• 한국한의학연구원논문집
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• 제6권1호
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• pp.123-132
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• 2000

Bone is continuously remodeled during adult life with the resorption of old bone by osteoclasts and its subsequent replacement by osteoblast. Bone homeostasis is maintained by a balance between activities of osteoblasts and osteoclasts, but an imbalance between resorption and formation results in bone diseases including osteoporosis. Osteoblasts line up on the bone surface, especially regions of new bone formation, lay down bone matrix (osteoid) in orderly lamellae and induce its mineralization. Thus, the increased activity of osteoblasts is helpful to treat and prevent osteoporosis. In this study, we examined whether 80% EtOH extract of yukmijiwhang-whan is capable of affecting osteoblast proliferation using human osteoblast-like cell line, MG-63 and Saos-2. In an in vivo experiment, extract of yukmijiwhang-whan was administered for 9 weeks to ovariectomized (OVX) rats. At necropsy, uterus weights were measured, and trabecular bone areas (TBAS) of tibia and the sixth lumbar vertebra were measured by bone histomorphology. The maximum cell proliferation of MG-63 caused by extract of yukmijiwhang-whan at $5\;{\times}\;10^{-6}\;mg/ml$ was approximately 115% compared with control. In Saos-2, cell proliferation was approximately 145% of control at $5\;{\times}\;10^{-4}\;mg/ml$ and maximum alkaline phosphatase (ALP) activity was approximately 143% of control at $5\;{\times}\;10^{-5}\;mg/ml$. In animal study, however, the tibia and lumbar TBAS of the yukmijiwhang-whan group did not increased than the OVX control group. In conclusion, the 80% EtOH extract of yukmijiwhang-whan increased proliferation of osteoblasts but did not prevent bone loss in OVX rats.

• 생명과학회지
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• 제16권6호
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• pp.925-931
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• 2006

골조직은 골아세포, 파골세포, 골세포 등으로 구성되며, 골개조시 여러 인자가 세포증식, 분화, 활성화 및 골대사 조절에 관여한다. 이때 조골세포의 활성은 골형성에 중요하므로, 본 연구에서는 MC3T3-E1 조골세포주를 이용하여 해조류로 조제한 알칼리의 ENA-A 이온수가 조골세포의 증식과 분화활성에 미치는 영향을 조사하였다. 먼저 ENA-A 이온수가 조골세포의 성장에 미치는 영향을 MTT 검색법으로 조사 한 결과, ENA-A 이온수 2% 처리시 대조군과 비교하여 134% 증가하여 조골세포에 대해 높은 성장률을 보였다. 또한 ALP 효소 활성에 미치는 영향을 9일간 배양하여 그 변화를 측정하였다. 결과, 농도 1, 2, 4, 8% 처리시 112, 150, 188%까지 ALP 활성을 증가시켰다. ENA-A 이온수는 다시 ALP 효소 염색법과 Alizarin Red 염색으로 조골세포의 ALP활성유도, 분화와 석회화 형성능을 재확인하였으며 골기질 유전자의 발현의 변화도 확인하였다. 이에 본 연구결과를 통해 각종 무기질 성분을 함유한 ENA 이온수가 조골세포의 분화시에 hydroxyapatite 형성에 영향을 줌으로써 골다공증 예방에 효과가 있을것으로 생각하여 이를 보고하는 바이다.

• 생명과학회지
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• 제27권4호
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• pp.456-463
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• 2017

본 연구는 3종의 한약재(감초(Glycyrrhizae radix), 황기(Astragali radix) 및 산약(Dioscorea rhizome)) 추출물의 에스트로겐 유사활성 및 조골세포 증식과 분화에 미치는 영향을 검토하고자 실험을 진행하였다. 에스트로겐 유사활성 측정을 위하여 인체 유방암 세포주인 MCF7 세포를 luciferase 유전자를 리포터로 사용하는 에스트로겐 반응성 리포터 시스템을 가지는 세포로 형질전환하여 사용하였다. 상기 세포를 이용하여 추출물의 에스트로겐 유사활성을 측정한 결과, 한약재 추출물이 음성대조군과 비교하여 1.11배~5.73배 높은 에스트로겐 유사활성을 나타내었다. 그 중 감초 추출물이 가장 높은 에스트로겐 유사활성을 보였다. 감초 추출물의 에스트로겐 유사활성은 추출물의 농도가 $50{\mu}g/ml$ 및 $500{\mu}g/ml$ 일 때 각각 $10^{-8}M$ 및 $10^{-7}M$ $17{\beta}-estradiol$과 거의 유사한 활성을 나타내었다. 조골세포주인 MC3T3-E1 세포를 이용한 실험에서는 감초 추출물의 농도가 $1{\sim}1,000{\mu}g/ml$ 범위일 때 독성을 나타내지 않았다. 황기 추출물은 조골세포에 있어서 유의적인 세포 증식률을 보이지 않았다. 그러나, 감초 및 산약 추출물은 각각 148% 및 133%의 최대 증식률을 보였다. 또한, 감초 추출물은 대조군과 비교하여 alkaline phosphatase(ALP) 활성이 증가하였으며, 추출물의 농도가 $100{\mu}g/ml$ 일 때 최대 122%의 ALP 활성을 나타내었다. 또한, Alizarin red S 염색법을 이용한 석회화 형성능 측정에서도 대조군과 비교하여 석회화가 증가하였다. 이러한 결과로 미루어 보아 감초 추출물이 골 형성 및 골다공증 예방 효과가 우수한 식품 소재인 것으로 사료된다.

• 대한치과보철학회지
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• 제52권3호
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• pp.211-221
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• 2014

목적: 기존의 SLA 표면을 높은 친수성을 가지는 표면으로 개질하고자 NaOH에 침적하는 방법이 SLA 표면 형상 및 특성에 어떤 영향을 미치는지 알아보고, 골모유사세포의 증식, 부착 및 분화에 어떤 영향을 미치는지 알아보고자 계획되었다. 재료 및 방법: Machined surface (대조군), SLA surface (SLA 군), SLA에 NaOH 처리한 표면(SLA/NaOH 군)의 각 시편을 제조하고 친수성을 극대화한 SLA/NaOH 군의 표면 특성을 평가하기 위해 표면성분(XPS), 표면 거칠기, 표면 접촉각 등을 평가하였다. 그 이후 MG-63 세포 배양 후 이번 실험에서 만든 표면들이 세포독성을 가지는지를 평가하고, WST assay를 통하여 세포 증식, F-actin 염색을 통하여 세포의 부착형태를 관찰하였다. 이 후 ALP assay를 통하여 세포 분화를 평가하였다. 각 군간 통계측정을 위해 ANOVA 후 다중비교를 하였다(P<.05). 결과: SLA/NaOH 군의 접촉각은 $5.59{\pm}1.13$도였다. 모든 군들은 MG-63 세포에 대해 세포독성을 가지지 않았다. 세포 부착 평가에서 SLA/NaOH 군에서 가장 높은 부착 정도를 보였고(P<.05), Machined 군과 SLA 군에서도 표면 거칠기가 높은 SLA군에서 더 높은 세포 부착정도를 확인할 수 있었다(P<.05). 배양 7일까지 모든 군에서 MG-63 세포의 증식이 점차 증가하였다. 모든 군에서 3일과 7일에 세포의 증식에서 유의할 만한 차이가 보였고, SLA/NaOH 군에서 가장 높은 세포증식을 보였다. ALP 활성도는 7일에서는 세 군 사이에 차이가 없었다. 하지만 14일에는 SLA/NaOH 군이 유의성 있는 증대를 보였다(P<.05). 결론: 본 연구를 통하여 NaOH를 처리하는 수화방식을 통해 SLA 표면을 변형시킴으로서 세포의 부착, 증식 및 분화를 촉진시켜 임플란트의 골유착을 증진시킬 수 있는 가능성을 확인하였다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.109-109
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• 2003

Taurine is present in a variety of tissue and exhibits many important physiological functions in many tissues. Although it is known that many tissues mediate taurine transport, its functions of taurine transport in bone have not been identified yet. In the present study, we investigated the expression of taurine transporter (TauT) and taurine uptake using mouse stromal ST2 cells and osteoblast-like MC3T3-El cells, which is bone related cells. Detection of TauT mRNA expression in these cells were performed by reverse transcription polymerase chain reaction (RT-PCR). The activity of TauT was assessed by measuring the uptake of ［$^3$H］taurine in the presence or absence of inhibitors. TauT mRNA was detected in these cells. ［$^3$H］Taurine uptake was dependent upon the presence of extracellular sodium, chloride and calcium ions, and inhibited by cold-taurine and ${\beta}$-alanine. These results suggest that taurine has biological functions in bone and some effect on the bone cells.

• 한국재료학회지
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• 제17권2호
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• pp.107-114
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• 2007

Porous Ti compacts were fabricated by spark plasma sintering (SPS) method and their in vitro and in vivo biocompatibilities were investigated. Alkaline phosphatase (ALP) activity representing the activity of osteoblast was increased when osteoblast-like MG-63 cells were cultured on the Ti powder surface. Some genes related to cell growth were over-expressed through microarray analysis. The porous Ti compact with 32.2% of porosity was implanted in the subcutaneous tissue of rats to confirm in vivo cytotoxicity. 12 weeks post-operation, outer surface and inside the porous body was fully filled with fibrous tissue and the formation of new blood vessels were observed. No inflammatory response was confirmed. To investigate the osteoinduction, porous Ti compact was implanted in the femur of NZW rabbits for 4 months. Active in-growth of new bone from the surrounded compact bone was observed around the porous body. From the results, The porous Ti compacts fabricated by spark plasma sintering might be available for the application of the stem part of artificial hip joint.

• Nutritional Sciences
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• 제8권4호
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• pp.242-249
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• 2005

It is well established that zinc plays an important role in bone metabolism and mineralization. The role of zinc in bone formation is well documented in animal models, but not much reported in cell models. In the present study, we evaluated zinc deficiency effects on osteoblastic cell proliferation, alkaline phosphatase activity and expression, and extracellular matrix bone nodule formation and bone-related gene expression in osteoblastic MC3T3-E1 cells. To deplete cellular zinc, chelexed-FBS and interpermeable zinc chelator TPEN were used. MC3T3-E1 cells were cultured in zinc concentration-dependent (0-15 ${\mu}M\;ZnCl_2$) and time-dependent (0-20 days) manners. MC3T3-E1 cell proliferation by MTT assay was increased as medium zinc level increased (p<0.05). Cellular Ca level and alkaline phosphatase activity were increased as medium zinc level increased (p<0.05). Alkaline phosphatase expression, a marker of commitment to the osteoblast lineage, measured by alkaline phosphatase staining was increased as medium zinc level increased. Extracellular calcium deposits measured by von Kossa staining for nodule formation also appeared higher in Zn+(15 ${\mu}M\;ZnCl_2$) than in Zn-(0 ${\mu}M\;ZnCl_2$). Bone formation marker genes, alkaline phosphatase and osteocalcin, were also expressed higher in Zn+ than in Zn-. The current work supports the beneficial effect of zinc on bone mineralization and bone-related gene expression. The results also promote further study as to the molecular mechanism of zinc deficiency for bone formation and thus facilitate to design preventive strategies for zinc-deficient bone diseases.

• Carbon letters
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• 제13권3호
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• pp.139-150
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• 2012

Poly-L-lactic acid (PLLA), PLLA/hydroxyapatite (HA), PLLA/multiwalled carbon nanotubes (MWNTs)/HA, PLLA/trifluoroethanol (TFE), PLLA/gelatin, and carbon nanofibers (CNFs)/${\beta}$-tricalcium phosphate (${\beta}$-TCP) composite membranes (scaffolds) were fabricated by electrospinning and their morphologies, and mechanical properties were characterized for use in bone tissue regeneration/guided tissue regeneration. MWNTs and HA nanoparticles were well distributed in the membranes and the degradation characteristics were improved. PLLA/MWNTs/HA membranes enhanced the adhesion and proliferation of periodontal ligament cells (PDLCs) by 30% and inhibited the adhesion of gingival epithelial cells by 30%. Osteoblast-like MG-63 cells on the randomly fiber oriented PLLA/TEF membrane showed irregular forms, while the cells exhibited shuttle-like shapes on the parallel fiber oriented membrane. Classical supersaturated simulated body fluids were modified by $CO_2$ bubbling and applied to promote the biomineralization of the PLLA/gelatin membrane; this resulted in predictions of bone bonding bioactivity of the substrates. The ${\beta}$-TCP membranes exhibit good biocompatibility, have an effect on PDLC growth comparable to that of pure CNF membrane, and can be applied as scaffolds for bone tissue regeneration.

• Journal of Periodontal and Implant Science
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• 제27권2호
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• pp.291-303
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• 1997

Periodontal ligament cells may have a role in the regulation of hard and soft periodontal tissues, but their specific function has not yet to be determined. To evaluate further their role in periodontal regeneration, they were examined for osteoblast-like behavior. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and as a measure of cell characterization, it was examined that the morphology, alkaline phosphatase activity, collagen synthesis, and immunocytochemistry for osteonectin, osteocalcin, and collagen type I. Healthy periodontal ligament cells has more osteoblastic-like cell property in alkaline phosphatase activity. and collagen synthesis than gingival fibroblast. Immunocytochemistry localization explained that calcitonin were expressed in periodontal ligament cells only, and osteonectin and type I collagen were produced in both cells simultaneously. This results indicate that the growth characteristics of periodontal ligament cells and gingival fibroblasts exhibit some differences in proliferative rates and biochemical synthesis. The differences may help to calrify the role such cells play in the regenearation of periodontal tissues.

• Archives of Plastic Surgery
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• 제37권4호
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• pp.340-345
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• 2010

Purpose: Recently, bioceramics have become popular as a substitute graft material for reconstruction of bony defect after trauma or tumor surgery. Among the bioceramic materials, hydroxyapatite (HA) is favored due to its biocompatibility. HA scaffold is composed of the interconnected reticular framework, macropores and micropores. Macropores play an important role in cell migration, nutrients supply and vascular ingrowth. On the other hand, a number of micropores less than $10{\mu}m$ form an irregular surface on HA scaffolds, which prevents the osteoblast from adhering and proliferating on the surface of HA scaffold. Methods: In this study, three different groups were designed for comparison. In the first group (group A), conventional method was used, in which HA pellet was applied without surface pretreatment. The second group (group B) was given a HA pellet that has been coated with crystalline HA solution prior to application. In the third group (group C), the same method was used as the second group, where the pretreated HA pellet was heated ($1250^{\circ}C$, 1 hour) before application. Osteoblast-like cells ($2{\times}10^4$/mL) were scattered onto every pellet, then they were incubated in 5% $CO_2$ incubator at $37^{\circ}C$ for twelve days. During the first three days, osteoblast cells were counted using the hemocytometer daily. ALP activity was measured on the 3, 6, 9 and 12 culture days using the spectrophotometer. Results: Under SEM, group A showed a surface with numerous micropores, and group B revealed more rough crystal surface. Group C revealed a fused crystal appearance and flattened smooth surface. In proliferation and ALP activity of osteoblast cells, group C showed better results compared to group B. Group A which lacks pretreatment of the surface showed less osteoblast proliferation and ALP activity than group C, but showed better results than group B. Conclusion: We found that crystallized HA with heat treatment method enhances the osteoblasts proliferation and differentiation on the surface of HA pellets.