Reflectance Confocal Microscopy Evaluation Research Articles

Non-invasive Assessment and Management of Folliculitis Decalvans by Trichoscopy and Reflectance Confocal Microscopy.

Folliculitis decalvans (FD) is a rare scarring alopecia mainly affecting middle-aged men, characterized by recurring episodes of follicular pustules, crusts, erythema, tufted hairs, and scars. This study investigates the effectiveness of reflectance confocal microscopy (RCM) compared to trichoscopy for diagnosing and monitoring FD. The study involved 24 Caucasian patients diagnosed with FD. Patients were examined using trichoscopy and RCM, with a focus on specific features like erythema and inflammatory cell distribution. A subgroup of 16 patients was followed up after 3 months of therapy. The reproducibility of RCM and trichoscopy was assessed using Cohen Kappa Test. RCM and trichoscopy consistently detected features such as tufted hairs, pustules, and perifollicular fibrosis. However, RCM provided more detailed insights into inflammatory activity and types of fibrosis, often overlooked by trichoscopy. It showed a reduction in vessels and inflammatory cells, which trichoscopy failed to detect. The concordance between RCM evaluations was excellent, indicating high reproducibility. RCM is effective in diagnosing and monitoring FD, offering detailed insights into inflammation and fibrosis. It complements trichoscopy, especially in aspects where trichoscopy is limited, such as precise measurement of inflammation. The study suggests that combining RCM with trichoscopy could enhance the accuracy of diagnosis and monitoring of FD, leading to tailored therapeutic approaches. Further studies with larger sample sizes and longitudinal designs are recommended to confirm these findings.

In Vivo Evaluation of Laser-Induced Optical Breakdown (LIOBS) by 1064-nm Nd:YAG Fractional Picosecond Laser With Reflectance Confocal Microscopy and Precise Histopathologic Correlation.

Picosecond lasers with a microlens array can cause laser-induced optical breakdown (LIOBS) and LIC (Intradermal laser-induced cavitation) within high-fluence areas. This study aimed to describe the clinical, reflectance confocal microscopy (RCM), histopathological findings, and the characteristics of vacuoles caused by LIOBS and LIC in individuals with skin types III and IV. This study was performed on six Chilean healthy volunteers, males and females, aged 35-65 years old with Fitzpatrick skin phototypes III-IV. The laser was applied in the inner proximal area of the nondominant arm. RCM evaluation was performed 24 h later; 48 h later, skin biopsies were performed on the laser-treated areas. Clinical, histological, and RCM findings were recorded. Every individual developed a 10 mm2 area of clinical erythema in the treated area. Under RCM, all six volunteers had hyporeflective spherical structures at the level of the epidermis, consistent with intraepidermal vacuoles. Histopathological evaluation revealed different sizes of vacuoles in both the epidermis and dermis. The LIOBS and LIC processes and the secondary production of vacuoles could be highly valuable for effective dermal remodeling treatment and aid in promoting the production of new collagen, elastic fibers, and growth factors that could improve skin texture. These structures were visible under RCM and histopathological evaluation.

Reflectance Confocal Microscopy and Dermoscopy of Facial Pigmented and Non-Pigmented Actinic Keratosis Features before and after Photodynamic Therapy Treatment

Actinic keratosis (AK), due to its widespread prevalence, as well as the possibility of progression to an invasive form of squamous cell carcinoma, requires treatment regardless of the clinical stage. New imaging techniques, such as in vivo reflectance confocal microscopy (RCM), significantly increase the accuracy of diagnosis and allow noninvasive evaluation of the therapeutic efficacy of the ongoing treatment. Our objective was to evaluate the prevalence of specific (video)dermoscopy and RCM features of pigmented and classical subtypes of AK before and after photodynamic therapy (PDT) treatment. We included patients with facial grade II AKs (25 pigmented, 275 non-pigmented) were included in the study. Skin lesions were evaluated by (video)dermoscopy and RCM at the baseline and three months after PDT. In classic AK, the most frequent dermoscopic findings were fine wavy vessels (96%), scale (92%), microerosions (48%), and “strawberry” pattern (36%), while pigmented AK was characterized mostly by “rhomboidal pattern” (80%), scale (60%), white globules (48%), “jelly sign”, and superficial pigmentation (40%). RCM’s most characteristic classic AK findings were abnormal honeycomb pattern in the spinous layer, epidermal inflammatory infiltrate, and solar elastosis that were present in 96% of lesions. Pigmented AKs presented mostly with dark central areas of parakeratosis (72%), mottled pigmentation (72%), dermal inflammatory infiltrate (64%), solar elastosis (60%), and abnormal honeycomb pattern in the spinous layer (56%). Dermoscopically, PDT resulted in complete disappearance of the “rhomboidal pattern” in both classical and pigmented AKs, “starburst pattern” and “jelly sign” in classical AKs, and inner gray halo, “rosette sign” and central crust in pigmented AKs. Three months after one PDT session, RCM evaluation showed mostly solar elastosis in both classical and pigmented AK subtypes, epidermal inflammatory infiltrate in classical AKs, and dermal inflammatory infiltrate in pigmented AKs. New noninvasive imaging techniques such as RCM and (video)dermoscopy can help practitioners better visualize the efficacy of the ongoing PDT treatment in either classical or pigmented AK subtypes.

Management of lentiginous melanoma with imiquimod assessed by reflectance confocal microscopy.

Management of lentiginous melanoma with imiquimod assessed by reflectance confocal microscopy.

In Vivo Reflectance Confocal Microscopy as a Response Monitoring Tool for Actinic Keratoses Undergoing Cryotherapy and Photodynamic Therapy.

Simple SummaryThe assessment of actinic keratoses (AKs) in prevention and therapeutic trials, as well as clinical practice, could significantly benefit from the incorporation of non-invasive imaging technology. Such technology has the potential to enhance the objective evaluation of clinical and subclinical AKs with the added advantage of sequential monitoring. In vivo reflectance confocal microscopy (RCM) allows for the non-invasive imaging of AKs at a cellular level. We aimed to establish an in in vivo RCM protocol for AK response monitoring, ultimately leading to more reliable characterization of longitudinal responses and therapy optimization.Reflectance confocal microscopy (RCM) presents a non-invasive method to image actinic keratosis (AK) at a cellular level. However, RCM criteria for AK response monitoring vary across studies and a universal, standardized approach is lacking. We aimed to identify reliable AK response criteria and to compare the clinical and RCM evaluation of responses across AK severity grades. Twenty patients were included and randomized to receive either cryotherapy (n = 10) or PDT (n = 10). Clinical assessment and RCM evaluation of 12 criteria were performed in AK lesions and photodamaged skin at baseline, 3 and 6 months. We identified the RCM criteria that reliably characterize AK at baseline and display significant reduction following treatment. Those with the highest baseline odds ratio (OR), good interobserver agreement, and most significant change over time were atypical honeycomb pattern (OR: 12.7, CI: 5.7–28.1), hyperkeratosis (OR: 13.6, CI: 5.3–34.9), stratum corneum disruption (OR: 7.8, CI: 3.5–17.3), and disarranged epidermal pattern (OR: 6.5, CI: 2.9–14.8). Clinical evaluation demonstrated a significant treatment response without relapse. However, in grade 2 AK, 10/12 RCM parameters increased from 3 to 6 months, which suggested early subclinical recurrence detection by RCM. Incorporating standardized RCM protocols for the assessment of AK may enable a more meaningful comparison across clinical trials, while allowing for the early detection of relapses and evaluation of biological responses to therapy over time.

Evaluation of diagnostic criteria of actinic keratosis through reflectance confocal microscopy.

The diagnosis of actinic keratosis (AK) is based on clinical evaluation and confirmed by histopathological analysis (HA). The challenge is to establish the correct diagnosis with a minimally invasive assessment. The aim of this study is to validate the analysis of AK by reflectance confocal microscopy (RCM), a cellular resolution, noninvasive imaging method and to determine the relevant parameters for diagnosis, compared to HA, by calculating the sensitivity (S), specificity (E), positive predictive value (PPV), and negative predictive value (NPV) of each criterion. Through clinical examination, 25 AKs were selected for dermoscopy and RCM evaluation followed by shaving excision for HA. Statistical analysis was done by hypothesis tests (McNemar for binary and Wilcoxon for continuous variables). There was no significant difference between RCM and HA for 5 of the 6 parameters analyzed. The criteria that were statistically relevant were as follows: parakeratosis (p-value 0.449690; S 90%; PPV 78.26%), hyperkeratosis (p-value 0.248213; S 87.5%; E 100%; PPV 100%; NPV 25%), dyskeratosis (p-value 0.617075; S 85.71%; E 75%; PPV 94.74%; NPV 50%), spinous layer keratinocyte atypia classified as mild, moderate or severe (P-value 0.145032) and inflammation in epidermis (P-value 1.000000; S 75%; E 20%; PPV 78.95%; NPV 16.67%). RCM could not adequately measure inflammation in dermis (P-value 0.013328), despite good sensitivity (68%) and PPV (100%). RCM proved to be an effective method for the diagnosis of AK, contributing to the selection of the most appropriate treatment option.

Concordance among in vivo reflectance confocal microscopy, trichoscopy, and histopathology in the evaluation of scalp discoid lupus.

Reflectance confocal microscopy (RCM) has been used for the evaluation of several inflammatory skin conditions, including skin discoid lupus erythematosus (DLE), and has been correlated with conventional histopathology (HP). However, RCM is not being widely used in trichology. Few reports and just preliminary data suggest the use of RCM as a complementary tool in alopecias. To correlate the major RCM features of scalp DLE with trichoscopy and HP findings of biopsy specimens obtained from the same lesions. This is an observational, analytical, and cross-sectional study involving 12 patients with a clinically established diagnosis of scalp DLE. Patients underwent global clinical photograph, trichoscopy, and RCM examination in the same site followed by two 4-mm punch biopsy specimens for HP analysis. Inter-methods agreement among RCM imaging, trichoscopy, and horizontal histopathology sections (HHS) were calculated using Cohen Kappa (k) statistics. Statistical analysis of the agreement between RCM and HP features disclosed an overall agreement similar to skin DLE. Seven of the eleven features evaluated had agreement superior to 75%. We also evaluated RCM features associated with three of their corresponding trichoscopic findings for further investigation of their agreement with HP. Statistical analysis showed an enhancement with agreement of 86% when the non-invasive techniques are used together. Consistent correlation between RCM and HP observed in our study supports the reliability of RCM in the diagnosis of scalp DLE. RCM may be considered a promising tool for scalp DLE microscopic evaluation and presents similar RCM features to DLE in other body sites. By associating clinical, trichoscopic and RCM evaluation, dermatologists will have a non-invasive arsenal for the assessment of hair and scalp disorders, benefiting patients.

Reflectance confocal microscopy for noninvasive examination of nonmelanocytic tumors and virus-associated skin lesions in organ transplant recipients.

Drug-induced immunosuppression is necessary to prevent rejection of the foreign organ in transplanted patients, but neoplastic and virus-associated skin diseases are frequent complications. Reflectance confocal microscopy (RCM) recently emerged as a promising tool for the early diagnosis of skin lesions. A total of 61 skin lesions, among them 20 basal cell carcinomas, six Bowen's diseases, 23 actinic keratoses, and 12 verrucae, were analyzed. All lesions were clinically evaluated followed by RCM evaluation by two independent dermatologists and histological examination. For the diagnosis of basal cell carcinoma, a sensitivity of 100% by both investigators (INV I+II) and a specificity of 100% by INV I and 80% by INV II were achieved. The sensitivity average rate for RCM features reached by both investigators ranged between 60% and 100%, and the specificity between 55% and 90%. For the diagnosis of actinic keratosis, a concordant sensitivity of 94.4% and a specificity of 80% (INV I) and 60% (INV II) were detected. The sensitivity average rate of specific RCM criteria ranged between 72.3% and 97.2%, whereas specificity ranged between 20% and 90%. Regarding verrucae, RCM confirmed the histological diagnosis with a sensitivity of 85.7% (INV I) and 100% (INV II), while specificity was 100% and 80%, respectively. Reflectance confocal microscopy resulted to be a reliable tool for the noninvasive diagnosis of neoplastic and virus-associated skin changes in organ transplant recipients. Nevertheless, given the frequency and diagnostic complexity of the hyperkeratotic lesions occurring post-transplantation, larger cohorts of patients are required to confirm and consolidate these findings.

Dermoscopy and confocal microscopy for different chemotherapy‐induced alopecia (CIA) phases characterization: Preliminary study

Chemotherapy-induced alopecia (CIA) affects 65% of patients receiving chemotherapy regimens and is often identified with the massive hair loss stage. Reflectance confocal microscopy (RCM) is a noninvasive technique used in alopecia assessment for disease characterization and state of activity. To describe RCM features of CIA in different timing and identify specific phases of alopecia development. A total of 16 patients treated with chemotherapy underwent dermoscopy and RCM evaluations four times during the observation: 2 and 4-6weeks after starting and 3 and 6months after the end of chemotherapy. Ten examinations for each stage were performed. Four phases of CIA have been identified. Initial hair loss showed specific dots not previously described, named CIA dots. massive hair loss phase was characterized by black dots (10/10 pt), CIA dots (8/10 pt) and hair shaft abnormalities. Three months after the end of chemotherapy, during the partial regrowth phase, 10/10 patients showed thin hair in regrowth and 8/10 presented black and yellow dots. At 6months, normal hair in regrowth appears in all patients (total regrowth phase). Chemotherapy-induced alopecia has to be considered as a dynamic process with specific phases characterized by distinctive dermoscopic and confocal features.

Reflectance confocal microscopy confirms residual basal cell carcinoma on clinically negative biopsy sites before Mohs micrographic surgery: A prospective study

Biopsy specimens from patients with basal cell carcinoma (BCC) can present to surgery with no clinically residual tumor, complicating treatment decisions. To evaluate reflectance confocal microscopy (RCM) for the assessment of residual BCC following biopsy. Consecutive patients with biopsy-proven BCC and no clinical evidence of residual tumor who had been referred for Mohs micrographic surgery were included. Biopsy sites were imaged with a handheld RCM device. On the basis of RCM evaluation, cases were labeled RCM positive or RCM negative. Mohs micrographic surgery was performed in all cases; margins and 15-μm serial vertical sectioning were evaluated. A total of 61 patients were included (mean age, 61.7years [standard deviation, 12.2years]; range, 37-87years); 60.7% were women. The mean lesion size was 5.1mm (range, 3-12mm); 73.8% of patients were positive on RCM, and 68.9% had residual BCC on histopathologic examination. The rates of RCM sensitivity, specificity, positive predictive value, and negative predictive value were 92.8%, 68.4%, 86.6%, and 81.2%, respectively. Three cases of BCC (high-risk, infiltrative, and basosquamous) were missed with use of RCM. When high-risk subtypes were excluded (n = 5), sensitivity and negative predictive value were both 100%. RCM can miss deep-seated residual tumor. RCM is a valuable tool for the evaluation of residual BCC following biopsy, with the potential to reduce unnecessary surgical procedures.

Effects of topical piroxicam and sun filters in actinic keratosis evolution and field cancerization: a two-center, assessor-blinded, clinical, confocal microscopy and dermoscopy evaluation trial

Background: Actinic keratosis (AK) is considered an “in situ” non-melanoma skin cancer induced by ultraviolet chronic exposure. Sunscreen and topical anti-inflammatory agents like diclofenac could improve the evolution of this kind of lesions. A topical product containing piroxicam 0.8% and sun filters (50 SPF) (ACTX) has been shown to be very effective in reducing AK lesions. So far, no data are available regarding the effects of this product on skin modifications evaluated by reflectance confocal microscopy (RCM) and dermoscopy at the lesion sites and on the skin around the lesions (field cancerization).Study aim: To evaluate in a two-center, assessor-blinded, prospective trial the effect of ACTX on AK number, RCM and dermoscopy parameter evolution of a target lesion in subjects with multiple AK lesions.Subjects and methods: A total of 54 subjects (42 men and 12 women; mean age 65 years) with AK lesions grade I–III located on the scalp (n = 36) or face (n = 18) were enrolled after their written informed consent. ACTX was applied twice daily on the face and scalp for six consecutive months. AK lesion count was performed at baseline and after 3 and 6 months. Lesion count was assessed in a blind fashion evaluating digital color high definition images performed at each visit and coded in a blinded fashion. RCM evaluations were performed at the same time-points. A dermoscopy evaluation was performed at baseline and after 6 months. RCM and dermoscopy were assessed on a pre-specified target lesion. The RCM severity score was used evaluating 11 items, examining stratum corneum, stratum granulosum, stratum spinous and dermal layers (maximum score 11 points). The dermoscopy score evaluated erythema, scaling and follicular plugs (from 0 to 4 for each item) and pigmentation (from 0 to 5).Results: Forty-nine subjects (90%) concluded the trial. At baseline, the mean (SD) number of AK lesions was 9.6 (5.2). AK lesions significantly decreased to 5.9 and to 5.6 after 3 and 6 months of ACTX treatment (p = .001; intention to treat analysis), representing a −42% reduction. A reduction of AK lesion numbers >50% in comparison with baseline was observed in 51% of subjects at month 6. New AK lesions appeared in five subjects (9%). The RCM mean (SD) severity score at baseline was 6.4 (2.0). ACTX treatment was associated with a progressive and significant (p = .002) reduction to 4.9 after 3 months and to 4.8 (2.3) at month 6 (a −25% reduction). The dermoscopy score at baseline was 5.5 (2) and it was reduced significantly (p = .007) to 4.5 (2) at the end of the study. The product was in general very well tolerated.Conclusion: A 6 month application of ACTX in subjects with AK lesions was associated with an improvement in AK lesion count and with a reduction in the RCM/dermoscopy severity scores of the target lesion.Trial registration number: ISRCTN22070974.

Long-term efficacy of a new medical device containing Fernblock® and DNA repair enzyme complex in the treatment and prevention of cancerization field in patients with actinic keratosis.

Actinic keratosis (AKs) are part of the cancerization field, a region adjacent to AKs containing subclinical and histologically abnormal epidermal tissue due to Ultraviolet (UV)-induced DNA damage. The photoproducts as consequence of DNA damage induced by UV are mainly cyclobutane pyrimidine dimers (CPDs). Fernblock® demonstrated in previous studies significant reduction of the number of CPDs induced by UV radiation. Photolyases are a specific group of enzymes that remove the major UV-induced DNA lesions by a mechanism called photo-reactivation. A monocentric, prospective, controlled, and double blind interventional study was performed to evaluate the effect of a new medical device (NMD) containing a DNA-repair enzyme complex (photolyases, endonucleases and glycosilases), a combination of UV-filters, and Fernblock® in the treatment of the cancerization field in 30 AK patients after photodynamic therapy. Patients were randomized into two groups: patients receiving a standard sunscreen (SS) andpatients receiving the NMD. Clinical, dermoscopic, reflectance confocal microscopy (RCM) and histological evaluations were performed. An increase of AKs was noted in all groups after three months of PDT without significant differences between them (p=0.476). A significant increase in the number of AKs was observed in SS group after six (p=0.026) and twelve months of PDT (p=0.038); however, this increase did not reach statistical significance in the NMD group. Regarding RCM evaluation, honeycomb pattern assessment after twelve months of PDT showed significant differences in the extension and grade of the atypia in the NMD group compared to SS group (p=0.030 and p=0.026, respectively). Concerning histopathological evaluation, keratinocyte atypia grade improved from baseline to six months after PDT in all the groups, with no statistically significant differences between the groups. Twelve months after PDT, p53 expression was significantly lower in the NMD group compared to SS group (p=0.028). The product was well-tolerated, with no serious adverse events reported. Our results provide evidence of the utility of this NMD in the improvement of the cancerization field and in the prevention of the development of new AKs.

Long-Term Efficacy of a New Medical Device Containing Fernblock® and DNA Repair Enzyme Complex in the Treatment and Prevention of Cancerization Field in Patients with Actinic Keratosis

Introduction: Actinic keratosis (AKs) are part of the cancerization field, a region adjacent to AKs containing subclinical and histologically abnormal epidermal tissue due to Ultraviolet (UV)-induced DNA damage. The photoproducts, “as consequence of DNA damage induced by UV”, are mainly cyclobutane pyrimidine dimers (CPDs). Fernblock® demonstrated in previous studies significant reduction of the number of CPDs induced by UV radiation. Photolyases are a specific group of enzymes that remove the major UV-induced DNA lesions by a mechanism called photo-reactivation. Methods: A monocentric, prospective, controlled, and double blind interventional study was performed to evaluate the effect of a new medical device (NMD) containing a DNA-repair enzyme complex (photolyases, endonucleases and glycosilases), a combination of UV-filters, and Fernblock® in the treatment of the cancerization field in 30 AK patients after photodynamic therapy (PDT). Patients were randomized into two groups: patients receiving a standard sunscreen (SS) and patients receiving the NMD. Clinical, dermoscopic, reflectance confocal microscopy (RCM) and histological evaluations were performed. Results: An increase of AKs was noted in all groups after three months of PDT without significant differences between them (p=0.476). A significant increase in the number of AKs was observed in SS group after six (p=0.026) and twelve months of PDT (p=0.038); however, this increase did not reach statistical significance in the NMD group. Regarding RCM evaluation, honeycomb pattern assessment after twelve months of PDT showed significant differences in the extension and grade of the atypia in the NMD group compared to SS group (p=0.030 and p=0.026, respectively). Concerning histopathological evaluation, keratinocyte atypia grade improved from baseline to six months after PDT in all the groups, with no statistically significant differences between the groups. Twelve months after PDT, p53 expression was significantly lower in the NMD group compared to SS group (p=0.028). The product was well-tolerated, with no serious adverse events reported. Conclusion: Our results provide evidence of the utility of this NMD in the improvement of the cancerization field and in the prevention of the development of new AKs.

Improving diagnostic sensitivity of combined dermoscopy and reflectance confocal microscopy imaging through double reader concordance evaluation in telemedicine settings: A retrospective study of 1000 equivocal cases.

BackgroundReflectance confocal microscopy (RCM) is an imaging device that permits non-invasive visualization of cellular morphology and has been shown to improve diagnostic accuracy of dermoscopically equivocal cutaneous lesions. The application of double reader concordance evaluation of dermoscopy-RCM image sets in retrospective settings and its potential application to telemedicine evaluation has not been tested in a large study population.ObjectiveTo improve diagnostic sensitivity of RCM image diagnosis using a double reader concordance evaluation approach; to reduce mismanagement of equivocal cutaneous lesions in retrospective consultation and telemedicine settings.Methods1000 combined dermoscopy-RCM image sets were evaluated in blind by 10 readers with advanced training and internship in dermoscopy and RCM evaluation. We compared sensitivity and specificity of single reader evaluation versus double reader concordance evaluation as well as the effect of diagnostic confidence on lesion management in a retrospective setting.ResultsSingle reader evaluation resulted in an overall sensitivity of 95.2% and specificity of 76.3%, with misdiagnosis of 8 melanomas, 4 basal cell carcinomas and 2 squamous cell carcinomas. Combined double reader evaluation resulted in an overall sensitivity of 98.3% and specificity of 65.5%, with misdiagnosis of 1 in-situ melanoma and 2 basal cell carcinomas.ConclusionEvaluation of dermoscopy-RCM image sets of cutaneous lesions by single reader evaluation in retrospective settings is limited by sensitivity levels that may result in potential mismanagement of malignant lesions. Double reader blind concordance evaluation may improve the sensitivity of diagnosis and management safety. The use of a second check can be implemented in telemedicine settings where expert consultation and second opinions may be required.

Reflectance confocal microscopy for the evaluation of sensitive skin.

Nowadays, the diagnosis for sensitive skin relies on subjective assessment or on the combination of subjective and objective evaluation. No quantitative evaluation is available. It could be expected that confocal microscopy imaging could be of interest to better define the condition. Total 166 healthy female subjects were recruited in this study. Firstly, all subjects completed the sensitive questionnaire. Then, the cutaneous structures were measured by the reflectance confocal microscopy (RCM) on the face and fossa cubitalia. The lactic acid sting test was conducted finally. According to the results of self-perception sensitive skin questionnaire and lactic acid stinging test to evaluate facial skin sensitivity the both positive subjects were regarded as sensitive skin group and both negative group as healthy control group. The results of RCM indicating that the proportion of 'disarranged honeycomb pattern' and 'spongiform edema' in the sensitive group and healthy control group were statistically different (P < 0.05), respectively; The following report 'damaged dermal papilla rings' was not a distinctive pattern, with no significant statistical difference (P > 0.05). The epidermal thickness was 38.88 ± 6.81 μm, healthy control group was 40.31 ± 9.37 μm in, respectively, sensitive skin group and healthy control group, there was no significant statistical difference between the two groups (P > 0.05). The honeycomb structure depth of sensitive group was 20.57 ± 4.86 μm. It was for 23.27 ± 6.38 μm, healthy control group the difference being statistically different between the two groups (P < 0.05). Based on the RCM results, 'epidermal honeycomb structure' and 'spongiform edema' may be used as new skin signs of RCM evaluation of sensitive skin effectively. Indeed, sensitive skin honeycomb structure depth was thinner compared with healthy control group. Such a specific pattern has good clinical and monitoring value for the further exploration. RCM could provide new data and patterns for the evaluation of sensitive skin.

Reflectance confocal microscopy as a useful diagnostic tool for monitoring of skin containing vascularized composite allograft rejection: A preliminary study on rats.

Vascularized composite allografts can undergo immune-mediated rejection, and skin biopsies are needed for monitoring of the transplant. However it is an invasive method, and requires processing time and pathological assessment. The purpose of this study is to use a new noninvasive monitoring method of the reflectance confocal microscopy (RCM) to determine severity of the allograft rejection on rats. Five groin flap allotransplantation were performed between 10 male Sprague-Dawley rats. Immunosuppressive therapy with cyclosporine A was given to the recipients during 10 days after surgery and was ended at the 10th postoperative days to allow acute transplant rejection. Following cessation of CsA, concomitant RCM evaluation and skin biopsy was performed every other day from each animal until total rejection of the allograft. Complete rejection of the allograft took nearly about 10 days and 4 or 5 RCM evaluation and skin biopsy was performed from each rat during this period. A total of 17 specimens were evaluated. A scoring system was developed based on the RCM findings. Skin biopsies were evaluated according to the Banff 2007 working classification criteria. RCM evaluation revealed epidermal irregularity and collagen destruction, however mild perivascular inflammation and degeneration of the basal epidermal layer were observed in early and late rejection period respectively with histopathologic evaluation. High correlation was found between the RCM scores and histopathologic grading. The RCM may be the useful tool to reduce the need for skin biopsy for monitoring of the skin containing vascularized composite allograft.

CD271 is expressed in melanomas with more aggressive behaviour, with correlation of characteristic morphology by in vivo reflectance confocal microscopy.

Melanoma is the most highly aggressive type of skin cancer. Its resistance to existing treatments and the rapid rise in incidence underscore the importance of acquiring a better understanding of melanomagenesis. To assess the impact of reflectance confocal microscopy (RCM) on the description of cell morphology, which may influence the growth pattern and changes with increasing tumour severity, correlating with biological aspects. A retrospective analysis of 30 primary melanomas in vivo, evaluated by RCM, to correlate cell morphology and cellular arrangement with a marker of melanoma progression (CD271) using immunohistochemical evaluations. Typical cells organized in dermal nests with peculiar in vivo confocal morphology result in melanoma with high malignancy and positivity to CD271. This architecture might be due to the presence of a type of cells, intrinsically predisposed to invasion, as a result of dedifferentiation programming, revealed by expression of the neural crest marker CD271. With the hypothesis that dedifferentiated cells would be strongly responsible for initiation of tumour development and progression, we propose that CD271 detection could be associated with RCM evaluation in order to detect more aggressive melanoma subtypes.

Reflectance Confocal Microscopy Features of Seborrheic Dermatitis for Plaque Psoriasis Differentiation

Background: Plaque psoriasis (PP) and seborrheic dermatitis (SD) are chronic inflammatory skin diseases with similar clinical and pathological features. Differential diagnosis can be difficult, especially when particular skin areas of the face are involved. Reflectance confocal microscopy (RCM) has been demonstrated to be useful for ‘real-time' diagnosis of skin inflammatory diseases. Objective: To define distinctive confocal criteria of SD and to evaluate the usefulness of this technique for noninvasive differential diagnosis with PP. Methods: A total of 40 patients affected by PP and 19 patients by SD involving the face were recruited and subjected to RCM evaluation. Univariate and adjusted odds ratios were calculated. Discriminant functions were used to plot ROC curves. Results: The results disclosed specific patterns for SD and PP. The following distinctive confocal features for SD have been identified: spongiosis, dermal inflammation and horizontal orientation of dilated blood vessels. Conclusion: SD has a specific and easily recognizable confocal pattern supporting clinical differentiation with PP.

Surveillance for treatment failure of lentigo maligna with dermoscopy andin vivoconfocal microscopy: new descriptors

Nonsurgical treatment (radiotherapy, imiquimod) is increasingly employed for the management of lentigo maligna (LM). While the diagnosis of LM remains difficult, the detection of treatment failure is even more challenging. To describe the sensitivity and specificity for the diagnosis of LM of individual features and methods using dermoscopy and in vivo reflectance confocal microscopy (RCM) to aid in the detection of treatment failure of LM following nonsurgical treatment. A retrospective study of dermoscopy and RCM images (blinded to the correlation with pathology) in patients with biopsy-confirmed LM who were undergoing nonsurgical treatment in two referral institutions - one in Sydney, Australia, and the other in Barcelona, Spain. Ninety-eight patients were treated nonsurgically for LM during the period 2006-2012. Thirty-one patients had abnormal dermoscopy or RCM evaluation, and had a biopsy that identified LM recurrence in 15 patients and nonmelanoma diagnoses in 16 patients (one Bowen disease, 15 solar changes). The diagnosis of treatment failure was difficult with dermoscopy, with a sensitivity of 80% and specificity of 56%, even with the interpretation of an expert. The best criterion was asymmetric hyperpigmented follicular openings, but this was present in only 47% of treatment failure LM. Isolated, very fine brown dots ('dust' appearance) correlated highly with the diagnosis of treatment failure LM (73% sensitivity and 88% specificity) and with pagetoid cells seen with RCM. The LM score, comprising six criteria, had a specificity of 94% and sensitivity of 100%. These methods and descriptors should help to manage the diagnosis of treatment failure.

In vivo reflectance confocal microscopy to monitor the response of lentigo maligna to imiquimod

Imiquimod has been used for treating lentigo maligna (LM) in selected cases when surgery is not an appropriate option because of functional or aesthetic impairment. Reflectance confocal microscopy (RCM) is a noninvasive method that has not been validated for monitoring the treatment of LM with imiquimod. We sought to evaluate the use of invivo RCM to accurately monitor the response of LM to nonsurgical treatment with topical imiquimod. Twenty patients with confirmed facial LM, not amenable to surgical treatment or radiation therapy, were included prospectively. Clinical evaluation was performed by dermoscopy, RCM, and histopathology. Patients applied imiquimod 5% for 8 weeks. The affected area was assessed using the previously described LM score on RCM, and target sample biopsies were performed to confirm or discard RCM findings. Fifteen of the 20 patients (75%) presented histologic tumor clearance. Confocal microscopy identified 70% of these responders with no false-negative results, and when compared with histopathology, there was no significant difference in evaluating the response to imiquimod. The impossibility of examining the entire lesion by means of histopathology is a limitation. Invivo RCM evaluation was useful in accurately monitoring the response of LM to nonsurgical treatment with topical imiquimod in patients when surgery is contraindicated.