Normal Scar Research Articles

β-Catenin-regulated myeloid cell adhesion and migration determine wound healing.

A β-catenin/T cell factor-dependent transcriptional program is critical during cutaneous wound repair for the regulation of scar size; however, the relative contribution of β-catenin activity and function in specific cell types in the granulation tissue during the healing process is unknown. Here, cell lineage tracing revealed that cells in which β-catenin is transcriptionally active express a gene profile that is characteristic of the myeloid lineage. Mice harboring a macrophage-specific deletion of the gene encoding β-catenin exhibited insufficient skin wound healing due to macrophage-specific defects in migration, adhesion to fibroblasts, and ability to produce TGF-β1. In irradiated mice, only macrophages expressing β-catenin were able to rescue wound-healing deficiency. Evaluation of scar tissue collected from patients with hypertrophic and normal scars revealed a correlation between the number of macrophages within the wound, β-catenin levels, and cellularity. Our data indicate that β-catenin regulates myeloid cell motility and adhesion and that β-catenin-mediated macrophage motility contributes to the number of mesenchymal cells and ultimate scar size following cutaneous injury.

Assessing quality of healing in skin: review of available methods and devices.

The process of wound healing is dynamic and takes place over months to years, during which there is a resolution of angiogenesis, continued wound contraction, and connective tissue remodeling. The outcome of this process is most commonly the formation of a scar, defined as a fibrous tissue replacing normal tissues destroyed by injury or disease. Scars often have a lowered or total loss of vital skin functions and imbue a large burden on both the patient and the health care system as a whole. Scar treatments are plentiful but are often unsatisfactory or inconsistent. No single treatment method has been universally adopted. To evaluate the clinical treatment as well as research focused on developing novel methods for scar management, objective studies of the progression of scar formation and the properties of mature scars are needed. Several parameters, including barrier function as well as mechanical and physiological properties, need to be taken into account when both categorizing and treating healing wounds and scars. To date, there is no available methodology that provides a comprehensive evaluation of a scar's properties. This review aims at presenting an overview of available scar assessment methods and devices, ranging from analysis of collagen properties in tissue biopsies to noninvasive methods for studies of mechanical parameters such as breaking strength and skin elasticity. In the cases where conclusive studies have been performed, the differences between normal skin and scar with respect to the above parameters are presented. Furthermore, this review highlights areas where the development of additional modalities are needed.

Immunohistochemical Evaluation of COX-1 and COX-2 Expression in Keloid and Hypertrophic Scar

Both keloids (KLs) and hypertrophic scars (HSs) are considered as dermal fibroproliferative diseases that differ clinically and histopathologically. Although several factors have been postulated in the etiopathogenesis of these conditions, there has been growing evidence to suggest the role of COXs in the pathogenesis of abnormal wound healing because of the reduction of formation of KL and HS in patients using nonsteroidal anti-inflammatory drugs and a COX-2 inhibitor. The aim of the present work is to evaluate the pattern and localization of COX-1 and COX-2 expression in KL and HS compared with surgical scars. COX-1 and COX-2 were analyzed on skin biopsies of 30 patients who presented with KL (15) and HS (15) and 10 normal surgical scars (controls). Both COX-1 and COX-2 were expressed not only in dermal components (fibroblasts, inflammatory cells, and endothelial cells) but also in keratinocytes of the overlying epidermis in the different studied scar lesions. The percentage of COX-1 expression increased progressively from surgical scar (40%) to HS (53.3%) to KL (100%) with a statistically significant difference (P = 0.002). COX-2 was expressed in 100% of surgical scars, 73.3% of HS and 86.7% of KL with the absence of significant differences (P > 0.05). The significant difference in COX-1 expression between HS and KL may refer to the presence of different pathways for the emergence of these diseases. The expression of COX-2 in all scars (normal or abnormal) indicates its active role as an inflammatory mediator. Keratinocytes play an active role in induction of scarring by up-regulation of inflammatory mediators, such as COX-1 and COX-2.

Expression and significance of sirtuin type 1 and forkhead box M1 in pathological scar

Objective To study the expression of sirtuin type 1 (SIRT1) and forkhead box M1 (FOXM1) gene in the pathological scars and to investigate its role and its probable mechanism in the pathogenesis of abnormal scar.Methods Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blotting was performed to detect the expression and distribution of mRNA and protein of SIRT1 and FOXM1 in normal skin (10 cases),normal scar (10 cases),and pathological scar (10cases),statistics was used to analyze the datas.Results The expression of SIRT1 mRNA and protein in the normal skin (10 cases),normal scar (10 cases),pathological scar (10 cases),were 1.0,1.187 982 ± 0.082 916,2.402 178 ± 0.178 598 ; 0.498 614 ± 0.056 342,0.534 159 ± 0.044 509,0.976 829 ±0.145 736.The expression of FOXM1 mRNA and protein in the normal skin (10 cases),normal scar (10 cases),pathological scar (10 cases),were 1.0,1.196 429 ±0.086 786,2.447 249 ±0.128 875; 0.469 935 ±0.089 373,0.552 653 ±0.078 293,0.915 367 ±0.114 097.The expression of SIRT1 and FOXM1 mRNA in pathologic scar were significantly higher than that in normal skin,normal scar (P ＜0.01) ; The expression of SIRT1 and FOXM1 protein in pathologic scar were all remarkably significant in comparison with normal skin and normal scar (P ＜ 0.05).Conclusion The result indicated that the expression of SIRT1 and FOXM1 was increased in pathological scar.Therefore,SIRT1 and FOXM1 overexpression may play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar. Key words: Sirtuin type 1 ; Forkhead box M1 ; Pathological scar

Impact of Myocardial Viability and Left Ventricular Lead Location on Clinical Outcome in Cardiac Resynchronization Therapy Recipients with Ischemic Cardiomyopathy

Cardiac resynchronization therapy (CRT) recipients with ischemic cardiomyopathy (ICM) have scar segments that may limit ventricular resynchronization and clinical response. The impact of myocardial viability at the left ventricular (LV) pacing site on CRT response is poorly elucidated. A retrospective cohort of 160 ICM patients with single photon emission computed tomography-myocardial perfusion imaging before device implantation were included. Coronary venous angiography and chest radiographs helped classify segmental location of LV lead (LVL). The primary outcome was a composite of heart failure (HF) hospitalization and mortality at 3 years, and secondary outcome was change in systolic function at 6 months. The patients were divided into groups based on the myocardial substrate at the site of LVL: LVL on or adjacent to (1) normal myocardium (LVL-N, n = 64), (2) segmental scar (LVL-S, n = 62), and (3) scar and ischemia (LVL-SI, n = 34). Upon follow-up, 75 (47%) patients reached primary endpoint with a higher incidence noted in LVL-S (60%), and LVL-SI (53%), compared to 31% in LVL-N (P = 0.004). Kaplan Meier method demonstrated poor event free survival for primary outcome in LVL-S (P = 0.002), and LVL-SI (P = 0.03). In Cox proportional hazard model, LVL-S (HR: 2.26, P = 0.004), and LVL-SI (1.9, P = 0.047) were independent predictors of primary outcome. In CRT recipients with ICM, scar and reversible ischemia in or adjacent to LV pacing site were independent predictors of HF hospitalization and death.

Clinical and endoscopic characteristics of drug-induced esophagitis.

To investigate clinical, endoscopic and pathological characteristics of drug-induced esophagitis. Data for patients diagnosed with drug-induced esophagitis from April 2002 to May 2013 was reviewed. Patients diagnosed with malignancy, viral or fungal esophagitis were excluded. Clinical, endoscopic and pathological characteristics of patients diagnosed with drug-induced esophagitis were analyzed. Seventy-eight patients were diagnosed with drug-induced esophagitis. Their mean age was 43.9 ± 18.9 years and 35.9% were male. Common symptoms were chest pain (71.8%), odynophagia (38.5%) and dysphagia (29.5%). The endoscopic location was in the middle third of esophagus in 78.2%. Endoscopic findings were ulcer (82.1%), erosion (17.9%), ulcer with bleeding (24.4%), coating with drug material (5.1%), impacted pill fragments (3.8%) and stricture (2.6%). Kissing ulcers were observed in 43.6%. The main causative agents were antibiotics and non-steroidal anti-inflammatory drugs. All the patients were treated with proton pump inhibitors (PPIs) or sucralfate, and the causative drugs were discontinued. Nineteen patients with drug-induced esophagitis were followed up with endoscopy and revealed normal findings, scars or healing ulcers. Drug-induced esophagitis mainly presents as chest pain, odynophagia and dysphagia, and may be successfully treated with PPIs and discontinuation of the causative drug. Kissing ulcers were observed in 43.6%.

Site-Specific Keloid Fibroblasts Alter the Behaviour of Normal Skin and Normal Scar Fibroblasts through Paracrine Signalling

Keloid disease (KD) is an abnormal cutaneous fibroproliferative disorder of unknown aetiopathogenesis. Keloid fibroblasts (KF) are implicated as mediators of elevated extracellular matrix deposition. Aberrant secretory behaviour by KF relative to normal skin fibroblasts (NF) may influence the disease state. To date, no previous reports exist on the ability of site-specific KF to induce fibrotic-like phenotypic changes in NF or normal scar fibroblasts (NS) by paracrine mechanisms. Therefore, the aim of this study was to investigate the influence of conditioned media from site-specific KF on the cellular and molecular behaviour of both NF and NS enabled by paracrine mechanisms. Conditioned media was collected from cultured primary fibroblasts during a proliferative log phase of growth including: NF, NS, peri-lesional keloid fibroblasts (PKF) and intra-lesional keloid fibroblasts (IKF). Conditioned media was used to grow NF, NS, PKF and IKF cells over 240 hrs. Cellular behavior was monitored through real time cell analysis (RTCA), proliferation rates and migration in a scratch wound assay. Fibrosis-associated marker expression was determined at both protein and gene level. PKF conditioned media treatment of both NF and NS elicited enhanced cell proliferation, spreading and viability as measured in real time over 240 hrs versus control conditioned media. Following PKF and IKF media treatments up to 240 hrs, both NF and NS showed significantly elevated proliferation rates (p<0.03) and migration in a scratch wound assay (p<0.04). Concomitant up-regulation of collagen I, fibronectin, α-SMA, PAI-1, TGF-β and CTGF (p<0.03) protein expression were also observed. Corresponding qRT-PCR analysis supported these findings (P<0.03). In all cases, conditioned media from growing marginal PKF elicited the strongest effects. In conclusion, primary NF and NS cells treated with PKF or IKF conditioned media exhibit enhanced expression of fibrosis-associated molecular markers and increased cellular activity as a result of keloid fibroblast-derived paracrine factors.

Estimation of citrus canker lesion size using hyperspectral reflectance imaging.

The Citrus industry has need for effective approaches to remove fruit with canker before they are shipped to selective international market such as the European Union. This research aims to determine the detectable size limit for cankerous lesions using hyperspectral imaging approaches. Previously developed multispectral algorithms using visible to near-infrared wavelengths, were used to segregate cankerous citrus fruits from other peel conditions (normal, greasy spot, insect damage, melanose, scab and wind scar). However, this previous work did not consider lesion size. A two-band ratio method with a simple threshold based classifier (ratio of reflectance at wavelengths 834 nm and 729 nm), which gave maximum overall classification accuracy of 95.7%, was selected for lesion size estimation in this study. The smallest size of cankerous lesion detected in terms of equivalent diameter was 1.66 mm. The effect of variation of threshold values and number of erosion cycles (applying morphological erosion multiple times to the image) on estimation of smallest detectable lesion was observed. It was found that small threshold values gave better canker classification accuracies, while exhibiting a lower overall classification accuracy. Meanwhile, higher threshold values portrayed the opposite tendency. The threshold value of 1.275 gave the optimum tradeoff between canker classification accuracy, overall classification accuracy and minimal lesion size detection. Increasing the number of erosion cycles reduced detection rates of smaller canker lesions, leading to the conclusion that a single erosion cycle gave the best size estimation results. The erosion kernel of the size 3 mm

Detection of decorin expression in keloid with fluorescent quantitative-PCR

Objective To detect the expression and content of decorin in fibroblasts of keloid to deeply reveal the mechenism and the role of decorin plays in scar formation.Methods Fibroblasts of keloid,normal scar and normal skin were cultured in vitro,and the morphology,activity,apoptosis of fibroblast were observed under light microscope and electron microscope; the mRNAs of decorin and TGF-β1 were detected and analyzed with real-time fluorescent quantitative-PCR (FQ-PCR).Results Fibroblasts of keloid showed irregular morphology,larger size and disorder arrangement.There were a large number of mitochondria,swelling rough endoplasmic reticulum,and euchromatin-rich in nucleus of fibroblasts,suggesting the protein synthesis of keloid fibroblast was very active.Compared with normal skin,the expression of decorin was significantly lower in keloid fibroblast; On the contrary,the expression of TGF-β1 was significantly higher in keloid fibroblast than in normal scar and normal skin.Conclusions Compared with normal skin,the expression of decorin in keloid fibroblast is significantly lower.Lower content of decorin in early stage of wound healing may induce weakly suppression of proliferation and synthesis of fibroblast,and up-regulate the activity of TGF-β1,which promotes the proliferation,migration and excessive collagen synthesis of the fibroblast of keloid.Thus,decorinis an suppressor factor of keloid formation. Key words: Keloid; Decorin; β1-transforming growth factor (TGF-β1) ; Fibroblast; fluorescent quantitative-PCR (FQ-PCR)

Electrical Homogenization of Ventricular Scar by Application of Collagenase

Background— Radiofrequency ablation for ventricular tachycardia is an established therapy. Use of chemical agents for scar homogenization represents an alternative approach. The purpose of this study was to characterize the efficacy of collagenase (CLG) application on epicardial ventricular scar homogenization. Methods and Results— Myocardial infarcts were created in Yorkshire pigs (n=6) by intracoronary microsphere injection. After 46.6±4.3 days, CLG type 2, type 4, and purified CLG were applied in vitro (n=1) to myocardial tissue blocks containing normal myocardium, border zone, and dense scar. Histopathologic studies were performed to identify the optimal CLG subtype. In vivo high-density electroanatomic mapping of the epicardium was also performed, and border zone and dense scar surface area and late potentials were quantified before and after CLG-4 application (n=5). Of the CLG subtypes tested in vitro, CLG-4 provided the best scar modification and least damage to normal myocardium. During in vivo testing, CLG-4 application decreased border zone area (21.3±14.3 to 17.1±11.1 mm 2 , P =0.043) and increased dense scar area (9.1±10.3 to 22.0±20.6 mm 2 , P =0.043). The total scar area before and after CLG application was 30.4±23.4 and 39.2±29.5 mm 2 , respectively ( P =0.08). Late potentials were reduced by CLG-4 application (28.8±21.8 to 13.8±13.1, P =0.043). During CLG-4 application (50.0±15.5 minutes), systolic blood pressure and heart rate were not significantly changed (68.0±7.7 versus 61.8±5.3 mmHg, P =0.08; 77.4±7.3 versus 78.8±6.0 beats per minute, P =0.50, respectively). Conclusions— Ventricular epicardial scar homogenization by CLG-4 application is feasible and effective. This represents the first report on bioenzymatic ablation of arrhythmogenic tissue as an alternative strategy for lesion formation.

Does Quiescence Exist in Subcutaneous Abscesses? A Case of a 10-Year-Old Abdominoplasty Abscess

Quiescence is the dormant state before the inevitable eruption of an infection. Several human diseases such as cancer may present with a relatively long period of inactivity, but soft tissue infections usually have a rapid progression. In this letter, we describe the rather uncommon clinical expression of an insidious subcutaneous abscess 10 years after abdominoplasty. It is our hope that this will be helpful in documenting this unique occurrence in the plastic surgery literature. A 61-year-old woman presented with tenderness in the infraumbilical region that was triggered after a blunt-force trauma (kick) in the abdomen 5 days before. Physical examination revealed normal periumbilical and abdominal scars (abdominoplasty sequelae), but the skin between the 2 scars was very firm and fixed to the underlying tissues, with associated localized inflammation (Figure 1). Apart from desquamation, no puncture point or other kind of wound was noted. The patient was a healthy woman and had no medical problems or treatment that could have compromised her immune response. Figure 1 Initial presentation. Note the skin deformation associated with desquamation and inflammation indicating an infectious subcutaneous collection. The patient had undergone an abdominoplasty 10 years before presentation that was complicated by a postoperative Staphylococcus aureus surgical site infection. The infection manifested an abscess of the infraumbilical region with no wound dehiscence; it was drained twice. No …

Destructive, multifocal squamous-cell carcinoma nodules on the cheecks and neck of an elderly woman with a dementia syndrome

Introduction: Squamous-cell carcinoma is the second most common form of skin cancer. It can develop on normal skin, actinic keratoses, leukoplakia, and burn scars. The tumor is characterized by remarkable variability at the macroscopic and histopathologic levels. Case report: A 93-year-old woman was admitted to nursing home with a diagnosis of dementia syndrome and squamous cell nodular carcinoma on cheeks and neck region. The physical examination revealed firm, black excrescences with irregular surfaces over both cheekbones, which were roughly the size of hazelnuts. Similar nodules were present with ulcers on other areas of the face. The patient was admitted to the day hospital twice for wide excision of the tumors in the zygomatic region. Later, the neck tumors were removed, and the wound was repaired with a rotation flap after careful control of bleeding. For the latter surgery, the patient was hospitalized for a few days in a geriatric ward to ensure optimal medical care and psychological support. The histological examination revealed ulcerated, well-differentiated squamous-cell carcinoma that extended down to the subcutaneous layer. Shortly after surgery, she returned to the nursing home to resume rehabilitation and group therapy.

Fibroblast KATP currents modulate myocyte electrophysiology in infarcted hearts

Cardiac metabolism remains altered for an extended period of time after myocardial infarction. Studies have shown fibroblasts from normal hearts express KATP channels in culture. It is unknown whether fibroblasts from infarcted hearts express KATP channels and whether these channels contribute to scar and border zone electrophysiology. KATP channel subunit expression levels were determined in fibroblasts isolated from normal hearts (Fb), and scar (sMI-Fb) and remote (rMI-Fb) regions of left anterior descending coronary artery (LAD) ligated rat hearts. Whole cell KATP current density was determined with patch clamp. Action potential duration (APD) was measured with optical mapping in myocyte-only cultures and heterocellular cultures with fibroblasts with and without 100 μmol/l pinacidil. Whole heart optical mapping was used to assess KATP channel activity following LAD ligation. Pinacidil activated a potassium current (35.4 ± 7.5 pA/pF at 50 mV) in sMI-Fb that was inhibited with 10 μmol/l glibenclamide. Kir6.2 and SUR2 transcript levels were elevated in sMI-Fb. Treatment with Kir6.2 short interfering RNA decreased KATP currents (87%) in sMI-Fb. Treatment with pinacidil decreased APD (26%) in co-cultures with sMI-Fb. APD values were prolonged in LAD ligated hearts after perfusion with glibenclamide. KATP channels are present in fibroblasts from the scar and border zones of infarcted hearts. Activation of fibroblast KATP channels could modulate the electrophysiological substrate beyond the acute ischemic event. Targeting fibroblast KATP channels could represent a novel therapeutic approach to modify border zone electrophysiology after cardiac injury.

Effects of TRAP-1-like protein (TLP) gene on collagen synthesis induced by TGF-β/Smad signaling in human dermal fibroblasts.

BackgroundHypertrophic scars are pathologic proliferations of the dermal skin layer resulting from excessive collagen deposition during the healing process of cutaneous wounds. Current research suggests that the TGF-β/Smad signaling pathway is closely associated with normal scar and hypertrophic scar formation. TRAP-1-like protein (TLP), a cytoplasmic protein, has been reported to efficiently regulate Smad2- and Smad3-dependent signal expression in the TGF-β pathway. The relationship between TLP and Type I/III collagen (Col I/III) synthesis explored in the present study provides an effective target for wound healing and gene therapy of hypertrophic scarring.ObjectiveTo investigate the effects of TLP on collagen synthesis in human dermal fibroblasts. MethodsLentiviral vectors encoding TLP was constructed to transfect fibroblasts derived from normal human skin. The expression of Col I/III and phosphorylation of Smad2 and Smad3 in fibroblasts were examined after TLP treatment. In addition, the comparison of TLP expression in normal skin tissues and in hypertrophic scar tissues was performed, and the effect of TLP on cell viability was analyzed by MTT assay.ResultsTLP expression in hypertrophic scar tissue was markedly higher than in normal skin tissue. The Real Time PCR and Western blot test results both revealed that the synthesis of Col I/III was positively correlated with the expression of TLP. TLP also facilitate Smad2 phosphorylation while, conversely, inhibiting Smad3 phosphorylation. TLP may play a cooperative role, along with the cytokine TGF-β1, in improving the overall cell viability of skin fibroblasts.ConclusionsTLP likely acts as a molecular modulator capable of altering the balance of Smad3- and Smad2-dependent signaling through regulation of phosphorylation, thus facilitating collagen synthesis in fibroblasts. Based on genetic variation in TLP levels in different tissues, these results suggest that TLP plays a key role in the process of TGF-β1/Smad3 signaling that contributes to wound healing and genesis of pathologic scars.

Differential scanning calorimetric examination of pathologic scar tissues of human skin

Keloid and hypertrophic scarring is a dermal fibroproliferative disorder characterized by increased fibroblast proliferation and excessive production of collagen. Excess scar formation occurs after dermal injury as a result of abnormal wound healing. Keloid formation has been ascribed to altered growth factor regulation, aberrant collagen turnover, genetics, immune dysfunction, sebum reaction, and altered mechanics. No single hypothesis adequately explains keloid formation. The thermal denaturations of pathologic human skin scar tissues were monitored by a SETARAM Micro DSC-II calorimeter. All the experiments were performed between 0 and 100 °C. The heating rate was 0.3 K min−1. DSC scans clearly demonstrated significant differences between the different types and conditions of samples (intact skin: T m = 54.8 °C and ΔH cal = 4.5 J g−1; normal scar: T m = 53.8 °C and ΔH cal = 4.2 J g−1; hypertrophic scar: T m = 54.2 °C and ΔH cal = 2.4 J g−1; keloid: T m = 52.9 °C and ΔH cal = 8.3 J g−1). The heat capacity change between native and denatured states of samples increased with the degree of structural alterations indicating significant water loosing. These observations could be explained with the structural alterations caused by the biochemical processes. With our investigations, we could demonstrate that DSC is a useful and well-applicable method for the investigation of collagen tissue of the human keloid and hypertrophic scar tissues. Our results may be of clinical relevance in the future, i.e., in the diagnosis of the two different pathologic scar formations, or in the choice of the optimal therapy of the disease.

Optical coherence tomography

Using optical coherence tomography (OCT) to image the larynx during diagnosis and treatment of a vast array of laryngeal disorders continues to develop along with innovative surgical techniques. Precise delineation of vocal fold-layered microstructure allows for better understanding of the impact of benign and malignant lesions on laryngeal function, and optical coherence is uniquely suited to provide detail of the interface between these lesions and adjacent normal tissue. This review provides the reader with a timely update on current clinical and research applications of OCT. Recent advances in OCT include adaptation of these modalities to obtain images of moving vocal folds and provide real-time information during procedures. Additionally, an augmented form of OCT called polarization-sensitive optical coherence tomography (PS-OCT) shows promise in further characterizing benign and malignant lesions and in delineating areas of normal pliability from scar. PS-OCT can quantify areas of increased collagen (scar) and three-dimensional PS-OCT imaging can visualize the extent and location of lesions within vocal folds. Imaging the layered microstructure of vocal folds to precisely delineate boundaries between normal pliability and scar provides valuable information during diagnosis and treatment of benign and malignant disorders. Current OCT imaging techniques are guiding phonomicrosurgical interventions and improving patient care.

In vivoassessment of human burn scars through automated quantification of vascularity using optical coherence tomography

In scars arising from burns, objective assessment of vascularity is important in the early identification of pathological scarring, and in the assessment of progression and treatment response. We demonstrate the first clinical assessment and automated quantification of vascularity in cutaneous burn scars of human patients in vivo that uses optical coherence tomography (OCT). Scar microvasculature was delineated in three-dimensional OCT images using speckle decorrelation. The diameter and area density of blood vessels were automatically quantified. A substantial increase was observed in the measured density of vasculature in hypertrophic scar tissues (38%) when compared against normal, unscarred skin (22%). A proliferation of larger vessels (diameter≥100 μm) was revealed in hypertrophic scarring, which was absent from normal scars and normal skin over the investigated physical depth range of 600 μm. This study establishes the feasibility of this methodology as a means of clinical monitoring of scar progression.

Comparative Evaluation of Agents on Dupuytrenʼs Contracture and Keloid Fibrosis

INTRODUCTION: Neoplasms are the result of abnormal cell proliferation. Dupuytren's contracture and keloid scar are characterized by fibroblast hyperproliferation and collagen deposition resulting in abnormal scar tissue. This similarity has lead to investigation of antiproliferative/antimetabolite agents used in the treatment of neoplasms to control/alleviate Dupuytren's contracture and keloid scar. While individual drug treatments show some decrease in contraction, there has not been a reported investigation comparing antiproliferative/antimetabolite substances and their proposed mechanism of action. The purpose of this experimental series is to comparatively evaluate respective levels of TGF-β1 and -β2, the rate and extent of contraction of fibroblasts derived from keloid scar and Dupuytren's contracture when treated with antiproliferative/antimetabolite agents. METHODS: Fibroblasts obtained from explants of Dupuytren's contracture and palmar fascia (control) as well as keloids and normal scar (control) were utilized. These fibroblast populated collagen lattices (FPCL) were exposed to 72 hours of non-cytotoxic doses of 5-Fluorouracil, Methotrexate, Paclitaxel, Tamoxifen, Mitomycin-C, and Bleomycin. Standardized photography was utilized to evaluate FPCL contraction. The supernatant of FPCL was analyzed using TGF-β1 and -β2 immunoassays. RESULTS: Dupuytren's and keloid fibroblasts' FPCLs exhibit increased contraction when compared with controls. These abnormal scar types showed a significant decrease of contraction after exposure to particular antiproliferative agents. TGF-β1 secretion did not result in a significant difference between abnormal scars when compared with controls. Various antiproliferative treatment agents on abnormal scar groups decreased the expression of TGF-β1compared to the untreated controls. Abnormal scar types had elevated TGF-β2 when compared with controls. Treatment of fibroblasts with all drugs tested resulted in significant down-regulation of TGF-β2 expression compared to untreated fibroblasts from these abnormal scar types. CONCLUSION: Non-cytotoxic doses of antiproliferative/antimetabolite agents used in this study decrease fibroblast contraction significantly when compared with untreated fibroblasts. These agents also cause a significant decrease in both TGF-β1 and -β2, a likely cause of decreased fibroblast contraction. Antiproliferative and antimetbolite agents, especially Paclitaxel, Methotrexate and Tamoxifen, are effective in vitro to limit fibroblast contraction. Further testing is warranted to provide evidence of their efficacy in vivo. DISCLOSURES: Funded in part with a grant from the Plastic Surgery Foundation. This material is the result of work supported with resources and the use of facilities at the Bay Pines V.A. Healthcare System. The information presented herein do not express the views of the Department of Veterans Affairs or the United States Government. Dr. Robson is a consultant for SteadMed Medical, Inc.

Site-specific immunophenotyping of keloid disease demonstrates immune upregulation and the presence of lymphoid aggregates

Keloid disease (KD) is a common fibroproliferative disorder of unknown aetiology. T cells and macrophages are increased in KD and are thought to contribute to its pathogenesis. However, while a link between inflammation and fibrotic disorders is well known for other disorders, it remains undetermined in KD. Systematically to immunophenotype the inflammatory infiltrate of KD in situ in a site-specific manner, and to compare this with normal skin and scar tissue. Sixty-eight keloid cases were screened for the presence of all three (intralesional, perilesional and extralesional) keloid-associated specific tissue sites. Subsequently, a complete set of 25 keloid biopsies (from different patients) was compared with normal skin (n = 11) and normal scar (n = 11) samples and subjected to systematic, site-specific quantitative immunohistomorphometry and histochemistry, using a range of immunological markers of B cells, T cells, macrophages, mast cells (MCs) and Langerhans cells. T cells, B cells, degranulated and mature MCs (coexpressing OX40 ligand) and alternative macrophages (M2) were all significantly increased in intralesional and perilesional KD sites compared with normal skin and scar tissue (P < 0·05). Additionally, 10 of 68 KD cases (15%) showed the presence of distinctive lymphoid aggregates, which resembled mucosa-associated lymphoid tissue (MALT). The increased number and activity of MCs and M2 may implicate inflammation in the fibrotic process in KD. The distinct KD-associated lymphoid aggregate resembles MALT, for which we propose the term 'keloid-associated lymphoid tissue' (KALT). It may perpetuate inflammatory stimuli that promote KD growth. KALT, MCs and M2 are promising novel targets for future KD therapy.

Use of a Novel Endoscopic Catheter for Direct Visualization and Ablation in an Ovine Model of Chronic Myocardial Infarction

Defining the arrhythmogenic substrate is essential for successful ablation of scar-related ventricular tachycardia. The visual characteristics of endocardial ischemic scar have not been described in vivo. The goal of this study was (1) to quantify the visual characteristics of normal tissue, scar border zone, and dense scar in vivo with the use of a novel endoscopic catheter that allows direct endocardial visualization and (2) to correlate visual attributes of myocardial scar with bipolar voltage. Percutaneous transient balloon occlusion (150 minutes) of the mid left anterior descending coronary artery was performed in an ovine model. Animals survived for 41.5±0.7 days. Detailed bipolar voltage maps of the left ventricle were acquired with the use of NavX. Video snapshots of the endocardium were acquired at sites distributed throughout the left ventricle. Visual tissue characteristics of normal (>1.5 mV), border (0.5-1.5 mV), and dense scar (<0.5 mV) were quantified with the use of image processing. Radiofrequency lesions (10-20 W, 30 seconds) were delivered under direct visualization. Mean white-threshold pixel area was lowest in normal tissue (189 969±41 478 pixels(2)), intermediate in scar border zone (255 979±36 016 pixels(2)), and highest in dense scar (324 452±30 152 pixels(2); P<0.0001 for all pairwise comparisons). Tissue whiteness, characteristic of scar, was inversely correlated with bipolar voltage (P<0.0001). During radiofrequency lesions, there was a significant increase in white-thresholded pixel area of the visual field after ablation (average increase, 85 381±52 618 pixels(2); P<0.001). Visual characteristics of chronic infarct scar in vivo observed with the use of a novel endoscopic catheter correlate with bipolar electrogram voltage. Irrigated radiofrequency lesions in normal endocardial tissue and postinfarction zone can be visualized and quantified with the use of image processing. This technology shows promise for visually based delivery of radiofrequency lesions for the treatment of scar-based ventricular tachycardia.