Successful Correction of Depressed Scars of the Forehead Secondary to Trauma and Morphea en coup de sabre by en bloc Autologous Dermal Fat Graft

Abstract Background Autologous fat grafting (AFG) has been employed in surgical practice as a filling method. However, controversies remain on the specifics of this technique. So far, few relevant experimental large animal studies have objectively assessed factors related to AFG integration. Objectives This study utilized an experimental, medium-sized animal model to compare the feasibility of AFG collected employing 2 different techniques with instruments of distinct thicknesses. Methods Twenty minipigs (Sus scropha domesticus) were subjected to AFG harvesting via en bloc resection utilizing 3- (Group I) and 5-mm-diameter (Group II) round punch blades (PBs) and liposuction (LS) with 3- (Group III) and 5-mm-diameter cannulas (Group IV). Both samples were grafted intramuscularly (biceps femoralis). Hematoxylin and eosin staining was employed to identify intact adipocytes, fat necrosis, fibrosis, inflammation, and oil cysts. Immunohistochemical staining (perilipin-A, tumor necrosis factor alfa, and cluster of differentiation number 31) was utilized to quantify the feasibility of adipocytes, tissue necrosis, and neoangiogenesis, respectively. Results Hematoxylin and eosin analysis showed that fat necrosis and histiocyte presence were significantly lower in the AFG harvested utilizing a PB than in LS. For perilipin-A, a statistical difference was observed between subgroups I and III (P = 0.001) and I and IV (P = 0.004). Instrument diameter had no effect on graft integration in comparisons between groups II and III (P = 0.059) and II and IV (P = 0.132). Conclusions In this experimental study, fat collected utilizing a PB demonstrated higher adipocyte viability than fat collected with LS. The diameter of the collection instruments, whether PB or LS, had no effect on graft integration.

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Single-Stage Orbital Socket Reconstruction Using the Oversized Dermis Fat Graft and the 22 mm Silicone Orbital Implant after an Extended Enucleation

Case Reports in Ophthalmological Medicine ◽

10.1155/2018/8954193 ◽

2018 ◽

Vol 2018 ◽

pp. 1-4

Author(s):

Biljana Kuzmanović Elabjer ◽

Mladen Bušić ◽

Daliborka Miletić ◽

Mirjana Bjeloš ◽

Borna Šarić ◽

...

Keyword(s):

Choroidal Melanoma ◽

Single Stage ◽

Long Term Results ◽

Fat Graft ◽

En Bloc ◽

Orbital Implant ◽

Absorbable Sutures ◽

Previously Treated ◽

Socket Reconstruction

We would like to present a surgical technique of orbital socket reconstruction using oversized dermis fat graft and 22 mm silicone orbital implant in a single-stage after extended enucleation in two patients with massive local recurrence of anteriorly located choroidal melanoma previously treated with endoresection. Orbital tissues en bloc were removed leaving conjunctival lining only at the fornices. Simultaneously, the 22 mm silicone sphere was implanted deeply into the orbit and covered with the oversized dermis fat graft of 30 mm in height and 35 mm in length with 20 mm of the fat thickness. The graft was sutured to the residual forniceal conjunctiva with interrupted 6/0 absorbable sutures overlapping conjunctiva with the graft edge for 2 mm to facilitate the epithelization. Epithelization was completed in two months, leaving well-formed fornices with good fitting of the prosthesis. The key point of orbital socket reconstruction after extended enucleation is to restore conjunctival lining prior to volume. Thus, whenever facing a massive volume and conjunctival lining loss, simultaneous insertion of the 22 mm silicone sphere deep into the orbit combined with oversized dermis fat graft is, in our opinion, the method of choice. It proved to be safe and effective with favourable long-term results.

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En Bloc Autogenous Fat Graft Harvest From the Anterior Iliac Crest

Journal of Oral and Maxillofacial Surgery ◽

10.1016/j.joms.2008.08.046 ◽

2009 ◽

Vol 67 (3) ◽

pp. 704-705 ◽

Cited By ~ 1

Author(s):

Harry Papadopoulos ◽

R. Scott Edwards

Keyword(s):

Iliac Crest ◽

Fat Graft ◽

En Bloc ◽

Graft Harvest

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Platinum Compounds as Stains for Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100051165 ◽

1974 ◽

Vol 32 ◽

pp. 230-231

Author(s):

S. K. Aggarwal ◽

P. McAllister ◽

R. W. Wagner ◽

B. Rosenberg

Keyword(s):

Electron Microscopy ◽

Hela Cells ◽

Uranyl Acetate ◽

Sarcoma 180 ◽

Platinum Compounds ◽

Kb Cells ◽

En Bloc ◽

Human Lymphoma ◽

Ultrathin Sections ◽

Blue Coloration

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.

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Cellular Contacts Within the Interhemal Membrane of the Rat Placenta at Term

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100050743 ◽

1974 ◽

Vol 32 ◽

pp. 146-147

Author(s):

William P. Jollie

Keyword(s):

Osmium Tetroxide ◽

Uranyl Acetate ◽

Capillary Endothelium ◽

En Bloc ◽

Aldehyde Fixation ◽

Chorioallantoic Placenta

By routine EM preparative techniques, the tissues which, collectively, separate maternal and fetal bloods in the fully formed chorioallantoic placenta of the rat have been shown to consist of three chorionic layers, or trophoblast, and a layer of allantoic capillary endothelium [Fig. 1]. Relationships between these layers are best demonstrated by special techniques, viz., cacodylate-buffered aldehyde fixation, collidine-buffered osmium tetroxide postfixation, and en bloc staining with uranyl acetate. By using this method on placentas at term, the cells of the outermost chorionic layer (Trophoblast 1) appear to be attached to each other by means of maculae adherentes which sometimes occur in clusters [Fig. 2].

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Localization of pancreatic enzymes in ultrathin frozen sections stained with metal labeled antibody

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081310 ◽

1978 ◽

Vol 36 (2) ◽

pp. 90-91

Author(s):

Kenjiro Yasuda

Keyword(s):

Fluorescent Antibody ◽

Pancreatic Enzymes ◽

Tissue Preparation ◽

Zymogen Granules ◽

Frozen Sections ◽

En Bloc ◽

Antibody Method ◽

Ultrathin Frozen Sections ◽

Fluorescent Antibody Method

Localization of amylase,chymotrypsinogen and trypsinogen in pancreas was demonstrated by Yasuda and Coons (1966), by using fluorescent antibody method. These enzymes were naturally found in the zymogen granules. Among them, amylase showed a diffuse localization around the nucleus, in addition to the zymogen granules. Using ferritin antibody method, scattered ferritin granules were also found around the Golgi area (Yasuda et al.,1967). The recent advance in the tissue preparation enables the antigen to be localized in the ultrathin frozen sections, by applying the labeled antibodies onto the sections instead of staining the tissue en bloc.The present study deals with the comparison of the localization of amylase and lipase demonstrated by applying the bismuth-labeled, peroxidase-labeled and ferritin-labeled antibody methods on the ultrathin frozen sections of pancreas, and on the blocks of the same tissue.

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Lead aspartate: a new en bloc stain for electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081036 ◽

1978 ◽

Vol 36 (2) ◽

pp. 34-35

Author(s):

J.R. Walton

Keyword(s):

Electron Microscopy ◽

Calcium Ion ◽

Enzyme Reaction ◽

Lead Citrate ◽

Reaction Products ◽

En Bloc ◽

Thin Sections ◽

Lead Salts ◽

Thin Sectioning ◽

High Alkalinity

In electron microscopy, lead is the metal most widely used for enhancing specimen contrast. Lead citrate requires a pH of 12 to stain thin sections of epoxy-embedded material rapidly and intensively. However, this high alkalinity tends to leach out enzyme reaction products, making lead citrate unsuitable for many cytochemical studies. Substitution of the chelator aspartate for citrate allows staining to be carried out at pH 6 or 7 without apparent effect on cytochemical products. Moreover, due to the low, controlled level of free lead ions, contamination-free staining can be carried out en bloc, prior to dehydration and embedding. En bloc use of lead aspartate permits the grid-staining step to be bypassed, allowing samples to be examined immediately after thin-sectioning.Procedures. To prevent precipitation of lead salts, double- or glass-distilled H20 used in the stain and rinses should be boiled to drive off carbon dioxide and glassware should be carefully rinsed to remove any persisting traces of calcium ion.

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Quantitative EM of gap junction distribution in mutant drosophila wing discs

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100077177 ◽

1983 ◽

Vol 41 ◽

pp. 720-721

Author(s):

J.S. Ryerse

Keyword(s):

Gap Junctions ◽

Growth Control ◽

Intercellular Junctions ◽

Wing Disc ◽

En Bloc ◽

Metabolic Cooperation ◽

Drosophila Wing ◽

Adult Wing ◽

Wing Discs ◽

Wing Pouch

Gap junctions are intercellular junctions found in both vertebrates and invertebrates through which ions and small molecules can pass. Their distribution in tissues could be of critical importance for ionic coupling or metabolic cooperation between cells or for regulating the intracellular movement of growth control and pattern formation factors. Studies of the distribution of gap junctions in mutants which develop abnormally may shed light upon their role in normal development. I report here the distribution of gap junctions in the wing pouch of 3 Drosophila wing disc mutants, vg (vestigial) a cell death mutant, 1(2)gd (lethal giant disc) a pattern abnormality mutant and 1(2)gl (lethal giant larva) a neoplastic mutant and compare these with wildtype wing discs.The wing pouch (the anlagen of the adult wing blade) of a wild-type wing disc is shown in Fig. 1 and consists of columnar cells (Fig. 5) joined by gap junctions (Fig. 6). 14000x EMs of conventionally processed, UA en bloc stained, longitudinally sectioned wing pouches were enlarged to 45000x with a projector and tracings were made on which the lateral plasma membrane (LPM) and gap junctions were marked.

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Ultrastructural comparison of prolactin adenomas of pituitary in men and women

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100103796 ◽

1988 ◽

Vol 46 ◽

pp. 346-347

Author(s):

R.C. Caughey ◽

U.P. Kalyan-Raman

Keyword(s):

Endoplasmic Reticulum ◽

Electron Microscope ◽

Phosphate Buffer ◽

Pituitary Adenomas ◽

Osmium Tetroxide ◽

Secretory Granules ◽

Uranyl Acetate ◽

En Bloc ◽

Men And Women ◽

Transmission Electron

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.

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