Scar Size Research Articles

Follistatin From hiPSC-Cardiomyocytes Promotes Myocyte Proliferation in Pigs With Postinfarction LV Remodeling.

When human induced pluripotent stem cells (hiPSCs) that CCND2-OE (overexpressed cyclin-D2) were differentiated into cardiomyocytes (CCND2-OEhiPSC-CMs) and administered to the infarcted hearts of immunodeficient mice, the cells proliferated after administration and repopulated >50% of the scar. Here, we knocked out human leukocyte antigen class I and class II expression in CCND2-OEhiPSC-CMs (KO/OEhiPSC-CMs) to reduce the cells' immunogenicity and then assessed the therapeutic efficacy of KO/OEhiPSC-CMs for the treatment of myocardial infarction. KO/OEhiPSC-CM and wild-type hiPSC-CM (WThiPSC-CM) spheroids were differentiated in shaking flasks, purified, characterized, and intramyocardially injected into pigs after ischemia/reperfusion injury; control animals were injected with basal medium. Cardiac function was evaluated via cardiac magnetic resonance imaging, and cardiomyocyte proliferation was assessed via immunostaining and single-nucleus RNA sequencing. Measurements of cardiac function and scar size were significantly better in pigs treated with KO/OEhiPSC-CM spheroids than in animals treated with medium or WThiPSC-CM spheroids. KO/OEhiPSC-CMs were detected for just 1 week after administration, but assessments of cell cycle activity and proliferation were significantly higher in the endogenous pig cardiomyocytes of the hearts from the KO/OEhiPSC-CM spheroid group than in those from the other 2 groups. Single-nucleus RNA-sequencing analysis identified a cluster of proliferating cardiomyocytes that was significantly more prevalent in the KO/OEhiPSC-CM spheroid-treated hearts (3.65%) than in the hearts from the medium (0.89%) or WThiPSC-CM spheroid (1.33%) groups at week 1. YAP (Yes-associated protein) protein levels and nuclear localization were also significantly upregulated in pig cardiomyocytes after treatment with KO/OEhiPSC-CM spheroids. Follistatin, which interacts with the HIPPO/YAP pathway, was significantly more abundant in the medium from KO/OEhiPSC-CM spheroids than WThiPSC-CM spheroids (30.29±2.39 versus 16.62±0.83 ng/mL, P=0.0056). Treatment with follistatin increased WThiPSC-CM cell counts by 28.3% over 16 days in culture and promoted cardiomyocyte proliferation in the infarcted hearts of adult mice. KO/OEhiPSC-CM spheroids significantly improved cardiac function and reduced infarct size in pig hearts after ischemia/reperfusion injury by secreting follistatin, which upregulated HIPPO/YAP signaling and proliferation in endogenous pig cardiomyocytes.

Feasibility of Periauricular Incision for Parotidectomy: A Retrospective Study.

Various incisions are used in parotid surgery, yet they often result in visible scarring on the neck and potential deformities. This study aimed to assess the efficacy and feasibility of utilizing a periauricular approach for excising parotid tumors irrespective of their location and size. This retrospective study involved 46 consecutive patients with parotid lesions, who underwent minimally invasive excision through a periauricular incision performed by a single surgeon between January 2022 and January 2024. Basic demographic data were recorded. Parameters such as the need for incision extension, total volume of drainage, length of hospital stay, postoperative complications, and final pathological diagnosis were also documented. The main trunk of the facial nerve was successfully identified in all patients, and the tumors were excised without requiring incision extension. Warthin tumor was the most common pathology (n=20, 43%). Three patients (6.5%) had malignant tumors. The inferior portion of the superficial lobe was the most frequent tumor location (n=18, 39%), with 11% of patients having deep lobe parotid tumors. Median operation time was 114 min (range, 90-170), tumor size was 2.8cm (range, 1-6.5cm), and drainage volume was 45mL (range, 10-80). Postoperative complications occurred in 7 patients, with sialocele formation being the most common (n=5, 11%). No serious complications like permanent facial paralysis or skin necrosis were observed. The periauricular approach is feasible for parotid tumors across all levels of the parotid gland. Its cosmetic advantage lies in the smaller scar size that does not extend into the neck.

CLINICAL PROFILE AND MANAGEMENT OF HYPERTROPHIC SCARS AT DR. SOETOMO GENERAL ACADEMIC HOSPITAL SURABAYA, INDONESIA

Highlights: Most hypertrophic scar patients were aged 17 to 25, mainly female, and the scars were mostly caused by burns on the face and upper extremities. Surgical techniques were the most common treatment, with most scars measuring 1 to 5 cm². Abstract: Introduction: Scars is a prevalent clinical concern that is often more complex than commonly perceived. Hypertrophic and keloid scars results from atypical wound healing responses following trauma, characterized by an excessive deposition of connective tissue. In Indonesia, there are remains a paucity of data on the clinical profile and management of hypertrophic scars. To address this gap, researchers conducted a descriptive study to systematically examine various variables associated with hypertrophic scars, aiming to enhance understanding and inform more effective management strategies. Methods: This research is a retrospective descriptive analysis utilizing secondary data from medical records of patients with hypertrophic scars treated at Dr. Soetomo General Academic Hospital in Surabaya during the 2019 – 2020 period. The data were collected and processed according to key variables, including age, gender, occupation, etiology, anatomical location of the hypertrophic scars, scar size, and therapeutic interventions. Results:The number of patients with hypertrophic scars at Dr. Soetomo General Academic Hospital, Surabaya, Indonesia, from 2019 to 2020 was 68. Most patients were aged 17 to 25 years. The gender distribution was nearly equal, with 50% male and 50.9% female patients. Among the patients, 12.5% and 43% were students. Burns caused 37.5% and 25% of cases. The most common scar locations were the face (25%) and upper extremities (25%). Most scars measured 1 to 5 cm² (25%). The most common treatment was surgical techniques (80% and 48%). Conclusion: Most hypertrophic scar patients were aged 17 to 25 years, predominantly female, and students. The primary cause of the scars was burns, with the scars most commonly located on the face and upper extremities. The majority of scars measured between 1 and 5 cm², and surgical techniques were the most widely used therapy.

Effect of ventricular fibrillation on infarct size after myocardial infarction: a translational study

Ventricular fibrillation (VF)-induced cardiac arrest frequently complicates ST-segment elevation myocardial infarction (STEMI). Although larger infarct sizes (IS) correlate with a higher risk of VF, the influence of VF itself on IS has remained poorly investigated. To address this knowledge gap, we analyzed the effect of VF on IS in patients and two experimental models. From a prospective cohort, 30 STEMI patients with VF were matched 1:2 with STEMI patients without VF on the common determinants of IS. The primary endpoint was IS, assessed using the 48-h area under the curve (AUC) for troponin. We also compared IS in pigs with/without spontaneous VF during STEMI (n = 15/group), and in an isolated rat heart model of myocardial infarction with/without electrically induced VF (n = 7/group). After matching, the patient characteristics, including the area at risk (AR), were similar. IS was 33% lower in the VF group compared to the control group (troponin AUC 1.6 [0.5–3.3] 106 arbitrary units vs. 2.4 [0.9–4.1] 106 arbitrary units; p < 0.05), but infarct scar size (assessed using MRI and ECG) did not differ between the groups at 1 and 6 months. In both experimental models, IS, expressed as a percentage of AR, was lower (p < 0.05) in the VF group than in the control group. When common determinants of IS are comparable, VF occurring prior to myocardial infarction reperfusion appears to be associated with smaller IS. Nevertheless, this finding, observed under specific experimental conditions and in a highly selected group of patients, was not associated with reduced infarct scar size.Registration (HIBISCUS-STEMI cohort): ClinicalTrials.gov NCT05794022.

Global, segmental, and layer-specific two-dimensional speckle tracking echocardiography immediately after acute myocardial infarction as a predictive tool to assess myocardial viability and scar size.

The identification of myocardial scar is key in clinical decision-making after acute myocardial infarction (AMI). However, the gold standard that is cardiac magnetic resonance imaging (CMR) encounters limitations in terms of availability. Two-dimensional speckle tracking echocardiography (2D-STE) may be an accessible alternative in detecting scar and assessing scar transmurality. We aim to evaluate the predictive value of 2D-STE, encompassing measures of global, segmental and layer-specific strain, with respect to myocardial viability and scar size at 6 months follow-up. In 43 patients admitted for primary AMI, we conducted a comparative analysis of strain parameters (including global longitudinal strain (GLS), segmental longitudinal strain (SLS), layer-specific GLS and SLS and the transmural strain gradient from endocardium to epicardium) in relation to conventional echocardiographic parameters at baseline in predicting for scar size and the transmurality index, as measured by CMR, 6 months post enrollment. We demonstrate a moderate correlation between both GLS and conventional echocardiographic parameters, and scar size as well as transmurality index. Wall motion score index exhibited superior predictive performance over GLS and left ventricular ejection fraction in anticipating scar formation. At a cut-off of -13.3% for any scar and -11.5% for transmural scar, SLS can predict scar formation. Layer-specific strain did not provide added predictive value. SLS, but not layer-specific strain, during admission after AMI is an easy and accessible quantitative tool for predicting scar formation and transmurality extent at 6 months follow-up. GLS correlates well with scar size, suggesting its potential utility as a predictive tool.

The de novo purine synthesis enzyme Adssl1 promotes cardiomyocyte proliferation and cardiac regeneration

There is a short window during which the neonatal heart has the proliferative capacity to completely repair damage, an ability that is lost in adulthood. Inducing proliferation in adult cardiomyocytes by reactivating cell cycle reentry after myocardial infarction (MI) improves cardiac function. De novo purine synthesis is a critical source of nucleotides for cell proliferation. Here, using loss- and gain-of-function genetic approaches, we explored the role of the muscle-specific de novo purine synthesis enzyme Adssl1 in cardiac regeneration. Deletion of Adssl1 in mouse neonatal hearts reduced cardiomyocyte proliferation and attenuated heart regeneration after apical resection. Conversely, cardiomyocyte-specific Adssl1 overexpression extended the postnatal regenerative window and induced robust cell cycle reentry after MI, which decreased fibrotic scar size and improved cardiac function. RNA sequencing analysis suggested that Adssl1 overexpression induced strong dedifferentiation and cell cycle entry. Moreover, LC-MS/MS analysis showed that Adssl1 overexpression was associated with increased amounts of purine metabolites, including inosine, which is in clinical use. Administration of exogenous inosine promoted cardiac repair after MI in adult mice. At a molecular level, the increase in purine metabolite production mediated by Adssl1 enhanced the activity of the proliferation-promoting mTORC1 pathway. Our study identifies a role for Adssl1 in supporting cardiomyocyte proliferation and cardiac regeneration.

Fibroblast-specific deletion of ERBB4 impairs cardiac regeneration in neonatal mice

Abstract Background Scarless regeneration of the neonatal mammalian heart is a remarkable but poorly understood process. Previous studies have shown that paracrine actions of neuregulin-1 (NRG1) are indispensable to stimulate cardiomyocyte proliferation in the injured neonatal heart, and are mediated by activation of the ERBB4 receptor. It remains unclear, however, whether NRG1 also contributes to other steps of neonatal cardiac regeneration, e.g. to the control of collagen synthesis by fibroblasts, hence contributing to scarless healing. Purpose To study temporal evolution of the infarct size during 21 days after myocardial infarction (MI) in neonatal fibroblast-specific Erbb4 knock-out (KO) and wild type (WT) mice. We hypothesized that scarless healing would be completed within 21 days in WT mice, and incomplete in KO mice. Methods Fibroblast-specific hemizygous Erbb4 KO (FB-Erbb4-KO) mice were generated (Erbb4F/+ Col1a2-Cre+, C57BL/6 background), because fibroblast-specific nullizygous Erbb4 KO were not viable. Validation of the KO was performed both on RNA (qPCR) and protein (western blot) level. MI was induced in one-day old FB-Erbb4-KO and WT littermates of both sexes by permanent ligation of the left anterior descending (LAD) coronary artery under hypothermic anaesthesia. Pups from both groups were euthanized at 4 (WT, n=9; KO, n=11), 7 (WT, n=12; KO, n=4), 10 (WT, n=11; KO, n=7) and 21 (WT, n=16; KO, n=8) days post-MI. Following dissection, hearts were formalin-fixed and paraffin embedded for histopathological evaluation. Transverse sections were cut at 4 distinct locations from the ligation site to the apex and stained with Masson’s Trichrome for infarct scar measurement. Genotyping was performed post-mortem and unblinded after data analysis. Results Both the transgenic mice model and the LAD ligation technique were successfully validated. No significant differences in the initial infarct scar size were observed between WT and FB-Erbb4-KO mice 4 days post-MI (p=0.6556). 7-days post-MI the infarct scar size was 6-fold smaller compared to 4 days post-MI in both FB-Erbb4-KO and WT mice, with a trend towards a larger infarct scar size in the FB-Erbb4-KO group compared to WT (p=0.0989). At day 10 post-MI, infarct scar size in the FB-Erbb4-KO group was significantly larger (p=0.0499) compared to WT. At 21 days post-MI, the infarct size had almost completely disappeared in the WT group, but remained clearly present in the FB-Erbb4-KO group (Fig. 1, p=0.0099). Infarct scar sizes at all timepoints are graphically shown in Fig. 2. Conclusion Fibroblast-specific deletion of the ERBB4 receptor causes incomplete resolution of the infarct scar in murine neonates 21 days after MI. While the initial infarct scar size 4 days after injury is the same, impaired scar resolution becomes progressively more evident towards day 21 post-MI. These findings indicate that scarless healing of the neonatal mammalian heart requires NRG1/ERBB4 signalling in fibroblasts.Fig. 1Fig. 2

Deep learning for quantification of myocardial scar and microvascular obstruction in late gadolinium enhancement cardiovascular magnetic resonance images

Abstract Objectives Detecting and measuring myocardial scar and microvascular obstruction (MVO) accurately following reperfusion therapy is clinically vital for patients with acute myocardial infarction (AMI). Current clinical practice relies on manual delineation of endocardial borders as well as hyper- and hypo-enhanced regions on late gadolinium enhancement (LGE) cardiovascular magnetic resonance (CMR) images for scar and MVO quantification, respectively. However, this approach is subjective, labor-intensive, and time-consuming. The objective of this study is to develop and assess a convolutional neural network (CNN)-based method for the automated quantification of LGE scar and MVO in AMI patients. Methods A total of 1836 LGE images from 252 patients with AMI were analyzed, with the cohort comprising 232 males and an average age of 56±9 years. Our approach, utilizing a combined architecture of cascaded YOLOv8 (You Only Look Once version 8) and nnU-Net, was implemented for the simultaneous segmentation of the left ventricular (LV) myocardium, scar, and MVO in LGE images. Independent datasets for training, validation, and testing were maintained in an 8:1:1 ratio. Performance evaluation was conducted by comparing the results to manual scar and MVO quantification, utilizing metrics such as the Dice similarity coefficient (DSC) and Bland-Altman analysis. Results The automated segmentation process took 0.05 seconds per image. The segmentation accuracy, as assessed by the DSC, yielded values of 0.96±0.04 (per-patient, n=25) and 0.96±0.05 (per-slice, n=184) for scar segmentation, and 0.86±0.17 (per-patient) and 0.89±0.15 (per-slice) for MVO segmentation. In per-patient analysis, the volumes of automatically segmented scar and MVO demonstrated strong agreement with manually derived results, with biases [limits of agreement] of -0.3 [-3.0, 2.4] cm³ and 0.1 [-0.2, 0.3] cm³, respectively. Furthermore, there was strong concordance between automatically and manually derived scar size as a percentage of LV mass (-0.2 [-3.1, 2.7] %) and MVO size as a percentage of scar (1.2 [-5.6, 8.0] %). Conclusions The findings demonstrate excellent agreement between our proposed automated deep-learning solution and clinical expert annotation, suggesting its potential as a predictive tool for prognosis and clinical outcomes in patients with AMI.

Molecular Scarring in Relapsing Psoriasis: Crosstalk of Keratinocytes and Immune Cells

Abstract Introduction/Objective Psoriasis is a chronic skin disorder with a tendency for relapse, characterized by immune- mediated mechanisms leading to papulosquamous lesions. Despite advances in treatment, many patients experience relapse, particularly in previously healed areas, referred to as “molecular scarring.” This scarring suggests a complex interaction between keratinocytes and immune cells, including tissue-resident memory T cells (TRMs), cytokines, and epigenetic modifications, contributing to the pattern-specific recurrence of psoriasis. Methods/Case Report After reviewing 31 articles on PubMed, we focused on the immunopathogenesis of psoriasis, examining the role of TRMs, the balance between regulatory T cells (Tregs) and Th17 cells, and the impact of various treatment modalities on disease recurrence. It evaluates the expression of TRM markers and the effectiveness of treatments such as anti-IL-17 agents and phototherapy in altering disease progression. The study also explores the genomic characteristics of psoriatic lesions post-treatment and the duration of clinical remission associated with different therapeutic approaches. Results (if a Case Study enter NA) The findings demonstrate a significant role of TRMs in psoriasis relapse, with environmental triggers reactivating these cells and leading to the production of inflammatory cytokines. Treatments targeting IL-17 showed a rapid decrease in TRM markers, particularly in the dermis, and were associated with different durations of disease remission based on the therapeutic approach. The study also found that the genomic profile of treated lesions does not fully normalize, with variations in gene expression linked to the type of treatment and possibly influencing the size of the molecular scar and relapse duration. Conclusion Psoriasis relapse is intricately linked to the presence of TRMs and the molecular scarring of previously healed lesions, influenced by various treatment modalities. The study underscores the importance of targeting TRMs and adjusting therapeutic strategies to extend the period of clinical remission and prevent the transition to more severe disease forms. Future research should focus on elucidating the detailed mechanisms of psoriasis relapse and developing targeted treatments to normalize the genomic profile of healed skin, ultimately improving patient outcomes and management of the disease.

Novel Fluorescence-Based Methods to Determine Infarct and Scar Size in Murine Models of Reperfused Myocardial Infarction.

Determination of infarct and scar size following myocardial infarction (MI) is commonly used to evaluate the efficacy of potential cardioprotective treatments in animal models. However, histological methods to determine morphological features in the infarcted heart have barely improved since implementation while still consuming large parts of the tissue and offering little options for parallel analyses. We aim to develop a new fluorescence technology for determining infarct area and area at risk that is comparable to 2,3,5-triphenyltetrazolium chloride (TTC) staining but allows for multiple analyses on the same heart tissue. For early and late time points following MI, we compared classical histochemical approaches with fluorescence staining methods. Reperfused MI was induced in male mice, the hearts were extracted 24 h, 7-, 21-, or 28-days later and fluorescently stained by combining Hoechst and phalloidin. This approach allowed for clear visualization of the infarct area, the area at ischemic risk and the remote area not affected by MI. The combined fluorescence staining correlated with the classic TTC/Evans Blue staining 24 h after MI (r = 0.8334). In later phases (>7 d) post-MI, wheat germ agglutinin (WGA) is equally accurate as classical Sirius Red (r = 0.9752), Masson's (r = 0.9920) and Gomori's Trichrome (r = 0.8082) staining for determination of scar size. Additionally, feasibility to co-localize fluorescence-stained immune cells in specific regions of the infarcted myocardium was demonstrated with this protocol. In conclusion, this new procedure for determination of post-MI infarct size is not inferior to classical TTC staining, yet provides substantial benefits, including the option for unbiased software-assisted analysis while sparing ample residual tissue for additional analyses. Overall, this enhances the data quality and reduces the required animal numbers consistent with the 3R concept of animal experimentation.

Evaluation of a Cosmetic Formulation Containing Arginine Glutamate in Patients with Burn Scars: A Pilot Study

Background: Patients with burn scars require effective treatments able to alleviate dry skin and persistent itching. Ion pairing has been employed in cosmetic formulations to enhance solubility in solvents and improve skin permeability. To evaluate the efficacy and safety of the cosmetic formula “RE:pair (arginine–glutamate ion pair)”, we analyzed scar size, itching and pain, skin barrier function, scar scale evaluation, and satisfaction in our study participants. Methods: A total of 10 patients were recruited, and the formula was used twice a day for up to 4 weeks. Results: Itching was significantly alleviated after 4 weeks of treatment (95% CI = −0.11–1.71) compared to before application (95% CI = 2.11–4.68). Transepidermal water loss (TEWL) showed an 11% improvement after 4 weeks (95% CI = 3.43–8.83) compared to before application (95% CI = 3.93–9.88), and skin coreneum hydration (SCH) showed a significant 41% improvement after 4 weeks (95% CI = 43.01–62.38) compared to before application (95% CI = 20.94–40.65). Conclusions: Based on the confirmation that RE:pair improves skin barrier function and relieves itching, it is likely to be used as a topical treatment for burn scars pending evaluation in follow-up studies (IRB no. HG2023-016).

In silico evaluation of cell therapy in acute versus chronic infarction: role of automaticity, heterogeneity and Purkinje in human

Human-based modelling and simulation offer an ideal testbed for novel medical therapies to guide experimental and clinical studies. Myocardial infarction (MI) is a common cause of heart failure and mortality, for which novel therapies are urgently needed. Although cell therapy offers promise, electrophysiological heterogeneity raises pro-arrhythmic safety concerns, where underlying complex spatio-temporal dynamics cannot be investigated experimentally. Here, after demonstrating credibility of the modelling and simulation framework, we investigate cell therapy in acute versus chronic MI and the role of cell heterogeneity, scar size and the Purkinje system. Simulations agreed with experimental and clinical recordings from ionic to ECG dynamics in acute and chronic infarction. Following cell delivery, spontaneous beats were facilitated by heterogeneity in cell populations, chronic MI due to tissue depolarisation and slow sinus rhythm. Subsequent re-entrant arrhythmias occurred, in some instances with Purkinje involvement and their susceptibility was enhanced by impaired Purkinje-myocardium coupling, large scars and acute infarction. We conclude that homogeneity in injected ventricular-like cell populations minimises their spontaneous beating, which is enhanced by chronic MI, whereas a healthy Purkinje-myocardium coupling is key to prevent subsequent re-entrant arrhythmias, particularly for large scars.

Microparticle Mediated Delivery of Apelin Improves Heart Function in Post Myocardial Infarction Mice.

Apelin is an endogenous prepropeptide that regulates cardiac homeostasis and various physiological processes. Intravenous injection has been shown to improve cardiac contractility in patients with heart failure. However, its short half-life prevents studying its impact on left ventricular remodeling in the long term. Here, we aim to study whether microparticle-mediated slow release of apelin improves heart function and left ventricular remodeling in mice with myocardial infarction (MI). A cardiac patch was fabricated by embedding apelin-containing microparticles in a fibrin gel scaffold. MI was induced via permanent ligation of the left anterior descending coronary artery in adult C57BL/6J mice followed by epicardial patch placement immediately after (acute MI) or 28 days (chronic MI) post-MI. Four groups were included in this study, namely sham, MI, MI plus empty microparticle-embedded patch treatment, and MI plus apelin-containing microparticle-embedded patch treatment. Cardiac function was assessed by transthoracic echocardiography. Cardiomyocyte morphology, apoptosis, and cardiac fibrosis were evaluated by histology. Cardioprotective pathways were determined by RNA sequencing, quantitative polymerase chain reaction, and Western blot. The level of endogenous apelin was largely reduced in the first 7 days after MI induction and it was normalized by day 28. Apelin-13 encapsulated in poly(lactic-co-glycolic acid) microparticles displayed a sustained release pattern for up to 28 days. Treatment with apelin-containing microparticle-embedded patch inhibited cardiac hypertrophy and reduced scar size in both acute and chronic MI models, which is associated with improved cardiac function. Data from cellular and molecular analyses showed that apelin inhibits the activation and proliferation of cardiac fibroblasts by preventing transforming growth factor-β-mediated activation of Smad2/3 (supporessor of mothers against decapentaplegic 2/3) and downstream profibrotic gene expression. Poly(lactic-co-glycolic acid) microparticles prolonged the apelin release time in the mouse hearts. Epicardial delivery of the apelin-containing microparticle-embedded patch protects mice from both acute and chronic MI-induced cardiac dysfunction, inhibits cardiac fibrosis, and improves left ventricular remodeling.

Association between in vitro susceptibility and clinical outcomes in fungal keratitis

PurposeThe purpose of this study was to assess the association between antifungal susceptibility as measured by minimum inhibitory concentration (MIC) and clinical outcomes in fungal keratitis.MethodsThis pre-specified secondary analysis of the Mycotic Ulcer Treatment Trial II (MUTT II) involved patients with filamentous fungal keratitis presenting to Aravind Eye Hospitals in South India. Antifungal susceptibility testing for natamycin and voriconazole was performed on all samples with positive fungal culture results according to Clinical and Laboratory Standards Institute Guidelines. The relationship between MIC and clinical outcomes of best-corrected visual acuity, infiltrate or scar size, corneal perforation, need for therapeutic penetrating keratoplasty, and time to re-epithelialization were assessed.ResultsWe obtained MIC values from 141 patients with fungal keratitis. The most commonly cultured organisms were Aspergillus (46.81%, n = 66) and Fusarium (44.68%, n = 63) species. Overall, there was no association between antifungal MICs and clinical outcomes. Subgroup analysis revealed that among Fusarium-positive cases, higher voriconazole MIC was correlated with worse three-month best-corrected visual acuity (p = 0.03), increased need for therapeutic penetrating keratoplasty (p = 0.04), and time to re-epithelialization (p = 0.03). No significant correlations were found among Aspergillus-positive cases. There were no significant correlations found between natamycin MIC and clinical outcomes among organism subgroups.ConclusionsDecreased susceptibility to voriconazole was associated with increased odds of requiring a therapeutic penetrating keratoplasty in Fusarium-positive cases. Susceptibility to natamycin was not associated with any of the measured outcomes.

A composite patch loaded with 2-Deoxy Glucose facilitates cardiac recovery after myocardial infarction via attenuating local inflammatory response

Local inflammatory microenvironment in the early stage of myocardial infarction (MI) severely impaired cardiac recovery post-MI. Macrophages play a pivotal role in this process. A classical glycolytic inhibitor, 2-Deoxy-Glucose (2-DG), has been found to regulate the excessive pro-inflammatory macrophage polarization in the infarcted myocardium. This study investigated the effect of 2-DG-loaded chitosan/gelatin composite patch on the infarct microenvironment post-MI and its impact on cardiac repair. The results showed that the 2-DG patch significantly inhibited the expression of inflammatory cytokines, alleviated reactive oxygen species (ROS) accumulation, repressed the proinflammatory polarization of macrophages, attenuated local inflammatory microenvironment in the ischemic hearts, as well as improved cardiac function, reduced scar size, and promoted angiogenesis post-MI. In terms of mechanism, 2-DG exerts anti-inflammatory effects through inhibiting the NF-κB signaling pathway and reducing the assembly and activation of the NLRP3 inflammasome. These findings suggest that 2-DG composite patch may represent a promising therapeutic strategy for cardiac repair after MI.

Predicting Late Gadolinium Enhancement of Acute Myocardial Infarction in Contrast-Free Cardiac Cine MRI Using Deep Generative Learning.

Late gadolinium enhancement (LGE) cardiac magnetic resonance (CMR) is a standard technique for diagnosing myocardial infarction (MI), which, however, poses risks due to gadolinium contrast usage. Techniques enabling MI assessment based on contrast-free CMR are desirable to overcome the limitations associated with contrast enhancement. We introduce a novel deep generative learning method, termed cine-generated enhancement (CGE), which transforms standard contrast-free cine CMR into LGE-equivalent images for MI assessment. CGE features with multislice spatiotemporal feature extractor, enhancement contrast modulation, and sophisticated loss function. Data from 430 patients with acute MI from 3 centers were collected. After image quality control, 1525 pairs (289 patients) of center I were used for training, and 293 slices (52 patients) of the same center were reserved for internal testing. The 40 patients (401 slices) of the other 2 centers were used for external testing. The CGE robustness was further tested in 20 normal subjects in a public cine CMR data set. CGE images were compared with LGE for image quality assessment and MI quantification regarding scar size and transmurality. The CGE method produced images of superior quality to LGE in both internal and external data sets. There was a significant (P<0.001) correlation between CGE and LGE measurements of scar size (Pearson correlation, 0.79/0.80; intraclass correlation coefficient, 0.79/0.77) and transmurality (Pearson correlation, 0.76/0.64; intraclass correlation coefficient, 0.76/0.63) in internal/external data set. Considering all data sets, CGE demonstrated high sensitivity (91.27%) and specificity (95.83%) in detecting scars. Realistic enhancement images were obtained for the normal subjects in the public data set without false positive subjects. CGE achieved superior image quality to LGE and accurate scar delineation in patients with acute MI of both internal and external data sets. CGE can significantly simplify the CMR examination, reducing scan times and risks associated with gadolinium-based contrasts, which are crucial for acute patients.

Efficacy of mesenchymal stem cell transplantation on major adverse cardiovascular events and cardiac function indices in patients with chronic heart failure: a meta-analysis of randomized controlled trials

BackgroundThe effects of mesenchymal stem cells (MSCs) on heart failure (HF) have been controversial. This study was conducted to investigate whether the transplantation of MSCs after HF could help improve clinical outcomes and myocardial performance indices.MethodsUsing a systematic approach, electronic databases were searched for randomized controlled trials (RCTs), which evaluated the transplantation of MSCs after HF. The outcomes owf interest included clinical outcomes and myocardial function indices. We also assessed the role of age, cause of heart failure, cell origin, cell number, type of donor (autologous/allogeneic), and route of cell delivery on these outcomes. Using the random-effects method, a relative risk (RR) or mean difference (MD) and their corresponding 95% confidence intervals (CI) were pooled.ResultsSeventeen RCTs including 1684 patients (927 and 757 patients in the intervention and control arms, respectively) were enrolled. The RR (95% CI) of mortality was 0.78 (0.62; 0.99, p = 0.04) in the MSC group compared to the controls. HF rehospitalization decreased in the MSC group (RR = 0.85 (0.71–1.01), p = 0.06), but this was only significant in those who received autologous MSCs (RR = 0.67 (0.49; 0.90), p = 0.008). LVEF was significantly increased among those who received MSC (MD = 3.38 (1.89; 4.87), p < 0.001). LVESV (MD = −9.14 (−13.25; −5.03), p < 0.001), LVEDV (MD = −8.34 −13.41; −3.27), p < 0.001), and scar size (standardized MD = -0.32 (-0.60; -0.05), p = 0.02) were significantly decreased. NYHA class (MD = −0.19 (−0.34; −0.06), p = 0.006), BNP level (standardized MD = −0.28 (−0.50; −0.06), p = 0.01), and MLHFQ (MD = −11.55 (−16.77; −6.33), p = 0.005) significantly decreased and 6-min walk test significantly improved (MD = 36.86 (11.22; 62.50), p = 0.001) in the MSC group. Trials were not affected by the participants’ etiology of heart failure, while trials with the autologous source of cells, MSC doses lower than 100 million cells, and intracoronary injection performed significantly better in some of the outcomes.ConclusionTransplantation of MSCs for ischemic or dilated heart failure patients may reduce all-cause mortality and improve clinical condition. Moreover, this treatment would improve left ventricular function indices and reduce scar size.Graphical

Intramyocardial immunomodulation with human CD16+ monocytes to treat myocardial infarction in pig: a blind randomized preclinical trial.

Human CD16+ monocytes (hCD16+ Ms) have proangiogenic properties. We assessed the feasibility, safety and efficacy of hCD16+ Ms in a porcine model of myocardial infarction (MI). A total of 27 female Large White pigs underwent MI with reperfusion and cardiac magnetic resonance (CMR). Five days later, animals received intramyocardial injections of hCD16+ Ms in saline (n = 13) or saline only (n = 14). hCD16+ Ms were selected from leucocyte cones. Feasibility/safety endpoints included injury at injected sites, malignant arrhythmias, cancer, haematoma, left ventricular (LV) dilatation, troponin release and downstream organ injury. Co-primary efficacy outcome included LV scar and ejection fraction (LVEF) at 30-day post-injections by CMR. Immunohistochemistry included neo-angiogenesis, fibrosis, markers of myofibroblast and inflammation. Four animals were excluded before injections due to untreatable malignant arrhythmias or lung injury. Median cell number and viability were 48.75 million and 87%, respectively. No feasibility/safety concerns were associated with the use of hCD16+ Ms. The LV scar dropped by 14.5gr (from 25.45 ± 8.24 to 10.8 ± 3.4 gr; -55%) and 6.4gr (from 18.83 ± 5.06 to 12.4 ± 3.9gr; -30%) in the hCD16+ Ms and control groups, respectively (p = 0.015). The 30-day LVEF did not differ between groups, but a prespecified sub-analysis within the hCD16+ Ms group showed that LVEF was 2.8% higher and LV scar 1.9gr lower in the subgroup receiving a higher cell dose. Higher tissue levels of neo-angiogenesis, myofibroblast and IL-6 and lower levels of TGF-β were observed in the hCD16+ Ms group. The use of hCD16+ Ms in acute MI is feasible, safe and associated with reduced LV scar size, increased tissue levels of neo-angiogenesis, myofibroblasts and IL-6 and reduced pro-fibrotic TGF-β at 30-day post-injections. A higher cell dose might increase the LVEF effect while reducing scar size, but this warrants validation in future studies.

Treating Scars After Burns With Pulsed Electric Fields in the Rat Model.

Reducing scar size after severe burn injuries is an important and challenging medical, technological, and social problem. We have developed a battery-powered pulsed electric field (PEF) device and surface needle electrode applicator to deliver PEFs to the healing dorsal burn wound in rats. The pulsed electric field was used to treat residual burn wounds caused by metal contact in rats starting 10 days after the injury for 4 months every 11 or 22 days for 4 months using varying time applied voltages at 250-350 V range, 400 mA current, 40 pulses, 70 μs duration each, delivered at pulse repetition frequency 10 Hz at 5 locations inside the wound. We found 40%-45% reduction in the scar size in comparison with untreated controls in both upper and lower dorsal locations on rats' backs 2 months after the last PEF application. We have not detected significant histopathological differences in the center of the scars besides the thickness of the newly generated epidermis, which was thicker in the PEF-treated group. We showed that minimally invasively applied PEFs through needle electrodes are effective method and device for treating residual burn wounds in the rat model, reducing the size of the resulting scars, without any adverse reaction.

Abstract We115: Modified mRNA Therapeutics Inhibits Cardiomyocyte Apoptosis and Induces Cardiac Protection in Ischemic Heart Diseases

Introduction: Cardiovascular diseases are the leading cause of mortality worldwide. After myocardial infarction (MI), there is a permanent loss of cardiomyocytes (CMs), and as the mammalian heart has limited regenerative capacity, it leads to Heart Failure. Extracellular vesicles (EVs) derived from induced pluripotent stem cells (iPSCs) have emerged as potential therapeutic agents, yet their precise mechanism of CM survival and cardiac repair remains elusive. Hypothesis: The myocardial delivery of hiPSC-EV-specific protein improves cardiac function and mice survival post-MI by inhibiting CM apoptosis and oxidative stress molecular pathways. Methods and Results: Our preliminary studies confirmed that hiPSC-EVs promote CM survival post-MI in mice, and through proteomic analysis, we identified a novel protein uniquely expressed in hiPSC-EVs. We showed that the myocardial delivery of hiPSC-EVs specific protein in the form of modRNA inhibited CM apoptosis and induced CM survival post-MI. This increase in the CM's survival by hiPSC-EVs specific protein expression was associated with reduced scar size, improved cardiac function, and mice survival 28 days post-MI. Furthermore, using the siRNA and small molecule inhibition of hiPSC-EVs specific protein, we found enhanced MI-induced CM apoptosis post-MI. Mechanistic insights reveal the hiPSC-EVs specific protein induced STAT3 pathway, while STAT3 pathway inhibition by small molecule repressed hiPSC-EVs specific protein induced CM survival in vitro . Additionally, our preliminary study shows this protein translocates to the nucleus post-MI or ischemic injury. Conclusion: Taken together, the myocardial injection of hiPSC-EVs specific protein through a highly therapeutic modRNA tool improves cardiac function by inhibiting CMs apoptosis, and reactivating cardiac repair post-injury. Moreover, our studies demonstrate that the modRNA (protein) approach in mouse models is potentially adaptable for mRNA therapeutics in a clinical setup.