• Title/Summary/Keyword: Human osteoblast

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Effects of enamel matrix derivative and titanium on the proliferation and differentiation of osteoblasts (법랑기질유도체를 도포한 타이태늄 표면에서 조골세포의 증식 및 분화)

  • Park, Sang-Hyun;Lee, In-Kyeong;Yang, Seung-Min;Shin, Seung-Yun;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.33 no.3
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    • pp.359-372
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    • 2003
  • Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.

Effect of Gallus gallus var. domesticus (Yeonsan ogolgye) Extracts on Osteoblast Differentiation and Osteoclast Formation (연산 오골계 물 추출물이 조골세포와 파골세포의 활성에 미치는 영향)

  • Yoo, Han-Seok;Chung, Kang-Hyun;Lee, Kwon-Jai;Kim, Dong-Hee;An, Jeung Hee
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.322-329
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    • 2015
  • The effects of water extracts of Gallus gallus var. domesticus (Yeonsan ogolgye, GD) on the activities of osteoblast differentiation and the restraint of osteoclast formation were investigated. The water extract of GD in the human osteoblast "MG-63" cell, was examined in relation to alkaline phosphatase (ALP) activity and alizarin red stains. In order to observe the effects of osteoclasts formation, we analyzed RAW 264.7 cell tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains. The ALP activity of the water extract of hen and cock flesh (3 years) were 133.8% and 129.6%, respectively. The ALP activity of flesh extracts was also higher than that of the skin extracts. Concerning the effects of age, the 3 years old flesh extracts had a higher activity than that of the one year old extracts. However the activity of the 3 years old skin extracts was lower than that of the one year old extracts. For gender conditions, the ALP activity of the hen extract was higher than that of the cock. The degree bone mineralization in the three years old hen flesh exhibited the highest rate, at 124.3%, amongst all the groups. The TRAP activity of the flesh extracts of the three years old cock revealed the lowest rate, at 31.8%, compared to the control. Our results demonstrate that the water extract of GD increases bone mineralization and osteoblast differentiation activity in MG-63 cells and enhances the inhibitory activity of bone-resorption in RAW 264.7 cells. In conclusion, the water extracts of GD seem to be effective in the prevention and treatment of bone related disorders.

Expression and Role of the System L Amino Acid Transporter in FOB Human Osteoblast Cells (사람 골모세포 FOB에서 아미노산 수송계 L의 발현 및 역할)

  • Kim, Chang-Hyun;Park, Joo-Cheol;Kim, Do Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.9
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    • pp.1367-1374
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    • 2005
  • Amino acid transporters play an important role in supplying nutrition to normal and cancer cells for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the $Na^+$-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-tyre amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In the present study, we have examined the expression and functional characterization of system L amino acid transporters in FOB human osteoblast cells. RT-PCR and western blot analysis have revealed that the FOB cells expressed LAT1, LAT2 together with their associating protein 4F2hc. The uptakes of $[^{14}C]_L$-leucine by FOB cells are $Na^+$-independent and almost completely inhibited by system L amino acid transporter selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). These results suggest that the transport of neutral amino acids including several essential amino acids for cellular nutrition into the FOB human osteoblast cells is mediated by system L amino acid transporters.

The Effect of Nicotine on the Proliferation and Differentiation of Normal Human Osteoblast at the Surface of Implants (임플란트 표면에서 배양된 정상인 조골세포의 증식 및 분화에 미치는 니코틴의 영향)

  • Ahn, Tae Woong;Lee, Chong Heon
    • The Korean Journal of Oral and Maxillofacial Pathology
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    • v.42 no.5
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    • pp.111-118
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    • 2018
  • Nicotine of tobacco component has a controversial impact in the clinical outcome of dental implants. Although numerous nicotine effects on bone healing around implants have been presented, it is rarely reported in vitro study about normal human osteoblast(NHost) from oral and maxillofacial area at the surface of implants. The purpose of the present study was to evaluate the effect of nicotine on the proliferation and differentiation response of NHost to plasmatic and salivary levels of nicotine reported in smokers at the surface of screw-type plasma-sprayed titanium implants. NHosts were seeded on the surface of titanium implants and cultured for 21 days in ${\alpha}-MEM$ supplemented with 10% FBS, 50mg/ml ascorbic acid, 5mM ${\beta}$-glycerophosphate and 100nM dexamethasone. Seeded implants were exposed to various nicotine concentration(0.05-0.5mg/ml) from 1 to 21 days, and characterized for cell morphology, proliferation, differentiation, alkaline phosphatase(ALP) activity and ionized calcium concentration(Cai) of medium. Continuous exposure to higher nicotine concentration(above 0.3mg/ml) induced a dose- and time-dependent vacuolation of the cytoplasm, and a tendency to detach from the implant surface. 0.05mg/ml(lower nicotine concentration) did not cause significant effects in the cell proliferation and ALP activity. 0.1-0.2mg/ml caused evident dose-dependent effects in increased cell proliferation, ALP activity and earlier onset of matrix mineralization at levels up to 0.2mg/ml, while a dose-dependent inhibitory effect at 0.3-0.5mg/ml. Cai concentration of control group was decreased at 14 days. Increased Cai concentration at 0.1-0.2mg/ml, decreased Cai concentration at 0.3mg/ml and no change at 0.5mg/ml during the culture period were seen. It suggested that nicotine concentration could paly an role in modulating NHost activity as a contributing factor associated with proliferation and differentiation of NHost at the surface of implants.

Increased Osteoblast Adhesion Densities on High Surface Roughness and on High Density of Pores in NiTi Surfaces

  • Im, Yeon-Min;Gang, Dong-U;Kim, Yeon-Uk;Nam, Tae-Hyeon
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.11a
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    • pp.39.1-39.1
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    • 2009
  • NiTi alloy is widely used innumerous biomedical applications (orthodontics, cardiovascular, orthopaedics, etc.) for its distinctive thermomechanical and mechanical properties such as shape memory effect, super elasticity, low elastic modulus and high damping capacity. However, NiTi alloy is still a controversial biomaterial because of its high Ni content which can trigger the risk of allergy and adverse reactions when Ni ion releases into the human body. In order to improve the corrosion resistance of the TiNi alloy and suppress the release of Ni ions, many surface modification techniques have been employed in previous literature such as thermal oxidation, laser surface treatment, sol-gel method, anodic oxidation and electrochemical methods. In this paper, the NiTi was electrochemically etched in various electrolytes to modify surface. The microstructure, element distribution, phase composition and roughness of the surface were investigatedby scanning electron microscopy (SEM), energy-dispersive X-ray spectrometry(EDS), X-ray diffractometry (XRD) and atomic force microscopy (AFM). Systematic controlling of nano and submicron surface features was achieved by altered density of hydro fluidic acid in etchant solution. Nanoscale surface topography, such as, pore density, pore width, pore height, surface roughness and surface tension were extensively analyzed as systematical variables.Importantly, bone forming cell, osteoblast adhesion was increased in high density of hydro fluidic treated surface structures, i.e., in greater nanoscale surface roughness and in high surface areas through increasing pore densities.All results delineate the importance of surface topography parameter (pores) inNiTi to increase the biocompatibility of NiTi in identical chemistry which is crucial factor for determining biomaterials.

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Effects of Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture on Osteoblast Differentiation and Osteoclast Formation (방사선 육종 차조기와 백출 복합물이 조골세포와 파골세포의 활성에 미치는 영향)

  • Sim, Boo-Yong;Ji, Joong-Gu
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.1
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    • pp.168-177
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    • 2021
  • The effects of the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture on the activities of osteoblast differentiation and the restraint of osteoclast formation were investigated. the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture in the human osteoblast "MG-63" cell, was examined in relation to alkaline phosphatase (ALP) activity and alizarin red stains. In order to observe the effects of osteoclasts formation, we analyzed RAW 264.7 cell tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains. In cytotoxicity testing, it was confirmed that apple extract is safe at a concentration of 50 ㎍/㎖ or less. The ALP activity and Bone calcification formation ability were the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture had a lower activity than that of control group. However the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture significantly reduced activity of TRAP in the RAW 264.7 osteoclastic cell generation and effectively Inhibited the TRAP(+) multinuclear cells. Thus, our results demonstrate that the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture enhances the inhibitory activity of bone-resorption in RAW 264.7 cells. In conclusion, the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture seem to be effective in the prevention and treatment of bone related disorders.

A STUDY ON A CULTURE OF HUMAN ALVEOLAR BONE CELLS (사람 치조골세포의 배양에 관한 연구)

  • Choi, Byung-Ho;Park, Jin-Hyung;Yoo, Jae-Ha
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.6
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    • pp.602-605
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    • 2000
  • Human alveolar bone cells were isolated from alveolar bone fragments obtained from normal individual undergoing third molar extractions. Alveolar bone fragments were cultured as explant. Cells began to migrate in the first $5{\sim}7$ day and were confluent in $5{\sim}7$ week. Matrix mineralization was observed by 4 week. Our studies utilize established protocols for the characterization of these cells as osteoblasts by means of alkaline phosphatase activity determination, identification of osteocalcin antigens, establishing the presence of cells expressing type I collagen and determining the ability of cells to produce calcification. Transmission electron microscopic observations confirmed the presence of a collagen matrix undergoing a mineralization process. This new model, using human alveolar bone cells, may provide a tool to investigate alveolar bone development and physiology and to set up new therapeutic approaches.

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EFFECTS OF CHITOSAN ON HUMAN OSTEOBLASTS (키토산이 조골세포에 미치는 영향)

  • Kim, Ki-Hong;Park, Young-Ju;Park, Jun-Woo;Lee, Yong-Chan;Cho, Byoung-Ouck;Ahn, Byoung-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.4
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    • pp.280-285
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    • 2002
  • As the result of the study concerning "bone inducibility of chitosan", 1. "BMP-2" was observed mainly through the test when the "osteoblast" is exposed to the "chitosan". The expression of BMP-2 was 542.63 times compared to control after 2 hours exposure and it was maintained 16.60 times till 24 hours. 2. The expression of BMP-4 was decreased compared to control during exposure. 3. The expression of BMP-7 revealed two peaks during exposure. 4. The expression of osteocalcin was increased in early phase, and then decreased. Although it is not clear whether the "chitosan" is clinically effective material as a "bone induction material", we could say that it has a function for bone induction. Further detailed study will be required.

BIOCHEMICAL CHARACTERISTICS OF HUMAN PERIODONTAL LIGAMENT CELLS IN VITRO (치주인대세포(齒周靭帶細胞)의 생화학적(生化學的) 특이성(特異性)에 대(對)한 연구(硏究))

  • Cho, Soung-Wook;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.22 no.1
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    • pp.273-283
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    • 1992
  • To find out the differences between periodontal ligament cells (PDL cells) and gingival fibroblast cells (GFB cells), alkaline phosphatase, a marker enzyme for osteoblast, was used to measure the activities and $^{45}CaCl_2$ isotope was used to find out cellular and release of $^{45}Ca$, a requisite for bone formation,. PDL cells and GFB cells from 1 to 5 passages were also measured in alkaline phosphatase activity assay. By the use of above methods, followings were concluded that the PDL cells and the GFB cells have characteristics that are different from each other. In that PDL cells showed large amount of calcium uptake and large amount of calcium release in initial stage, they seem to possess characteristics which are similar to osteoblast-like cells. 1. The PDL cells, in contrast to the gingival fibroblast, showed exceedingly high alkaline phosphatase activity which was highest at the second passage, decreasing thereon. But gingival fibroblasts cells showed no distinct differences in alkaline phosphatase activity as the passage were elapsed. 2. For both PDL cells and GF cells, the $^{45}Ca$ uptake was greatest at 2 hours period. The PDL cells showed higher measuring than GFB cells through out the whole time period. 3. Whereas the GFB cells showed slow increase of $^{45}Ca$ release as time relapsed, the PDL cells showed rapid increase of $^{45}Ca$ release.

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