Keloid Correlation Between Lymphatic Microvessel Density And Growth Of Invasive

Objective：Malignant tumors present invasive growth, and this pattern ofgrowth is related with lymphatic microvessel density, therefore, in order tobetter understand the pathogenesis of keloid, especially whether its showinginvasive growth is associated with lymphatic microvessel density.Thisexperiment by testing different parts of keloid lymphatic microvessel densityfor the first time, to explore the keloid lymphatic microvessel density andcorrelation of invasive growth.Methods：110cases of keloid were obtained from the Department of Burn and PlasticSurgery inpatient surgery, the Third Hospital of Hebei Medical University, andwere diagnosed by clinical and history: formed after trauma, or no specificincentives, and more than1year history, the skin lesions beyond the originalwound, invasive, accompanied by pain and itching. All patients did not anytreatment before operation. Peripheral normal skin of keloid that were checkedfor normal skin by naked eye derived from "cat ears"; Normal human skinderived from patients who had no keloid undergoing skin grafting operation,or extra normal skin when we cleaned the wound. Proliferation of keloid is themost prominent part, dark red, glossy or uneven surface; Invasion of keloid isthe junction with normal skin, sometimes out of "crab foot" around is invasive.All specimens were fixed in the4%formalin.2Keloids were divided into two groups in accordance with the standard:proliferation part and invasion part. All specimens were trimmed2mmthickness of lamellar structure. Specimens were included in the dewateringboxes. Then, specimens were paraffin-embed after dehydration and trimming.3All specimens of four groups were cut5μm thickness, routine HE stainedwas executed,observed by light microscope and pictures capture for observed the difference between four-group specimens.4All specimens of four groups were cut5μm thickness, and were stained byD2-40immunohistochemical staining which specific lymphatic microvesselendothelial cell. We take positive coloring as a lymphatic vessel which singleendothelial cells, cell clusters, or tube-like structure. Each slice was observedlymphatic microvessel distribution by a low-microscopic, selection aftersuperficial dermal layer at high magnification, then randomly selected10regional to count lymphatic microvessel.5Use statistical software SAS V8.0, four sets of data are in line with thenormality tests, mean±standard deviation (x±s). Three sets of data whichassociated with the keloid separately test normality and homogeneity ofvariance tests, resulting in three sets of data are not up to standard, thenmultiple dependent samples nonparametric tests, and respectively compared tobetween the two groups.Results:1Macropathology: Keloids significantly thicken, non-uniform size, colorred, irregular shape, texture hard, a little fat under the base, and proliferationparts thicken significantly than invasion parts.2HE staining: Proliferation of keloid structure performance: Epidermisbecome thinner, papillary dermal layer structures are disappeared, skinadenexal is compromised, no hair follicles, sweat glands, sebaceous glandstructures；Superficial layer fibers of reticular layer of dermis are parallel array,but deeper layer fibers are thick and disorder. Invasion parts of keloidperformance: Epidermis are thicker than proliferation parts, papillary dermallayers are appeared, fibers of reticular layer are thinner than proliferation part,disorder, and increased content of capillaries. Tissue structures of marginalnormal skin of keloid are similar to patients with non-keloid normal skin.3Immunohistochemical staining: D2-40is expressed specificitly in thecytolymph and epicyte of lymphatic endothelial cells, and lymphaticmicrovessels are observed around the capillaries sometimes. Lymphaticmicrovessel of keloid mainly located in the papillary dermal layer and superficial layer fibers of reticular layer of dermis. The number of thelymphatic microvessels of proliferation part are smaller than invasive part;Shape of lymphatic microvessels of invasive part is irregular, some lockingstatus. However, lymphatic microvessels of marginal normal skin are regular.Lymphocytes occasionally are observed in the lymphatic microvessel. Thenumbers of lymphatic microvessels of every group are counted by the expertsof pathology, then calculated the lymphatic microvessels of every field.Statistical analysis shows that lymphatic microvessels of invasive part ofkeloid is (27.315±7.430), proliferation (17.375±6.221), margin normal skin(12.563±1.803) and normal skin of well area of keloid of non-keloid patients(11.071±2.645).4Statistical analysis result which multiple related sample nonparametricrank sum test of three groups of keloid-related data (χ2=16.800, P<0.001). Sowe think that there are significant differences between lymphatic microvesseldensity of three group, and invasive part is the higher than others. There aresignificant differences between proliferation part and margin normal skin(t=2.276, P=0.049<0.05), so we can prove that lymphatic microvessel densityof proliferation part is higher than margin normal skin. However, there are nosignificant between margin normal skin of keloid and normal skin of well areaof keloid of non-keloid patients (t=-1.389, P=0.185>0.05).Conclusions：Lymphatic endothelial marker D2-40is used to calculate lymphaticmicrovessels density in different parts of keloid, displaying lymphatic vesseldensity of invasive part of the keloid are obviously highest than proliferationpart and marginal normal skin, which show that the growing of lymphaticmicrovessels of keloid participate and contribute to the formation of keloid, asinvasive growth of keloid offers another theory.