• Title/Summary/Keyword: $TGF{\beta}$

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Biological Effect of Platelet Rich Plasma on the Initial Attachment, Proliferation and Cellular Activity of Osteoblast (혈소판 농축혈장이 조골세포의 초기부착과 증식 및 활성에 미치는 생물학적 영향)

  • Park, Sang-Il;Lim, Sung-Bin;Kim, Jung-Keun;Chung, Chin-Hyung
    • Journal of Periodontal and Implant Science
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    • v.31 no.3
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    • pp.513-529
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    • 2001
  • For reconstruction of the bony defect, various artificial substitutes were developed. Among them, there has been a study of calcium phosphate coated bone substitutes for increasing attachment of osteoblasts in vivo. The purpose of this study was to evaluate the effects of serum and platelet-rich plasma (PRP) on calcium phosphate coated culture plate for the initial attachment, proliferation and activity of osteoblasts. After sampling the blood from white rats and concentrating by centrifugation, the amount of attachment of PDGF-BB and $TGF-{\beta}$ on the calcium phosphate coated culture plate was measured. Cultured HOS and ROS 17/2.8 cell was measured on attachment level and proliferation rate of osteoblasts. Alkaline phosphatase activity of HOS and ROS 17/2.8 cell was measured for studying on the activating rate of osteoblast. 1. Counting the amount of platelets of seperated plasma and PRP, the average number of platelets was 177,003 $cell/{\mu}l$ in plasma, and 1,656,062 $cell/{\mu}l$ in PRP, which was about 9 times as high as in plasma. 2. Amount of PDGF-BB deposited at calcium phosphate coated plate had increased by the total amount of plasma and PRP on the culture plate, whereas $TGF-{\beta}$had been deposited on the plate only when treated by $50{\mu}{\ell}$ of PRP(p<0.01). 3. After plating serum and PRP for 3 hours, we attached with HOS and ROS17/2.8 cell for 1 hour and 4 hours. There were no significant difference of the attachment between serum and control group, whereas there were significantly difference of the attachment between depositioning of PRP and control group. 4. After attaching plasma and PRP for 3 hours, cell number has much increased when HOS and ROS17/2.8 cell had been cultured for 48 hours(p<0.05). 5. After attaching plasma and PRP for 3 hours, concentration of alkaline-phosphatase has increased when HOS and ROS17/2.8 cell had been cultured for 48 hours(p<0.01). These results suggested that PRP affected on initial cell attachment rather than proliferation and activation of osteoblasts at calcium phosphate coated plate.

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Angiogenesis in newly regenerated bone by secretomes of human mesenchymal stem cells

  • Katagiri, Wataru;Kawai, Takamasa;Osugi, Masashi;Sugimura-Wakayama, Yukiko;Sakaguchi, Kohei;Kojima, Taku;Kobayashi, Tadaharu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.39
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    • pp.8.1-8.8
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    • 2017
  • Background: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor $(TGF)-{\beta}1$, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. Methods: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. Results: The concentrations of IGF-1, VEGF, and $TGF-{\beta}1$ in MSC-CM were $1515.6{\pm}211.8pg/mL$, $465.8{\pm}108.8pg/mL$, and $339.8{\pm}14.4pg/mL$, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. Conclusions: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

The Effects of various Regeneration techniques on Bone Regeneration around Dental Implant (수종의 재생 술식 시행이 매식체 근원심부의 골재생에 미치는 영향)

  • Lee, Myung-Ja;Lim, Sung-Bin;Chung, Chin-Hyung;Hong, Ki-Seok
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.383-399
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    • 2005
  • The successful implantation necessitate tissue regeneration m site of future implant placement, there being severe bone defect. Therapeutic approaches to tissue regeneration in the site have used bone grafts, root surface treatments, barrier membranes, and growth factors, the same way being applied to periodontal tissue regeneration. Great interest in periodontal tissue regeneration has lead to research in bone graft, guided-tissue regeneration, and the administration of growth factors as possible means of regenerating lost periodontal tissue. The blood component separated by centrifuging the blood is the platelet-rich plasma. There are growth factors, PDGF, $TGF{beta}1$, $TGF{beta}2$ and IGF in the platelet-rich plasma. The purpose of this study was to study the histopathological correlation between the use of platelet-rich plasma and the healing of bone defect around implant fixture site. Implant fixtures were inserted and graft materials were placed into the left femur of in the experimental group, while the control group received only implant fixtures. In the first experimental group, platelet-rich plasma and BBP xenograft were placed at the implant fixture site, and the second experimental group had platelet-rich plasma, BBP xenograft, and the e-PTFE membrane placed at the fixture site. The degree of bone regeneration adjacent to the implant fixture was observed and compared histopathologically at 2, 4, and 8 weeks after implant fixture insertion. The results of the experiment were as follows: 1. Bone remodeling in acid etched surface near the implant fixture of all experimental groups was found to be greater than new bone formation. 2. Bone remodeling in acid etched surface distant to the implant fixture of all experimental groups was decreased and new bone formation was not changed. 3. Significant new bone formation in machined surface near the implant fixture of bothl experimental groups was observed in 2 weeks. 4. New bone formation in machined surface distant to the implant fixture of both experimental groups was observed. Bone remodeling was significant in near the implant fixture and not in distant to the implant fixture. The results of the experiment suggested that the change of bone formation around implant. Remodeling in machined surface distant to the implant fixture of both experimental groups, and new bone formation and remodeling near the implant fixture were significant.

THE COMPARISON OF GENE EXPRESSION FROM HUMAN DENTAL PULP CELLS AND PERIODONTAL LIGAMENT CELLS (사람 치수 세포와 치주 인대 세포의 유전자 발현에 관한 비교 연구)

  • Hyoun, So;Park, Sang-Hyuk;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.34 no.5
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    • pp.430-441
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    • 2009
  • The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA micro array assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well. According to this study, the results were as follows: 1. From the micro array assay, 51 genes were more expressed (2 fold) from PC than PDLC. 2. RT-PCR confirmed that ITGA4 and TGF ${\beta}2$ were more expressed in PC than in PDLC 3. From the micro array assay, 19 genes were more expressed (2 fold) from PDLC than PC. 4. RT-PCR confirmed that LUM, WISP1. and MMP1 were more expressed in PDLC than in PC. From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.

Wound Healing Effect of Low Molecular PDRN on Experimental Surgical Excision Rat Model (저분자화된 Polydeoxynucleotide (PDRN)의 흰쥐에 대한 외과적 창상 치유 효과)

  • Yun, Jong-Kuk;Yoon, Hye-Eun;Park, Jeong-Kyu;Kim, Mi Ryeo;Kim, Dae-Ik
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.401-411
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    • 2015
  • This study was performed to investigate the wound healing effect of skin regeneration cosmetics utilizing low molecular weight Polydeoxynucleotide (PDRN). High purity PDRN was prepared from salmon testes poly-deoxy-ribonucleotide through protein and toxin removal process and molecular weight reduction. In order to evaluate the wound healing effect of PDRN in SD rats, 4 sites of dorsal skin of each animal were excised by using biopsy punch and $500{\mu}L$ of test solution was topically applied once daily for 4 weeks. The tissue changes were observed for every week during the application periods. After applying the PDRN to the wound, the skin was cut flower and contraction of the wounds more quickly, and the coating of PDRN in the wound area was reduced significantly as compared to the positive control group $Fucidin^{(R)}$ applied. The microscopic observation of stained tissue showed that a positive control was most rapid in re-epithelialization ability followed by the PH group, PDRN group, HA group. In addition, transforming growth factor ($TGF-{\beta}$) and vascular endothelial growth factor (VEGF), such as in the growth factor was similar to the results of staining of tissue lesions. In conclusion, it is determined that the low molecular weight PDRN has the therapeutic effect to the wound, and could be used as a functional material of cosmetics and medical industries.

Effect of MSE on Activity and Molecular Biology in Rat Induced Liver Injury (간 손상으로 유도된 랫드에서 MSE가 활성 변화와 분자생물학에 미치는 영향)

  • Yea, Chun-Jung;Lee, Tae-Jong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.2
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    • pp.520-526
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    • 2019
  • The purpose of this experiment is to investigate the feasibility of extracts (sample extract, MSE) mixed with hot - water extracts as health functional nutritional foods for prevention and treatment of liver damage. The experimental method was to investigate the effects of carbon tetrachloride on liver damage in rats by 60% of liver cirrhosis level. The effect of mixed extracts (MSE) was investigated by measuring the physiological functions, biochemical functions, and molecular biologic TGF - ${\beta}$ and P53 levels in liver - injured rats. with MSE doses administered differently(0.50ml, 0.75ml). A total of 30 rats were used for normal group, control group, positive control group, experimental group 1, and experimental group 2. The rats were given 6 times a week for 6 weeks at a fixed time once a day. The results of the experiment were statistically significantly higher in the control group than in the normal group and the positive control group, and statistically significantly lower in the experimental group (1 and 2) than the control group. There were some significant differences in the comparison between the experimental groups (1 and 2), but no significant changes were found. In conclusion, the results of this study are expected to be useful as functional food, and the variation of physiological activity and the degree of liver molecular biotechnology will be used for the research of the same field.

Effects of PGA-LM on CD4+CD25+foxp3+ Treg Cell Activation in Isolated CD4+ T Cells in NC/Nga Mice (NC/Nga 생쥐에서 분리한 T 세포에서 foxp3+ 세포 활성화에 대한 PGA-LM의 효과)

  • Jang, Soon-Nam;Kim, Kum-Lan;Kang, Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.160-169
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    • 2009
  • Poly-$\gamma$-glutamic acid ($\gamma$-PGA) was mixed natural flora of Bacillus subtilis, contaminated from cooked soybeans. Also, it was performed to find out the antiallergic activity by using NC/Nga mice, in vitro. The $\gamma$-PGA (PGA-HM : PGA-high molecular weight), Molecular weight 300 kDa, was decomposed and made PGA-LM (PGA-low molecular weight) which has molecular weight below 30 kDa by sonication. Therefore, it was same result between PGA-HM and PGA-LM, and reported PGA-LM as basic result. We found that PGA-LM contains antiallergic efficacy that inhibit B cells and Th2 cells activation from isolated CD4+T cells in NC/Nga atopic dermatitis model mice, and not show a cytotoxicity in the hFCs. To investigate the effects of these PGA-LM in vitro, isolation of splenic B cell and CD4+ T cells in atopic dermatitis mice were used. To elucidate the role of PGA-LM in anti-CD40+ interleukin-4 (IL-4)-mediated B-cell activation, showed that the capacity of B cells to expression IL-$1\beta$, IL-6, and TNF-$\alpha$ mRNA down-regulated, and IL-10 mRNA up-regulation by PGA-LM treatment, but it had no effect on TGF-$\beta$ expression. In addition to CD4+IFN-$\gamma$+ and CD4+CD25+foxp3+, the functions of PGA-LM in the development of the CD4+CD25+foxp3+ and CD4+IFN-$\gamma$+cells, the phenotype and functions of PGA-LM induced CD4+CD25+foxp3+, and CD4+IFN-$\gamma$+cells in CD4+T cells. These results suggested that PGA-LM could change cytokine production and generate CD4+CD25+foxp3+ Tregs in NC/Nga mice, and may be effective for immunotherapy in patients with AD.

Study on Effect of Saengbal-eum-II(Shēngfà-yĭn-II)on Hair Regrowth Promotion in C57BL/6 Mice (생발음(生髮飮)-II 피부도포가 C57BL/6 마우스의 육모촉진에 미치는 효과)

  • Han, Ae-Ri;Sohn, Nak-Won;Chung, Seok-Hee;Kim, Sung-Su;Song, Mi-Yeon
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.4
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    • pp.95-113
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    • 2009
  • Objectives : Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) is a hair care product which is composed of ten plant extracts used in oriental medicine. This study was carried out to investigate the effects of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) on hair regrowth and cytokine changes in a shaving model of C57BL/6 mice. Materials and Methods : Five-week-old mice were acclimated for 1 week at a temperature between $21-23^{\circ}C$, 40-60% relative humidity, and 12h of a light/dark cycle before beginning of the experiment. There were three experimental groups including 50% ethanol (EtOH, control), a positive control of 3% Minoxidil, and 30% Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) in 50% ethanol in 18 female mice. The test compounds were topically treated once a day over 12 days. The hair regrowth was photographically and histologically determined during the experimental period of 12 days. Revelation of EGF, $TGF-{\beta}1$ and IL-6 in hair follicle were also determined using immunohistochemistry. In addition to that, IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue were determined using enzyme-linked immunosorbent assay(ELISA). Results : Hair regrowth in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups was promoted earlier and faster than the control group. Concentrations of hairs and thick-hair ratio in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups were promoted than the control group. EGF was moderately positive in hair follicle of 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups, but negative in the control group. $TGF-{\beta}1$ was not significantly difference between the groups. IL-6 in hair follicle of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group was negative, but weakly positive in 3% Minoxidil and control group. IL-6 and $IL-1{\beta}$ in skin tissue were significantly decreased in Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group, but there was not significantly decreased in 3% Minoxidil and control group. $TNF-{\alpha}$ in skin tissue was significantly decreased in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups. Conclusions : These results suggest that Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) has hair growth promoting activity and it can be used for treatment of alopecia. And these effects relate to EGF revelation of hair follicle and a decrease IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue.

Role of Cordycepin and Adenosine on the Phenotypic Switch of Macrophages via Induced Anti-inflammatory Cytokines

  • Shin, Seul-Mee;Moon, Sun-Hee;Park, Yoon-Hee;Kwon, Jeong-Hak;Lee, Seung-Jeong;Lee, Chong-Kil;Cho, Kyung-Hae;Kim, Kyung-Jae
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.255-264
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    • 2009
  • Background: Chronic low grade inflammation is closely linked to type II diabetes, obesity, and atherosclerosis. Macrophages play a key role in the regulation of pro- or anti-inflammatory actions at the lesion sites of disease. Components of cordyceps militaris, cordycepin and adenosine, have been used for the modulation of inflammatory diseases. The effects of cordycepin in the modulation of macrophages have yet to be be elucidated. We investigated the effects of cordycepin and adenosine on the morphological changes of macrophages under the inflammatory condition of LPS and an anti-inflammatory condition involving high concentrations of adenosine. Methods: We confirmed the mRNA levels of the M1/M2 cytokine genes through RT-PCR and morphological change. Results: LPS-activated macrophages returned to their inactivated original shape, i.e., they looked like naive macrophages, through the treatment with high concentrations of cordycepin ($40{\mu}g/ml$). LPS and adenosine activated macrophages also returned to their original inactivated shapes after cordycepin treatment; however, at relatively higher levels of cordycepin than adenosine. This change did not occur with relatively low concentrations of cordycepin. Adenosine down-regulated the gene expression of M1 cytokines (IL-$1{\beta}$, TNF-${\alpha}$) and chemokines (CX3CR1, RANTES), such as cordycepin. Additionally, M2 cytokines (IL-10, IL-1ra, TGF-${\beta}$) were up-regulated by both cordycepin and adenosine. Conclusion: Based on these observations, both cordycepin and adenosine regulated the phenotypic switch on macrophages and suggested that cordycepin and adenosine may potentially be used as immunomodulatory agents in the treatment of inflammatory disease.

Hydrogen Peroxide Promotes Epithelial to Mesenchymal Transition and Stemness in Human Malignant Mesothelioma Cells

  • Kim, Myung-Chul;Cui, Feng-Ji;Kim, Yongbaek
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3625-3630
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    • 2013
  • Reactive oxygen species (ROS) are known to promote mesothelial carcinogenesis that is closely associated with asbestos fibers and inflammation. Epithelial to mesenchymal cell transition (EMT) is an important process involved in the progression of tumors, providing cancer cells with aggressiveness. The present study was performed to determine if EMT is induced by $H_2O_2$ in human malignant mesothelioma (HMM) cells. Cultured HMM cells were treated with $H_2O_2$, followed by measuring expression levels of EMT-related genes and proteins. Immunohistochemically, TWIST1 expression was confined to sarcomatous cells in HMM tissues, but not in epithelioid cells. Treatment of HMM cells with $H_2O_2$ promoted EMT, as indicated by increased expression levels of vimentin, SLUG and TWIST1, and decreased E-cadherin expression. Expression of stemness genes such as OCT4, SOX2 and NANOG was also significantly increased by treatment of HMM cells with $H_2O_2$. Alteration of these genes was mediated via activation of hypoxia inducible factor 1 alpha (HIF-$1{\alpha}$) and transforming growth factor beta 1 (TGF-${\beta}1$). Considering that treatment with $H_2O_2$ results in excess ROS, the present study suggests that oxidative stress may play a critical role in HMM carcinogenesis by promoting EMT processes and enhancing the expression of stemness genes.