Early Detection of Steroid-Induced Femoral Head Necrosis using 99mTc-Cys-Annexin V-based Apoptosis Imaging in a Rabbit Model

Abstract Background At present, the early diagnosis of femoral head necrosis mainly relies on MRI, and most early patients are difficult to make an accurate diagnosis. Therefore, to investigate the early diagnostic value of 99mTc-Cys-Annexin V SPECT imaging were compared with MRI in rabbit models of steroid-induced femoral head necrosis. Methods The rabbit models of steroid-induced femoral head necrosis were established by intravenous injection of horse serum and gluteal muscle injection of methylprednisolone in of 5-month-old healthy New Zealand white rabbits. 99mTc-Cys-Annexin V SPECT imaging and MRI were performed at 2nd week, 4th week, and 6th week after modeling. After that, histopathology was used to verify the success of modeling. Apoptosis was detected by transmission electron microscopy and TUNEL. Results At 2 weeks after the injection of hormone, 99mTc-Cys-Annexin V SPECT image showed abnormal radioactive uptake in the bilateral femoral head. And over time, the radioactivity concentration was more obvious, and the ratio of T/NT (target tissue/non-target tissues) was gradually increased. In the SPECT imaging at each time point, T/NT ratio of the model group was significantly higher than that of the control group (P < 0.01); at 4 weeks after the injection of hormone, MRI showed an abnormal signal of osteonecrosis. At 2, 4, and 6 weeks after hormone injection, apoptosis was observed by TUNEL and transmission electron microscopy. Conclusions 99mTc-Cys-Annexin V SPECT imaging can diagnose steroid-induced femoral head necrosis earlier than MRI, and has potential application value for non-invasively detecting early and even ultra-early stage of femoral head necrosis.

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Early Detection of Steroid-Induced Femoral Head Necrosis using 99mTc-Cys-Annexin V-based Apoptosis Imaging in a Rabbit Model

10.21203/rs.3.rs-68831/v1 ◽

2020 ◽

Author(s):

Xiaolong Wang ◽

Jianbo Li ◽

Da Man ◽

Rui Liu ◽

Jianmin Zhao

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Femoral Head ◽

Rabbit Model ◽

Annexin V ◽

Spect Imaging ◽

Femoral Head Necrosis ◽

Control Group ◽

Target Tissue ◽

Transmission Electron

Abstract Background At present, the early diagnosis of femoral head necrosis mainly relies on MRI, and most early patients are difficult to make an accurate diagnosis. Therefore, to investigate the early diagnostic value of 99mTc-Cys-Annexin V SPECT imaging were compared with MRI in rabbit models of steroid-induced femoral head necrosis. Methods The rabbit models of steroid-induced femoral head necrosis were established by intravenous injection of horse serum and gluteal muscle injection of methylprednisolone in of 5-month-old healthy New Zealand white rabbits. 99mTc-Cys-Annexin V SPECT imaging and MRI were performed at 2nd week, 4th week, and 6th week after modeling. After that, histopathology was used to verify the success of modeling. Apoptosis was detected by transmission electron microscopy and TUNEL. Results At 2 weeks after the injection of hormone, 99mTc-Cys-Annexin V SPECT image showed abnormal radioactive uptake in the bilateral femoral head. And over time, the radioactivity concentration was more obvious, and the ratio of T/NT (target tissue/non-target tissues) was gradually increased. In the SPECT imaging at each time point, T/NT ratio of the model group was significantly higher than that of the control group (P < 0.01); at 4 weeks after the injection of hormone, MRI showed an abnormal signal of osteonecrosis. At 2, 4, and 6 weeks after hormone injection, apoptosis was observed by TUNEL and transmission electron microscopy. Conclusion s 99mTc-Cys-Annexin V SPECT imaging can diagnose steroid-induced femoral head necrosis earlier than MRI, and has potential application value for non-invasively detecting early and even ultra-early stage of femoral head necrosis.

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Early detection of steroid-induced femoral head necrosis using 99mTc-Cys-Annexin V-based apoptosis imaging in a rabbit model

Molecular Medicine ◽

10.1186/s10020-020-00248-1 ◽

2020 ◽

Vol 26 (1) ◽

Author(s):

Xiaolong Wang ◽

Jianbo Li ◽

Da Man ◽

Rui Liu ◽

Jianmin Zhao

Keyword(s):

Femoral Head ◽

Photon Emission ◽

Rabbit Model ◽

Annexin V ◽

Spect Imaging ◽

Femoral Head Necrosis ◽

Terminal Deoxynucleotidyl Transferase ◽

Control Group ◽

Emission Computed Tomography ◽

Target Tissue

Abstract Background At present, the early diagnosis of femoral head necrosis mainly relies on Magnetic resonance imaging (MRI), and most early patients are difficult to make an accurate diagnosis. Therefore, to investigate the early diagnostic value of 99mTc-Cys-Annexin V Single-photon emission computed tomography (SPECT) imaging were compared with MRI in rabbit models of steroid-induced femoral head necrosis. Methods The animal model of steroid-induced femoral head necrosis (SIFHN) was established in 5-month-old healthy New Zealand white rabbits by injecting horse serum into ear vein and methylprednisolone into gluteal muscle, the purpose of modeling is to simulate the actual clinical situation of SIFNH. 99mTc-Cys-Annexin V SPECT imaging and MRI were performed at 2nd week, 4th week, and 6th week after modeling. After that, histopathology was used to verify the success of modeling. Apoptosis was detected by transmission electron microscopy (TEM) and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). Results At 2 weeks after the injection of hormone, 99mTc-Cys-Annexin V SPECT image showed abnormal radioactive uptake in the bilateral femoral head. And over time, the radioactivity concentration was more obvious, and the ratio of T/NT (target tissue/non-target tissues, which is the ratio of femoral head and the ipsilateral femoral shaft) was gradually increased. In the 99mTc-Cys-Annexin V SPECT imaging at each time point, T/NT ratio of the model group was significantly higher than that of the control group (P < 0.01); at 4 weeks after the injection of hormone, MRI showed an abnormal signal of osteonecrosis. At 2, 4, and 6 weeks after hormone injection, apoptosis was observed by TUNEL and TEM. Conclusions 99mTc-Cys-Annexin V SPECT imaging can diagnose steroid-induced femoral head necrosis earlier than MRI, and has potential application value for non-invasively detecting early and even ultra-early stage of femoral head necrosis.

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Early Detection of Steroid-Induced Femoral Head Necrosis using 99mTc-Cys-Annexin V-based Apoptosis Imaging in a Rabbit Model

10.21203/rs.3.rs-68831/v2 ◽

2020 ◽

Author(s):

Xiaolong Wang ◽

Jianbo Li ◽

Da Man ◽

Rui Liu ◽

Jianmin Zhao

Keyword(s):

Femoral Head ◽

Photon Emission ◽

Rabbit Model ◽

Annexin V ◽

Spect Imaging ◽

Femoral Head Necrosis ◽

Terminal Deoxynucleotidyl Transferase ◽

Control Group ◽

Emission Computed Tomography ◽

Target Tissue

Abstract BackgroundAt present, the early diagnosis of femoral head necrosis mainly relies on Magnetic resonance imaging (MRI), and most early patients are difficult to make an accurate diagnosis. Therefore, to investigate the early diagnostic value of 99mTc-Cys-Annexin V Single-photon emission computed tomography (SPECT) imaging were compared with MRI in rabbit models of steroid-induced femoral head necrosis.MethodsThe animal model of steroid-induced femoral head necrosis (SIFHN) was established in 5-month-old healthy New Zealand white rabbits by injecting horse serum into ear vein and methylprednisolone into gluteal muscle, the purpose of modeling is to simulate the actual clinical situation of SIFNH. 99mTc-Cys-Annexin V SPECT imaging and MRI were performed at 2nd week, 4th week, and 6th week after modeling. After that, histopathology was used to verify the success of modeling. Apoptosis was detected by transmission electron microscopy (TEM) and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). ResultsAt 2 weeks after the injection of hormone, 99mTc-Cys-Annexin V SPECT image showed abnormal radioactive uptake in the bilateral femoral head. And over time, the radioactivity concentration was more obvious, and the ratio of T/NT (target tissue/non-target tissues, which is the ratio of femoral head and the ipsilateral femoral shaft) was gradually increased. In the 99mTc-Cys-Annexin V SPECT imaging at each time point, T/NT ratio of the model group was significantly higher than that of the control group (P < 0.01); at 4 weeks after the injection of hormone, MRI showed an abnormal signal of osteonecrosis. At 2, 4, and 6 weeks after hormone injection, apoptosis was observed by TUNEL and TEM. Conclusions99mTc-Cys-Annexin V SPECT imaging can diagnose steroid-induced femoral head necrosis earlier than MRI, and has potential application value for non-invasively detecting early and even ultra-early stage of femoral head necrosis.

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Noise-induced gastric lesions: a light and electron microscopy study of the rat gastric wall exposed to low frequency noise

Arquivos de Gastroenterologia ◽

10.1590/s0004-28032012000100014 ◽

2012 ◽

Vol 49 (1) ◽

pp. 82-88 ◽

Cited By ~ 4

Author(s):

Jorge Fonseca ◽

José Martins-dos-Santos ◽

Pedro Oliveira ◽

Nuno Laranjeira ◽

Artur Aguas ◽

...

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Gastric Wall ◽

Low Frequency ◽

Collagen Iv ◽

Control Group ◽

Frequency Noise ◽

Low Frequency Noise ◽

Transmission Electron ◽

Significant Difference

CONTEXT: Only a few studies evaluated the digestive alterations caused by low frequency noise (LFN) and most focused only on mucosal alterations. OBJECTIVES: To investigate the morphological injury of LFN-exposed gastric wall, beyond the epithelial layer. METHODS: Wistar rats were exposed to low frequency noise (LFN), during increasing periods, 1 to 13 weeks. A control group was kept in silence. Gastric specimens were studied using: (i) light microscopy with hematoxylin-eosin and immunostaining for collagens; (ii) transmission electron microscopy; (iii) morphometry allowing statistical analysis. RESULTS: Submucosa of all LFN-exposed animals exhibit increased thickness with fibrous proliferation. Transmission electron microscopy showed massive collagen deposition. Immunostaining identified collagen IV as responsible for the increased thickness. Morphometry allowed the demonstration of a significant difference of thickness between control and exposed groups. Vascular alterations included: i) intima proliferation and thickening, rupture of the internal elastic lamina, thrombotic changes; ii) thickening of the media; iii) after 9 weeks of LFN-exposure, we found new formed vessel presenting tortuous and twisted. There is a significant difference of arterial wall thickness between control and exposed groups. CONCLUSIONS: Deeper layers of gastric wall undergo alterations, including fibrosis of the submucosa caused by collagen IV deposition, an early marker of neoangiogenesis. Vascular alterations included thickening and thrombotic phenomena, but also images of newly formed vessels. This study suggests that, at least in the stomach, LFN-induced fibrosis could be linked with neoangiogenesis.

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Effect of Chaihu Shugan Powder-Contained Serum on Glutamate-Induced Autophagy of Interstitial Cells of Cajal in the Rat Gastric Antrum

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/7318616 ◽

2019 ◽

Vol 2019 ◽

pp. 1-7 ◽

Cited By ~ 1

Author(s):

Ren-Qian Tan ◽

Zhi Zhang ◽

Jing Ju ◽

Jiang-Hong Ling

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Interstitial Cells Of Cajal ◽

Underlying Mechanism ◽

Control Group ◽

Motility Disorders ◽

Transmission Electron ◽

Cells Of Cajal ◽

Bcl2 Expression ◽

Gi Motility

Gastrointestinal (GI) motility disorder is caused by excessive autophagy of the interstitial cells of Cajal (ICC). Chaihu Shugan Powder (CSP) is a traditional Chinese medicine with therapeutic benefits in GI motility disorders; however, the underlying mechanism of its therapeutic effect in GI disorders, especially autophagy of ICC, remains unclear. Thus, this study investigated the effects of CSP-contained serum on glutamate-induced autophagy in rat gastric ICC, exploring its underlying mechanism. In vitro cultured rat stomach ICC were identified by fluorescence microscopy and then stimulated with glutamate (5 mmol/L) for 3 h to establish the autophagy model. These cells were then treated with 10% CSP-containing serum or the autophagy inhibitor 3-methyladenine (3-MA; 5 mmol/L) for 24 h. The control group was cultured with only 10% serum containing physiological saline. The viability of ICC was measured by the CCK-8 assay. The ultrastructure and autophagosomes of ICC were observed using transmission electron microscopy. LC3 expression was detected by immunofluorescence, and LC3, Beclin1, Bcl2, and PI3KC3 expression was detected by western blot analysis. Transmission electron microscopy showed abundant endoplasmic reticulum, mitochondria, and other organelles in the control group, whereas the cells in the autophagy model control group had clear autophagic vacuoles, which were not apparent in both CSP and 3-MA groups. ICC viability was significantly increased by CSP and 3-MA interventions (P < 0.01), accompanied by a decrease in LC3 fluorescence (P < 0.01). Moreover, the expression levels of LC3II/I, Beclin1, and PI3KC3 were significantly decreased (all P < 0.01) with CSP and 3-MA treatment, while Bcl2 expression level was higher than that of the model group (P < 0.01). Thus, CSP can reduce autophagic damage by enhancing Bcl2 expression and downregulating the expression of LC3, Beclin1, and PI3KC3 to protect ICC. These results highlight the potential of CSP in the treatment of GI motility disorders.

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Phenethyl Isothiocyanate (PEITC) Regulates Autophagy in Chronic Lymphocytic Leukemia.

Blood ◽

10.1182/blood.v120.21.2906.2906 ◽

2012 ◽

Vol 120 (21) ◽

pp. 2906-2906

Author(s):

Jemimah Adams ◽

R Gitendra Wickremasinghe ◽

Archibald G Prentice ◽

Jonathan C. Strefford ◽

Andrew Duncombe ◽

...

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Annexin V ◽

Lymphocytic Leukemia ◽

Phenethyl Isothiocyanate ◽

Autophagosome Formation ◽

Double Membrane ◽

Transmission Electron ◽

Membrane Bound ◽

Autophagy Pathway

Abstract Abstract 2906 Chronic Lymphocytic leukemia (CLL) is currently incurable using conventional therapies. CLL cells can evade killing by various therapeutic strategies. However the precise mechanisms are currently unknown. Autophagy is regulated by a complex system of proteins, and is used by both normal and malignant cells as a protective mechanism against cellular stress induced by starvation, hypoxia, reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress. In malignant cells autophagy was shown to promote tumorigenesis and/or resistance to chemotherapy. Therefore we hypothesized that autophagy may play a role in CLL biology. Autophagy can also promote cell death when stress signals are elevated above a particular threshold for a prolonged period of time. In this study we investigated the basal expression levels of autophagy specific genes and the effect of autophagy specific inhibitors (Bafilomycin, 3-methyladenine and hydroxychloroquine) and inducers (Phenethyl isothiocyanate) on CLL survival. Phenethyl isothiocyanate (PEITC) is about to enter clinical trials for CLL (NCT00968461). We have investigated induction of components of the autophagic pathway following treatment of CLL cells in vitro with a range of chemical inhibitors. Immunoblotting was carried out to investigate components of the autophagy pathway using phosphorylation state-specific and pan-reactive antibodies. Bafilomycin (BAF), 3-methyladenine (3-MA) and hydroxychloroquine (HCQ) toxicity towards CLL samples were evaluated by Annexin V/PI staining, MTT assay and immunoblotting for cleavage of the caspase 3 substrate poly(ADP ribose) polymerase (PARP) from its 116KDa to its 85KDa form. PEITC was used at concentrations between 2.5 and 25μM to investigate its effect on signaling. Autophagy was quantitated by immunoblotting of LC3-I and LC3-II. Lipidation of LC3 from LC3-I to LC3-II is a surrogate marker of autophagy and is essential for autophagasome formation. Immunoblotting was also performed for ATG3, ATG5 and ATG7, key components of the autophagy pathway. Monodansylcadaverine (MDC) was used with immunofluorescence and FACS analysis to investigate increases in autophagasome formation. Transmission electron microscopy (TEM) was used to confirm double membrane bound autophagosomes. Co-immunoprecipitation was used to evaluate if Beclin-1 was sequestered by Bcl-2 preventing autophagy. Its release from Bcl-2 enables Beclin-1 to interact with other autophagy specific proteins and initiates autophagasome formation. LC3-I was lipidated to LC3-II (p=0.019) and ATG3 (p=0.021) was upregulated to a greater extent in CLL samples compared with normal B-cell controls at basal levels. This suggested that autophagy was active to a greater extent in CLL samples compared with normal individuals. In addition Beclin was dissociated from Bcl-2 in CLL samples indicating that autophagy was active. Autophagy appears to be a pro-survival mechanism in untreated CLL cells as inhibiting basal levels of autophagy with autophagy inhibitors BAF (50–200nM), 3-MA (5–10mM) and hydroxychlorquine (5–10μM) resulted in CLL apoptosis as shown by MTT, Annexin V/PI analysis and PARP cleavage. Interestingly augmenting autophagy was also capable of inducing apoptosis in CLL samples. Treatment with PEITC caused an increase in punctate staining using MDC which is suggestive of autophagosome formation. We went on to determine that PEITC further induced LC3-II lipidation using immunoblotting and showed a substantial increase in overall LC3 protein expression. PEITC also induced the expression of ATG3, a key protein in the autophagy pathway. We then evaluated autophagosome formation using TEM (Figure 1). Our data showed greater numbers of autophagosomes in the PEITC treated samples compared to the untreated controls. Therefore autophagy in CLL sits on a knife-edge, such that perturbations that either increase pro- death or decrease pro-survival autophagy signals can result in CLL cell death, depending on the duration and intensity of the signal. Figure 1. Transmission electron microscopy of CLL cells CLL cells were treated with 10μM PEITC. Double membrane bound organelles were found in the CLL cells after treatment which were not present in the no addition control (depicted by the arrows). These organelles are autophagsomes. Magnification (left picture) ruler is 500nM, (right picture) ruler is 100nM Figure 1. Transmission electron microscopy of CLL cells . / CLL cells were treated with 10μM PEITC. Double membrane bound organelles were found in the CLL cells after treatment which were not present in the no addition control (depicted by the arrows). These organelles are autophagsomes. Magnification (left picture) ruler is 500nM, (right picture) ruler is 100nM Disclosures: No relevant conflicts of interest to declare.

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Effect of the Radioprotector WR2721 on Irradiation-Induced Injury to Ciliated Cells of Eustachian Tube

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348949210100504 ◽

1992 ◽

Vol 101 (5) ◽

pp. 395-402 ◽

Cited By ~ 9

Author(s):

Timothy T. K. Jung ◽

Young Min Park ◽

David Panossian ◽

Douglas Weeks ◽

Stanley K. Miller ◽

...

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Eustachian Tube ◽

Control Group ◽

Ciliated Cells ◽

Transmission Electron ◽

Irradiation Injury ◽

Intraperitoneal Dose ◽

Serous Otitis Media ◽

Experimental Group

Our previous studies revealed that injury to the ciliated cells of the eustachian tube may be the primary cause of irradiation-induced serous otitis media. The purpose of this study was to investigate the effects of the radioprotector WR2721 on irradiation-induced injury to ciliated cells of the eustachian tube (ET) in chinchillas. Twelve chinchillas were divided into two groups: the control group and the experimental group, which was pretreated with a single intraperitoneal dose of the radioprotector S-2-[3-aminopropylamino]ethylphosphorothioic acid (WR2721) 400 mg/kg. The two groups were exposed to 30 Gy of 13-MeV electrons in a single fraction to the area of the bullae and nasopharynx. Ciliary dysfunction was tested and ciliated cells of the ET were examined by scanning and transmission electron microscopy. Pretreatment with WR2721 was found to protect ciliated cells of the ET from irradiation injury.

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Ultrastructural lesions and immunohistochemical analysis of Bcl-2 protein expression in the kidney of chickens with experimental ochratoxicosis

Acta Veterinaria Hungarica ◽

10.1556/avet.2013.021 ◽

2013 ◽

Vol 61 (3) ◽

pp. 344-353 ◽

Cited By ~ 4

Author(s):

Carmen Solcan ◽

Dorina Timofte ◽

Viorel Floristean ◽

Stuart Carter ◽

Gheorghe Solcan

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Ochratoxin A ◽

Broiler Chickens ◽

Immunohistochemical Analysis ◽

Control Group ◽

Transmission Electron ◽

Apoptotic Marker ◽

Convoluted Tubules ◽

Nephrotoxic Effect

A study was conducted to evaluate the nephrotoxic effect of ochratoxin A (OTA) in broiler chickens. Forty Ross 308 broilers (6 days old) were divided into two groups: one group received daily, by gavage, ochratoxin A at a daily dose of 50 μg/kg body weight for up to 21 days, while the control group received only diluent (sunflower oil). After 21 days, the chickens were euthanised and the kidneys removed for analysis by histopathology and immunohistochemistry to detect an anti-apoptotic marker (Bcl-2), and by transmission electron microscopy. Macroscopically the kidneys were enlarged, showing degeneration and gout deposits. Histologically, glomerulonephrosis and tubulonephrosis were common lesions in all chicks. In two of the five chicks exposed to OTA for 21 days, focal tubular cell proliferation, multiple adenoma-like structures and Bcl-2-positive epithelial cells were identified in layers of the renal papilla and in convoluted tubules. Transmission electron microscopy of the proximal convoluted tubules identified abnormal forms of mitochondria. The nephrotoxic effect of ochratoxicosis in chickens is probably due to carcinogenic changes induced in the epithelial tissues.

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Hepatotoxic effects of melamine exposure from the weaning period in rats: a flow cytometric, electron microscopic, and histopathologic study

Toxicology Research ◽

10.1093/toxres/tfab022 ◽

2021 ◽

Author(s):

Zuleyha Erisgin ◽

Hasan Serdar Mutlu ◽

Yavuz Tekelioglu ◽

Engin Deveci ◽

Ugur Seker

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Periodic Acid ◽

Flow Cytometric Analysis ◽

Control Group ◽

Albino Rats ◽

Histopathological Analysis ◽

Periodic Acid Schiff ◽

Flow Cytometric ◽

Transmission Electron

Abstract This study aims to investigate the effects of melamine exposure from the weaning period (21st postnatal days in rats) on liver tissue. Female Wistar albino rats (n = 18) were divided into three groups. About 0.1-ml saline was applied to the control group by gavage for 21 days from the postnatal 21st day. The second group was taken 50-mg/kg melamine (in 0.1-ml saline) and the third group was taken 75-mg/kg melamine (in 0.1-ml saline) p.o. On the postnatal 45th day, all rats were sacrificed under anesthesia. Then, liver tissues were cut into three parts and two of them placed in neutral formalin for histopathological and flow cytometric analysis, and one of them placed in 2.5% glutaraldehyde. Histopathological analysis was performed with hematoxylin & eosin, Masson trichrome, periodic acid Schiff stained sections, and also with transmission electron microscopy. Apoptosis (Annexin V positivity) was analyzed by flow cytometry. According to histopathological analysis, hepatocyte damage, sinusoidal dilatation, and inflammatory cell infiltration significantly increased in both melamine groups compared with the control group. Apoptosis significantly increased in the 50 and 75-mg melamine groups compared with the control group. In the results of transmission electron microscopy analysis, there was abnormal chromatin distribution in the hepatocyte nuclei, loss in the cristae of the mitochondria, and organelle loss in large areas in the cytoplasm in both melamine exposure groups. As result, melamine exposure from the weaning period causes liver damage with increasing doses.

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Effect of Electrocautery on Endothelial Integrity of the Internal Thoracic Artery: Ultrastructural Analysis with Transmission Electron Microscopy

Texas Heart Institute Journal ◽

10.14503/thij-13-3658 ◽

2014 ◽

Vol 41 (5) ◽

pp. 484-490 ◽

Cited By ~ 2

Author(s):

Burak Onan ◽

Mehmet Yeniterzi ◽

Ismihan Selen Onan ◽

Burak Ersoy ◽

Suheyla Gonca ◽

...

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Endothelial Cells ◽

Internal Thoracic Artery ◽

Ultrastructural Changes ◽

Control Group ◽

Elastic Tissue ◽

Cytoplasmic Organelles ◽

Transmission Electron ◽

Sharp Dissection

The internal thoracic artery (ITA) is typically harvested from the chest wall by means of conventional electrocautery. We investigated the effects of electrocautery on endothelial-cell and vessel-wall morphology at the ultrastructural level during ITA harvesting. Internal thoracic artery specimens from 20 patients who underwent elective coronary artery bypass grafting were investigated in 2 groups. The ITA grafts were sharply dissected with use of a scalpel and clips in the control group (n=10) and were harvested by means of electrocautery in the study group (n=10). Each sample was evaluated for intimal, elastic-tissue, muscular-layer, and adventitial changes. Free flow was measured intraoperatively. Light microscopic examinations were performed after hematoxylin-eosin and Masson's trichrome staining. Transmission electron microscopy was used to evaluate ultrastructural changes in the endothelial cells and vessel walls of each ITA. In the sharp-dissection group, the endothelial surfaces were lined with normal amounts of original endothelium, endothelial cells were distinctly attached to the basal lamina, cytoplasmic organelles were evident, and intercellular junctional complexes were intact. Conversely, in the electrocautery group, the morphologic integrity of endothelial cells was distorted, with some cell separations and splits, contracted cells, numerous large cytoplasmic vacuoles, and no visible cytoplasmic organelles. The subendothelial layer exhibited disintegration. Free ITA flow was higher in the sharp-dissection group (P=0.04). The integrity of endothelial cells can be better preserved when the ITA is mobilized by means of sharp dissection, rather than solely by electrocautery; we recommend a combined approach.

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