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New System Delivering Microwaves Energy for Inducing
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Global Dermatology, Asan 2017 2 in which it has been considered the months following treatment. properties of some tissues related to the absorption of. Temperature monitoring was performed, microwaves energy in different models using. superficially by the thermometric infrared camera, equivalent phantom and ex vivo measurements The. vision system and internally in the subdermal fat layer. most of experimental evaluations available in the, at 7mm depth by sterile thermometric glass fibre. literature related to fat reduction have been, The epidermal temperature in the treated area was.
performed in vitro conditions which only in few cases. always observed in a safety rage of 15 25 C while at. could represent what and how happens in vivo in the. 4 to 7 mm in depth was of 50 C Figure 1A and 1B, complex dynamics inside tissues and organs in the. individuals In this paper we present morphological. observations on the subcutaneous adipose tissue, following microwaves irradiation of the skin. A controlled electromagnetic field 2 45, GHz perpendicularly applied to the skin is selectively. absorbed by the subdermal fat layer thanks to the, dielectric properties of the different tissues epidermis. dermis fat crossed by the applied EMF The high, controlled emission of the EMF can establish a perfect.
coupling only in the presence of a fat layer due to the. absorption characteristics of this tissue at the, established frequency. In the present in vivo study on an animal, model 7 minutes thermal exposure to 50 C. histological and ultrastructural evidences show, conservation without damage to the epidermis and. dermis layers while the subdermal fat is modified, through a series of molecular processes stimulating a. Figure 1 A and B are showing the tip of the temperature probe. massive delivery of fat droplets and the beginning of arrow by ultrasound A and the temperature measured inside the. auto adipolysis hypodermal layer B respectively C and D correlation of. ultrasound image C and biopsy sample D tissue layers The. different layers are clearly identifiable Epidermis and Dermis E. D Subcutis Hypodermal Adipose Tissue Fascia Muscle. Material and Methods, In each time step biopsy samples were.
collected immediately immersed in the fixative, A mathematical model was used to correlate solutions and processed both for light microscopy and. the frequency of the electromagnetic field and the transmission electron microscopy. dielectric properties of the tissues to induce localised. hyperthermia only in the subcutaneous fat layer The. non invasive pseudo transcutaneous electromagnetic Light microscopy. field Coolwaves by Onda DEKA Florence Italy Biopsy samples were immediately immersed. was applied to thermally induce adipocytes damage in a 4 paraformaldehyde sodium phosphate buffer. During the treatment applying a transmitting solution for 24 hrs and then processed dehydration. handpiece on the surface of the skin the paraffin embedding and sectioning for light. electromagnetic field energy was delivered into the microscopy Sections were stained with Haematoxylin. subcutaneous adipose tissue SAT of a Vietnamese and Eosin and other sections were stained with. pig this study on the animal model was carried out in Picrosirius Red for collagen staining. full compliance with international guidelines for safety. and compliance with their use with a 50 000 J total. dose delivered in 7 minutes over a cutaneous area 15 Electron microscopy. x 15 cm The handpiece used was maintained at a, temperature of 5 C through a specific cooling system Fixation was performed by immersion of. biopsy samples in a 2 5 glutaraldehyde EM grade, The experimental plan was designed on the 2 paraformaldehyde in 0 1M sodium cacodylate. basis of a single irradiation session and for each skin buffer solution pH 7 3 for 6 hours at 4 C After. region treated biopsy samples were collected and washing in the same buffer samples were post fixed. processed for light and electron microscopy owing to for 2 h in osmium tetroxide 1 33 in 0 1 M s collidine. the following steps T0 before treatment as control buffer dehydrated in a graded series of ethanol 30. T1 immediately after treatment T2 6h following 50 70 80 95 100 propylene oxide and. treatment T3 1 month following treatment and T4 2 finally embedded in epoxy resin Epon 812 Semithin. 2992 https www id press eu mjms index, Fioranelli et al New System Delivering Microwaves Energy for Inducing Subcutaneous Fat Reduction. 0 2 m and ultrathin 40 60 nm sections were Small blood vessels were also identifiable At. obtained at the ultramicrotome Reichert Ultracut S the electron microscope Figure 2B the very thin. provided with a diamond knife Semithin sections were peripheral cytoplasm appeared to contain dispersed. stained with Toluidine blue and ultrathin sections organelles as mitochondria rare profiles of the. previously collected on 200 m mesh copper grids endoplasmic reticulum and small vesicles involved. were counterstained with lead citrate and uranyl realistically in the physiological transport of materials. acetate A Zeiss EM 902 transmission electron lipids included responsible of the correct. microscope operating at 80 kV with an objective homeostatic processes across the cytoplasm to and. aperture of 30 60 m was used for direct from the interstitial connective tissue. observation Electron micrographs were recorded on, At time T1 immediately the following.
Kodak 4489 Electron Image film and finally digitised. treatment small single or multiple vesicles were, with an Epson Perfection V750 Pro scanner at 1200. visible at the light microscope in the peripheral, cytoplasm of adipocytes Figure 3A. At time T2 6 hrs after treatment many, vesicles containing material very similar to the big. lipidic content of the adipocyte were detectable inside. Results adipocyte peripheral cytoplasm In ultrathin sections. observed at the electron microscope the whole, peripheral cytoplasm was completely occupied by. The description of our observations will follow vesicles containing a relatively electron transparent. the time sequence of samples collection using the material This feature appears particularly evident in. most suitable presentation of our findings to better the oblique sections of the inner surface of the. understand the functional dynamics peripheral cytoplasm as a holey structure Figure 3B. Before sample collection it has been used In a relatively thick section at the electron microscope. ultrasound imaging to define the thickness of the we observed blebbing structures at the surface of. different structural layers particularly the depth and adipocytes Figure 3C projecting part of their lipidic. extension of the subcutaneous tissue as the target of content towards the interstitial connective tissue. the microwave energy transfer Figure 1C and, compare it with biopsy sample Figure 1D.
At time T0 defining the control samples, collected before the treatment microscopic. morphology of adipocytes appeared normal spherical. in shape with the whole volume of cells appearing, filled with a homogeneous content surrounded by a. very thin layer of peripheral cytoplasm where an, elongated and flattened nucleus was well identifiable. Figure 2A All around adipocytes and between them, a loose connective tissue was appreciable with some. cells fibroblasts and an extracellular matrix, constituted by collagen fibrils relatively dispersed.
inside a faint ground substance Figure 2A, Figure 3 T1 T2 T3 A T1 immediately the following treatment. Semithin section at the light microscope from epoxy resin. embedding stained with toluidine blue Inside the thin peripheral. cytoplasm of adipocytes some vesicles with the content similar to. the big lipid content of cells are visible B T2 6 hrs following. treatment at the electron microscope the peripheral cytoplasm of. adipocytes shows numerous vesicles occupying the whole. thickness of the cytoplasm In the oblique sections the internal. cytoplasmic surface appears fully of vesicles C T3 6 hrs following. Figure 2 T0 Control A Semithin section at the light microscope treatment the high magnification at the electron microscope. from epoxy resin embedding stained with toluidine blue Adipocytes represents the superficial blebbing as a direct extrusion of a lipid. show peripheral nucleus N with dispersed chromatin a very thin droplet asterisk through peripheral cytoplasm from the inside of an. peripheral layer of cytoplasm and a homogeneous lipidic content adipocyte left to the outside in the interstitial connective tissue. Around them elongated fibroblasts and bundles of collagen fibres ICT Arrows mark the thin peripheral cytoplasm of the adipocyte. are also observable B Ultrastructure of the very thin peripheral. cytoplasmic of an adipocyte and inside it two elongated. mitochondria were observable A continuous basal lamina close to. the plasma membrane and collagen microfibrils in the surrounding Further some cytological details of. connective tissue is also visible The homogeneous fat is adipocytes involved in these mechanisms have been. constituting the inner content of the adipocyte, observed at the electron microscope as mitochondrial. Open Access Maced J Med Sci 2019 Sep 30 7 18 2991 2997 2993. Global Dermatology, swelling with few disorganised cristae and. interruptions of the inner membrane interruptions of. the plasma membrane dilations of the endoplasmic, reticulum data not shown The whole of these. features suggests adipolysis via necrotic processes. At time T3 1 month after treatment, observing transverse sections of the wall of.
adipocytes numerous vesicles were evident inside an. electron dense cytoplasm Some of them appeared as, invaginations of the inner side of the peripheral. cytoplasm with a content continuous with the big, lipidic sphere of the adipocyte others appeared as. openings towards the interstitial connective tissue. These features are suggesting a mechanism of endo, exo cytosis of lipids from the inside of the adipocyte to. the outside towards the interstitial connective tissue. At time T3 in some areas of the biopsies an, inflammatory infiltrate was detectable This infiltrates. appeared constituted by numerous cells penetrating. the interstitial tissue singularly or grouped encircling. single adipocytes Figure 4A, The infiltrate is constituted mainly by.
monocytes distributing free in the interstitial, connective tissue or close contact with adipocytes. covering their external surface Single monocytes, appeared provided with cytoplasmic processes. projecting free in the interstitial connective tissue or. directly contacting free vesicles released by, adipocytes The first contact of monocytes with lipids. released by necrotic adipocytes is realistically, suggested by a direct contact representing the result. of the find me action of the extracellular lipid, structures towards monocytes and the starting of the.
Figure 4 T3 1 month following treatment A Light microscopy of a. mechanism eat me stimulating the phagocytic action Crown Like Structure constituted by a crown of single monocytes. of macrophages The following step in the mechanism joined between them by thin cytoplasmic processes distributed all. of adipolysis observed at the same time T3 after 1 around an adipocyte These monocytes appear to contain in the. month from treatment is represented by the formation cytoplasm small droplets similar to the content of the adipocyte. Realistically these droplets are the result of phagocytosis due to. of Crown Like Structures as the most typical feature macrophage activity B The smaller adipocyte appears completely. CLS are constituted by single monocytes closely surrounded by a multinucleated structure originated by the fusion of. related one with the other forming a pluricellular single monocytes forming a single syncytial macrophage The small. structure directly encircling adipocytes Figure 4A dimensions of the adipocyte are realistically due to stimulated. phagocytosis of the fat of necrotic adipocyte Semithin sections. Inside these cells small droplets appear very similar from epoxy resin embedding toluidine blue stained C In the high. to the adipocyte content suggesting the starting of a left part of the electron micrographs red arrow a portion of the big. phagocytosis mechanism In a following step of the lipidic content of the adipocyte is directly contacting the interstitial. mechanism suggested by images single monocytes tissue The remnant part of this content is directly contacting the. plasma membrane of a macrophage small black arrows which. forming the crown around adipocyte fuse their has already internalised in his cytoplasm numerous lipid droplets. cytoplasm forming a unique big cytoplasm with many The lack of the peripheral cytoplasm of the adipocyte demonstrates. nuclei inside syncytium all around adipocytes Figure that this adipocyte has completed the process of cellular death. 4B realistically for a more effective action of Furtherly the basal membrane far from the adipocyte appears. highly disorganised inside interstitial tissue asterisks N. phagocytosis of the content of these cells and the macrophage nucleus. cytoplasmic residuals of the same necrotic, adipocytes Remark in Figure 4B the small. dimensions of the represented adipocyte due to the The surface of this cell appears expanded in. action of fat removal by macrophages At the electron numerous and complex superficial projections towards. Nicola Zerbinati1 Edoardo d Este2 Antonia Icaro Cornaglia3 Federica Italy 12Clinic for Psychiatric Disorders Dr Laza Lazarevic Belgrade Serbia Citation Zerbinati N d Este E Cornaglia AI Riva F Farina A Calligaro A Gallo G Perrotta ER Protasoni M Bonan P Vojvodic A Fioranelli M Van Thuong N Lotti T Tirant M Vojvodic P New System Delivering Microwaves Energy

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