2.Improved cell adhesion to ion beam-irradiated biodegradable membranes.
Yong Moo LEE ; Yoon Jeong PARK ; Seung Jin LEE ; Young KU ; In Chul RHYU ; Soo Boo HAN ; Sang Mook CHOI ; Chong Pyoung CHUNG
The Journal of the Korean Academy of Periodontology 1998;28(4):601-610
Ion irradiation is a very promising tool to modify the chemical structure and physical properities of polymers. This study was aimed to evaluate the cellular adhesion to ion beam-irradiated surface of biodegradable poly-llactide(PLLA) membrane. The PLLA membrane samples were irradiated by using 35 KeV Ar+ to fluence of 5x10(13), 5x10(14) and 5x10(15)ion/cm2. Water contact angles to control and each dose of ion beam-irradiated PLLA membranes were measured. Cultured fetal rat calvarial osteoblasts were seeded onto control and each dose of ion beam-irradiated PLLA membranes and cultured. After 24 hours, each PLLA membranes onto which osteoblasts attached were examined by scanning electron microscopy(SEM). Osteoblasts were removed from each PLLA membrane and then, the vitality and the number of cells were calibrated. Alkaline phosphatase of detached cells from each PLLA membranes were measured. Ion beam-irradiated PLLA membranes showed no significantly morphological change from control PLLA membranes. In the measurement of water contact angle to each membrane, the dose range of ion beam employed in this study reduced significantly contact angles. Among them, 5x10(14) ion/cm2 showed the least contact angle. The vitalities of osteoblastes detached from each membranes were confirmed by flow cytometer and well attached cells with their own morphology onto each membranes were observed by SEM. A very strong improvement of the cell adhesion and proliferation was observed for ion beam-irradiated surfaces of PLLA membranes. 5x10(14)ions/cm2 exhibited the most strong effect also in cellular adherence. ALPase activities also tended to increase in ion beam-irradiated membranes but statistical differences were not found. These results suggested that ion beam irradiation is an effective tool to improve the adhesion and spreading behaviour of the cells onto the biodegradable PLLA membranes for the promotion of membrane-tissue integration.
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3.DNA ploidy and Cellular Proliferation Activity in Experimentally Induced Malignant Fibrous Histiocytoma.
Ji Shin LEE ; Jong Tae PARK ; Sang Woo JUHNG ; Hong Ran CHOI ; Kyu Hyuk CHO
Korean Journal of Pathology 1993;27(3):205-216
To fine out the changes of DNA ploidy and cellular proliferation activity during carcinogenesis and evaluate correlation between flow cytometrically determined S-phase fraction and proportion of proliferation cell nuclear antigen(PCNA, PC10) immunoreactive cells, the authors studied on malignant fibrous histocytoma induced by intra-articular injection of 9, 10-dimethy1-1, 2-benzanthracene(DMBA) in the rats. Forty Wistar rats were used. The results obtained were as follows. 1) Firstly, tumors were palpated 5 weeks after the last injection of DMBA and formed in 27 rats at sacrificed. Histologically, these lesions showed storiform, indicative of malignant fibrous histiocytoma. 2) Three cases of DNA aneuploidy were observed at 4 and 5 months after the last injection of DBMA and one of them, which was DNA diploidy at main mass, was found at daughter mass. 3) Flow cytometrically determined S-phase fraction and proportion of PCNA(PC10) immunoreactive cells in malignant fibrous histiocytoma induced by DMBA were much higher than in control groups and slightly increased according to sequential changes after formation of mass. The comparison of flow cytometrically determined S-phase fraction and proportion of PCNA(PC10) immunoreactive cells showed significant correlation(r=0.6092, p<0.001). Above results strongly suggest that ploidy pattern may evolve into aneuploid type during the development of tumor and proliferation activity increases during the carcinogenesis.
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4.Capillary Plexus and Vasa of the Rat Lung as Revealed by Scanning Electron Microscope of Corrosion Casts.
Kun Young KWON ; Kam Rae CHO ; Sang Pyo KIM ; Kwan Kyu PARK ; Eun Sook CHANG
Korean Journal of Pathology 1993;27(1):11-18
The pulmonary microvasculatures of rats were studied by injection replicas prepared from Mercox. This medium enabled us to easily obtain consistent, stable, and complete injected replicas of the pulmonary vasculature. In order to investigate the three dimensional structures of the tributaries of the bronchial artery, such as the capillary plexus and vasa vasorum, we performed a scanning electron microscopic(SEM) study of the vascular casts, using Mercox CL-2B as a media. The alveolar capillaries revealed hexagonal or pentagonal rings of vascular networks. In some areas, the vascular rings composed a square network, The bronchial tree was supplied by the bronchial arteries which form a coarse capillary extending as far as the terminal bronchioles. Occasionally the capillary plexus was connected with adjacent capillary networks in and around the alveolar walls. The walls of the pulmonary artery revealed only a single layer of the vasa vasorum, but those of the pulmonary vein were surrounded by more complicated and well developed vasa vasorum than the pulmonary arterial side. The mean diameters of the venous vasa vasorum are greater than the arterial vasa vasorum.
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6.Light Electron Microscopic Study in Rat Livers Following Cadmium Chloride Administration.
Kwan Kyu PARK ; Young Ho KIM ; Kun Young KWON ; Eun Sook CHANG ; Moo Ung CHANG
Korean Journal of Pathology 1992;26(1):28-39
This study was carried out to investigate the light and electron microscopic findings of the livers of rats after an intraperitoneal injection of cadmium chloride. The Sprague-Dawley rats were intraperitoneally injected with cadmium chloride dissolved in water, once a day for three days. These animals were sacrificed at 1, 3, 8, and 24 hr after the last injection. Control groups of the rats were also sacrificed in the same manner. The liver was extirpated and examined by both light and electron microscopy. The results obtained are as follows: The parenchyma of the liver shows focal neutrophilic infiltration and spotty necrosis. The hepatocytes show fatty change, ballooning degeneration, swelling of the endoplasmic reticulum and mitochondria, increase numbers of secondary lysosomes and residual bodies. Focal patic venules and sinusoids of the liver are congested. The Kupffer cells are increase in number. Therefore, it can be concluded that the cadmium is directly acted to hepatocytes resulting in cellular injuries and deposits in the fat droplets of the cytoplasm of the hepatocytes, not Ito cells as previously suggeted.
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7.A Scanning Electron Microscopic Study on Microvascular Changes in the Monocrotaline-induced Rat Lung by Corrosion Casting Method.
Korean Journal of Pathology 1995;29(5):644-659
To investigate the microvascular changes in primary pulmonary hypertension, the lungs of 24 Sprague-Dawley rats were treated by an intraperitoneal injection of 2% monocrotaline(MCT) solution and then examined with scanning electron microscopy(SEM) after microvascular corrosion casting. Histologic examination revealed significant medial thickening in the small to medium-sized pulmonary arteries. Scanning electron microscopic findings of the normal lungs showed two kinds of microvascular structures. One showed a well-fortned three-dimensional basket structure of uniform flat-tubular alveolar capillaries, which were connected to each other in a T or Y shape or at right angles. The other revealed a two-dimensional reticular sheet of round tubular branches mainly in the bronchial artery-supplying regions. The MCT-treated groups(remodelling) showed apparent changes in both kinds of microvasculatures in comparison to the normal group but the more prominent change was found in Lbe bronchial artery microvasculature showing the dense thick encasement around large pulmonary arteries. Alveolar microvasculature of the pulmonary artery revealed individually enlarged angular appearance, with generally deformed alveolar architecture. Quantitatively, the significant enlargement of diameter and intercapillary distance appeared in both microvasculatures of MCT-induced rat lungs, but the density was increased only in the bronchial artery microvasculature. In conclusion, our three-dimensional microvascular study of the MCT-treated rat lungs demonstrates a new morphologic finding of vascular remodeling in primary puhnonary hypertension, which is thought to play an important vascular role in the pathogenesis in addition to interstitial fibrosis.
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8.Cell Mediated Immunity in Tubulointerstitial Nephritis of Rats.
Korean Journal of Pathology 1995;29(5):634-643
To investigate the tubular major histocompatibility complex(MHC) expression and inflammatory phenotypes in tubulointerstitial nephritis, Lewis rats were inununized with azobenzen-earsonate-tyrosine in complete Freund adjuvant and challenged either foot pad or kidney, either by subcapsular injection or by ex vivo perfusion. The rats were sacrificed 2, 3, 5, 10 and 15 days after antigenic challenge. Foot pad swelling was significant at the antigenic challenge site (151.8 vs 6.8 x 10(-2) mm) at 24 hours. Tubulointerstitial nephritis was induced by both methods and the inflammatory infiltrate which first appeared on day 2, became prominent at day 5, then gradually subsided in ex vivo perfused rats, while inflannnation started on day 3 in subcapsular injected rats. The major site of inflammation was in the cortex and outer stripe of the outer medulla, with predominance of mononuclear cells throughout the course. The inflammatory cells showed mainly OX8 and ED1 positivity with OX19, W3/25 and CD5 positivity in minority. RT1B expression was diffuse in the cytoplasm of proximal tubules at day 2 and 5. These results suggest the involvement of cell mediated immunity in this experimental model, and the possibility that tubular epidielial cells process antigen and then become targets in immune injury.
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9.The Effects of Immunosuppressant and Immunostimulant on the Splenic Cell Subset of Rats Having Undergone Experimentally Induced Septal Fibrosis of Liver.
Mee Young SOL ; Joon Yeon KIM ; Sun Kyoung LEE
Korean Journal of Pathology 1995;29(5):572-583
Although there have been many reports about the importance of the spleen's role in hepatic fibrogenesis, the exact mechanism is still uncertain. The author designed this study to evaluate splenic function on hepatic fibrogenesis. The degree of hepatic fibrosis and the population of splenocyte subsets were studied in the experimental animal model with fibrosis produced by injecting normal swine serum intra-peritoneally into Sprague-Dawley rats. The animals were divided into three groups; group A was subjected to injection of swine serum only, group B swine serum and complete Freund's adjuvant and group C swine serum and cyclosporin A. The experimental hepatic fibrogenesis by swine serum was augumented by coinjection with the adjuvant and inhibited by cyclosporin A. The study of the splenocyte subset revealed increased percentages of spienic B cell and CD4+ cell and a decreased percentage of CD8+ cell, and these changes of splenocyte subset were also augumented by the adjuvant and inhibited by cyclosporin A. The percent of monocytes was not significantly altered, although a tendancy of early decrease by the adjuvant was noted.
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10.The Effects of Excitatory Amino Acids and Their Receptors on Neuronal Damage of Rat Brain in Hypoxic-Ischemic Encephalopathy.
Korean Journal of Pathology 1995;29(5):545-562
Since the role of excitatory amino acids such as glutamate and aspartate and their receptors mediating cellular injury through various mechanisms were known in hypoxic-ischemic injury and associated diseases of central nervous system, blocking agents for transmitter release or receptors have been tried to reduce the cellular damages and subsequent sequelae experimentally. Several in vitro studies suggested two kinds of glutamate neurotoxicity: (1) rapid toxicity due to influx of sodium or chloride with resultant cellular edema and consequent damage, which is associated with N-methyl-D-Aspartate(NMDA) as well as non-NMDA receptors, (2) calcium mediated delayed toxicity associated mainly with NMDA receptor. This study was conducted to investigate the role of rapid toxicity in hypoxic-ischemic injury. Early lesions of 30 minutes to 24 hours after hypoxic-ischemic insult were examined by autoradiography with radiolabelled glutamate and kainitic acid (KA) as well as light and electron microscopy. Late changes were evaluated on formaldehyde-acetic acid-methanol(FAM) fixed brain 1 week after the insult. Cornus ammonis(CA) l of hippocampus showed the highest density of NMDA receptors, which was decreased constantly from 2 hours to 24 hours. In contrast, CA3 of hippocampus showed the highest density of KA receptors, which was the lowest at 6 hour and increased thereafter. Light microscopic examination showed the worst changes during 30 minutes to 6 hours. After 1 week, most of the cases showed degeneration of neurons and CAI and CA3 did not show the difference. Electron microscopic examination showed marked degenerative changes of neurons as well as neuropils starting from 30 minutes after the insult. In conclusion, rapid toxicity mediated by non-NMDA(KA) receptor seen in CA3 lead to permanent damage in 1 week old lesion.
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