Normal Scar Research Articles

A Signature of Genes Featuring FGF11 Revealed Aberrant Fibroblast Activation and Immune Infiltration Properties in Keloid Tissue.

Keloid is a fibroproliferative disorder in the skin, which manifested with extensive deposition of collagen and extracellular matrix. Its etiology remains a mystery and its recurrence rate remains high despite combinative treatment regimens. Current hypotheses of its pathogenesis centered on the role of inflammatory processes as well as immune infiltration in the microenvironment. However, there are a lot of discrepancies when it comes to the verification of certain well-recognized pathways involved in the dysfunctional fibroblast. Further exploration and characterization are required to reveal the driving force and even leading genes responsible for keloid formation. In this study, we provided supportive evidence of the immunologic nature of keloids distinct from normal fibroblasts and physiological scars by incorporating multiple available expressional profiles in the Gene Expression Omnibus (GEO). Through differential analyses and functional analyses, we identified a set of genes that successfully captures the dissimilarities between keloid lesions and nonlesions. They were differentially regulated in keloid samples and had opposite behavior in exposure to hydrocortisone. A key signature of six genes featuring FGF11 not only was highly correlated with significantly dysregulated fibroblast activation but also reflected various levels of immune cell infiltration. FGF11, in particular, revealed the heterogenous immunologic nature of keloid lesions. This study further supported that aberrant fibroblast was one of the main contributing factors and shed some light on investigating immune properties in future studies.

P-747 Prevalence of Caesarean scar defect (CSD) and clinical impact on reproductive outcomes in patients undergoing in vitro fertilization (IVF)

Abstract Study question Can sonohysterography-detected CSD have a deleterious effect on reproductive outcomes in women with secondary infertility undergoing IVF? Summary answer A 68% prevalence of CSD has been estimated in the study population. CSD significantly reduces the ongoing pregnancy rates in women undergoing IVF cycles. What is known already here is limited and conflicting evidence about the relation between Caesarean section, CSD and IVF treatment outcomes. Notably, most of the previous studies did not investigate the presence of CSD in infertile women with a history of Caesarian delivery. Furthermore, even if considered, the diagnosis of CSD is commonly based on transvaginal ultrasound assessment. Given the lower sensitivity of this method compared to saline infusion sonohysterography, an underestimation of the real effect is plausible. Study design, size, duration Retrospective cohort study of 122 women with a history of Caesarean delivery and an indication to IVF for secondary infertility referred to an Italian public assisted reproductive centre between 1 January 2016 and 30 April 2021. Participants/materials, setting, methods Women aged 18-43 with secondary infertility and a history of Caesarean delivery, tested for CSD with a saline infusion sonohysterography were included. Women with congenital uterine anomalies, intra-cavitary uterine pathologies, adenomyosis or previous myomectomy were excluded. CSD was defined as an anechoic indentation on the cesarean scar at the midsagittal plane, with a depth ≥ 2 mm. Main results and the role of chance Among the 122 women included, 83 (68%) were diagnosed with CSD by saline infusion sonohysterography. One hundred and fourteen patients underwent at least one IVF cycle: 76 (case group) had a CSD, 38 (control group) had a normal Caesarean scar. Groups were similar in patients and treatment characteristics. Clinical pregnancy rate was 43% in the CSD group and 71% in the control group (odds ratio [OR] 0.31; 95% confidence interval [CI], 0.14-0.72; p = .006). Ongoing pregnancy rates were 33% and 58%, respectively (OR, 0.34; 95% CI, 0.15-0.76; p = .015). Similar results were obtained after adjusting for potential confounders on the regression analysis. Ectopic pregnancy and miscarriage rate along with obstetric and neonatal complications were similar in both groups. In a subgroup-analysis comparing women with CSD who achieved pregnancy and those who did not, pregnant patients had a greater residual myometrial thickness compared to those who did not get pregnant, even if this finding did not reach statistical significance (5.3 mm versus 4.4 mm, p .07). Limitations, reasons for caution Retrospective nature of the study does not permit to infer a causal association between CSD and the reduced chance of pregnancy. Our results are representative of an infertile population but generalization needs further investigation. The sample size was too small to reveal the potential effect on obstetric and neonatal outcomes. Wider implications of the findings In infertile patients with a previous Caesarean section, the presence of CSD is more common than previously thought. The presence of CSD might significantly reduce the IVF success rate. This finding is important in relation to the rising of Caesarean section rates and the possible application of CSD surgical repair. Trial registration number not applicable

Macrophages of the M1 and M2 types play a role in keloids pathogenesis.

The pathophysiology of keloid formation is unknown, however, macrophages are thought to play a role in keloid formation. Understanding the mechanism(s) of keloid development might be crucial in developing a new treatment regimen for keloids. The aim of this study was to understand possible status of M1 and M2 type macrophages in the pathogenesis of keloid. Thirty cases of Keloid tissues were selected according to our inclusion and exclusion criteria, as well as 30 normal scars, were enrolled in our study as a control group. An excisional biopsy was harvested and ELISA was done on keloid tissue and normal scar samples, with CD68, the surface marker for M1 and CD163 representing M2. The results revealed the low expression of M1 (CD68) in keloid tissue meanwhile high levels of M1 were detected in normal scars. We also detected that higher tissue expression of M2 (CD163) was significantly associated with keloid cases when compared to low M2 expression in the control group. An important finding that was discovered during our study is that the M1 and M2 are significant predictors of keloid. Every increase of 1 ng/mL in M1 decreases the risk of keloid by 0.99 while every increase of one unit in M2 increases the risk of keloid by 2.01. This study concluded that the keloid formation could be a result of an abnormal response to tissue injury where there is an excessive entry of inflammatory cells into the wound, including macrophages and that the keloid incidence might be related to a decrease in M1 and an increase in M2.

536 Objective Quantification of Hypertrophic Scar and Donor Scar between 2 to 7 months Post-burn Injury

IntroductionVery few objective scar evaluations have been conducted with the burn survivor population, which limits our knowledge of the clinical recovery profile of hypertrophic scars (HSc) and donor site scars (D), thus having an impact on rehabilitation intervention and treatment prioritization. The purpose of this study was to prospectively quantify the thickness, pliability, erythema and pigmentation of post-burn HSc, donor sites (normal scar) and normal skin in different anatomical locations between 2 and 7 months post-burn using objective instrumentation. A secondary objective was to compare this data with anatomic-specific normative data using the same objective instrumentation.MethodsSkin characteristics of HSc, D and N in 44 burn survivors were measured at 2, 3, 4, 5, 6 and 7 months post-burn using validated instrumentation: high-frequency ultrasound for thickness, Cutometer® to measure pliability and Mexameter® to measure erythema and pigmentation. Up to five sites were assessed on the same participant if their scar was located on the upper extremity (UE), lower extremity (LE) and trunk. A mixed model two-way analysis of variance was used to investigate the differences in means between sites at each time point and between time points at each site.ResultsThe results revealed that HSc sites were thicker than D and N at all time points and UE and trunk HSc were thicker than LE HSc at 7 months post-burn, pliability of trunk HSc did not improve over time, and UE HSc was more erythematous at 7 months compared to other anatomical sites whereas D erythema decreases from 2 to 7 months.ConclusionsScar management treatments should prioritize the UE and trunk sites which developed HSc during the first two months post-burn and continues to vary significantly from normal scar and normal skin at 7 months. Furthermore, these results provide preliminary evidence that the recovery profile of HSc varies at different anatomical sites and that thickness is the characteristic that distinguishes HSc from normal scar and normal skin.

609 Available non-invasive skin probes distinguish between normal skin and hypertrophic scar but not laser-treated scar

IntroductionSkin fibrosis is the most under-studied fibrotic condition, and as such, there are limited treatment options for hypertrophic scar (HTS). HTS is difficult to study because scars improve over time, thus making the distinction between natural improvements and interventions difficult. In addition, clinical, histologic, cellular, and molecular outcome metrics are not agreed upon among providers/researchers. It was hypothesized that a set of non-invasive skin probes would be able to distinguish between HTS and normal skin (NS), but not between HTS before and after treatment with fractional ablative laser, despite histologic-level evidence showing improvement with treatment.MethodsWounds were created and HTS were allowed to form (n=8 scars). At Day 77, HTS and NS (n=4) were assessed with non-invasive skin probes measuring elasticity and trans-epidermal water loss (TEWL). HTS were then treated with CO2 fractional ablative laser (FLSR) at 70 mJ, 250Hz, and 1% density. The same data, sample collection, and FLSR treatment was carried out at weeks 1, 2, 3 and 4. Formalin-fixed biopsies were processed and stained with H&E, Herovici, Masson’s Trichrome, and Picrosirius red stains. Collagen type and architecture were qualitatively evaluated. Rete ridge ratio (RRR) was calculated from H&E stains.ResultsAll stains distinguished between HTS and NS with ease. Herovici stain showed differences in collagen architecture and collagen type after treatment with FLSR. Pre laser treatment, elasticity was different between HTS and NS (290.50±32.90 N/m vs. 90.92±5.61 N/m, p< 0.001). However, elasticity did not change after FLSR at 1, 2, 3, or 4 weeks (248.17±48.54, 215.54±44.68, 295.29±37.5, 290.79±30.17, p >0.05). RRR was different between HTS and NS (1.22±0.11 vs. 1.51±0.03, p=0.08). FLSR induced a significant increase in RRR in HTSs (pre=1.3±0.1 vs. week 4=1.9±0.1, p< 0.05). Pre-treatment, TEWL was different between HTS and NS (12.96±1.99 g/m2h vs. 8.72±1.11 g/m2h, p=0.09). However, TEWL did not change after FLSR at 1, 2, 3, or 4 weeks (15.95±1.99, 20.35± 3.15, 15.9±1.96, 14.01±2.22, p >0.05).ConclusionsOutcome metrics that non-invasively measure the qualities of scar are critical for HTS research to evaluate treatment effectiveness. Currently available technologies distinguish between NS and HTS well. However, they are inadequate to distinguish scars pre- and post-treatment despite evidence at the histologic level that changes occur. New technologies should be developed that are able to more effectively demonstrate changes to HTS after treatment.

Managing Wound Healing with a High-Risk Patient: A Case Report

Wound healing is a complex, multi-step process. This process begins immediately after skin damage. The outcome of wound healing depends on the quality of each stage of this process: a normal or pathological scar. Violation of wound healing entails a decrease in the function of scar tissue as well as aesthetic dissatisfaction with the patient. This problem is especially important in aesthetic surgery. Patients who have come for beauty feel frustration, obtaining pathological scars. We have been dealing with the problem of wound healing after plastic surgery for about 10 years. Our approach includes the assessment of the risk of pathological wound healing and the treatment of high-risk patients. The risk assessment includes historical data on wound healing, signs of connective tissue dysfunction (especially patients with connective tissue dysplasia), and genetic polymorphisms of genes responsible for the structure of the components of the extracellular matrix of the skin. In the future, patients with a high risk of pathological scarring can be prescribed treatment after surgery. This article presents a clinical case in which we demonstrate our approach.

Integrated Analysis of Single-Cell and Spatial Transcriptomics in Keloids: Highlights on Fibrovascular Interactions in Keloid Pathogenesis

Studies on the etiopathogenesis of keloids mostly have focused on fibroblasts and their dysfunction. In this study, two cutting-edge technologies, single-cell RNA sequencing and spatial transcriptomics, were applied to uncover the underlying pathophysiology of keloids. Keloid tissue samples and normal skin control data were analyzed as well as those of patient-matched keloid and normal mature scar. Single-cell RNA sequencing revealed cellular heterogenicity such as fibroblasts, endothelial cells (ECs), and myofibroblasts within the keloids. Spatial transcriptomics results showed that disease-associated fibroblasts were enriched in the deeper keloid areas, mostly located around the spots with endothelial transcripts. Mesenchymal activation was observed in keloid ECs, characterized by dysregulation of TGF-β/SMAD signaling. Colocalization of mesenchymal and vascular markers through multiplex immunofluorescence suggested mesenchymal activation of keloid ECs. Cell‒cell interaction analysis identified a significant network between keloid fibroblasts and ECs, and this cellular crosstalk was supported by colocalization analysis of spatial transcriptomics. This study depicted the cellular landscape of keloids at a single-cell resolution as well as the integration of single-cell and valuable spatial data of keloids using spatial transcriptomics and multiplex immunofluorescence technologies. Our findings suggested a potential role of fibrovascular communication and mesenchymal activation of ECs that might be involved in the keloid pathogenesis.

A comparison of proliferation levels in normal skin, physiological scar and keloid tissue

Proliferation is an important characteristic of life, and many signaling pathways participate in this complicated process. The MAPK/Erk pathway is a classic pathway in cell proliferation. In this study, expression levels of key factors in the MAPK/Erk pathway were measured to assess the proliferation level among normal skin, physiological scar, and keloid tissue. Thirty patients were selected randomly from the Department of Plastic Surgery at Peking Union Medical College Hospital from January 2019 to December 2020. Histological appearance and fiber tissue content were observed by Hematoxylin and eosin staining and Masson staining. Expression levels of key factors in the MAPK/Erk pathway (ATF2, c-Jun, c-Myc, p38 and STAT1) and relative proteins (HIF-1α and PCNA) in tissues were detected by immunohistochemistry and analyzed as the percentage of positively stained cells in both the tissue epidermis and dermis. Western blot was used for quantitative analysis of the above factors. In results, keloid tissue showed a significantly higher fiber and less cell content. In the immunohistochemical result, higher expression of key factors was observed in the epidermis than in the dermal layer, and the expression of all factors was increased remarkably in keloid tissue. In western blot analysis, all factors (except STAT1) showed higher expression in keloid tissue. In our former research, keloid showed similar apoptosis level as physiological scar and normal skin. On combining our former conclusion and results in this study, an imbalance condition between the high proliferation level and normal apoptosis level may lead to the growth characteristics of keloid.

A Methylome and Transcriptome Analysis of Normal Human Scar Cells Reveals a Role for FOXF2 in Scar Maintenance

Scars are maintained for life and increase in size during periods of growth such as puberty. Epigenetic changes in fibroblasts after injury may underpin the maintenance and growth of scars. In this study, we combined methylome and transcriptome data from normotrophic mature scar and contralateral uninjured normal skin fibroblasts to identify potential regulators of scar maintenance. In total, 219 significantly differentially expressed and 1,199 significantly differentially methylated promoters were identified, of which there were 12 genes both significantly differentially methylated and expressed. Of these, the two transcription factors, FOXF2 and MKX, were selected for further analysis. Immunocytochemistry and qPCR suggested that FOXF2 but not MKX had elevated expression in scar fibroblasts. Using RNA sequencing, FOXF2 knockdown was shown to significantly reduce the expression of extracellular matrix‒related genes, whereas MKX did not appear to affect similar pathways. Finally, FOXF2 knockdown was also shown to significantly decrease collagen I production in scar and keloid fibroblasts. This study provides insights into the maintenance of normotrophic scar, suggesting that FOXF2 is an important regulator of this process. Targeting genes responsible for maintenance of scar phenotype may ameliorate scar appearance and improve patient outcomes in the future.

Integrated bioinformatics analysis of core regulatory elements involved in keloid formation

BackgroundKeloid is a benign fibro-proliferative dermal tumor formed by an abnormal scarring response to injury and characterized by excessive collagen accumulation and invasive growth. The mechanism of keloid formation has not been fully elucidated, especially during abnormal scarring. Here, we investigated the regulatory genes, micro-RNAs (miRNAs) and transcription factors (TFs) that influence keloid development by comparing keloid and normal scar as well as keloid and normal skin.MethodsGene expression profiles (GSE7890, GSE92566, GSE44270 and GSE3189) of 5 normal scar samples, 10 normal skin samples and 18 keloid samples from the Gene Expression Omnibus (GEO) database were interrogated. Differentially expressed genes (DEGs) were identified between keloid and normal skin samples as well as keloid and normal scar samples with R Project for Statistical Computing. Gene Ontology (GO) functional enrichment analysis was also performed with R software. DEG-associated protein–protein interaction (PPI) network was constructed by STRING, followed by module selection from the PPI network based on the MCODE analysis. Regulatory relationships between TF/miRNA and target genes were predicted with miRnet and cytoscape. Core regulatory genes were verified by RT-qPCR.ResultsWe identified 628 DEGs, of which 626 were up-regulated and 2 were down-regulated. Seven core genes [neuropeptide Y(NPY), 5-hydroxytryptamine receptor 1A(HTR1A), somatostatin (SST), adenylate cyclase 8 (ADCY8), neuromedin U receptor 1 (NMUR1), G protein subunit gamma 3 (GNG3), and G protein subunit gamma 13 (GNG13)] all belong to MCODE1 and were enriched in the “G protein coupled receptor signaling pathway” of the GO biological process category. Furthermore, nine core miRNAs (hsa-mir-124, hsa-let-7, hsa-mir-155, hsa-mir-26a, hsa-mir-941, hsa-mir-10b, hsa-mir-20, hsa-mir-31 and hsa-mir-372), and two core TFs (SP1 and TERT) were identified to play important roles in keloid formation. In the TF/miRNA-target gene network, both hsa-mir-372 and hsa-mir-20 had a regulatory effect on GNG13, ADCY8 was predicted to be target by hsa-mir-10b, and HTR1A and NPY were potentially by SP1. Furthermore, the expression of core regulatory genes (GNG13, ADCY8, HTR1A and NPY) was validated in clinical samples.ConclusionsGNG13, ADCY8, NPY and HTR1A may act as core genes in keloid formation and these core genes establish relationship with SP1 and miRNA (hsa-mir-372, hsa-mir-20, hsa-mir-10b), which may influence multiple signaling pathways in the pathogenesis of keloid.

Systemic Fibrocyte Levels and Keloid Expression of the Chemoattractant CXCL12 Are Upregulated Compared With Patients With Normal Scar.

Fibrocytes are bone marrow mesenchymal precursors with a surface phenotype compatible with leukocytes, fibroblasts, and hematopoietic progenitors that have been shown to traffic to wound healing sites in response to described chemokine pathways. Keloids are focal fibrotic responses to cutaneous trauma characterized by disordered collagen, which may be associated with elevated systemic fibrocyte levels and/or wound bed chemokine expression. Blood specimens from patients with longstanding keloids and those who form grossly normal scars were assayed by fluorescence activated cell sorting analysis for fibrocytes (CD45+, Col I+). The expression of the fibrocyte chemotactic cell surface marker CXCR4, intracellular markers of fibroblast differentiation (pSMAD2/3), and plasma levels of the CXCR4 cognate CXCL12 were compared. Keloid specimens and grossly normal scars were excised, and local expression of CXCL12 was assayed. Keloid-forming patients demonstrated a significantly greater number of circulating fibrocytes (17.4 × 105 cells/mL) than control patients (1.01 × 105 cells/mL, P = 0.004). The absolute number of fibrocytes expressing CXCR4 was significantly greater (P = 0.012) in keloid-forming patients. Systemic CXCL12 levels were insignificantly greater in keloid-forming patients than controls. Keloid specimens had significantly greater CXCL12 expression (529.3 pg/mL) than normal scar (undetectable). Systemic fibrocyte levels and the CXCR4/CXCL12 biologic axis responsible for fibrocyte trafficking to areas of regional fibrosis were both upregulated in patients who form keloids compared with controls. Keloids persistently expressed CXLC12, which serves both as the main chemoattractant for fibrocytes and a downstream mediator for local inflammation, suggesting a role for this biologic axis in keloid formation and possibly recurrence.

Fibroblasts: Heterogeneous Cells With Potential in Regenerative Therapy for Scarless Wound Healing.

In recent years, research on wound healing has become increasingly in-depth, but therapeutic effects are still not satisfactory. Occasionally, pathological tissue repair occurs. Influencing factors have been proposed, but finding the turning point between normal and pathological tissue repair is difficult. Therefore, we focused our attention on the most basic level of tissue repair: fibroblasts. Fibroblasts were once considered terminally differentiated cells that represent a single cell type, and their heterogeneity was not studied until recently. We believe that subpopulations of fibroblasts play different roles in tissue repair, resulting in different repair results, such as the formation of normal scars in physiological tissue repair and fibrosis or ulcers in pathological tissue repair. It is also proposed that scarless healing can be achieved by regulating fibroblast subpopulations.

Performance of a machine-learning algorithm for fully automatic LGE scar quantification in the large multi-national derivate registry

Abstract Funding Acknowledgements Type of funding sources: Other. Main funding source(s): J. Schwitter receives research support by “ Bayer Schweiz AG “. C.N.C. received grant by Siemens. Gianluca Pontone received institutional fees by General Electric, Bracco, Heartflow, Medtronic, and Bayer. U.J.S received grand by Astellas, Bayer, General Electric. This work was supported by Italian Ministry of Health, Rome, Italy (RC 2017 R659/17-CCM698). This work was supported by Gyrotools, Zurich, Switzerland. Background Late Gadolinium enhancement (LGE) scar quantification is generally recognized as an accurate and reproducible technique, but it is observer-dependent and time consuming. Machine learning (ML) potentially offers to solve this problem. Purpose to develop and validate a ML-algorithm to allow for scar quantification thereby fully avoiding observer variability, and to apply this algorithm to the prospective international multicentre Derivate cohort. Method The Derivate Registry collected heart failure patients with LV ejection fraction &amp;lt;50% in 20 European and US centres. In the post-myocardial infarction patients (n = 689) quality of the LGE short-axis breath-hold images was determined (good, acceptable, sufficient, borderline, poor, excluded) and ground truth (GT) was produced (endo-epicardial contours, 2 remote reference regions, artefact elimination) to determine mass of non-infarcted myocardium and of dense (≥5SD above mean-remote) and non-dense scar (&amp;gt;2SD to &amp;lt;5SD above mean-remote). Data were divided into the learning (total n = 573; training: n = 289; testing: n = 284) and validation set (n = 116). A Ternaus-network (loss function = average of dice and binary-cross-entropy) produced 4 outputs (initial prediction, test time augmentation (TTA), threshold-based prediction (TB), and TTA + TB) representing normal myocardium, non-dense, and dense scar (Figure 1).Outputs were evaluated by dice metrics, Bland-Altman, and correlations. Results In the validation and test data sets, both not used for training, the dense scar GT was 20.8 ± 9.6% and 21.9 ± 13.3% of LV mass, respectively. The TTA-network yielded the best results with small biases vs GT (-2.2 ± 6.1%, p &amp;lt; 0.02; -1.7 ± 6.0%, p &amp;lt; 0.003, respectively) and 95%CI vs GT in the range of inter-human comparisons, i.e. TTA yielded SD of the differences vs GT in the validation and test data of 6.1 and 6.0 percentage points (%p), respectively (Fig 2), which was comparable to the 7.7%p for the inter-observer comparison (n = 40). For non-dense scar, TTA performance was similar with small biases (-1.9 ± 8.6%, p &amp;lt; 0.0005, -1.4 ± 8.2%, p &amp;lt; 0.0001, in the validation and test sets, respectively, GT 39.2 ± 13.8% and 42.1 ± 14.2%) and acceptable 95%CI with SD of the differences of 8.6 and 8.2%p for TTA vs GT, respectively, and 9.3%p for inter-observer. Conclusions In the large Derivate cohort from 20 centres, performance of the presented ML-algorithm to quantify dense and non-dense scar fully automatically is comparable to that of experienced humans with small bias and acceptable 95%-CI. Such a tool could facilitate scar quantification in clinical routine as it eliminates human observer variability and can handle large data sets.

Single-cell RNA-seq reveals fibroblast heterogeneity and increased mesenchymal fibroblasts in human fibrotic skin diseases

Fibrotic skin disease represents a major global healthcare burden, characterized by fibroblast hyperproliferation and excessive accumulation of extracellular matrix. Fibroblasts are found to be heterogeneous in multiple fibrotic diseases, but fibroblast heterogeneity in fibrotic skin diseases is not well characterized. In this study, we explore fibroblast heterogeneity in keloid, a paradigm of fibrotic skin diseases, by using single-cell RNA-seq. Our results indicate that keloid fibroblasts can be divided into 4 subpopulations: secretory-papillary, secretory-reticular, mesenchymal and pro-inflammatory. Interestingly, the percentage of mesenchymal fibroblast subpopulation is significantly increased in keloid compared to normal scar. Functional studies indicate that mesenchymal fibroblasts are crucial for collagen overexpression in keloid. Increased mesenchymal fibroblast subpopulation is also found in another fibrotic skin disease, scleroderma, suggesting this is a broad mechanism for skin fibrosis. These findings will help us better understand skin fibrotic pathogenesis, and provide potential targets for fibrotic disease therapies.

Cardiac reparative therapy with adipose graft transposition procedure reduces slow conduction areas in a chronic myocardial infarction swine model

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Insituto de Salud Carlos III BACKGROUND Cardiac regenerative therapy is a promising treatment for patients with ischemic heart disease, but there are some concerns on the potential increased risk of arrhythmic events following specific cell therapies. Adipose graft transposition procedure (AGTP) is a cardiac reparative therapy consisting in transposing a vascularized adipose flap from the autologous pericardium and placing it over the epicardial scar area and has demonstrated to reduce infarct size and improve the left ventricular ejection fraction in preclinical and human studies. Specific electrophysiological properties of the scar, (i.e. slow conduction velocity (CV)) have been identified as key features of ventricular tachycardia (VT) isthmuses. PURPOSE To assess the effect of the AGTP on VT inducibility and the electrophysiological properties of the post-MI scar with ultra-high density (UHD) mapping. METHODS A left circumflex artery (first marginal branch) MI was induced in 10 Landrace X Large White pigs by delivering 1-3 coils. Two weeks post-MI, all subjects underwent baseline left ventricular endocardial UHD mapping during right ventricular pacing with 64-electrode basket mapping catheter, as well as electrophysiological study (EPS) to test for VT inducibility. Following the mapping, subjects were allocated 1:1 to AGTP or sham group. UHD mapping and EPS were repeated 30 days post-treatment (6 weeks after MI). Voltage and activation maps were analyzed off-line with self-customized Paraview-based software. Voltage cut-offs of 1.5 and 0.5mV (bipolar) defined normal tissue, border zone (BZ) and dense scar, respectively, and 6.7mV for unipolar. Conduction velocity (CV) was determined for every pair of contiguous points and areas of similar CV were quantified for every 0.2m/s steps (for up to 4 m/s). RESULTS There were no differences between groups with regard of dense scar, BZ an low unipolar voltage areas. The AGTP group had a significant reduction of the size of slow CV (&amp;lt;0.2 m/s) areas, compared to the sham group in whom it increased (-4.1 ± 1.7 vs. +2.4 ± 1.6 mm2, p = 0.028)(Figure). There were no differences in the size of other ranges of CV. EPS did not induce VT in any subject at baseline, and only in 1 of the sham group at the follow-up EPS. CONCLUSIONS Cardiac reparative therapy with AGTP of post-MI scar reduced the size of slow conduction areas and could provide a protective effect against arrhythmic events in ischemic heart disease. Abstract Figure.

A CASE STUDY ON MANAGEMENT OF KELOID WITH VIRECHANA

Keloid refers to irregularly shaped, thickened, firm, hypertrophic, pink or red lesion associated with tenderness, pain and pruritis which is generally due to fibroblast dysfunction. Wound healing is a complex process involving fibroblasts, profibroblast along with overproduction of type 1 procollagen which takes about 3 months to one year for its manifestation. Where the skin restores the lost tissues by scar formation and the new cell is supplied by new vasculature and incorporates new connective tissues. There is a change in the organization of the components from that of the dermis. Some develop it more often due to preceding injury, skin trauma, due to acne, folliculitis, burns, body piercings, surgical wounds or sometimes without a cause and this deregulation makes it difficult to cure and is associated with high rate of recurrence. Normal scars seem to have negative feedback mechanism where the fibroblasts are mobilized to repair their cutaneous defect. This negative feedback mechanism is defective in keloid formation. It has an effect on the aesthetic beauty concept which can create a mental agony too. Thus, keeping the versatility of Shodhana (purificatory measure) in mind, an attempt was done to manage the condition with Shodhana and marked changes were noted in the condition. Keywords: Keloid, Vrana granthi, Shodhana,

Keloid tissue analysis discredits a role for myofibroblasts in disease pathogenesis.

Myofibroblasts, renowned for their contractility and extracellular matrix production, are widely considered the key effector cells for nearly all scars resulting from tissue repair processes, ranging from normal scars to extreme fibrosis. For example, it is often assumed that myofibroblasts underpin the characteristics of keloid scars, which are debilitating pathological skin scars lacking effective treatments because of a poor understanding of the disease mechanisms. Here, we present primary and published transcriptional and histological evidence that myofibroblasts are not consistently present in primary keloid lesions, and when alpha-smooth muscle actin (αSMA)-positive cells are detected, they are not greater in number or expressing more αSMA than in normal or hypertrophic scars. In conclusion, keloid scars do not appear to require αSMA-positive myofibroblasts; continuing to consider keloids on a quantitative spectrum with normal or hypertrophic scars, with αSMA serving as a biomarker of disease severity, is hindering advancement of understanding and therapy development.

How to Manage the Large Nonpedunculated Colorectal Polyp

How to Manage the Large Nonpedunculated Colorectal Polyp

Association between MeCP2 and Smad7 in the pathogenesis and development of pathological scars

To explore the relationship between methylated binding protein 2 (MeCP2) and mothers against decapentaplegic homolog 7 (Smad7) in the pathogenesis and development of pathological scars. Immunohistochemistry, Western blot and real-time polymerase chain reaction (RT-PCR) were used to detect the expression of MeCP2 in different types of human scars and hypertrophic scars at different growth times. The methylation status of Smad7 gene promoter in different scar tissues was determined by methylation-specific PCR. After transfection with MeCP2-siRNA (small interfering RNA) in human keloid fibroblasts, MTT assay was used to assess the proliferation activity of keloid fibroblasts, while RT-PCR and Western blot assays were used to detect the expression levels of MeCP2, transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA), phospho-Smad2 (p-Smad2) and Smad7. MeCP2 was mainly expressed in the nucleus of fibroblasts. The mRNA and protein levels of MeCP2 were significantly higher in keloids than in hypertrophic scars, normal scars and normal skin (p<.05). The expression level of MeCP2 in hypertrophic scars during the growth period of <6 months was markedly higher than that of >6 months (p<.05). The methylation level of Smad7 was significantly higher in keloids compared to normal skin. After MeCP2 silencing, the proliferation rate of human keloid fibroblasts was decreased, the mRNA and protein levels of Smad7 were increased, and the expression levels of TGF-β1, α-SMA and p-Smad2 were decreased (p<.05). MeCP2 and Smad7 play an important role in formation of pathological scars. During keloid formation, MeCP2 weakens the inhibitory effect of Smad7 on p-Smad2/3 by downregulating the expression of Smad7, which in turn promotes fibrosis and scar hyperplasia.

Attenuation corrected-optical coherence tomography for quantitative assessment of skin wound healing and scar morphology.

Imaging the structural modifications of underlying tissues is vital to monitor wound healing. Optical coherence tomography (OCT) images high-resolution sub-surface information, but suffers a loss of intensity with depth, limiting quantification. Hence correcting the attenuation loss is important. We performed swept source-OCT of full-thickness excision wounds for 300 days in mice skin. We used single-scatter attenuation models to determine and correct the attenuation loss in the images. The phantom studies established the correspondence of corrected-OCT intensity (reflectivity) with matrix density and hydration. We histologically validated the corrected-OCT and measured the wound healing rate. We noted two distinct phases of healing-rapid and steady-state. We also detected two compartments in normal scars using corrected OCT that otherwise were not visible in the OCT scans. The OCT reflectivity in the scar compartments corresponded to distinct cell populations, mechanical properties and composition. OCT reflectivity has potential applications in evaluating the therapeutic efficacy of healing and characterizing scars.