• Journal of Biomedical Engineering Research
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• v.36 no.1
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• pp.1-6
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• 2015

This study aims to design a monolayer cell adsorption apparatus that would help to produce high-quality slides for Liquid-Based Cytology (LBC) of an early cancer diagnosis for human bodies. The LBC collects exfoliated cells of human bodies and spreads the cells on the slides. Through processes of dyeing and cytological examination, the LBC screens for cancers in early stage. In this study, both of a cell suction module and a cell adsorption module, which are the key elements of the monolayer cell adsorption apparatus, were developed, and using those modules, the study set, first, conditions to help both GYN and NON-GYN apply principal cells without de-endothelialization before conducting its own analysis on the utility. As a results, for GYN, apparatus was determined to be able to produce high-quality slides under the condition of 4 and for NON-GYN, the apparatus would come up with other slides of high-quality under the condition of 2. The study carried out a repetitive test on selected conditions and proved 96% of the repetitive success rate. By the results of what has been learned so far, the study presents that the apparatus has a possibility to replace device from South Korea as one of those other currently-applied systems to run the LBC and that the system will also present a new paradigm for cancer diagnosis as it makes a contribution to the improvement in the LBC.

• Archives of Reconstructive Microsurgery
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• v.13 no.2
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• pp.101-106
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• 2004

Purpose : This study evaluated the effect of VEGF in the arterial anastomosis by using light and electron microscopy. Marerials and method : Rats underwent femoral arterial end-to-end anastomosis after transection and topical VEGF treatment. The proximal and distal segments of the femoral arteries was drenched with 1 drop of VEGF $(100ng/100{\mu}l/bottle)$. and when half of the repair was finished, the other 1 drop was drenched and then the repair was continued to complete the anastomosis. Gross and histologic characteristics of arterial wall were assessed after 3 days, 1, 3 and 5 weeks. In the control group, normal saline solution instead of VEGF was dropped with the same method in the anastomosis. Results : The histologic findings of the arterial wall were the vascular remodeling with the infiltration of inflammatory cells at early stages and the tissue fibrosis at lately stages in the anastomotic sites of the control and the VEGF-treated groups. The scanning electron microscopic results were; (1) the anastomotic sites were covered by many irregular cells with long cytoplasmic processes at the early stages. (2) After 1 week, endothelial cells started to cover the anastomotic sites. (3) After 3 weeks, the anastomotic sites were partially covered by endothelial cells in the control group. (4) After 5 weeks, the anastomotic sites were completely covered by endothelial cells in the control and VEGF-treated groups. (5) In the VEGF-treated group, the anastomotic site was completely covered by endothelial cells which directed parallel to longitudinal axis of arteries after 3 weeks. Conclusion : Topical VEGF maintained luminal integrity by decreasing fibrosis and increasing re-endothelialization. These findings suggest that topical VEGF may be a promising new strategy to enhance healing and improve the outcome of vascular anastomosis.

• Journal of Chest Surgery
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• v.29 no.4
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• pp.373-380
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• 1996

Prevention of thromboembolism is the most important task in the development of bioconpatible small caliber artificial vascular graft. In normal vessels, vascular endothelial cells maintain homeosatsis by secreting numerous factors. The aim of this study is to develope a method which Improves biocompatibility of small caliver polyurethane graft using endothelial cell culture technique, and ev luate the efTectiveness of extracelluar matrix for endothelization which was produced by cultured fibroblast. Methods ; Multiporous polyurethane tube of 3 mm diameter, 0.3 mm thickness was manufactured for vascular graft. Three mongrel dogs were intubated and internal jugular veins removed. Extracelluar matrix produced by cultured flbrobast which was obtained from dog's internal jugular vein were coated to the polyurethane graft. Then, endothelial cells extracted from Jugular vein were cultured and fixed on the extracelluar matrix layer of vascular graft. Endothelial cell coated vascular grafts were implanted to the carotid arteries of experimental dogs as interposed autograft. Implanted grafts were removed after 3 and 6 weeks. As a control, PTFE graft was interposed on carotid artery. These experiments demonstrated that extracelluar matrix produced by fibroblast can afford a base for endothelial cell linings of polyurethane graft. Although thrombosis were developed on autografted en othelial cell coated graft, 33% opening was noticed, and showed less adhesion to adjacent tissue layer. These findings suggest that fiboblast produced extracelluar matrix which can be used for edothelial cell lining vascular graft, and by improving the cultured endothelial cell function, there will be a new modality for reducing thrombosis on small vascular graft.

• Journal of Chest Surgery
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• v.41 no.6
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• pp.679-686
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• 2008

Background: Cellular remnants in the bioprosthetic heart valve are known to be related to a host's immunologic response and they can form the nidus for calcification. The extracellular matrix of the decellularized valve tissue can also be used as a biological scaffold for cell attachment, endothelialization and tissue reconstitution. Thus, decellularization is the most important part in making a bioprosthetic valve and biological caffold. Many protocols and agents have been suggested for decellularization, yet there ave been few reports about the effect of a treatment with hypotonic solution prior to chemical or enzymatic treatment. This study investigated the effect of a treatment with hypotonic solution and the appropriate environments such as temperature, the treatment duration and the concentration of sodium dodecylsulfate (SDS) for achieving proper decellularization. Material and Method: Porcine aortic valves were decellularized with odium dodecylsulfate at various concentrations (0.25%, 0.5%), time durations (6, 12, 24 hours) and temperatures ($4^{\circ}C$, $20^{\circ}C$)(Group B). Same the number of porcine aortic valves (group A) was treated with hypotonic solution prior to SDS treatment at the same conditions. The duration of exposure to the hypotonic solution was 4, 7 and 14 hours and he temperature was $4^{\circ}C$ and $20^{\circ}C$, respectively. The degree of decellularization was analyzed by performing hematoxylin and eosin staining. Result: There were no differences in the degree of decellularization between the two concentrations (0.25% 0.5%) of SDS. Twenty four hours treatment with SDS revealed the best decellularization effect for both roups A and B at the temperature of $4^{\circ}C$, but there was no differences between the roups at $20^{\circ}C$. Treatment with hypotonic solution (group A) showed a better ecellularization effect at all the matched conditions. Fourteen hours treatment at $4^{\circ}C$ ith ypotonic solution prior to 80S treatment revealed the best decellularization effect. The treatment with hypotonic solution at $20^{\circ}C$ revealed a good decellularization effect, but his showed significant extracellular matrix destruction. Conclusion: The exposure of porcine heart valves to hypotonic solution prior to SDS treatment is highly effective for achieving decellularization. Osmotic treatment with hypotonic solution should be considered or achieving decellularization of porcine aortic valves. Further study should be carried out to see whether the treatment with hypotonic solution could reduce the exposure duration and concentration of chemical detergents, and also to evaluate how the structure of the extracellular matrix of the porcine valve is affected by the exposure to hypotonic solution.

• Journal of Chest Surgery
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• v.29 no.1
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• pp.7-13
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• 1996

The conventional glutaraldehyde (GA) fixation method of tissue valves is considered to be responsible for accelerated valve degeneration. The release of toxic GA from the valve tissue is believed to limit endothelial cell (EC) ingrowth. Removal of toxic GA by reaction with L-glutamic acid and storage in a Paraben solution may offer good EC growth. To investigate the conditions for endothelialization of tissue valves, the growth properties of ECs on the conventionally and alternatively treated pericardial tissue were compared. Conventional preparation included zero-pressure fixation for 72 hours in phosphated-buffered saline (PBS) solution containing 0.5% GA at 4$^{\circ}C$ and storage into PBS containing 0.2% GA(group I). Alternatively treated pericardial tissues were divided into three postfixation treatment groups : (1) storage in PBS solution containing Paraben(group II), (2) treatment with PBS containing 8$^{\circ}C$ L-glutamic acid(PH 7.35) and storage in PBS solution containing Paraben (g oup III), (3) treatment with L-glutamic acid dissolved in distilled water (PH 3.5) (group IV). Pericardial tissue were transferred into the 24-well plate after storage for 4 weeks. ECs were harvested enzymatically from the bovine pulmonary artery and grown to confluence on culture flask surfaces. Detached ECs by trypsin were incubated into the each well of the 24-well plate including test pericardial tissues. Cells were detached by trypsin, 1, 2, 3, 5, 7 days after incubation and counted on the hemacytometer. Cell viability test was performed by frypan-blue exclusion method. Acute cell death in the group I were found even after prolonged washing. The group II showed prolonged cell survival compared with the group I. Both group III and group IV showed better cell growth than group II. There was no statistically significant difference between group III and group IV method in terms of EC growth. This results suggest that treatment by L-glutamic ac id and storage in a Paraben solution be a promising approach for improvement of durability of GA-treated tissue valves.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.13 no.1
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• pp.16-29
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• 1991

The occurrence of thrombotic occlusion & endothelial injuries at the site of anastomosis have been considered as major problems in microvascular surgery. The purpose of this study was to ascertain whether a end-in-end(sleeve, telescope) anastomosis compared favorably with end-to-end anastomosis in healing procedures on the endothelium and to study the possibility of clinical application in end-in-end method. The microvascular anastomoses have been performed with end-in-end method in the femoral arteries of 20 rats group, also with end-to-end method on the same arteries of another 20 rats group, and then the four anastomosed vessels in subdivided groups on each group were taken after period of 1, 3, 7, 14, 21 days following by anastomoses for scanning electron microscope observation. The results were as following: 1. The patency rate was 90% in the end-in-end group and 85%in the end-to-end group and late thrombus occurred at 7, 14 days on both groups, which suggested -consistent monitoring of patency was required for two weeks at least. 2. Platelet aggregates at the site of anastomosis began to organize on post-operation 3 days and in the end-in-end group, the initially decreased lumen of inserted vessel was gradually increasing on 7 days due to atrophy of the medial layers. 3. Re-endothelialization was completed between 7 and 14 days in end-in-end group, whereas between 14 and 21 days in end-to-end group.

• Journal of Chest Surgery
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• v.36 no.12
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• pp.887-893
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• 2003

External stimuli increases intracellular (IC) $Ca^{2+}$, which increases extracellular (EC) $K^{+}$. To verify $K^{+}$ effects on the vascular contraction, we performed an experiment using mouse aortic endothelial cell. Meterial and Method: We examined the mouse aortic contractility changes as we measured the IC $Ca^{2+}$ change and ionic current by using the voltage clamp technique under different conditions such as: increasing EC $K^{+}$, removing endothelial cell, giving L-NAME (N-nitro-L-arginine methyl ester) which suppress nitric oxide formation, Ouabain which control N $a^{+}$ - $K^{+}$ pump and N $i^{2+}$ which repress N $a^{+}$-C $a^{2+}$ exchanger Result: When we increased EC $K^{+}$ from 6 to 12 mM, there was no change in aortic contractility. Aorta contracted with more than 12 mM of EC $K^{+}$. Ace-tylcholine (ACh) induced relaxation was inhibited with EC $K^{+}$ from 6 to 12 mM, but was not found after de-endothelialization or L-NAME treatment. ATP or ACh increased IC $Ca^{2+}$ in cultured endothelium. After maximal increase of IC $Ca^{2+}$, increasing EC $K^{+}$ from 6 to 12 mM made IC $Ca^{2+}$ decrease and re-decreasing EC $K^{+}$ to 6 mM made IC $Ca^{2+}$ increase. Ouabain and N $i^{2+}$ masked the inhibitory effect of endothelium dependent relaxation by increased EC $K^{+}$. Conclusion: These data indicate that increase in EC $K^{+}$ relaxes vascular smooth muscle and reduces $Ca^{2+}$ in the endothelial cells which inhibit endothelium dependent relaxation. This inhibitory mechanism may be due to the activation of N $a^{+}$- $K^{+}$ pump and N $a^{+}$-C $a^{2+}$ exchanger. $a^{+}$-C $a^{2+}$ exchanger.r.

• Journal of Chest Surgery
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• v.39 no.4 s.261
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• pp.261-268
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• 2006

Background: Current vascular prostheses are still inadequate for reconstruction of small-diameter vessels. Autologous pericardium can be a good alternative for this purpose as it already possesses good blood compatibility and shows a mechanical behavior similar to that of natural arteries. However, the clinical use of autologous pericardial tissue as a small-diameter vascular graft has limitations due to mixed outcomes from uncertain biological behavior and difficulty to gain reliable patency results in animal experiments. To study this issue, we implanted fresh and glutaraldehyde-treated autologous pericardium as small-diameter arterial grafts in dogs, and compared their time-related changes histologically. Material and Method: As a form of 5mm-diameter arterial graft, one pair of autologous pericardial tissue was used for comparison between the glutaraldehyde-treated and the glutaraldehyde-untreated grafts in the bilateral carotid arteries in the same dog. The patency of the grafts were evaluated at regular intervals with Doppler ultrasonography. After the predetermined periods of 3 days, 2 weeks, 1 month, 3 months and 6 months, the grafts in each animal were explanted. The retrieved grafts were processed for light and electron microscopic analyses following gross observation. Result: Of 7 animals, 2 were excluded from the study because one died postoperatively due to bleeding and the other was documented as one side of the grafts being obstructed. All 10 grafts in the remaining 5 dogs were patent. Grossly, a variable degree of thromboses were observed in the luminal surfaces of the grafts at 3 days and 2 weeks, despite good patency. Pseudointimal smooth blood-contacting surfaces were developed in the grafts at f month and later. By light microscopy, mesothelial cell layers of the pericardial tissue were absent in all explanted grafts. Newly formed endothelial cell layers on the blood-contacting surface were observed in both the glutaraldehyde-treated and fresh grafts at 3 months and later. The collagen fibers became degraded by fragmentation in the fresh graft at 1 month and In the glutaraldehyde-treated graft at 3 months. At 6 months, the collagen layers were no longer visible in either the glutaraldehyde-treated or fresh grafts. By electron microscopy, a greater amount of coarse fibrin fibers were observed in the fresh grafts than in the glutaraldehyde-treated grafts and, more compact and well-arrayed layers were observed in the glutaraldehyde-treated grafts than in the fresh grafts. Conclusion: The glutaraldehyde-treated small-diameter pericardial arterial grafts showed a better endothelialization of the blood-contacting surface and a slower fragmentation of the collagen layers than the fresh grafts, although it has yet to be proven whether these differences are so significant as to affect the patency results between the groups.