• Journal of Periodontal and Implant Science
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• 제33권4호
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• pp.585-597
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• 2003

Porphyromonas gingivalis has been implicated as an important periodontophathic bacterium in the etiology and progression of periodontal diseases. It has been reported that P.gingivalis may mediate periodontal destruction not only directly through its virulence factors, but also indirectly by including complex host mediated inflammatory reponses. The purpose of this study was t o evaluate the effects of P.gingivalis on the bone formation and resorption by osteoblasts. For this purpose, after determining the concentration below which sonicated P.gingivalis extracts (SPEs) have no cytotoxicity on mouse calvarial primary osteoblastic (POB) cells, we investigated the effects of SPEs on the alkaline phosphatase (ALP) activity, matrix metalloproteinase (MMP) expression (MMP-2, -9, 13), and prostaglandin $E_2$ ($PGE_2$) release in POB cells by treatment with SPEs below that concentration. The results were as follows; 1. SPEs showed no cytotoxic effect on POB cells up to a concentration of 1 ${\mu}m$/ml. 2. The treatment with SPEs reduced ALP activity in a dose-dependent manner in POB cells, In addition, when we investigated the effect of SPEs (1 ${\mu}m$/ml) on ALP activity for different exposure periods, statistically significant inhibition of ALP activity was shown at 2 days of exposure, and further significant inhibition occurred by extending the periods of exposure. 3. The treatment with SPEs stimulated the gene expression of MMP-9 in POB cells. 4. The pre-treatment with SPEs increased the amount of $PGE_2$ released in POB cells. In summary, the present study shows that P.gingivalis could inhibit osteogenesis and stimulate bone resorption not only by reducing ALP activity but also by increasing MMP-9 mRNA expression in osteoblasts, possibly through an endogenous $PGE_2$ pathway. In addition, our results suggest that if P.gingivalis affects osteoblasts in early differentiation stage, such effects by P. gingivalis could be irreversible.

• 대한한의학방제학회지
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• 제25권2호
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• pp.223-251
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• 2017

Objectives : A lot of experiment results of Yugmijihwangtang(YM) are reported in various kinds of journals. Many of them report on the new effects that are not recorded in the traditional medical texts. So it is necessary to take it into consideration that newly reported effects could be of help to clinical practice, because this process of comparison of Donguibogam and scientific experiment results will have basis to lead into the evidence based medicine. Methods : We compared the effects of in Donguibogam and the experiment results of YM. Results : The effects of YM in Donguibogam are to replenish essence and marrow, and to treat red wen, fatigue, treat hypouresis, urinary sediment, urinary urgency, hematuria, hydrocephalus, speech and movement retardation, yin-deficiency, diabetes mellitus, nonalcoholic fatty liver, melanoma, disability to see near and far sight, tinnitus, hearing loss, alopecia, angiogenesis, cough, cough at night, trachyphonia, and, infantile convulsion. The experiment results of YM since 2000 in both Korea and China are to inhibit atopic dermatitis, renal interstitial fibrosis, anti-oxidant, emphysema, stress, glomerulosclerosis, diabetic nephropathy, chronic glomerulonephritis, hemorrhage, plantar sweating, dermal aging, kidney aging, bone loss, breast cancer, pathological myocardial cell, primary liver cancer, thrombosis, osteoporosis, intrauterine growth retardation, chronic renal failure, IgA nepropathy, slow cerebral development, and hippocampal tissue lesions on the one hand, and to help bone formation, renin-angiotensin- aldosterone system, cerebral recovery, cognitive function and expression, osteoblast proliferation and differentiation, learning and memory, cold-tolerance and oxygen deficit-tolerance and anti-fatigue, endometrial formation, humoral and cell-mediated immunity, immune regulation effect, Hypothalamus-Pituitary-Ovary Axis, and spermatogenesis, on the other hand. Conclusion : When we compared the effects of YM with the experiment results of YM, there existed a considerable gap between them. So, from now on, it is expected that a great effort and consideration are needed to solve these gaps from an academic and clinical point of view.

• Journal of Periodontal and Implant Science
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• 제33권1호
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• pp.79-89
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• 2003

치주질환은 세균감염에 의해 치조골이 파괴되는 염증성질환으로서 치아상실의 주된 원인이다. Treponema denticola는 성인성 치주염의 병소에서 자주 발견되는 세균으로서 부착능 및 단백분해효소생성능과 같은 독성 인자가 밝혀져 치주조직 파괴에 있어서 중요성이 강조되어 왔다. 골개조는 조골세포의 골형성및 파골세포에 의한 골흡수의 균형에 의하여 유지되며 치주염시 야기되는 치조골파괴는 조골세포 및 파골세포 기능의 불균형에 의하여 야기되는 것으로 설명되고 있다. 골세포에 대한 영향으로서 T. denticola는 파골세포의 형성을 촉진시키는 것으로 보고되었으나 조골세포에 대한 영향은 아직 밝혀져 있지 않다. 따라서 본 연구에서는 T. denticola가 골형성에 미치는 영향을 알아보고자 마우스의 두개골세포로부터 조골세포를 분리한 후 T. denticola분쇄액으로 처리하여 본 세균이 조골세포의 alkaline phosphatase(ALPase) 활성, 석회화결절 형성 및 Prostaglandin $E_2\;(PGE_2)$ 생성에 미치는 영향을 평가하였다. ALPase활성은 p-nitrophenylphosphate분해능, 석회화결절형성은 Von Kossa 염색법, 그리고 PGE2의 농도는 효소면역측정법으로 측정하였다. T. denticola분쇄액 (2.5 ug/ml)은 마우스 두개골세포의 ALPase활성을 억제하였으며 석회화결절의 형성을 감소시켰다. 또한 동일한 농도의 균분쇄액은 마우스 두개골세포의 $PGE_2$ 생산을 증가시켰다. 균분쇄액과 prostaglandin의 합성억제제인 indomethacin으로 세포를 동시에 처리한 경우 T .denticola분쇄액에 의한 $PGE_2$의 생산은 감소되었으나, ALPase의 활성억제에는 변화가 없었다. 균분쇄액을 열처리하여 마우스 두개골세포에 처리하였을 때에도 ALPase의 활성이 억제되는 것에는 변함이 없었다. 이러한 결과는 T. denticola의 구성성분 중 열에 안정한 물질이 prostaglandin과 무관한 경로를 통해 조골세포의 분화를 억제함을 시사하며 이와 같은 T. denticola에 의한 골형성억제가 치주염시 야기되는 치조골 파괴에 관여할 수 있을 것으로 생각된다.

• 대한치과교정학회지
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• 제25권3호
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• pp.333-339
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• 1995

[ $1,25-(OH)_2D_3$ ]가 치주인대세포의 증식에 미치는 영향과 조골세포 기능을 나타내는 지표이며 골의 석회화 과정에 관여하는 alkaline phosphatase의 활성도에 미치는 영향을 관찰하여 아직 확실히 밝혀져 있지 않은 치주인대조직에 대한 $1,25-(OH)_2D_3$의 생물학적 기능을 알아보고자 교정 치료 목적으로 발거된 치아로부터 치주인대세포를 배양하였다. $1,25-(OH)_2D_3$를 첨가하여 24시간 후 [${^3}H$]thymidine으로 DNA를 표지하여 세포의 증식을 관찰하였으며 $1,25-(OH)_2D_3$가 치주인대세포에서 조골세포의 표지효소인 alkaline phosphatase의 활성도에 미치는 영향을 24시간 또는 6일간 $1,25-(OH)_2D_3$ 처리후 측정한 결과 100nM농도의 $1,25-(OH)_2D_3$은 치주인대세포의 증식을 유의하게 증가시켰으며 10nM에서는 차이를 보이지 않았다. $1,25-(OH)_2D_3$을 24시간 첨가시 10nM에서는 ALP활성도는 $80.8\pm31.4$nmol로 서 대조군 $38.5\pm5.3$nmol에 비해 유의하게 증가하였으며 또한 $1,25-(OH)_2D_3$을 6일동안 첨가하였을때에도 10nM에서 $106.7\pm23.0$nmol로 대조군 $29.3\pm1.0$nmol에 비해 유의하게 증가되었다. 이상의 결과를 종합하면 $1,25-(OH)_2D_3$이 치주인대세포의 증식 및 세포기능을 시사하는 결과로 사료된다.

• Imaging Science in Dentistry
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• 제36권4호
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• pp.189-198
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• 2006

Purpose: To investigate the in vitro response of MC3T3-E1 osteoblastic cells to X-ray in the presence and absence of 2 deoxy-D-glucose (2-DG) and quercetin (QCT). Materials and Methods: The MC3T3-E1 cells were cultured in an ${\alpha}-MEM$ supplemented with 5 mM 2-DG or $10{\mu}M$ QCT and then the cells were incubated for 12 h prior to irradiation with 2, 4, 6, and 8Gy using a linear accelerator (Mevaprimus, Germany) delivered at a rate of 1.5 Gy/min. At various times after the irradiation, the cells were processed for the analyses of proliferation, viability, cytotoxicity, and mineralization. Results: Exposure of the cells to X-ray inhibited the tritium incorporation, 3-(4, 5-dimethylthiazol-2yl-)-2, 5-diphenyl tetrazolium bromide (MTT)-reducing activity, and alkaline phosphatase (ALP) activity, and caused cytotoxicity and apoptosis in a dose-dependent manner of the X-ray. This effect was further apparent on day 3 and 7 after the irradiation. RA+2-DG showed the decrease of DNA content, cell viability, and increase of cytotoxicity rather than RA. ALP activity increased on day 7 and subsequently its activity dropped to a lower level. 2-DG suppressed the calcium concentration, but visual difference of number of calcified nodules between RA and RA+2-DG was not noticed. RA+QCT showed the increase of DNA content, cell viability, but decrease of cytotoxicity and subG1 stage cells in the cell cycle, and increased calcified nodules in von Kossa staining rather than the RA. ALP activity showed significant increases on day 7 and subsequently its activity dropped to a lower level. Conclusion: The results showed that the 2-DG acted as a radiosensitizing agent and QCT acted as a radiosensitizing agent respectively in the irradiated MC3T3-E1 osteoblast-like cells.

• Journal of Periodontal and Implant Science
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• 제35권1호
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• pp.133-152
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• 2005

1. 목적 Prostaglandin은 치주질환과 관련된 국소적 골 대사에 중요한 역할을 한다. ${\Delta}^{12}PGJ_2$는 생체 내에서 혈장의 존재 하에 형성되는 천연 $PGD_2$ 대사산물이며 peroxisome- proliferator에 의해 활성화되는 감마 수용체 (PPAR ${\Gamma}$)에 대해 높은 친화성을 갖는 리간드로서 핵 수용체군에 속하는 전사조절인자이다. 이 연구의 목적은 골화 과정에서 ${\Delta}^{12}PGJ_2$의 역할을 규명하기 위해, 조골세포주의 증식과 분화에 미치는 영향과 그에 관련된 세포기전을 조사하는 데에 있다. 2. 방법 인간 골육종세포주인 Saos-2 (ATCC.HTB 85)와 쥐의 조골세포주 (MC3T3-E1)를 배양한 후 실험군에 농도가 각각 $10^{-5}$, $10^{-6}$, $10^{-7}$, $10^{-8}$, $10^{-9}$ 몰인 ${\Delta}^{12}PGJ_2$와 ciglitazone (합성 PPAR 감마 길항체)를 첨가하였다. 조골세포에서 PPAR 감마의 발현을 관찰하기 위해 역전사효소-중합효소연쇄반응(RT-PCR)을 특정한 primer를 이용하여 시행하였다. 세포 증식은 1일, 2일, 3 일째에 MIT 분석법으로 측정하였고, 2 일째에 알칼리성 인산효소 (ALPase) 생산을 측정하였다. 위의 결과에서 얻은 적정한 농도에서 다양한 조골세포 분화의 표지자들-제 1 형 교원질, 알칼리성 인산효소, osteopontin 및 bone sialoprotein-에 대한 간이 정량적 역전사효소-중합효소연쇄반응 (semiquantitative RT-PCR)을 실시하였으며 골결절 형성에 대한 효과를 알아보고자 석회화 분석도 시행하였다. 3. 결과 ${\Delta}^{12}PGJ_2$와 ciglitazone 모두 Saos-2 세포주의 증식을 촉진시켰다 .$10^{-8}$ 몰의 ${\Delta}^{12}PGJ_2$와 $10^{-6}$몰의 ciglitazone을 첨가한 실험군을 대조군과 비교했을 때, 시간에 비례하여 세포 증식률이 증가되었다. 알칼리성 인산효소의 활성화 검사에서도 증식률에서와 유사한 결과를 보여주었다. 간이 정량적 RT-PCR에서는 ${\Delta}^{12}PGJ_2$로 처리한 군의 경우 제 1 형 교원질, 알칼리성 인산효소, osteopontin, 그리고 bone sialoprotein의 상대적 mRNA 수준이 유의하게 높았다. 석회화 분석에서는 MC3T3-E1 세포를 $10^{-6}$ 몰의 ${\Delta}^{12}PGJ_2$로 처리한 군과 $10^{-5}$ 몰의 ciglitazone으로 처리한 군에서 현저한 골결절 형성을 보였다. 이러한 결과들은 ${\Delta}^{12}PGJ_2$가 유용한 골 유도물질이 될 수 있으며 또한 그 작용기전이 PPAR 감마-의존형 경로와 연관되어 있음을 보여준다.

• 한국발생생물학회지:발생과생식
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• 제18권4호
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• pp.197-212
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• 2014

Osteosarcoma (OS) is one of the most common malignant primary bone tumors and NF-${\kappa}B$ appears to play a causative role, but the mechanisms are poorly understood. OS is one of the pleomorphic, highly metastasized and invasive neoplasm which is capable to generate osteoid, osteoclast and osteoblast matrix. Its high incidence has been reported in adolescent and children. Cell signal cascade is the pivotal functional mechanism acquired during the differentiation, proliferation, growth and survival of the cells in neoplasm including OS. The major limitation to the success of chemotherapy in OS is the development of multidrug resistance (MDR). Answers to all such queries might come from the knock-in experiments in which the combined approach of miRNAs with NF-${\kappa}B$ pathway is put into use. Abnormal miRNAs can modulate several epigenetical switching as a hallmark of number of diseases via different cell signaling. Studies on miRNAs have opened up the new avenues for both the diagnosis and treatment of cancers including OS. Collectively, through the present study an attempt has been made to establish a new systematic approach for the investigation of microRNAs, bio-physiological factors and their target pairs with NF-${\kappa}B$ to ameliorate oncogenesis with the "bridge between miRNAs and NF-${\kappa}B$". The application of NF-${\kappa}B$ inhibitors in combination with miRNAs is expected to result in a more efficient killing of the cancer stem cells and a slower or less likely recurrence of cancer.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제29권6호
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• pp.379-391
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• 2003

The present study was undertaken to evaluate bone regenerative capacity around titanium screw implants placed in irradiated rat's tibiae. At one week after single 15-Gy dose irradiation, miniaturized titanium screw implants were inserted into anterior aspect of the upper tibia of rats weighing 200-250g. Seventy rats were involved: 35 rats were control and 35 rats radiation group. The rats were killed at different intervals as 1, 2, 3, 4, 6, 8, 12 weeks after implantation for histologic observation, histomorphometric analysis and immunohistochemical study with fibronectin and CD34 antibody. 1. Histologically, various stages of bone maturation and ossification can be seen at 4 weeks and regenerated bone close to edges demonstrates more advanced calcification, and network of new bone are well formed at 12 weeks in non-irradiated group. In contrast, active bone formation with increased contact of newly formed bone to implant surface was noted at 4 weeks and a significant amount of new bone formation and bone-implant contact is oberved at 12 weeks in irradiated group. 2. Histomorphometrical analysis confirmed these histologic findings. A significant difference in implant-bone contact and bone density was measured between the control and radiation group. Mean MBD was 62.2% in control group and 27.5% in radiation group, mean MBIC was 86.6% in control group and 47.7% in radiation group, and mean TBIC was 87.3% in control group and 45.6% in radiation group at 12 weeks after implantation. 3. In immunohistochemical study with fibronectin and CD34, radiation reduced hematopoietic progenitor cells severely and disturbed differentiation of osteoblast in bone marrow. The results of this study revealed bone healing capacity around implant after radiation therapy was severely impaired and irradiation reduces the capacity for osseointegration of titanium implants. Many factors including radiation dose, period between radiation and implantation, bone quality, time elapse between first and second surgery, type of prosthetics and hyperbaric oxygen therapy must be considered carefully in postradiation implantation.

• Journal of Periodontal and Implant Science
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• 제36권3호
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• pp.745-755
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• 2006

The purpose of this study was to investigate the effects of irradiated frozen allogenic bone(IFAB) on the cell proliferation and differentiation of human fetal osteoblasts. Human fetal osteoblasts(hFOB1) were cultured to examine the cellular proliferation for 3 days and 5 days with $1mg/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$, $1{\mu}g/m{\ell}$, $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of IFAB, and to compare the ALP synthesis to control groups for 3 days with DMEM/F-12 1:1 Mixture and $1mg/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$, $1{\mu}g/m{\ell}$, $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of IFAB. To compare the calcium accumulation, hFOBl cultured for 23 days were quantified and photographed. The cellular proliferation of hFOBls treated with IFAB was increased at 5 days to control(p<0.05). The activity of ALP in hFOBls treated with $100ng/m{\ell}$ IFAB was significantly increased at 5 days(p<0.05). A quantified calcium accumulation in hFOBl was significantly increased at $100ng/m{\ell}$, $10ng/m{\ell}$ of IFAB(p<0.05). In the present study, we found that IFAB playa important role of bone formation in the early stage. There was considered that IFAB could be used in the bone graft material.

• 구강회복응용과학지
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• 제29권2호
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• pp.141-151
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• 2013

티타늄 산화막을 나노단위에서 변형시키는 방법은 다공성 표면을 강화하는 내부적 접근과 나노입자를 피복하는 외부적 접근으로 나눌 수 있다. 나노표면은 나노튜브, 나노피트, 나노노듈 및 다형구조 등 다양한 형태를 지닌다. 형성방법 및 형성재료에 따라 다른 표면이 생성되지만, 현재까지 표준화된 형성방법은 없다. 나노표면을 분석해 보면 마이크론 단위의 표면구조에는 영향을 미치지 않으며 전기화학적으로 안정적이다. 나노표면은 세포독성이 거의 없으며 조골세포의 증식과 분화를 모두 촉진하고, 섬유모세포의 증식을 저해하여 연조직 개재를 감소시키는 효과를 가진다. 또한 세포 및 단백질과 유사한 크기 및 형태를 가지기 때문에 조직과의 친화성이 우수하여 골유착을 증진시킨다. 하지만 그 작용이 미치는 범위는 극히 제한되어 있기 때문에 골조직과의 거리가 있는 경우에는 효과가 미미하다. 마이크론 단위의 표면과는 달리 나노표면은 광촉매효과로 인한 항균작용을 가지지만 지속시간이 짧아 실제 임상에서의 적용효과는 의문시 된다. 하지만 마이크론 단위의 표면거칠기가 가지는 단점을 배제할 수 있어 다양한 가능성을 가지기 때문에 더 많은 연구가 필요하다.