• Journal of Periodontal and Implant Science
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• v.23 no.1
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• pp.77-96
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• 1993

This in vitro study was undertaken to observe whether citric acid application aids the attachment and proliferation of human periodontal ligament cells to the root surfaces of periodontally diseased teeth. The roots were prepared so that the comparison could be made among the control healthy root surface, citric acid demineralized and non-demineralized root planted surfaces. Prior to the cell attachment experiment, each groups were prepared for scanning electron microscopic (SEM) examinations of root surface morphology, All specimens were fixed with phosphate buffered glutaraldehydes, postfixed with phosphate buffered osmium tetraoxide and stained with phosphate buffered tannic acid. dehydrated in ethanol, critical point dried, sputter coated with gold and examined under the SEM. In the cell attachement experiment, human cultured periodontal ligament cells at concentration to $4.5{\times}\;10^4\;cells/ml$ were seeded in each culture well which contained prepared roots and incubated for 30min 1, 2, 6, 12 and 24 hours at 37, 5% $CO_2$air incubator. Than the specimens were prepared for SEM examination using, the same methods as described above. In the cell proliferation experiment, $5{\times}\;10^4\;cells/ml$ cells were seeded incubated with the specimens for 6 hours. Then, all of the specimens were moved into fresh culture well and incubated for 24, 48, and 72 hours. The cell counting was done after trypsinization, under light microscope. The results were as follows. When viewed the surface morphology prior to the cell attachment, the non acid treated root planed surface displayed scaling striation and occasional bacteria and calculus. The citric acid treated specimens displayed little debris on the surface and funnel shaped orifices of dentinal tubules. There were no apparent differences in the morphology of cells attached to the control and experiment groups. However, in initial attachement, there was a slight more enhanced appearance in attachment in citric acid treated groups than other root surfaces. After 6 hours of incubation, most of the cells initiated the alteration of cell morphology from ovoid to spindle shapes. After 24 hours of incubation, most of the cells displayed proliferated appearance and connected with each other via numerous processes. In the cell proliferation experiments, there were statistically significant increased number of cells in citic acid treated groups than other groups.

• Journal of Yeungnam Medical Science
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• v.13 no.1
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• pp.116-133
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• 1996

The biocompatibility of the titanium has been estabilished through various experimental studies such as cell culture toxicity test, pyrogen test, mutagen test and others. In order to confirm biocompatibility after fabrication of titanium and to clarify the difference between the bone reaction after insertion of the lathed titanium rods and the bone reaction after insertion of the finished and polished rods, both rods were implanted into the proximal femur of a rabbit. Histologic reactions in the bone were observed according to the ASTM standards at the intervals of 6 weeks, 12 weeks and 26 weeks after implantation. The result were as follows : In 6 weeks after implantation of lathed titanium rods, inflammatory reactions, such as minimal degree infiltration of polymorphonuclear leukocytes and lymphocytes were observed in all cases. This was thought to he caused by surgical trauma. However, inflammatory cell infiltration was not seen after implantation of polished and finished rods in all cases. The cellular infiltration and the histologic reaction of the hone after implantation of lathed group were significantly more pronounced than those after implantation of the finished group. In 12 weeks after implantation of lathed rods, two of four cases revealed a minimal degree of cellular infiltration. No inflammatory cell infiltration was demonstrated after implantation of the finished group. The cellular infiltration and histologic reaction seemed to be more pronounced in the lathed group, but they were not significant statistically. At 26 weeks after implantation of the lathed and finished group, there was no cellular infiltration in both groups. New bone formation was observed up 26 weeks, and no difference between lathed titanium rods and finished titanium rods were apparent. Mild bone necrosis was observed in 1 case out of 11 cases in which lathed titanium rods were implanted. Bone necrosis was not observed in the finished titanium rod group. Fibrosis was observed in both groups, but differences were not significant between the experimental groups. In the lathed titanium rods group and the shorter interval group, inflammatory cell infiltration was significantly higher. Finished titanium rods and longer interval groups had markedly decreased tendences in histologic reaction ratings. As a conclusion, although certificated titanium might be safe to use, difference of biocompatibility were observed depending on the method of surface finish. By identifying biocompatibility as a long-term standardized animal study, we can develop progressed internal fixation device that is safe for human beings.

• The Korean Journal of Malacology
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• v.24 no.1
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• pp.41-50
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• 2008

The histochemical characteristics and ultrastructure of the mantle in the spiny top shell, Batillus cornutus were described using light and electron microscopy. The simple epidermal layer wrapped on the top and bottom of the centrally located connective tissue. And then the epidermal layer were divided into the outer epidermal layer near a shell and the inner epidermal layer closed to the visceral mass. The connective tissue layer was composed of the collagen fiber muscularfiber bundle and hemolymph sinus. Mucous cells in the apical mantle contained acid and neutral mucopolysaccaride, and acidic carboxylated mucopolysaccaride in the mid and marginal mantle. The mantle thickness, epidermal layer thickness and hemolymph sinus area displayed a trend of reduction from the marginal zone to the apical zone. From TEM observation, it was possible to distinguish epithelium, ciliated cell, absorptive cell and secretory cell in the epidermal layer. The epithelia were columnar and the nucleus was elliptical. The free surface were covered with microvilli. The lateral membranes of epithelium was con nected with neighboring cells by the zonular occludens, zonular adherens and membrane interdigitation. Ciliated cell on free surface had cilia and microvilli, and numerous mitochondria in the apical cytoplasm. In the epidermal layer, it observed 2 type cells having absorptive function. The absorptive cells were columnar in shape, and contained microvilli, pinocytotic vesicles, mitochondria and lysosomes of various electron density. Secretory cells can be divided into four types (A, B, C, D) depending on the cell shape and characteristics of secretory granules. These cells were unicellular glands and had similar characteristics to previously reported on the mantle of the gastropod and bivalves.

• Journal of Chest Surgery
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• v.42 no.4
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• pp.409-417
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• 2009

Background: Glutaraldehyde-fixed heterografts are prone to calcification after long-term implantation in human, and this is one of the limiting factors for the longevity of the heterografts used in cardiovascular surgery. The aim of the study was to evaluate the anticalcification effect of an ethanol and amino acids treatment on glutaraldehyde-fixed bovine pericardium. Material and Method: Bovine pericardial tissues were divided into 5 groups. Group 1 consisted of tissues fixed with glutaraldehyde, group 2 consisted of commercially available bovine pericardial valve tissues (Carpentier-Edwards PERIMOUNT), group 3 consisted of glutaraldehyde-fixed tissues treated with ethanol, group 4 consisted of glutaraldehyde-fixed tissues treated with ethanol and L-glutamic acid, and group 5 consisted of glutaraldehyde-fixed tissues treated with ethanol and homocysteic acid. The tissue microstructure was examined by light and electron microscopy. Tissue samples of each group were implanted into rat subcutaneous tissue for 3 $\sim$ 4 months and the calcium contents were measured after harvest. Result: The collagen fibers appeared to be well preserved in all the groups. The calcium contents of groups 2, 3, 4 and 5 (13.46$\pm$11.74, 0.33$\pm$0.02, 0.39$\pm$0.08 and 0.42$\pm$0.06 $\mu$g/mg, respectively) were all significantly lower than that of group 1 (149.97$\pm$28.25 $\mu$g/mg) (p<0.05). The calcium contents of groups 3, 4 and 5 were all significantly lower than that of group 2 (p<0.05). Conclusion: Treatment with ethanol alone or in combination with amino acids (L-glutamic acid or homocysteic acid) strongly prevented the calcification of glutaraldehyde-fixed bovine pericardium.

• Childhood Kidney Diseases
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• v.2 no.2
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• pp.138-144
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• 1998

Purpose : The use of cyclosporine and mitomycin in various immunologic or neoplastic disorders has been known to cause wide-ranged nephrotoxic effects including thrombotic microangiopathy. However, the mechanism of nephrotoxicity of these drugs has not been studied adequately, so that present experimental study has been undertaken to find out whether these drugs can cause direct damage to the kidney and to clarify the pathogenetic mechanism of nephrotoxic effect of these drugs. Materials and methods : Sprague-Dawley rats weighing 250-300 gm were used for experimental animals and unilateral renal perfusion technique, modified from the method described by Hoyer et al was used. Isolation of left kidney from systemic circulation was made by clamping aorta and left renal vein and a hole was punctured in the anterior wall of the left renal vein. Cyclosporine (2.5 mg in 4 ml solution) and mitomycin (1.6 mg in 4ml solution) were infused through left renal artery and normal saline was used in control rats. Forty-eight hours after infusion of the drugs, animals were sacrificed and left kidney removed and processed for histologic examination. Total ischemic time of left kidney was less than 15 minutes: Results : Cyclosporine-perfused group showed severe swelling of glomerular endothelial ceil along with swelling of glomerular epithelial cell and interstitial vascular endothelial cell. Mitomycin-perfused group also showed severe swelling of glomerular endothelial and epithelial cells. And in addition to these findings, they demonstrated platelets aggregation, swelling and degranulation of platelets and fibrin accumulation in some of the capillaries, indicating occurrance of thrombotic microangiopathy. Conclusion : present experiment indicates that cyclosporine and mitomycin can cause direct toxic injury to renal endothelial cell. And this direct toxic damage to endothelial cell seems to be an important initiating event for the development of thrombotic microangiopathy.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.2
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• pp.318-332
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• 2000

The purpose of this study was to investigate the effects of demineralized freeze-dried bone (DFDB) on mechanically exposed pulp of dog by evaluating the pulpal inflammation and healing process, formation of dental hard tissue, and structural changes of fibroblasts of the remaining pulp tissue. Teeth of 4 dogs, weighing 10kg, were used in this study. Class V cavities were prepared followed by exposed the pulp tissue mechanically by sterilized round bur. In control group, exposed pulps were capped with calcium hydroxide paste followed by sealed with IRM. In experimental groups, the exposed pulps of one group were capped with the collagen and those of the other group were capped with DFDB. All cavities were sealed with same manor as control group. The animals were sacrificed at the intervals of 3, 7, 14, and 28 days for histopathlogic evaluation. The specimens were observed by the light microscope and trans-electron microscope. The results were as follows: 1. Pulp necrosis was not observed in all groups. Inflammatory response was disappeared from 1 week in control group and group 2. But it was not disappeared until 2 weeks and also irregular arrangement of odontoblasts was showed at the lateral walls of root canal just beneath the amputated site of the pulp in group 1. 2. Dentinal bridge was formed incompletely at 2 weeks but it was formed completely at 4 weeks in control group. Odontoid tissue was also found in control group at 4 weeks from treatment. Amputated site of pulp was encapsulated with fibrous tissue and odontoblast and dentinal bridge was not found in group 1. Preodontoid tissue and reparative dentin which were formed by odontoblast differentiated around DFDB were found, but dentinal bridge was not found in group 2. 3. Cell with large basophillic-stained nuclei infiltrated to amputated site and DFDB at 1 week from treatment in control group and group 2. They were found more in group 2 than in control group. Odontoblasts arranged more regularly and reparative dentin was found more as time elapsed. 4. Dentin-formative odontoblasts which showed ultramicrostructure of cytoplasm with polarized nucleus, rEM, Golgi complex, secretory granules, secretion of organic matrix in control of group and group 2. In regards to above results, the demineralized freeze-dried bone(DFDB) induce odontoblastic differentiation and further come up to the dentin formation in amputated pulp.

• Investigative Magnetic Resonance Imaging
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• v.18 no.1
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• pp.34-42
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• 2014

Purpose : To evaluate the correlation of lesion-to-normal ratio (LNR) of signal intensity from double inversion recovery MR imaging and total choline-containing compound (tCho) resonance from single voxel MR spectroscopy in breast cancers. Materials and Methods: Between August 2008 and December 2009, 28 patients who were diagnosed as breast cancer and had undergone both double inversion recovery (DIR) MR imaging and MR spectroscopy (MRS) were included in this study. The signal intensities of the lesion (L) and ipsilateral normal breast tissue (N) were measured in region of interest of each breast cancer in DIR and contrast enhance MR image (CE-T1WI) to calculate the LNR value for each technique. MRS was performed using single-voxel MR spectroscopy. The height, width and area of tCho resonance were compared with each LNR of DIR and CE-T1WI. We used Pearson's correlation coefficient(r) for correlation analysis and the significance level was p=0.05. Results: There was no statistically significant correlation between LNR of CE-T1WI and height (r=-0.322, p=0.094), width (r=-0.233, p=0.232) and area (r=-0.309, p=0.109) of MRS tCho. There was no statistically significant correlation between LNR of DIR and height (r=0.067, p=0.735), width (r=-0.287, p=0.139) and area (r=0.012, p=0.953) of MRS tCho, either. The Pearson's correlation coefficient was 0.186 between LNRs of CET1WI and DIR (p=0.344). Conclusion: There was no statistically significant correlation between LNR of DIR and relative amount of tCho resonance of MRS.

• Journal of Periodontal and Implant Science
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• v.35 no.1
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• pp.217-234
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• 2005

혈소판 농축 혈장은 구강과 안면부 재건수술에 새로이 사용되는 유용한 첨가물이다. 혈소판은 상처 치유과정에서 매우 중요하며, 혈소판은 상처부위에 빠르게 도달하여 응고를 형성한다. 그리고 다양한 성장인자를 분비한다. 이러한 성장인자는 골의 형성과 혈관의 증가, 골 이식재의 치유에 관여하는 것으로 생각된다. 본 연구의 목적은 실험 동물을 통하여 혈소판 농축 혈장에 함유된 혈소판의 정량화를 통한 성장인자 함유량을 추정하고, 방사선학적, 조직학적 평가를 통해 혈소판 농축 혈장이 초기의 골형성에 미치는 영향에 대한 평가를 하는데 있다. 15마리의 가토 두개골에 6mm trephine bur(외경 8mm)를 이용하여 경뇌막의 손상을 주지 않도록 하면서 4개의 결손부를 형성하였다. 각각의 두개골 결손부는 $Bio-Oss^{(R)}$만 이식한 군, PRP만 이식한 군, PRP와 $Bio-Oss^{(R)}$를 혼합하여 이식한군, 그리고 아무것도 이식하지 않은 군을 대조군으로 설정하였다. 각각의 재료를 이식한 후 비흡수성 차폐막($Tefgen^{(R)}$)을 위치시키고 흡수성 봉합사로 일차봉합을 시행하였다. 각 군 당 술 후 1, 2, 4주의 치유기간을 설정하였다. 동물을 희생시키고 두개골을 절제하였다. 먼저 방사선학적인 골 밀도 측정을 시행하고, 조직학적 평가를 위해 통법에 따라 조직 표본을 제작한 후 광학현미경으로 관찰하였다. 또한 가토 귀 변연정맥에서 채취한 10 ml의 혈액을 원심분리하여 혈소판 함유량을 평가하여 다음과 같은 결과를 얻었다. 1. 혈소판 농축 혈장은 일반 혈액에 비해 약 4.02배 많은 수의 혈소판이 함유되어 있었다. 2. 방사선적인 평가에서 1, 2, 4주 사이에 대조군과 비교하여 $Bio-Oss^{(R)}$에 PRP를 이식한 군에서 골의 밀도는 큰 차이를 보이고 있다(p<0.01). 하지만, 동일한 시기에 PRP만 이식한 군과 대조군의 차이는 발견할 수 없었으며 (p>0.05), $Bio-Oss^{(R)}$만 이식한 군과 $Bio-Oss^{(R)}$에 PRP를 이식한 군의 차이 또한 발견할 수 없었다(p>0.05). 3. 조직학적 평가에서 모든 이식재는 시간이 경과할수록 골 형성이 증가함을 알 수 있었다. 대조군에 비해 PRP만 이식한 군에서 더 두꺼운 섬유성 결합을 보이고 있다. 대조군과 PRP만 이식한 군과 비교해 $Bio-Oss^{(R)}$와 $Bio-Oss^{(R)}$에 PRP를 혼합 이식한 군에서 골의 형성이 더 진행됨을 알 수 있었다. $Bio-Oss^{(R)}$에 PRP를 혼합 이식한 군이 $Bio-Oss^{(R)}$만 이식한 군에서보다 더 많은 신생골 형성을 관찰할 수 있다. 이상의 결과에서 가토의 두개골 결손부에 $Bio-Oss^{(R)}$에 PRP를 혼합 이식하였을 경우 결손부의 초기 골 형성을 촉진 할 수 있음을 시사하였다.

• Applied Microscopy
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• v.36 no.3
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• pp.141-153
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• 2006

The purpose of this study is to find out how soft contact lens multi-purpose solution (MPS), often used for medical treatment, effects the inhibition on cell growth and research the result of using MPS, suspected to demage eye cells, on rabbit eye's corneal epithelium and endothelium tissue. In this treatise, $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea) had been selected among the MPS. After culturing L929 roil line, cell growth inhibition rate was measured by MTT assay, and by making Hematoxylin and Eosin stain specimen. the morphology was observed by optical microscope. In the In vivo experiment,9 white rabbit eyes (18 eyes) were classified into 3 groups. The experimental group is left eyes (9 eyes) of rabbit, and MPS were dropped; however. the control group, the right eyes (9 eyes), were only used a saline solution without preservatives. After the dropping within the period, the cornea surface of rabbit eyes were stained by Rose bengal and observed. To figure out the changes of the corneal epithelium and endothelium tissue scanning electron microscopy (SEM) has been used. As the result, the rate of cell growth inhibition was 54%. 73% and 36%, respectively. Morphological changes represented that the shape has been changed into oval or round shape and those are not considered as a common formation of L929 cell line. When it comes to staining Rose bengal, each experiment group was stained red which is not shown in controls. The polygonal mosaic pattern of a corneal epithelium was disturbed in the picture taken by SEM; furthermore, the shape of the corneal endothelium was irregular. In conclusion, as we consider antimicrobial effect and the safety on living cells, it is necessary that we should improve concentration of preservatives and study continuously to develop a new preservatives without a toxic effect on the cornea surface.

• Applied Microscopy
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• v.36 no.3
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• pp.165-172
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• 2006

Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.