Assessment of VEGF and EGFR in the study of angiogenesis of eyelid carcinomas.

A tumor represents an abnormal tissue growth that can arise from any ocular structure, such as eyelids, muscles or the optic nerve. At the eyelids, there are two main tumor types: basal cell carcinoma and squamous cell carcinoma. Angiogenesis plays a crucial role in growth, invasion and metastasis processes of any tumor. It is well known the fact that without new vessels formation tumors cannot exceed 1-2 mm diameter. Immunohistochemical analysis has been performed on 43 cases of primary carcinomas of the eyelid, diagnosed between 2010 and 2014 in the Laboratory of Pathological Anatomy of the University Emergency County Hospital of Craiova, Romania. Biological material was represented by surgical resection samples, coming from the Clinic of Ophthalmology the anteriorly named Hospital. Within the immunohistochemical study, we have evaluated epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) expression in a group of 43 cutaneous carcinomas of the eyelid, depending on the type and differentiation grade of the tumor. Of the 43 samples, 23 came from patients with eyelid basal cell carcinoma and 20 came from patients with eyelid squamous cell carcinoma. In our study, EGFR and VEGF immunoexpression was superior for squamous cell carcinomas, compared to basal cell carcinomas, fact that was statistically significant. Regarding squamous cell carcinomas, the immunoexpression of these two markers was superior in moderate÷poor differentiated forms, compared to well differentiated forms, fact that was statistically significant. The markers used in this study were found to be associated with the acquisition of aggression and angiogenic phenotypes by analyzed carcinomas.

Investigation of vasculogenic mimicry in sebaceous carcinoma of the eyelid.

PURPOSE: Vasculogenic mimicry (VM) is a newly proposed pattern of tumour angiogenesis that has been identified in some malignancies and is associated with poor prognosis. The purpose of this study was to investigate whether sebaceous carcinomas of the eyelid exhibit VM and to determine whether these fluid-conducting patterns are associated with clinicopathologic features, the number of microvessels and the levels of endothelial growth factor (VEGF) and matrix metalloprotease-2 (MMP-2) in tumours. METHODS: Forty paraffin-embedded samples of sebaceous carcinoma of the eyelid were collected, along with complete clinical and pathologic data for all the cases. Tissue sections were stained for CD34, periodic acid and Schiff (PAS), VEGF and MMP-2. VM was identified by the presence of PAS-positive and CD34-negative loops lined by tumour cells. The VM status of tumour samples was compared with the clinical and pathological data using statistical tests. The levels of VEGF, MMP-2 and the number of microvessels were compared between patients with and without VM. RESULTS: VM was detected in 14 of 40 (35%) tumour samples. The existence of VM in tumours was associated with tumour size (p=0.007) and recurrence (p=0.021). The number of microvessels was lower in tumours with VM (13.03+/-4.02 versus 22.99+/-7.72; p<0.0001). The staining index of MMP-2 was higher in tumours with VM (27.43, range: 0-5.3) compared to tumours without VM (16.77, range: 0-2.7; p=0.004). However, there was no difference in the expression of VEGF between groups with and without VM (p=0.244). CONCLUSIONS: Vasculogenic mimicry is present in sebaceous carcinoma of the eyelid making it an unfavourable prognosis sign. MMP-2 is associated with VM formation in sebaceous carcinoma of the eyelid.

Repair of inner canthus surgical wound using superior palpebral artery based advancement flap.

BACKGROUND: Small defects of the inner canthus can be difficult to repair with an aesthetically satisfactory result, due to the difficulty in matching skin types and the resulting donor site scars. METHODS: We describe a technique which we have employed for repairing skin defects around the inner canthus, using a cutaneous advancement flap from the upper eyelid, based on the superior palpebral artery. RESULTS: Complete excision of the lesion was possible, and a cosmetically acceptable result was obtained. CONCLUSIONS: By using this technique, skin is moved within one aesthetic unit, thus maximising skin type matching and cosmetic results. In addition, the technique creates a donor site which can be easily hidden within the natural creases of the upper lid, and the arterial supply maximises the chances of flap survival.

Natural history of periocular capillary haemangiomas: changes in internal blood velocity and lesion volume.

AIM: Although the clinical characteristics of childhood periocular capillary haemangiomas are well known, serial measurements of blood velocity and lesion size are unknown. This investigation was designed to measure the changes in maximum blood velocity and estimated size of lesion in children with capillary haemangioma not requiring active intervention. STUDY DESIGN: Retrospective case-note review for a cohort of children with capillary haemangioma involving the eyelid and orbit. PATIENTS AND METHODS: Children with periocular capillary haemangioma, under the care of the Orbital unit at Moorfields Eye Hospital between 1996 and 2005, were monitored clinically and with repeated ultrasonographic examination. Volume estimates were calculated as an ovoid based on the three maximum orthogonal measurements for the haemangioma, and blood velocity was assessed by Colour Flow Mapping, Colour Doppler Energy Imaging, and Spectral Doppler techniques using a Sequoia 512 Acuson scanner. RESULTS: Twenty-four children (12 boys) had initial assessment by 18 months of age, and the haemangioma increased in size in 14/24 (58%), the increase being between 4 and 931% of initial volume estimate. The largest measured size for an individual haemangioma appears inversely related to the child's age at measurement, this mirroring a similar trend in measurements for the maximum blood velocity. Blood velocity measurements also tend to decrease with time, the peak velocity being before 1 year of age in the majority (15/24; 62%). In many children, both volume estimates and blood velocities show a cyclic variation-this occurring with increasing intervals between the maxima, before a final decay in both parameters. Although, for the whole group, there was no correlation (correlation coefficient=0.29) between estimated size and measured blood velocity, some individual children showed a significant correlation between the two parameters. The age at maximum blood velocity appeared to precede the age at maximum volume in most children, and in many there was an orbital anomaly detectable on ultrasonographic examination, even with complete clinical resolution of the haemangioma. CONCLUSIONS: Ultrasonographic examination of periocular capillary haemangiomas show that these lesions have a very high blood velocity in feeding vessels-about 2-3 orders of magnitude greater than normal capillary beds-and that the velocity and volume of such lesions undergo a cyclic variation during their natural history. Evidence suggests that both velocity and volume decrease with time, although often not returning to zero on ultrasonography (unlike the clinical resolution of the lesions). In most children, blood velocity peaks before volume estimates and this might suggest that decreasing perfusion leads to later tissue atrophy and involution of the haemangioma.

Gene transfer of a soluble receptor of VEGF inhibits the growth of experimental eyelid malignant melanoma.

PURPOSE: To determine the effect of adenovirus-mediated gene transfer of a soluble receptor of vascular endothelial growth factor (VEGF) on the growth of experimental eyelid malignant melanoma. METHODS: An adenovirus vector encoding a soluble VEGF receptor/flt-1 (Adflt-ExR) was constructed. The bovine retinal endothelial cells (ECs) were incubated in a culture medium of 293E1 cells infected by means of an adenovirus vector or uninfected (control), which contained human recombinant VEGF, and the [3H]thymidine uptake was tested. The experimental eyelid malignant melanoma was induced by the injection of B16 melanoma cells (4 x 10(6) cells) into the right upper eyelid of BALB/c nu/nu mice, and the size of the tumor was recorded for 3 weeks after tumor cell injection. The effect of Adflt-ExR was examined in three ways. Model 1: B16 cells were infected by Adflt-ExR beforehand (at a multiplicity of infection [MOI] of 10) and injected into the eyelid. Model 2: Adflt-ExR was injected into pre-established B16 cell-induced eyelid malignant melanoma. Model 3: Adflt-ExR was injected into the femoral muscle of mice before B16 cell injection into the eyelid, and the remote effect was evaluated. An adenovirus vector bearing the LacZ gene (AdLacZ) or phosphate-buffered saline was used as a control. The amount of VEGF and the flt-ExR protein was measured by sandwich enzyme-linked immunosorbent assay (ELISA). Vascularization was evaluated by counting the number and the size of the vessels. RESULTS: The supernatant of Adflt-ExR-transfected cells clearly inhibited VEGF-induced bovine retinal EC proliferation in vitro. In models 1 and 2, the tumor growth in Adflt-ExR-treated mice was significantly lower than that of controls (P < 0.05). In model 3, no significant difference was found (P = 0.14). The molar ratio of VEGF/flt-ExR protein was clearly low in the tumors of Adflt-ExR-treated mice in models 1 and 2 (P < 0.01) but not in model 3 (P > 0.05). In vessel density, the tumors in Adflt-ExR-treated mice had fewer vessels than tumors in control animals in models 1 and 2 (P < 0.05). CONCLUSIONS: Adenovirus-mediated gene transfer of a soluble form of VEGF receptor (flt-1) gene inhibited the growth of the experimental eyelid malignant melanoma. This method may be useful as an antiangiogenic therapy for eyelid malignant melanoma.

Pleomorphic fibroma of the eyelid.

BACKGROUND: This report describes the pathology of an unusual variant of fibroma of the eyelid. The soft ovoid tumour was excised from the centre of the left lower lid in an 84-year-old patient had been growing slowly for several years. METHODS: The tumour was studied by conventional histology, immunohistochemistry and electron microscopy. RESULTS: The tumor was formed by fibroblasts of spindle and multinucleate type, and the collagenous stroma contained mast cells and blood vessels which exhibited minor inflammatory changes. With the immunohistochemical methods the only positive marker was for vimentin. CONCLUSION: On morphological grounds it was possible to distinguish this extremely rare tumour from other mesenchymal tumours. The presence of distinctive multinucleate giant cells makes the term pleomorphic fibroma an appropriate name for this type of lesion.