• 제목/요약/키워드: Regenerative Medicine

검색결과 392건 처리시간 0.029초

수종의 생약제제가 hFOB1의 염기성 인산분해 효소 활성에 미치는 영향 (Effects of Several Natural Medicines on Alkaline Phosphatase Activity in hFOB1)

  • 장길용;현아나;김윤상;유형근;신형식
    • 동의생리병리학회지
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    • 제16권5호
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    • pp.1042-1047
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    • 2002
  • Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential for periodontal tissues. Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have been traditionally used as medicines for treatment of bone disease in Korea. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity in human fetal osteoblast cell line (hFOB1) with several natural medicines. hFOB1 added DMEM/F-12 were cultured with dexamethasone as a positive control, and with each natural medicine. ALP activity was measured by spectrophotometer for enzyme activity and naphthol AS-Bl staining was performed for morphometry. All of the natural medicines induced a higher ALP activity compared to negative control, especially, Cortex Eucommiae increased an ALP activity in all experimental groups (p<0.05). In naphthol AS-Bl staining, all of the natural medicines of this study increased the stained area compared to negative control. Especially, Cortex Eucommiae and Eupoly phaga showed statistical significance compared to negative control (p<0.05). These results indicate that Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have an inducing ability of ALP synthesis on osteoblasts.

자궁축농증 있는 개에서의 심한 백혈병성 반응 (Extreme Leukemoid Reaction in a Dog with Pyometra)

  • 한재익;장혜진;전귀호;강현구;나기정
    • 한국임상수의학회지
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    • 제26권6호
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    • pp.619-621
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    • 2009
  • 개에서 백혈병성 반응은 흔하지는 않다. 이 것은 백혈구가 많이 증가하여 백혈병성 반응을 보이는 증례이다. 10년령의 암컷 말티즈로 화농성의 혈액섞인 질 분피물이 있었다. 전체혈구계산에서 백혈병성 반응($976\times10^9$ cells/l)을 나타내고 비 재생성 빈혈이 있었다. 혈액도말검사에서 증가된 백혈구는 호중구와 단핵구만 나타났다. 방사선 및 초음파검사에서 자궁은 확장되고 복합적인 액체가 차있는 것이 확인되었다. 자궁절제술 이후에 E. coli가 자궁내 액체로부터 분리되었다. 저자의 조사에 의하면 국소감염의 예에서 이와 같이 극도의 백혈병성 변화를 보이는 것은 처음이다.

석화채(石花菜) 추출물 도포가 마우스의 심재성 2도 화상에 미치는 영향 (Effects of Skin Application of Gelidium amansii Extract on Deep Second-degree Burns in Mice)

  • 김동현;홍승욱
    • 대한한의학회지
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    • 제31권2호
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    • pp.36-47
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    • 2010
  • Objective: The purpose of this study was to investigate the effects of skin application with Gelidium amansii extract on skin with deep second degree burns in mice. Methods: BALB/c mice were divided into four groups: normal (NOR) group; burn-elicited mice (CON) group, Silmazine-treated mice after burn elicitation (ST) group, and Gelidium amansii-extract treated mice after burn elicitation (GT) group. To examine the skin recovery effect after burn, changes of burn area, angiogenesis and histologic structure were analyzed. To measure effect of edema regulation, matrix metalloproteinase-9 (MMP-9) was analyzed. To estimate the skin regenerative & stable effect, 5-bromo-2'-deoxyuridine (BrdU) and substance P were analyzed. Results: 2 weeks later, 1. The size of burn area decreased in the GT and ST groups more than the CON group. 2. Alleviation of angiogenesis appeared in the GT and ST groups more than in the CON group. 3. Blood clot, epithelial cell hyperplasia, and inflammatory cell infiltration declined in the GT and ST groups more than in the CON group. 4. MMP-9, BrdU, and substance P positive reaction decreased in the GT and ST groups more than in the CON group 5. In the comparative study, the GT group was superior to the ST group. Conclusion: The skin application of Gelidium amansii extract could lessen skin damage by the medium of regulation MMP-9 activation. This skin stabilization was induced in mice with deep second degree burns.

The Effect of Acupuncture in Promoting Neurogenesis and Angiogenesis after Middle Cerebral Artery Occlusion in Rats

  • Lee, Hong Min;Nam, Sang Soo;Kim, Yong Suk
    • Journal of Acupuncture Research
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    • 제30권3호
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    • pp.1-13
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    • 2013
  • Objectives : This study was performed to choose more effective neuro-protective acupuncture point and to verify the effect of acupuncture in promoting neurogenesis and angiogenesis as a result of its neuro-vasculo-regenerative effect in middle cerebral artery occlusion model in rats. Methods : By TTc staining we chose the most effective acupuncture point with neuro-protection. We randomly divided into four groups: Such as (1) sham group(with sham-operation), (2) sham+acupuncture group(with sham-operation), (3) middle cerebral artery occlusion group, (4) MCAO+AT group. Acupuncture procedure was performed for four days. Total RNA was extracted using TRIzol reagent, according to the manufacturer's instructions, and was purified using an RNAeasy mini kit. Immuno-histochemistry was performed using primary antibody mouse anti-BrdU, NeuN, Dcx, and VEGF. Results : We found that $ST_{36}$ had the more neuroprotective effect than $LI_{11}$ and $SP_3$. The microarray analysis revealed that 54 genes were more expressed neurogenesis pathway in MCAO+AT group compared with MCAO group(fold changes greater than or equal to twofold change). 11 genes were more expressed angiogenesis pathway. And 7 genes were more expressed VEGF pathway. Immuno-histochemistry revealed that cell proliferation, cell migration and cell maturation were increased. Conclusions : This study demonstrated that acupuncture on $ST_{36}$ had neuro-protective and neuro-restorative effect in ischemic brain injuries. And its mechanism might be related to promote neurogenesis and angiogenesis. These results suggest that acupuncture have potential benefits for the treatment of ischemic stroke.

카사이 수술 후 장기생존환자의 간 자기공명영상 소견 (MRI Findings of Long-term Survivals after Kasai Portoenterostomy)

  • 정지광;정은영;박우현;최순옥
    • Advances in pediatric surgery
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    • 제18권1호
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    • pp.12-17
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    • 2012
  • The purpose of this study is to analyse clinical impact of specific MRI findings in liver in patients of long-term survivors after Kasai portoenterostomy (KPE). Twenty-eight patients who were underwent KPE were followed up more than 5 years. Macro-regenerative nodule (MRN) and beaded-duct dilatation (BDD) were considered as important findings in liver MRI. The association between these findings in MRI and clinical indicator, serum bilirubin level and history of cholangitis were evaluated. Sixteen patients (57.1%) were shown MRN in liver MRI. Therewere 14 patients(50%) whose MRI showed BDD. Serum total and direct bilirubin were 3.6 mg/dL and 1.8 mg/dL respectively in positive MRN group whereas 1.4 mg/dL and 0.7 mg/dL in negative MRN group (p = 0.427). Serum total and direct bilirubin level were 4.2 mg/dL and 2.1 mg/dL in patients with BDD negative group compare to 1.1 mg/dL and 0.5 mg/dL in BDD positive group (p = 0.281). The odds ratio to have cholangitis in the patient with MRN was 2.3 and 0.53 in patient with BDD in their MRI findings. MRN in liver MRI may suggest high bilirubin level and more chance to have cholangitis, but the findings of BDD may related to low bilirubin level and less change to have cholangitis.

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Platelet-Rich Plasma Enhances Proliferation and Migration and Inhibits Inflammatory Processes in Canine Chondrocytes

  • Kim, Dongyub;Jeong, Seong Mok;Kwon, Youngsam;Yun, Sungho
    • 한국임상수의학회지
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    • 제36권4호
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    • pp.200-206
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    • 2019
  • This study was performed to assess the anti-inflammatory and cartilage regenerative effects of platelet-rich plasma (PRP) on canine chondrocytes. Proliferation and migration assays under both normal and lipopolysaccharide (LPS)-induced inflammatory conditions were performed with various concentrations of PRP (1% to 10%). The expression levels of genes related to osteoarthritis were evaluated in the following groups: PRP group, LPS group and LPS + PRP group. mRNA expression levels were detected using real-time polymerase chain reaction (RT-PCR). Proliferation assays showed significantly enhanced proliferation in all PRP-treated groups compared with the no serum group. Compared with 10% fetal bovine serum (FBS), PRP concentrations above 3% in the normal condition and 1% to 7% PRP in the LPS-induced inflammatory condition were found to significantly promote chondrocyte proliferation. In the normal condition, all PRP-treated groups showed significantly increased cell migration compared with the no serum group. Chondrocyte migration was decreased with LPS-induced inflammation, but PRP treatment resulted in significantly enhanced migration compared with the other groups in this condition. According to RT-PCR, the LPS + PRP group showed significantly higher levels of COL1A1, IL-6, aggrecan and lower levels of $TNF-{\alpha}$, MMP-1, MMP-3 mRNA expression compared to the LPS group. The results of this study suggest that PRP application can enhance the proliferation and migration of canine chondrocytes and improve canine articular cartilage regeneration.

Dieckol Attenuates Microglia-mediated Neuronal Cell Death via ERK, Akt and NADPH Oxidase-mediated Pathways

  • Cui, Yanji;Park, Jee-Yun;Wu, Jinji;Lee, Ji Hyung;Yang, Yoon-Sil;Kang, Moon-Seok;Jung, Sung-Cherl;Park, Joo Min;Yoo, Eun-Sook;Kim, Seong-Ho;Ahn Jo, Sangmee;Suk, Kyoungho;Eun, Su-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • 제19권3호
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    • pp.219-228
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    • 2015
  • Excessive microglial activation and subsequent neuroinflammation lead to synaptic loss and dysfunction as well as neuronal cell death, which are involved in the pathogenesis and progression of several neurodegenerative diseases. Thus, the regulation of microglial activation has been evaluated as effective therapeutic strategies. Although dieckol (DEK), one of the phlorotannins isolated from marine brown alga Ecklonia cava, has been previously reported to inhibit microglial activation, the molecular mechanism is still unclear. Therefore, we investigated here molecular mechanism of DEK via extracellular signal-regulated kinase (ERK), Akt and nicotinamide adenine dinuclelotide phosphate (NADPH) oxidase-mediated pathways. In addition, the neuroprotective mechanism of DEK was investigated in microglia-mediated neurotoxicity models such as neuron-microglia co-culture and microglial conditioned media system. Our results demonstrated that treatment of anti-oxidant DEK potently suppressed phosphorylation of ERK in lipopolysaccharide (LPS, $1{\mu}g/ml$)-stimulated BV-2 microglia. In addition, DEK markedly attenuated Akt phosphorylation and increased expression of $gp91^{phox}$, which is the catalytic component of NADPH oxidase complex responsible for microglial reactive oxygen species (ROS) generation. Finally, DEK significantly attenuated neuronal cell death that is induced by treatment of microglial conditioned media containing neurotoxic secretary molecules. These neuroprotective effects of DEK were also confirmed in a neuron-microglia co-culture system using enhanced green fluorescent protein (EGFP)-transfected B35 neuroblastoma cell line. Taken together, these results suggest that DEK suppresses excessive microglial activation and microglia-mediated neuronal cell death via downregulation of ERK, Akt and NADPH oxidase-mediated pathways.

Highly efficient genome editing via CRISPR-Cas9 ribonucleoprotein (RNP) delivery in mesenchymal stem cells

  • A Reum Han;Ha Rim Shin;Jiyeon Kweon;Soo Been Lee;Sang Eun Lee;Eun-Young Kim;Jiyeon Kweon;Eun-Ju Chang;Yongsub Kim;Seong Who Kim
    • BMB Reports
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    • 제57권1호
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    • pp.60-65
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    • 2024
  • The CRISPR-Cas9 system has significantly advanced regenerative medicine research by enabling genome editing in stem cells. Due to their desirable properties, mesenchymal stem cells (MSCs) have recently emerged as highly promising therapeutic agents, which properties include differentiation ability and cytokine production. While CRISPR-Cas9 technology is applied to develop MSC-based therapeutics, MSCs exhibit inefficient genome editing, and susceptibility to plasmid DNA. In this study, we compared and optimized plasmid DNA and RNP approaches for efficient genome engineering in MSCs. The RNP-mediated approach enabled genome editing with high indel frequency and low cytotoxicity in MSCs. By utilizing Cas9 RNPs, we successfully generated B2M-knockout MSCs, which reduced T-cell differentiation, and improved MSC survival. Furthermore, this approach enhanced the immunomodulatory effect of IFN-r priming. These findings indicate that the RNP-mediated engineering of MSC genomes can achieve high efficiency, and engineered MSCs offer potential as a promising therapeutic strategy.

Optimization of Procedure for Efficient Gene Transfer into Porcine Somatic Cells with Lipofection

  • Kim, D.Y.;McElroy, S.L.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권5호
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    • pp.648-656
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    • 2008
  • The objective of this study was to establish conditions for transfection of a foreign gene into somatic cells using cationic lipid reagents and to evaluate the effects of transfection on in vitro development of somatic cell nuclear transfer (SCNT) embryos. Green fluorescent protein (GFP) gene was used as a foreign gene and a non-transfected somatic cell was utilized as a control karyoplast. Monolayers of porcine cells were established and subsequently transfected with a GFP-expressing gene (pEGFP-N1) using three types of transfection reagents (LipofectAMINE PLUS, FuGENE 6 or ExGen500). Donor cells used for SCNT included transfected fetal or adult fibroblasts and oviduct epithelial cells, either serum-fed or serum-starved. Oocytes matured in vitro for 42 h were reconstructed with either transfected or non-transfected porcine somatic cells by electric fusion and activation using a single DC pulse of 1.8 kV/cm for $30{\mu}s$ in $Ca^{2+}$ and $Mg^{2+}-containing$ 0.26 M mannitol solution. Reconstructed oocytes were subsequently cultured in NCSU-23 medium for 168 h and the developmental competence and cell number in blastocyst were compared. There were no significant differences (P>0.05) in fusion, cleavage rates or development to the blastocyst stage between non-transfected, transfected, serum-fed and serum-starved cells. However, the rates of GFP-expressing blastocysts were higher in the FuGENE 6 group (71.4%) among transfection reagents and in the fetal fibroblasts group (70.4%) for donor cells. These results indicate that fetal fibroblasts transfected with FuGENE 6 can be used as donor cells for porcine SCNT and that GFP gene can be safely used as a marker of foreign genes in porcine transgenesis.

Bone regenerative effect of recombinant human bone morphogenetic protein-2 after cyst enucleation

  • Hwang, Doo Yeon;On, Sung Woon;Song, Seung II
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제38권
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    • pp.22.1-22.6
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    • 2016
  • Background: The aim of this study is to quantitatively evaluate the effect of rhBMP-2 for repair of bone defects after cyst enucleation using the osteogenesis index (OI). Methods: Under general anesthesia, 10 patients (12 lesions) underwent oral or maxillofacial surgery for cyst enucleation. Postoperatively, 12 lesions were divided into two groups: group A (six lesions) was treated with absorbable collagen sponge (ACS) in combination with rhBMP-2, and group B (six lesions) was treated with ACS alone. After 3 months, cone-beam computed tomographic scans were obtained to measure changes in the volume of the lesions. We then calculated the OI of each group at two different Hounsfield units to determine any statistically significant difference between these two groups (Mann-Whitney U test). Results: As tested at the level of new bone, the mean OI was 72.37 % in group A and 55.08 % in group B -a statistically significant difference (p = 0.041). As tested at the level of mature bone, the mean OI was 27.47 % in group A and 18.88 % in group B, but the difference was not statistically significant (p = 0.394). Conclusions: The application of rhBMP-2 after maxillofacial cyst enucleation accelerated new bone formation in the bone defects. Thus, the use of rhBMP-2 in combination with ACS may be considered an alternative to conventional bone grafting in some patients with postoperative bone defects.