• Applied Biological Chemistry
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• 제34권2호
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• pp.134-141
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• 1991

대두의 조직배양에서 배양기간중 생화학적 대사산물의 변화와 특성을 조사하기 위하여 개화후 15일된 미숙자엽을 채취하여 기내에서 3주간 배양하였다. 이때의 배양체를 embryogenic callus(EC)와 non-embryogenic callus(NEC)로 구분하였다. EC의 일부는 다시 3주간 계 대배양하여 root forming cultures(RFC)와 shoot forming cultures(SFC)로 구분하였으며, EC의 또 다른 일부는 원형질체의 분리에 사용되었으며, 분리된 원형질체는 4주간 배양하였다. 이때 유기된 배양체를 protoplasts로부터 유기된 embryogenic callus(PEC)와 non-embryogenic callus(PNEC)로 구분하였다. 각각의 배양체에 대하여 단백질 및 그 아미노산조성을 조사한 결과, 아미노산의 조성 은 NEC와 PNEC에서보다 EC와 PEC에 서 methionine의 함량이 현저히 낮은 반면, phenylalanine의 함량이 높았다. 단백질의 양상은 EC에서는 18KD, NEC에서는 22KD 정도에서 차이가 났다. 또한 각각의 배양체에 대한 peroxidase 동위 효소의 활성을 조사한 결과, EC와 PEC에서는 peroxidase isozyme A(piA)의 활성이 높게 나타났으며, RFC와 SFC에서는 peroxidase isozyme B(piB)의 활성이 높았다.

• Journal of Periodontal and Implant Science
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• 제28권1호
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.

• 대한화학회지
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• 제37권8호
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• pp.723-729
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• 1993

탄소분말과 Nujol oil의 carbon paste 혼합물에 1-(2-pyridylazo)-2-naphthol (PAN)을 섞어서 Cu(Ⅱ)이온에 감응하는 수식전극을 제작하였다. Cu(II) 이온이 포함된 완충용액에 전극을 담구어 전위를 가하지 않고 PAN-수식전극의 표면에 Cu(II) 이온을 석출시킨 후, 전해액으로 옮겨 일정시간 동안 일정 전위에서 환원시켰다. 그런 다음 양의 방향으로 전위를 주사함으로써 좋은 전압전류파를 얻을 수 있었다. 전극표면을 산용액에 담금으로써 재생시킬 수 있었다. 전극의 재현성은 석출/측정/재생의 순으로 5회 반복실험하여 조사한 결과 상대표준편차는 6.1${\%}$였다. 시차펄스 전압전류법으로 조사한 경우 2.0 ${times}$ 10$^{-7}$M에서 1.0 ${times}$ 10$^{-6}$ M의 농도 범위에 걸쳐 직선성이 성립했으며, 검출한계는 6.0 ${times}$ 10$^{-8}$ M이었다. Cu(II) 이온을 정량하는데 있어서 EDTA와 oxalate 이온을 제외한 다른 공존이온의 영향을 별로 받지 않았다.

• Applied Biological Chemistry
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• 제42권1호
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• pp.12-19
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• 1999

Biopolymer를 생산하는 Bacillus subtilis K-1과 유당 이용능이 있는 Bacillus coagulans를 융합시켜 획득된 융합주에 의해 생산된 biopolymer의 특성 변화, 생산성 향상의 연구의 일환으로 두균주의 융합을 시도하였고 융합주의 특성을 조사하였다. 원형질체 융합은 33% PEG(Mw. 6,000), 1% PVP, 10 mM $CaCl_2$이 함유된 fusion fluid(FF)에서 5분간 반응하였고, 항생제가 함유된 선별배지에 중층 후, 3일간 배양했을 때 Bacillus subtillus coagulans 변이주 들간의 융합빈도는 $4.6{\times}10^{-5}{\sim}1.8{\times}10^{-7}$이었다. 융합주의 세포특성 검토에서 분리된 모든 융합주의 segregation 비는 $1{\sim}6%$로 나타나 유전적으로 안정하였고, 각 균주가 요구하는 영양 요구성 marker와 항생제의 함유배지에서 생육이 확인되었다. 주요 융합주의 DNA함량은 모균주보다 $1.6{\sim}2.7$배 높게 나타났다. 융합주의 전자현미경 관찰에서두 모균주는 간상체로서 두꺼운 세포벽과 이중막을 확인할 수 있었고 원형질체 형성과정에서 팽융된 세포에서 구형의 원형질체 방출이 이루어지고, 원형질체 융합과정 및 형성된 융합체에서 융합체내 전형적인 vesicle이 확인되었으며 모균주보다는 큰 형태의 세포를 확인할 수 있었다.

• Journal of Acupuncture Research
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• 제23권5호
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• pp.69-77
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• 2006

Background : Mesenchymal stem cells (MSCs) are present in most of the tissue matrix, taking part in their regeneration when injury or damage occurs. The aim of this study was to investigate the presence of cells with pluripotential characteristics in human subchondral bone and the capacity of these cells to differentiate to osteoblast. Methods : Human subchondral bone were digested with collagenase. Isolated cells were cultured with a-MEM, 15% FBS, 10-8M dexamethasone and 50 ng/mL ascoric acid. Cells from 0 day(isolated cells), 7 day (first subculture) and 14 days (third subculture) were used to carry out phenotypic characterization experiments flowcytometry analysis with 11 monoclonal antibodies) and osteogenic differentiation experiments. Osteogenic differentiation of cells was assessment by quantification of bone extracellular matrix components by following analysis: alkaline phosphatase(ALP) stains to detect ALP activity, RT-PCR and western blot to detect osteocalcin (OCN), osteopontin (OPN) and type I collagen(Col I), and Alizarin red stains to detect calcium deposition. Results : Flowcytometry analyses showed that in our population more than 98% of cells were positive for MSC markers: SH-2(CD105, 99%), CD29 (95%), CD73 (95%). Cells were negative for hematopoietic markers (CD11b, CD34, and CD45). Furthermore, cells showed positive stain to multipotent markers such as CDl17 (c-kit) (15.1%), and CD166 (74.9%), and cell adhesion molecules such as CD54 (78.1%) and CD106 (63.5%). The osteogenic specific marker analyses showed that the culture of these cells for 7 and 14 days stimulates ALP, OCN, OPN and Col I synthesis by RT-PCR and Western blot analysis. Also, after 14 days in the culture of MSCs induces mineralization by Arizarin red stain. Conclusion : In this work, we demonstrated a new and efficient method for osteoblastic differentiation of human subchondral bone stem cells. As MSCs takes part in reparative processes of adult tissues, these cells could play an important role in osteogenesis.

• 대한화학회지
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• 제37권10호
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• pp.881-887
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• 1993

코발트(II) 이온의 정량을 위해 ${\iota}$-sparteine(SP)으로 변성된 탄소 반죽 전극(carbon paste electrode : CPE)을 제작하였다. 수용액 중의 코발트 이온을 CPE에 처리시킨${\iota}$-sparteine과 반응시켜서 착화합물 상태로 전극표면에 석출시킨 후, 이 착물을 전압-전류법을 사용하여 정량하였다. 또한 변성된 CPE의 전극반응을 순환 전압-전류법을 사용하여 아세트산/아세트산염 완충용액에서 조사하였다. 한번 사용한 전극은 산용액으로 처리하여 5회 이상 재사용할 수 있었다. CPE에 흡착된 Co(II)-SP 착물의 산화환원 전위는 0.17V / 0.27V였으며 산화피이크는 0.17V의 환원피이크를 지나야 나타나는 피크이다. 이는 전극표면의 SP와 용액내의 Co(II)이온이 착물을 형성하여 흡착되고, 이 전극표면에 형성된 착물이 산화환원되어 나타나는 피이크이다. 펄스차이 전압-전류법을 사용할 경우 $2{\times}10^{-7}$M 농도까지 직선적으로 감응함을 알 수 있었으며, 그 때의 상대표준 편차는 ${\pm}$5.6%이었다. 이 경우 검출한계는 $1.0{\times}10^{-7}$M이었다. 리간드와 착물을 형성할 것으로 예상되는 여러 금속이온에 대한 방해작용을 조사하였다.

• Journal of Periodontal and Implant Science
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• 제36권1호
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• pp.1-14
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• 2006

The purpose of the present study was to evaluate the histologic results of bone cavities that were surgically created in the calvaria of rabbit and filled with $HA/{\beta}-TCP$ composite powders, which had been developed in Korea (Dentium, Korea). Ten young adult rabbits were used. Four defects were surgically produced in calvaria of each rabbit. Each rabbit was anesthetized with Ketamine-HCI (5 mg/kg, Yuhan Cor. Korea) and Xylazine-HCI (1.5 ml/kg, Yuhan Cor. Korea)). An incision was made to the bony cranium and the periosteum was reflected. Using a trephine bur (external diameter: 8 mm, 3i, USA), 4 'through-and-through' bone defects were created with copious irrigation, and classified into 4 groups: control group: no graft materials, experimental group I: normal saline + graft materials: experimental group II: venous blood + graft materials: experimental group III: graft materials only. The defects were randomly filled with graft materials. The defects were closed with resorbable suture material. At the end of the surgical procedure, all animals received a single intramuscular injection of antibiotics Gentamicin (0.1 mg/kg, Dae Sung Microb. Korea). Rabbits were sacrificed with phentobarbital (100 mg/kg) intravenously at 1-, 2-, 4-, 6- and 8-week after. Specimens were treated with hydrochloric acid decalcifying solution (Fisher Scientific, Tustin, CA) and sectioned by bisecting the 8 mm diameter defects. The histologic specimens were prepared in the general method with H & E staining at 6 ${\mu}m$ in thickness. The results were as follows; 1. New bone formation showed from after 2-week of surgery in defect area. As time lapsed, lots of new bone formation and mature bones showed. 2. Histologically, degree of new bone formation could not be discerned among the experimental groups. But, for experimental group II, lots of cells gathered around graft materials after 1-week of surgery, new bone formed slightly faster and than the others at 1-week after. For experimental group I, a few inflammatory finding showed around graft material at after 1-week and after 2-week of surgery. 3. No bone formation did show for control group. Based on histologic results, the new $HA/{\beta}-TCP$ composite powders appeared to act as a scaffolding material for regeneration of osseous defects.

• Molecules and Cells
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• 제41권6호
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• pp.582-590
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• 2018

Endothelial progenitor cells (EPCs) and outgrowth endothelial cells (OECs) play a pivotal role in vascular regeneration in ischemic tissues; however, their therapeutic application in clinical settings is limited due to the low quality and quantity of patient-derived circulating EPCs. To solve this problem, we evaluated whether three priming small molecules (tauroursodeoxycholic acid, fucoidan, and oleuropein) could enhance the angiogenic potential of EPCs. Such enhancement would promote the cellular bioactivities and help to develop functionally improved EPC therapeutics for ischemic diseases by accelerating the priming effect of the defined physiological molecules. We found that preconditioning of each of the three small molecules significantly induced the differentiation potential of $CD34^+$ stem cells into EPC lineage cells. Notably, long-term priming of OECs with the three chemical cocktail (OEC-3C) increased the proliferation potential of EPCs via ERK activation. The migration, invasion, and tube-forming capacities were also significantly enhanced in OEC-3Cs compared with unprimed OECs. Further, the cell survival ratio was dramatically increased in OEC-3Cs against $H_2O_2$-induced oxidative stress via the augmented expression of Bcl-2, a pro-survival protein. In conclusion, we identified three small molecules for enhancing the bioactivities of ex vivo-expanded OECs for vascular repair. Long-term 3C priming might be a promising methodology for EPC-based therapy against ischemic diseases.

• 식물조직배양학회지
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• 제27권3호
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• pp.213-217
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• 2000

기내에서 무균적으로 생장시킨 오이 유묘의 경단조직과 하배축을 배양하여 캘러스 분화 및 생장에 대해 조사했다. 오이 경단조직을 생장조절물질이 첨가하지 않은 배지에서는 유식 물체로 분화하였으나 2,4-D가 0.5∼2 mg/L 첨가된 MS배지에서는 비배발생캘러스와 점액성캘러스가 유도되었고 배발생 캘러스는 분화되지 않았다. 그러나 유묘 육성시 암조건에서 발아 후 5∼7일 된 개체의 하배축을 5∼10mm크기로 절단하여 2,4-D가 1.0 mg/L이 첨가된 MS배지에 50일간 광조건에서 배양했을 때에는 배발생캘러스와 배상체가 분화되었다. 아울러 하배축 유래 비배발생캘러스 중 백색캘러스는 45.1％가 발근된 반면에, 노랑색캘러스는 불과 0.6%만 발근되었으며. 조단백질의 함량은 백색캘러스보다 노랑색캘러스가 월등히 많았다. 배발생캘러스와 배상체에서 발달된 소식물체는 순화과정을 거쳐 비닐하우스 내의 고형배지경에서 양액재배로 정상적인 과실을 수확하였다.

• Journal of Periodontal and Implant Science
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• 제51권6호
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• pp.374-385
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• 2021

Purpose: The aim of this study was to evaluate the effects of locally delivered 1% alendronate (ALN) gel used as an adjunct to non-invasive periodontal therapy. Methods: Ligature-induced periodontitis was performed in 96 rats. The ligature was tied in the cervical area of the mandibular left first molar. The animals were randomly divided into 4 groups: 1) NT, no treatment; 2) SRP, scaling and root planning; 3) SRP/PLA, SRP followed by filling the periodontal pocket with placebo gel (PLA); and 4) SRP/ALN, SRP followed by filling the periodontal pockets with 1% ALN gel. Histomorphometric (percentage of bone in the furcation region [PBF]) and immunohistochemical (receptor activator of nuclear factor-κB ligand, osteoprotegerin, and tartrate-resistant acid phosphatase) analyses were performed. Data were statistically analyzed, with the threshold of statistical significance set at P≤0.05. Results: The SRP, SRP/PLA, and SRP/ALN groups presented a higher PBF than the NT group (P≤0.01) at 7, 15, and 30 days. The SRP/ALN group presented a higher PBF than the SRP/PLA group in all experimental periods, as well as a higher PBF than the SRP group at 15 and 30 days. No differences were observed in the immunohistochemical analyses (P>0.05 for all). Conclusions: Locally delivered 1% ALN gel used as an adjunct to SRP enhanced bone regeneration in the furcation region in a rat model of experimental periodontitis.