.

Purpose: To investigate the 3-months outcomes of patients who underwent intraoperative intravitreal injection of Ozurdex for proliferative diabetic retinopathy (PDR). Methods: This is a prospective randomized controlled clinical trial (ChiCTR2100043399). Seventy-one patients with PDR who had indications for surgery without intravitreal injection history within 3 months preoperatively were enrolled. Patients were randomly divided into three groups based on the medicine injected intraoperatively: Ozurdex, Conbercept, and Control group. The primary outcome is the best-corrected visual acuity (BCVA) within 3 months postoperatively. The secondary outcomes include the intraocular pressure (IOP), mean sensitivity, central retinal thickness and vessels perfusion. Results: The BCVA and the mean sensitivity improved in the three groups (F = 130.8, P < 0.0001; F = 34.18, P < 0.0001), but there was no statistical difference among the three groups (F = 0.858, P = 0.552; F = 0.964, P = 0.452). The IOP was no significant differences among the three groups within 3 months postoperatively (F = 0.881, P = 0.533). Compared with the other two groups, central retinal thickness (CRT) and outer retinal layer (ORL) thickness decreased significantly in patients of the Ozurdex group (F = 3.037, P = 0.008; F = 2.626, P = 0.018), especially in the diabetic macular edema (DME) patients (F = 2.761, P = 0.0164; F = 2.572, P = 0.0240). In macular region, superficial vascular plexus (SVP), intermediate capillary plexus (ICP) and deep capillary plexus (DCP) perfusion were not shown statistical difference at 3 months postoperatively in the all three groups compared with 1 day postoperatively (P > 0.05). Conclusion: Compared with the other two groups, anatomical outcomes was improved significantly in Ozurdex group for DR patients. Ozurdex may help to improve the visual acuity and visual sensitivity, and there is no significant difference in the change of IOP and microvascular improvement. Clinical Trial Registration: This trial is registered with the Chinese Clinical Trial Registry (http://www.chictr.org.cn, registration number ChiCTR2100043399).

The predictive accuracy of Barrett toric calculator using measured posterior corneal astigmatism derived from swept source-OCT and Scheimpflug camera.

PURPOSE: To compare the performance of Barrett toric calculator incorporated with measured posterior corneal astigmatism (PCA) derived from IOL Master 700 and Pentacam HR versus predicted PCA. METHODS: The predicted residual astigmatism using Barrett toric IOL calculator with predicted PCA, measured PCA from IOL Master 700 and measured PCA from Pentacam were calculated with the preoperative keratometry and intended IOL axis with modification. The vector analysis was performed to calculate the mean absolute prediction error (MAE), the centroid of the prediction error and the percentage of eyes with a prediction error within ±0.50 D, ±0.75 D, and ±1.00 D. RESULTS: In 57 eyes of 57 patients with mean age of 70.42 ± 10.75 years, the MAE among the three calculation methods were 0.59 ± 0.38 D (Predicted PCA), 0.60 ± 0.38 D (Measured PCA from IOL Master 700) and 0.60 ± 0.36 D (Measured PCA from Pentacam) with no significant difference, either in the whole sample, the WTR eyes and the ATR eyes (F = 0.078, 0.306 and 0.083, p = 0.925, 0.739 and 0.920, respectively). Measured PCA obtained from IOL Master 700 resulted in one level reduction (from Tn to Tn-1) in 49.12% eyes in cylindrical model selection, while measured PCA obtained from Pentacam resulted in one level reduction of toric model selection in 18.18% eyes. CONCLUSION: The present study suggested that the incorporation of measured PCA values derived from IOL Master 700 and Pentacam produce comparable clinical outcome with the predicted PCA mode in Barrett toric calculator.

Comparison of two swept-source optical coherence tomography devices, a Scheimpflug camera system and a ray-tracing aberrometer in the measurement of corneal power in patients with cataract.

PURPOSE: To assess the differences and similarities in the corneal curvature obtained by two swept-source optical coherence tomography (SS-OCT) devices, Scheimpflug imaging system and one ray tracing aberrometer in patients with cataracts. Moreover, this study aimed to compare the differences in posterior corneal (PK), total corneal (TK) and true net power (TNP) measurements among the IOLMaster 700, CASIA2, and Pentacam. METHODS: A total of 200 eyes of 200 patients (116 female, 58%) were enrolled in this study, with a mean age of 65.9 ± 9.5 years. The flattest (Kf), steepest (Ks), and mean cornal powers (Km), J0, and J45 were obtained using two SS-OCT-based biometric devices, one rotating camera system and one ray-tracing aberrometer. The PK, TK and TNP values were also measured using these devices. To evaluate the differences and similarities between the devicves, the Friedman test, Pearson correlation coefficient (r), intraclass coefficient correlation (ICC) and Bland‒Altman plots with 95% limits of agreement (LoA) were used, and boxplots and stacked histograms were generated to describe the distributions of the data. RESULTS: There were no significant differences between the IOLMaster 700 and Pentacam for any of the keratometry values. Additionally, there were no significant differences between the IOLMaster 700 and iTrace in evaluating J0 and J45. Bland‒Altman plots revealed relatively wide LoA widths, almost larger than 1 diopter for the keratometry values and almost larger than 0.5 diopter for J0 and J45 values among the four devices. In terms of PK and TK values, significant differences and low ICCs were found among the three devices. CONCLUSIONS: Although strong correlations and good agreement were found among the IOLMaster700, CASIA2, Pentacam and iTrace for Kf, Ks, Km and J0, J45, it seems that the measurements should not be used interchangeably because of the wide LoA widths and the presence of significant differences among the devices. Similarly, due to significant differences and low ICCs, the PK, TK and TNP values obtained by IOLMaster 700, CASIA2, and Pentacam should not be used interchangeably.

The impact of posterior corneal astigmatism on the surgical planning of toric multifocal intraocular lens implantation.

Purpose: To investigate the influence of posterior corneal astigmatism on the prediction accuracy of toric multifocal intraocular lens (IOL) calculation. Methods: The keratometric astigmatism measured by Lenstar LS 900 (KCAL), keratometric astigmatism (KCAP) and total corneal astigmatism (TCA) measured by Scheimpflug camera (Pentacam HR) were documented and analyzed accordingly. Three deduction models using different parameters were compared. Model 1: KCAL â€‹+ â€‹keratometric corneal surgically induced astigmatism (KCSIA, 0.30 D @ 50°); Model 2: KCAP â€‹+ â€‹KCSIA); Model 3: TCA â€‹+ â€‹total CSIA (TCSIA, 0.23 D @ 50°). The prediction errors of each model as the difference vector between the actual and the intended residual astigmatism were compared. Results: Seventy-six eyes implanted with toric multifocal IOLs were included in this study. The vector differences of the actual KCSIA and TCSIA were statistically significant in the total sample and against-the-rule (ATR) subgroup (both P â€‹< â€‹0.05). Model 1 deduced the smallest mean values of prediction error, while that of Model 3 were smaller than that of Model 2, both in the total sample and the ATR subgroups (all P â€‹< â€‹0.05). Meanwhile, in the total sample and ATR subgroups, the centroid vector magnitudes of Model 3 were smaller than that of Model 1 (0.31 â€‹± â€‹0.76 D and 0.39 â€‹± â€‹0.76 D). Conclusions: The calculation of toric multifocal IOL should be individualized especially in the ATR eyes for the impact of PCA on the estimation of the preoperative corneal astigmatism and the CSIA.

Comparison of two objective methods for measuring postoperative toric intraocular lens orientation under the natural pupil.

PURPOSE: To evaluate and compare the accuracy of iTrace and CASIA2 in measuring the postoperative orientation of toric intraocular lens (IOL) without mydriasis. SETTING: Tianjin Medical University Eye Hospital, Tianjin, China. DESIGN: Prospective cohort study. METHODS: Patients with SN6AT toric IOLs implanted after cataract surgery were enrolled. 1 month after surgery, the toric IOL orientation were measured by iTrace and CASIA2 in non-mydriatic, semi-dark conditions. Then, the toric axis was directly reviewed using the slit-lamp under full mydriasis. Axis measurement differences between each of the 2 devices and the slit-lamp, described as their relative differences (RDs), were calculated and compared. The percentage of RDs within 5 degrees, within 10 degrees and greater than 30 degrees were analyzed. RESULTS: 77 eyes of 70 patients were included. Generally, the mean toric axis measurement RDs of CASIA2 and iTrace were 9.24 ± 10.53 degrees and 13.89 ± 15.47 degrees respectively ( P = .04). For CASIA2 (72 eyes), 54.17% (39), 72.22% (52), and 4.17% (3) of eyes had RDs within 5 degrees, within 10 degrees and greater than 30 degrees, compared with 40.00% (28), 61.43% (43) and 12.86% (9) for iTrace (70 eyes). The 95% limits of agreements of CASIA2 with slit-lamp was narrower than that of iTrace with slit-lamp. The median RD of CASIA2 was significantly smaller in eyes with pupil ≥4 mm under dark condition compared with eyes with pupil <4 mm ( P = .03). CONCLUSIONS: CASIA2 demonstrates greater precision in measuring toric IOL orientation under non-mydriatic conditions compared with iTrace. Moreover, the accuracy of CASIA2 is enhanced in cases of pupil >4 mm.

Plasmin-Induced Lens Epithelial Cells Detachment for the Reduction of Posterior Capsular Opacification.

Purpose: We sought to evaluate the efficacy and safety of plasmin injection in the capsular bag during the cataract operation for the prevention of posterior capsule opacification. Methods: Thirty-seven anterior capsular flaps taken from phacoemulsification surgery were immersed in either 1 µg/mL plasmin (plasmin group, n = 27) or phosphate-buffered saline (control group, n = 10) for 2 minutes and photographed after fixation and nuclear staining to compare the numbers of residual lens epithelial cells. In the animal experiments, the plasmin solution was injected into the capsular bag and remained for 5 minutes during hydrodissection or after lens extraction. The degree of posterior capsular opacity of the rabbits at 2 months were photographed by slit lamp biomicroscopy. In HLE-B3 cell culture, the cell detachment rate, proliferation, and apoptosis after the plasmin digestion were analyzed. Results: The residual lens epithelial cell numbers on the capsule after plasmin treatment were 168 ± 190.7/mm2 in the 1 µg/mL plasmin group, which was significantly lower than that of the control (1012 ± 798.8/mm2; P < 0.0001). In a rabbit model, the treatment of plasmin resulted in a significantly clearer posterior capsule compared with that of the control group at 2 months postoperatively. Conclusions: This study suggested that plasmin injection can induce effective lens epithelial cell detachment, which could be a promising adjunctive treatment to further improve the success rate in posterior capsule opacification prevention. Translational Relevance: Plasmin injection for lens epithelial cell detachment could significantly decrease the number of residual lens epithelial cells. This approach could be a promising treatment incorporating the current treatment approach to further improve the success rate in posterior capsule opacification prevention.

Mesenchymal-Stem-Cell-Based Strategies for Retinal Diseases.

Retinal diseases are major causes of irreversible vision loss and blindness. Despite extensive research into their pathophysiology and etiology, pharmacotherapy effectiveness and surgical outcomes remain poor. Based largely on numerous preclinical studies, administration of mesenchymal stem cells (MSCs) as a therapeutic strategy for retinal diseases holds great promise, and various approaches have been applied to the therapies. However, hindered by the retinal barriers, the initial vision for the stem cell replacement strategy fails to achieve the anticipated effect and has now been questioned. Accumulating evidence now suggests that the paracrine effect may play a dominant role in MSC-based treatment, and MSC-derived extracellular vesicles emerge as a novel compelling alternative for cell-free therapy. This review summarizes the therapeutic potential and current strategies of this fascinating class of cells in retinal degeneration and other retinal dysfunctions.

Characteristics of antibiotic resistance genes in full-scale anaerobic digesters of food waste and the effects of application of biogas slurry on soil antibiotic resistance genes.

The fate of antibiotic resistance genes (ARGs) in full-scale anaerobic digestion (AD) of food waste (FW) and in the soil applied with biogas slurry has not been fully understood. In this study, 12 targeted ARGs and intI1 in FW, intermediate product, and biogas slurry from three full-scale AD were analyzed. The results showed that subcritical water pretreatment was an effective method for ARG attenuation, by which the absolute abundance of total targeted ARGs was removed by 99.69%. The predominant ARGs (ermB, tetM, and tetW) in FW were removed more than 99% after subcritical water pretreatment. The result of field experiments with biogas slurry as fertilizer showed that the absolute abundance of several ARGs (sul2, tetM, blaOXA-1, blaTEM) and intI1 accumulated significantly compared to the control group (CK) during three consecutive growth stages of the rice. The detected abundance of ARGs in paddy field soil increased from 190.50 (CK) to 8.87 × 104 copies/g (wet weight) (soil) during tillering stage, and increased from 4102.65 (CK) to 4.38 × 104 copies/g (wet weight) (soil) during heading time. Biogas slurry improved the soil nutrients (TN, AN, TP, and AP); meanwhile, the concentrations of total salt and Cl- increased. Network analysis indicated that 28 genera were the possible hosts of ARGs; variation partitioning analysis (VPA) indicated that microbial communities (contribution 59.30%) were the main factors that affected the fate of ARGs and intI1.

Current development and perspectives of anaerobic bioconversion of crop stalks to Biogas: A review.

As one of the most abundant biomass resources, crop stalks are great potential feedstock available for anaerobic digestion (AD) to produce biogas. However, the specific physical properties and complex chemical structures of crop stalks form strong barriers to efficient AD bioconversion. To overcome these problems, many efforts have been made over the past few years. This paper reviewed recent research in the evolving field of anaerobic bioconversion of crop stalks and was focused on three critical aspects affecting AD performance: various pretreatment methods and their effects on the improvement of crop stalk biodegradability, determination of specific AD operation parameters for crop stalks, and development of AD technologies. Finally, recommendations on the future development of crop stalk AD were proposed.

Human umbilical cord-derived mesenchymal stem cells treatment for refractory uveitis: a case series.

AIM: To evaluate therapeutic outcomes of human umbilical cord-derived mesenchymal stem cells (HUC-MSCs) treatment in patients with refractory uveitis. METHODS: A retrospective and noncomparative review was performed on four patients with refractory uveitis from December 2013 to December 2017. HUC-MSCs were administered intravenously at a dose of 1×106 cells/kg. Clinical response, relapse rate, change of visual acuity, and other metrics were evaluated. RESULTS: All four patients presented with responses to HUC-MSCs treatment, with three males and one female. The numbers of uveitis attacks per year after the HUC-MSCs treatment (0, 2, 0, 0 respectively) all decreased compared with the numbers before the treatment (3, 6, 4, 4 respectively). The oral steroid and immunosuppressive agents were tapered in all patients without recrudescence of ocular inflammation, and three patients discontinued their oral medicine at the last visit. The best corrected visual acuity (BCVA) of 3 patients was improved to varying degrees, and the BCVA of 1 patient remained at 20/20 (Snellen chart) from the first to the last consultation. CONCLUSION: The study provides an effective therapy of HUC-MSCs in maintaining remission in patients affected by uveitis refractory to previous immunosuppressant treatments.

Effects of bentonite on antibiotic resistance genes in biogas slurry and residue from thermophilic and mesophilic anaerobic digestion of food waste.

Anaerobic digestion (AD) processes of food waste (FW) have potential risk on environments due to the prevalence and dissemination of antibiotic resistance genes (ARGs). This study investigated the effect of bentonite on methane production and the abundance of ARGs in biogas slurry and residue during AD of FW. Results showed that methane production increased by 68.52% and 56.79% with 3 g/L and 5 g/L of bentonite in mesophilic and thermophilic digestion, respectively. Adding 5 g/L of bentonite effectively reduced the genes of ermB, ermF, tetQ, tetX, sul1, sul2 and intI1 with a range of 80.82% - 100.00% in biogas residue under mesophilic reactor. The abundance of ARGs and intI1 in biogas residue were lower than in slurry under both temperatures with 5 g/L of bentonite. Statistical analysis indicated that bentonite and temperature were main driver factors which could impact ARGs by influencing the abundance and structure of microbial communities.

Effects of nanoscale zero-valent iron on the performance and the fate of antibiotic resistance genes during thermophilic and mesophilic anaerobic digestion of food waste.

The effects of nanoscale zero-valent iron (nZVI) on the performance of food waste anaerobic digestion and the fate of antibiotic resistance genes (ARGs) were investigated in thermophilic (TR) and mesophilic (MR) reactors. Results showed that nZVI enhanced biogas production and facilitated ARGs reduction. The maximum CH4 production was 212.00 ±â€¯4.77 ml/gVS with 5 g/L of nZVI in MR. The highest ARGs removal ratio was 86.64 ±â€¯0.72% obtained in TR at nZVI of 2 g/L. nZVI corrosion products and their contribution on AD performance were analyzed. The abundance of tetracycline genes reduced significantly in nZVI amended digesters. Firmicutes, Chloroflexi, Proteobacteria and Spirochaetes showed significant positive correlations with various ARGs (p < 0.05) in MR and TR. Redundancy analysis indicated that microbial community was the main factor that influenced the fate of ARGs. nZVI changed microbial communities, with decreasing the abundance bacteria belonging to Firmicutes and resulting in the reduction of ARGs.

Metabolomic insights into polyhydroxyalkanoates production by halophilic bacteria with acetic acid as carbon source.

A metabolomics method was established to analyze changes of intracellular metabolites and study the mechanism for enhancing polyhydroxyalkanoates production by halotolerant bacteria, Bacillus cereus strain HY-3, using acetic acid as carbon source. Maximum poly(3-hydroxybutyrate) (PHB) contents for the medium with 0.5 g/L and 5.0 g/L of acetic acid were 41.0 ± 0.415% and 49.2 ± 1.21%. Principal components analysis revealed clear metabolic differences in different growth stages and different concentrations of carbon source. According to statistical analysis, 3-hydroxybutyrate (3-HB), serine, threonine, malate, and pyruvate were determined as potential biomarkers for PHB production. Moreover, metabolic pathways analysis indicated that high level of 3-HB in death phase was due to the limitation of carbon source. Metabolism of glycine, serine, and threonine was influential pathway for PHB production among amino acid metabolisms. High levels of organic acids from the TCA cycle could stimulate the carbon source flux into PHB biosynthetic pathway.

Author Correction: Effects of Mesenchymal Stem Cell-Derived Exosomes on Experimental Autoimmune Uveitis.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.

Effects of Mesenchymal Stem Cell-Derived Exosomes on Experimental Autoimmune Uveitis.

We previously demonstrated that mesenchymal stem cells (MSCs) ameliorated experimental autoimmune uveoretinitis (EAU) in rats. Recently, MSC-derived exosomes (MSC-Exo) were thought to carry functions of MSCs. In this study, we tested the effect of local administration of human MSC-Exo on established EAU in the same species. Rats with EAU induced by immunization with interphotoreceptor retinol-binding protein 1177-1191 peptide were treated by periocular injections of increasing doses of MSC-Exo starting at the disease onset for 7 consecutive days. The in vitro effects of MSC-Exo on immune cell migration and responder T cell proliferation were examined by chemotactic assays and lymphocyte proliferation assays, respectively. We found that MSC-Exo greatly reduced the intensity of ongoing EAU as their parent cells by reducing the infiltration of T cell subsets, and other inflammatory cells, in the eyes. Furthermore, the chemoattractive effects of CCL2 and CCL21 on inflammatory cells were inhibited by MSC-Exo. However, no inhibitory effect of MSC-Exo on IRBP-specific T cell proliferation was observed. These results suggest that MSC-Exo effectively ameliorate EAU by inhibiting the migration of inflammatory cells, indicating a potential novel therapy of MSC-Exo for uveitis.

Exosomes derived from MSCs ameliorate retinal laser injury partially by inhibition of MCP-1.

Although accumulated evidence supports the notion that mesenchymal stem cells (MSCs) act in a paracrine manner, the mechanisms are still not fully understood. Recently, MSC-derived exosomes (MSC-Exos), a type of microvesicle released from MSCs, were thought to carry functional proteins and RNAs to recipient cells and play therapeutic roles. In the present study, we intravitreally injected MSCs derived from either mouse adipose tissue or human umbilical cord, and their exosomes to observe and compare their functions in a mouse model of laser-induced retinal injury. We found that both MSCs and their exosomes reduced damage, inhibited apoptosis, and suppressed inflammatory responses to obtain better visual function to nearly the same extent in vivo. Obvious down-regulation of monocyte chemotactic protein (MCP)-1 in the retina was found after MSC-Exos injection. In vitro, MSC-Exos also down-regulated MCP-1 mRNA expression in primarily cultured retinal cells after thermal injury. It was further demonstrated that intravitreal injection of an MCP-1-neutralizing antibody promoted the recovery of retinal laser injury, whereas the therapeutic effect of exosomes was abolished when MSC-Exos and MCP-1 were administrated simultaneously. Collectively, these results suggest that MSC-Exos ameliorate laser-induced retinal injury partially through down-regulation of MCP-1.

CD73 Pathway Contributes to the Immunosuppressive Ability of Mesenchymal Stem Cells in Intraocular Autoimmune Responses.

Mesenchymal stem cells (MSCs) exhibit a potent immunomodulatory capacity and have been applied to treat diseases such as graft versus host disease and severe autoimmune diseases. However, the mechanism underlying their immunosuppressive effect is not yet completely understood. Here, we investigated the role of the CD73/adenosine pathway in immune modulation by MSCs using a mouse model of experimental autoimmune uveitis (EAU). Moreover, we examined the in vitro modulatory effect of MSCs mediated through the CD73/adenosine pathway in human and mouse T cells. We found that the severity of EAU was significantly attenuated by MSCs; however, most therapeutic effects of MSCs were lost by pretreatment with a CD73 inhibitor. The inhibitory mechanism of MSCs might be contributed by CD73 on MSCs that cooperated with CD39 and CD73 on activated T cells to produce adenosine, resulting in inhibition of T-cell proliferation. Furthermore, MSCs increased the expression of CD73 on CD4(+) T cells, and transforming growth factor-ß1 (TGF-ß1) was the only tested cytokine that contributed to upregulation of CD73. Hence, our study demonstrates that the CD73/adenosine pathway involves the immunomodulatory function of MSCs in autoimmune responses.