• Journal of Korean Neurosurgical Society
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• v.63 no.2
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• pp.163-170
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• 2020

Objective : Milk fat globule-epidermal growth factor VIII (MFG-E8) may play a key role in inflammatory responses and has the potential to function as a neuroprotective agent for ameliorating brain injury in cerebral infarction. This study aimed to determine the role of MFG-E8 in brain injury in the subacute phase of cerebral ischemia in a rat model. Methods : Focal cerebral ischemia was induced in rats by occluding the middle cerebral artery with the modified intraluminal filament technique. Twenty-four hours after ischemia induction, rats were randomly assigned to two groups and treated with either recombinant human MFG-E8 or saline. Functional outcomes were assessed using the modified Neurological Severity Score (mNSS), and infarct volumes were evaluated using histology. Anti-inflammation, angiogenesis, and neurogenesis were assessed using immunohistochemistry with antibodies against ionized calcium-binding adapter molecule 1 (Iba-1), rat endothelial cell antigen-1 (RECA-1), and bromodeoxyuridine (BrdU)/doublecortin (DCX), respectively. Results : Our results showed that intravenous MFG-E8 treatment did not reduce the infarct volume; however, the mNSS test revealed that neurobehavioral deficits were significantly improved in the MFG-E8-treated group than in the vehicle group. Immunofluorescence staining revealed a significantly lower number of Iba-1-positive cells and higher number of RECA-1 in the periinfarcted brain region, and significantly higher numbers of BrdU- and DCX-positive cells in the subventricular zone in the MFG-E8-treated group than in the vehicle group. Conclusion : Our findings suggest that MFG-E8 improves neurological function by suppressing inflammation and enhancing angiogenesis and neuronal proliferation in the subacute phase of cerebral infarction.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.4
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• pp.308-315
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• 2004

Aim: The aim of this study was to investigate the mechanism of promoted skin wound healing in skin defects with primary cultured oral mucosal keratinocytes. Materials and methods: Thirty adult female nude mice weighing $20{\pm}2g$ were used for the experiment. Primary cultured and suspended oral mucosal keratinocytes, labeled with BrdU, were scattered onto $1.5cm{\times}1.5cm$ sized full thickness skin defects in the experimental group(N=15), and no grafts were placed the control group(N=15). They were sacrificed at 3 days, 1 week and 2 weeks after the treatment respectively. Histological examination of each wounds were performed to review the healing progress on measuring the length from the wound margin to regenerating epithelial front. The role of keratinocytes were assessed by double immunohistochemical staining with Anti-BrdU and Anti-cytokeratin AE1/3. Results: In the experimental group the wound was completely covered with regenerating epithelia in 2 weeks, but partially regenerated in the control group. The immunohistochemical studies unexpectedly reveal that most of regenerating epithelial cells were induced from marginal epithelium of the margin, not from the scattered keratinocytes. Conclusion: We could successfully confirm that graft of primary cultured oral mucosal keratinocytes promotes the regeneration of skin defects.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.438-457
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• 1995

The origin of fibroblasts, their proliferative activity and roles in the early stages of periodontal regeneration were investigated in order to better understand the periodontal healing process in furcation defects of the beagle dog after guided tissue regeneration. Newly divided cells were identified and quantitated by immunolocalization of bromodeoxyuridine (BrdU) injected 1 hour prior to sacrificing the animals. The results were as follows :1. During periodontal healing in horizontal furcation defect, three different stages, namely the granulation tissue, connective tissue, and bone formation stages, were identified on the basis of major types of cells and tissue. 2. In the early stages of periodontal regeneration, both the remaining periodontal ligament and alveolar bone compartment were the major sources. 3. The majority of BrdU-labeled fibroblasts were located at the following areas ; 1) the coronal zone of the defect in case of the connective tissue fanned on the root surface. 2) the area within an 400 ${\mu}m$ distance from the remaining bone level in case of the periodontal ligament. 3) the area within an 100 ${\mu}m$ distance from the bone surface in case of areas of active bone formation.4. The highly proliferative fibroblasts adjacent to bone surface played a major role in the formation of osteoblast precursor cells, whereas both paravascular and endosteal cells played a minor role in new bone formation, In conclusion, it was suggested that the fibroblasts in the remaining periodontal ligament and bone will play a major role in periodontal regeneration, whereas both paravascular and endosteal cells will play a minor role in new bone formation.

• The Journal of Internal Korean Medicine
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• v.34 no.4
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• pp.412-427
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• 2013

Objectives : This study was carried out to compare the effects of Eejin-tang, Hyangsaeejin-tang and Naeso-san extracts on indomethacin-induced gastric mucosal lesions in mice. Methods : Experimental mice were divided into six groups. The normal group had no gastro-inflammation. In the control group, gastro-inflammation was elicited by indomethacin. Misoprostol, Eejin-tang, Hyangsaeejin-tang and Naeso-san group were those in which misoprostol, Eejin-tang extract, Hyangsaeejin-tang extract and Naeso-san extract were administered after gastro-inflammation is elicited. This study examined the anti-inflammation effects and distribution of mucus secreting cells, zonula occludin-1 (ZO-1), heat shock protein (HSP) 70, periodic acid-schiff reaction stain (PAS), peanut agglutinin (PNA), cyclooxygenase-1 (COX-1), 5-bromo-2'-deoxyuridine (BrdU), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$) p65, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Results : 1. The hemorrhagic erosion and damaged mucus secreting cell, the positive reaction HSP70 increased in the control group, but decreased in the Eejin-tang, Hyangsaeejin-tang and Naeso-san groups. 2. The positive reaction of ZO-1, PAS, PNA, COX-1 and BrdU decreased in the control group, but increased in the Eejin-tang, Hyangsaeejin-tang and Naeso-san groups. 3. The positive reaction of NF-${\kappa}B$ p65, iNOS and COX-2 increased in the control group, but decreased in the Eejin-tang, Hyangsaeejin-tang and Naeso-san groups. Conclusions : Among the three extracts, the effects were in the order of Naeso-san, Hyangsaeejin-tang and Eejin-tang group, Naeso-san being the most effective.

• The Journal of Korean Medicine
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• v.28 no.2 s.70
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• pp.166-184
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• 2007

Objectives : This study was conducted to investigate the effects of Daegeum-eumja, Igwi-tang, and Sihosogan-san on gastric mucosal lesions induced by alcohol, indomethacin, and burn-stress in mice. Methods : Experimental mice were divided into six groups. The normal group (NOR) did not receive any treatment to elicit gastropathy. In the control group (GE), gastropathy was elicited by alcohol, indomethacin, and stress. In the misoprostol group (MS), misoprostol was administered after gastropathy was elicited by alcohol, indomethacin, and stress. In the Daegeum-eumja group (DG), Daegeum-eumja was administered after gastropathy was elicited by alcohol, indomethacin, and stress. In the Igwi-tang group (IW), Igwi-tang was administered after gastropathy was elicited by alcohol, indomethacin, and stress. In the Sihosogan-san group (SH), Sihosogan-san was administered after gastropathy waselicited by alcohol, indomethacin, and stress. The effects on gastric mucosal lesions were evaluated by the morphological change of gastric mucosa, the anti-oxidant effect, HSP 70, $NF-{\kappa}B$ p65, $I{\kappa}B$, COX-1, PNA, BrdU, and iNOS. Results : Hemorrhage erosion, HSP70, and $NF-{\kappa}B$ in the DG, IW and SH groups decreased more than that of the control. The $I{\kappa}B$, COX-1, PNA, BrdU, and iNOS in the DG, IW, and SH groups increased more than that of the control. DG showed the most effect against gastric mucosal lesions induced by alcohol; IW against gastric mucosal lesions induced by indomethacin; and SH against gastric mucosal lesions induced by burn-stress. Conclusions: Daegeum-eumja, Igwi-tang, and Sihosogan-san extracts have excellent effects on gastric mucosal lesions induced by alcohol, indomethacin, and burn-stress, respectively.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.1
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• pp.16-21
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• 2014

Micro needle roller therapy has been used for cosmetic purposes, such as reducing skin winkles and improving elasticity of skin. It is claimed that micro needle roller therapy has potentials for connective tissue regeneration by facilitating collagen synthesis. Therefore, there seems to be a possibility that connective tissue regenerating potential of micro needle roller therapy could influence the hair growth cycle. This study, we investigated the hair growth-promoting effects of micro needle roller therapy. C57BL/6 mice were devided into three groups as follows: normal saline-treated, minoxidil-treated, and micro needle roller therapy-received group. Hair growth activity was evaluated by handscopic and microscopic observations. Sections of dorsal skin were stained with hematoxylin and eosin. Expression of BrdU, FGF, and VEGF was detected by immunohistochemical staining. Micro needle roller therapy enhanced the development of hair follicle during anagen. Immunohistochemical analysis revealed that micro neeld roller therapy incresed the expression of BrdU and FGF in the hair follicles of C57BL/6 mice. Furthermore, micro needle roller therapy upregulated mRNA expression of VEGFR-2, FGF-2, EGF - growth factors that play a central role in hair follicle development during anagen. These results suggest that Micro needle roller therapy can potentially be used for the treatment of alopecia.

• The Journal of Korean Medicine
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• v.31 no.2
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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 Journal of Korean Academy of Prosthodontics
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• v.45 no.6
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• pp.787-794
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• 2007

Statement of problem. Surface microgrooves on Ti substrata have been shown to alter the expression of genes responsible for various biological activities of cultured fibroblasts. However, their effect on enhancing cell proliferation is not yet clear. Purpose. The purpose of this study was to determine the dimension of surface microgrooves on Ti substrata that enhances proliferation and alters gene expression of cultured human gingival fibroblasts. Material and methods. Commercially pure Ti discs with surface microgrooves of monotonous $3.5{\mu}m$ in depth and respective 15 and $30{\mu}m$ in width were fabricated using photolithography and used as the culture substrata in the two experimental groups in this study (TiD15 and TiD30), whereas the smooth Ti was used as the control substrata (smooth Ti group). Human gingival fibroblasts were cultured on the three groups of titanium substrata and the proliferation, DNA synthesis, and gene expression of theses cells were analyzed and compared between all groups using XTT assay, BrdU assay, and reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. Results. From the XTT assay at 48 h incubation, the proliferation of human gingival fibroblasts in TiD30 was significantly enhanced compared to that in smooth Ti and TiD15. The results from the BrdU assay showed that, at 24 h incubation, the DNA synthesis was significantly enhanced in TiD30 compared to that in smooth Ti. In RT-PCR, increase in the expression of PCR transcripts of fibronectin, CDK6, $p21^{cip1}$ genes was noted at 48h incubation. Conclusion. Surface microgrooves $30{\mu}m$ in width and $3.5{\mu}m$ in depth on Ti substrata enhance proliferation and alter gene expression of cultured human gingival fibroblasts.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.299-304
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• 2008

Monocytic cells in myeloid lineage are known for latent site of HCMV Previous studies have suggested that HCMV regulates cell cycle progression in a variety of cells, but studies in monocytic cells are limited. In this study, we attempted to understand cell cycle changes after HCMV infection in the monocytic cell lines. Flow cytometric analyses using propidium iodide revealed that the proportion of G0-G1 phase was increased and the proportion of S phase decreased in HCMV-infected THP-1 cells, but not in HL-60 cells. BrdU-incorporation assay supported that cell proliferation was inhibited in HCMV-infected THP-1 cells by inhibition of de novo DNA synthesis. Western blot analysis revealed that p21, inhibitor of cell cycle progression from G1 phase to S phase, was induced in HCMV-infected THP-1 cells but not in HL-60 cells. Thus, HCMV inhibited cell pro-liferation by arresting the cell cycle at G0-G1 phase through induction of p21 protein in promocytic THP-1 cells.

• Biomolecules & Therapeutics
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• v.23 no.4
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• pp.313-319
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• 2015

P7C3 and its derivatives, 1-(3,6-dibromo-9H-carbazol-9-yl)-3-(p-tolylamino)propan-2-ol (1) and N-(3-(3,6-dibromo-9H-carbazol-9-yl)-2-hydroxypropyl)-N-(3-methoxyphenyl)-4-methylbenzenesulfonamide (2), were previously reported to increase neurogenesis in rat neural stem cells (NSCs). Although P7C3 is known to increase neurogenesis by protecting newborn neurons, it is not known whether its derivatives also have protective effects to increase neurogenesis. In the current study, we examined how 1 induces neurogenesis. The treatment of 1 in NSCs increased numbers of cells in the absence of epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2), while not affecting those in the presence of growth factors. Compound 1 did not induce astrocytogenesis during NSC differentiation. 5-Bromo-2'-deoxyuridine (BrdU) pulsing experiments showed that 1 significantly enhanced BrdU-positive neurons. Taken together, our data suggest that 1 promotes neurogenesis by the induction of final cell division during NSC differentiation.