• 대한의용생체공학회:의공학회지
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• 제27권3호
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• pp.125-130
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• 2006

The number of the living endothelial cells and the shape of those are very import clinical parameters for the evaluation of the quality of cornea. In this paper, we developed the automated endothelial cell counting and shape analysis algorithm for a confocal microscope. Since, the endothelial images from the confocal microscope has a non-uniform illumination and low contrast between cell boundaries and cell bodies, it is very difficult to segment the cells from the endothelial images. To cope with these difficulties, we proposed the new two stage image processing algorithm. At first stage algorithm, we used a high-pass filter and histogram equalization to compensate the non-uniform brightness pattern and a morphological filter and a watershed method are applied to detect the boundary of cells. From this stage, we could count the number of cells in an endothelial image. At second stage algorithm, we used a Voronoi diagram method to classify the shape of cells. This cell shape analysis and the percent of hexagonal cells are very sensitive in detecting the early endothelium damage. To evaluate the performance of the proposed system, we p개cessed seven endothelial images obtained using a confocal microscope. The proposed system correctly counted 95.5% cells and classified 92.0% of hexagonal cell shapes. This result is better than any others in this research area.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.235-247
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• 2002

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권3호
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• pp.325-340
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• 2008

The purpose of this study was to evaluate the artificial dental plaque by Streptococcus mutans on 4 different implant surfaces. In this study, the specimens were divided into 4 groups according to implant surface treatment. Uncoated implant group(n=5) which has an uncoated, smooth surfaced implant(Osstem, Korea), SLA implant group(n=5) which has an sandblasted large grit and acid-etched surface implant(Bicon, USA). Oxidized implant group(n=5) which has an oxidized surfaced implant (Osstem, Korea), and RBM implant group(n=5) which has resorbable blasting media(RBM) surfaced implant(Osstem, Korea). Acquired pellicle by human saliva and dental plaque by Streptococcus mutans were made on each implant surface. To analyze the plaque condition on implants surfaces, cell count and optical density were taken as a microbiologic method, and SEM(Scanning Electronic Microscope) findings was also taken for evaluation of surface condition. The following results were obtained. 1. Cell counting results of artificial dental plaque were Uncoated group($658.0{\pm}102.0$), RBM group($878.0{\pm}170.0$), SLA group ($946.0{\pm}42.0$), Oxidized group($992.0{\pm}40.0$), and there was difference between Oxidized group and Uncoated implant group(p<0.05). In case of modified cell counting results by v/w% were RBM group($197.8{\pm}45.2$), Oxidized group($207.04{\pm}8.34$), Uncoated group($261.6{\pm}40.6$), SLA group($315.4{\pm}14.0$), and there was difference between RBM group and SLA group(p<0.05). 2. Optical density results of artificial dental plaque after ultrasonic treatment was that there was difference among groups, and optical density of RBM group was higher than that of Uncoated group(p<0.05). In case of modified optical density results by v/w%, there was difference among groups, and the modified optical density of Uncoated group and SLA group was higher than those of Oxidized group and RBM group(P>0.05). 3. SEM findings of artificial dental plaque on the surfaces of implant as follows; there were artificial dental plaque on the surfaces of all test implants. Streptococcus mutans and by-product were observed at 10,000 times magnified condition on all test implants. Adhesion area of artificial dental plaque was about 1/2 of total surface after 24 hours incubate at $37^{\circ}C$. These results showed that there were differences among implant surfaces on the growth of Streptococcus mutans, and bacteria and by-product were covered about 1/2 area of total implant surfaces at 24 hours incubate at $37^{\circ}C$.

• Journal of Periodontal and Implant Science
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• 제34권4호
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• pp.733-746
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• 2004

Bisphosphonates는 파골세포에 의한 골흡수를 방지하는 물질로 알려져 있으며 임상에서 널리 쓰이고 있다. 그 중 Alendronate는 Aminobisphosphonates의 한 종류로 non-aminobisphosphonates인 etidronate보다 100-1000배 더 강한 효과를 보이는 것으로 알려져 있다. 본 실험의 목적은 백서두개골 결손부의 골재생을 실험모델로 하여 alendronate의 국소투여 효과를 알아보는 것으로 액체의 흡수성과 골전도성이 우수한 것으로 알려진 collagen membrane을 사용하여 결손부 양측에 alendronate와 physiologic saline을 각각 적용하여 1주, 2주, 4주의 조직학적 치유양상, 파골세포활성도, 경도를 평가하였다. 조직학적 치유양상은 1주째 collagen membrane에 의한 염증성 침윤이 나타났으며 2주째부터 골성회복이 관찰되었고 4주째 완전한 골성회복을 보여 각주별 실험군, 대조군 공히 유사한 양상을 보였고 실험에 사용한 $200{\mu}g$의 용량은 조직학적으로 관찰할만한 골재생의 향상을 위해서는 부족한 용량으로 사료되는 바이다. TRAP(+) cell은 1주째 대조군에 비해 실험군에서 유의하게 적은 수를 보였으며(p<0.01) 2주와 4주째는 유의한 차이를 나타내지 않았고 경도측정에서는 2주째 대조군에 비해 실험군에서 유의하게 높은 경도를 보였으며 4주째 유의한 차이를 나타내지 않았다(p<0.05). 이상의 실험에서 alendronate는 골조직 치유과정의 초기에 파골세포의 활성도와 경도에 다소 영향을 미친 것으로 사료되며 향후 골조직 재생을 위한 임상적용에 응용을 위해서는 치유과정을 더욱 향상시킬 수 있는 추가적인 연구가 필요하리라 사료된다.

• 대한소아치과학회지
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• 제50권3호
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• pp.347-359
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• 2023

본 연구는 탈구된 치아의 저장 매체로서 polydeoxyribonucleotide (PDRN)의 적합성을 평가하고자 하였다. HBSS와 10, 25, 50, 100 ㎍/mL 농도의 PDRN 용액과 수돗물에 저장된 인간 치주인대 세포의 생존율을 측정하기 위해 Cell Counting Kit-8 assay와 Live/Dead assay를 시행하였다. 또한, PDRN의 항염증 효과를 평가하기 위한 NO 검출 및 qRT-PCR 실험을 진행하였다. 100 ㎍/mL 농도의 PDRN 용액에 저장된 치주인대 세포의 생존율이 다른 용액보다 유의하게 높았다(p < 0.01). 또한, 100 ㎍/mL 농도의 PDRN 용액은 유의하게 NO의 생산을 줄였다(p < 0.0001). 그리고, HBSS 용액에 비하여 50 및 100 ㎍/mL 농도의 PDRN 용액에서 유의하게 tumor necrosis factor α, interleukin (IL)-4, IL-6, 그리고 IL-10의 발현이 낮았다(p < 0.01). 이 연구를 통해 PDRN은 치주인대 세포에 세포 보존 및 항염증 효과를 가진 것으로 밝혀졌다. 이 연구는 효과적인 탈구치 저장매체의 개발을 위한 향후 추가적인 실험의 기반이 될 수 있을 것이라 생각한다.

• 한국식품위생안전성학회지
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• 제26권3호
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• pp.187-191
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• 2011

Platelet derived growth factor (PDGF)-BB is one of the most potent vascular smooth muscle cell(VSMC) proliferative factors, and abnormal VSMC proliferation by PDGF-BB plays an important role in the development and progression of atherosclerosis. The aim of this study was to assess the effect of YP 12, a newly synthesized obovatol derivative, on the proliferation of PDGF-BB-stimulated rat aortic VSMCs. The anti-proliferative effects of YP 12 on rat aortic VSMCs were examined by direct cell counting and by using $[^3H]$ thymidine incorporation assays. It was found that YP 12 potently inhibited the growth of VSMCs. The pre-incubation of YP 12 (1-4 ${\mu}M$) significantly inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated rat aortic VSMCs in a concentration-dependent manner. In accordance with these findings, YP 12 revealed blocking of the PDGF-BB-inducible progression through G0/G1 to S phase of the cell cycle in synchronized cells. Whereas, YP 12 did not show any cytotoxicity in rat aortic VSMCs in this experimental condition by WST-1 assay. These results also show that YP 12 may have potential as an anti-proliferative agent for the treatment of restenosis and atherosclerosis.

• 한국가금학회지
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• 제27권1호
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• pp.63-71
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• 2000

The testis is an extremely heterogeneous organ, containing numerous compartments types. Morphometric studies were performed of 3 avian species (pigeon, pheasant and chicken) to determine volume density absolute volume, numerical density, total number of serminiferous tubule components, and sperm production, especially those related to the Sertoli cell, and to make comparisons among the species. Volume density of seminiferous tubule components per testis was determined by point counting method. Testis volume and sperm production were measured by routine techniques. Numerical density (the number of cells per unit volume of testis) of seminiferous tubule components per testis was determined by morphometry (Floderus method). The volume density of seminiferous tubules per testis was 91.58, 92.18 and 94.21% in pigeon, pheasant, and chicken, respectively. The volume density of spermatogonium, spermatocyte, spermatid, spermatozoon, and Sertoli cell did not produce significant changes in the three species. The absolute volume of spermatogonium, spermatocyte, spermatid, and Sertoli cell showed significant changes in the three species (p<0.05). The average volume of Sertoli cell ranged from 758.34(pheasant) to 1,212.9 ㎛$^3$(chicken) and was not significantoy different in the three species(p>0.05). The number of Sertoli cells per testis showed significant differences in the three species : 34.52 $\times$10(sup)6, 186.82$\times$10(sup)6, 810.62$\times$10(sup)6 in pigeon, pheasant, and chicken, respectively(p<0.05). The sperm production was significantly different in the three species : 3,018$\times$10(sup)6, 993.9$\times$10(sup)6, and 8.9$\times$10(sup)6 in chicken, pheasant, and pigeon, respectively(p<0.05). These results suggest that number of Sertoli cells may be more important than Sertoli cell size in explaining the difference in sperm production among the three species.

• 한국전기전자재료학회논문지
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• 제21권1호
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• pp.12-17
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• 2008

In this paper, an I-V characteristics of bypass diode has been studied by counting the shading effect in photovoltaic module. The shadow induces hot spot phenomenon in PV module due to the increase of resistance in the current path. Two different types of PV module with and without bypass diode were fabricated to expect maximum output power with an increasing shading rate of 5 % on the solar cell. Temperature distribution is also detected by shading the whole solar cell for the outdoor test. From the result, the bypass diode works properly over 60 % of shading per cell with constant output power. Maximum power generation in case of solar cell being totally shaded with bypass diode decreases 41.3 % compared with the one under STC(Standard Test Condition). On the other hand, the maximum output power of the module without bypass diode gradually decreases by showing hot spot phenomenon with the increase of shading ratio on the cell and finally indicates 95.5 % of power loss compared with the output under STC. Finally the module temperature measured increases around $10^{\circ}C$ higher than that under STC due to hot-spots which come from the condition without bypass diode. It has been therefore one of the main reasons for degrading the PV module and shortening the durability of the PV system.

• Asian-Australasian Journal of Animal Sciences
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• 제13권2호
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• pp.161-166
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• 2000

This study was designed to determine the effects of the insulin-like growth factor (IGF-1) and estradiol $17-{\beta}$ on the in vitro proliferation of stromal vascular cell from Hanwoo omental, subcutaneous, intermuscular and intramuscular adipose tissues. Cells were cultured in M199+20% newborn calf serum and the proliferation of cells was measured by direct microscopic cell counting and change of genomic DNA amount. Cell numbers increased slightly over the first 72 hour of culture and then increased greatly, regardless of adipose tissue depots. In IGF-1 treatment, the number of omental preadipocytes maintained highest level from the beginning to the 20th day of culture. However, in estradiol-$17{\beta}$ treatment, those tended to be lower than the control from the beginning of culture and significantly lower at the 24th day. When IGF-1 was added to subcutaneous preadipocytes, the numbers of cells were higher from 11th day than those from other treatments, although there was no statistical significance. For intermuscular preadipocytes treated with IGF-1, its numbers were significantly (p<0.05) higher at 11th day, and in the other days it showed a similar tendency to those of the subcutaneous tissue. In this experiment, preadipocytes were taken from 24 month old fully matured steers and the highest proliferation rate was shown in intramuscular tissue followed by those of subcutaneous preadipocytes. Addition of $5{\mu}M$ estradiol-$17{\beta}$ to the growth medium failed to promote the replication of Hanwoo preadipocytes, as indicated by direct cell counts and total genomic DNA content. As the culture period proceeded, the amounts of DNA were increased, but the patterns of increment were not consistent with the results of cell numbers.

• Biomolecules & Therapeutics
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• 제18권1호
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• pp.56-64
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• 2010

Pentoxifylline (PTX) has been used for the local reduction of fat tissue in the clinical setting. However, its safety and efficacy have not been proven. The aim of this study was to evaluate the effects of PTX on cell lines established from fat tissue. Newly cultured human preadipocytes and adipocytes from subcutaneous abdominal fat in addition to purchased human lung fibroblasts and keratinocytes were treated with PTX at different concentrations. Cell viability was determined using the Cell counting kit (CCK)-8 assay and lipolysis was evaluated using an Elisa kit. DNA fragmentation, Western blot analysis, Hoechst and Propidium Iodide (PI) staining and fluorescence activated cell scanning analysis were performed to confirm apoptosis. The viability of adipocytes, preadipocytes, keratinocytes and fibroblasts was markedly decreased at concentrations of PTX above 20 mM. Apoptosis was induced at concentrations of PTX over 40 mM in all cell lines. Lipolysis was increased by 60% at concentrations of PTX of 20 mM compared to the control. In conclusion, the results of this study showed that 20 mM of PTX induced lipolysis. At concentrations over 20 mM, PTX reduced the viability of all cells studied including: adipocytes, preadipocytes, fibroblasts and keratinocytes, in a non-specific manner.