• Title/Summary/Keyword: enzyme-linked immunosorbent assay (ELISA)

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Effect of Intraoperative Glucose Fluctuation and Postoperative IL-6, TNF-α, CRP Levels on the Short-term Prognosis of Patients with Intracranial Supratentorial Neoplasms

  • Liu, Tie-Cheng;Liu, Qi-Ran;Huang, Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10879-10882
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    • 2015
  • Objective: To investigate the effect of intraoperative glucose fluctuation and postoperative interlukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), C-reactive protein (CRP) levels on the short-term prognosis of patients with intracranial supratentorial neoplasms. Materials and Methods: Eighty-six patients undergoing intracranial excision were selected in The Second Hospital of Jilin University. According to the condition of glucose fluctuation, the patients were divided into group A (glucose fluctuation <2.2 mmol/L, n=57) and group B (glucose fluctuation ${\geq}2.2mmol/L$, n=29). Glucose was assessed by drawing 2 mL blood from internal jugular vein in two groups in the following time points, namely fasting blood glucose 1 d before operation ($T_0$), 5 min after anesthesia induction ($T_1$), intraoperative peak glucose ($T_2$), intraoperative lowest glucose ($T_3$), 5 min after closing the skull ($T_4$), immediately after returning to intensive care unit (ICU) ($T_5$) and 2 h after returning to ICU ($T_6$). 1 d before operation and 1, 3 and 6 d after operation, serum IL-6 and TNF-${\alpha}$ levels were detected with enzyme-linked immunosorbent assay (ELISA), and CRP level with immunoturbidimetry. Additionally, postoperative adverse reactions were monitored. Results: There was no statistical significance between two groups regarding the operation time, anesthesia time, amount of intraoperative bleeding and blood transfusion (P>0.05). The glucose levels in both groups at $T_1{\sim}T_6$ went up conspicuously compared with that at $T_0$ (P<0.01), and those in group B at $T_2$, $T_4$, $T_5$ and $T_6$ were significantly higher than in group A (P<0.01). Serum IL-6, TNF-${\alpha}$ and CRP levels in both groups 1, 3 and 6 d after operation increased markedly compared with 1 d before operation (P<0.01), but the increased range in group A was notably lower than in group B (P<0.05 or P<0.01). Postoperative incidences of hypoglycemia, hyperglycemia and myocardial ischemia in group A were significantly lower than in group B (P<0.05), and respiratory support time obviously shorter than in group B (P<0.01). Conclusions: The glucose fluctuation of patients undergoing intracranial excision is related to postoperative IL-6, TNF-${\alpha}$ and CRP levels and those with small range of glucose fluctuation have better prognosis.

Effect of Jaeumganghwa-tang on Production and Secretion of Respiratory Mucus (자음강화탕(滋陰降火湯)이 호흡기 점액의 생성 및 분비에 미치는 영향)

  • Cheon, Jin Hong;Min, Sang Yoen;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.30 no.2
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    • pp.31-46
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    • 2016
  • Objectives In this study, the effects of Ja-eum-gang-hwa-tang (JGT) on the increase in airway epithelial mucosubstances of rats and ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells were investigated. Materials and Methods Hypersecretion of airway mucus was produced by exposure of $SO_2$ to rats for 3 weeks. The effect of orally-administered JGT for 2 weeks on increased epithelial mucosubstances from tracheal goblet cells of rats was assessed by using histopathological analysis after staining the epithelial tissue with Hematoxylin-eosin and PAS-alcian blue. Possible cytotoxicity of JGT was assessed by investigating the potential damage on kidneys and liver functions by measuring serum GOT/GPT activities and serum BUN concentration of rats and the body weight gain during experiment. Also, the effect of JGT on ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of JGT and treated with ATP ($200{\mu}M$) or PMA ($10ng/ml$) or EGF ($25ng/ml$) or TNF-${\alpha}$ (0.2 nM) for 24 hrs to assess the effect of JGT both on ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production by using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). Results (1) JGT decreased the amount of intraepithelial mucosubstances of trachea of rats. (2) JGT did not show any renal and hepatic toxicities, and did not affect body weights either. (3) JGT significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions from NCI-H292 cells. (4) JGT inhibited EGF-, and PMA-induced expression levels of MUC5AC gene in NCI-H292 cells. However, ATP- and TNF-${\alpha}$-induced MUC5AC gene expression levels were not affected in NCI-H292 cells. Conclusions The result from the present study suggests that JGT might control the production and gene expression of airway mucin observed in various respiratory diseases which accompanied by mucus hypersecretion. Also, JGT did not show liver toxicity or impact on kidney functions. The effect of JGT should be further studied by using animal experimental models which can show proper pathophysiology of airway diseases.

Effects of Four Oriental Medicines on Secretion and Gene Expression of Mucin in Airway Epithelial Cells (가미청폐탕, 가감정기탕, 가미사물탕, 가미이중탕이 호흡기 뮤신의 분비 및 뮤신 유전자의 발현에 미치는 영향)

  • Lee, Hyun Sook;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.31 no.2
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    • pp.1-13
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    • 2017
  • Objectives In this study, the author intended to investigate whether Gami-cheongpetang (GCP), Gagam-jeongkitang (GJG), Gami-samooltang (GSM) and Gami-ijoongtang (GIJ) significantly affect in vivo (animal model) and in vitro (cultured cells) mucin secretion and MUC5AC gene expression in airway epithelial cells. Methods For in vivo experiment, the author induced hypersecretion of airway mucin in rats by introducing SO2 for 3 weeks. Enzyme-linked immunosorbent assay (ELISA) was used to assess the effects of orally-administered GCP, GJG, GSM and GIJ in vivo mucin secretion from tracheal goblet cells of rats after 1 week. Also, the effects of the agents on TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Possible cytotoxicities of the agents were assessed by examining the rate of survival and proliferation of NCI-H292 cells. Results (1) GCP and GJG significantly inhibited hypersecretion of in vivo mucin, although GSM and GIJ did not affect hypersecretion of in vivo mucin; (2) GCP and GJG significantly increased in vitro mucin secretion from cultured HTSE cells. However, GSM and GIJ did not affect in vitro mucin secretion from HTSE cells; (3) GCP and GJG significantly inhibited the expression levels of EGF-induced MUC5AC gene in NCI-H292 cells. However, GSM and GIJ increased the expression levels of EGF-induced MUC 5AC gene in NCI-H292 cells; (4) GCP, GJG, GSM and GIJ did not significantly inhibit the survival and proliferation of NCI-H292 cells. Conclusions These results suggest that GCP, GJG, GSM and GIJ can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects of GCP, GJG, GSM and GIJ with their components should be further investigated by using animal experimental models that simulate the diverse pathophysiology of pulmonary diseases.

Determination of Appropriate Sampling Time for Job Stress Assessment: the Salivary Chromogranin A and Cortisol in Adult Females

  • Hong, Ran-Hi;Yang, Yun-Jung;Kim, Sang-Yon;Lee, Won-Young;Hong, Yeon-Pyo
    • Journal of Preventive Medicine and Public Health
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    • v.42 no.4
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    • pp.231-236
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    • 2009
  • Objectives : This study was conducted to determine the appropriate sampling time of the salivary stress markers, chromogranin A (CgA) and cortisol as objective indices of job stress assessment in adult females. Methods : The subjects were 20${\sim}$39-year-old women (13 office workers, 11 sales-service workers, and 11 college students) who were eligible for the study and free of acute and chronic medical conditions. Salivary CgA and cortisol levels were determined by enzyme-linked immunosorbent assay (ELISA). Saliva samples were collected (2 $m{\ell}$ each) at 7:00, 8:00, 10:30, 12:00, 17:30, and 22:30 on a typical day. Salivary CgA and cortisol levels, according to sampling time, were compared among the three groups using general linear model. The full version of the Korean Occupational Stress Scale (KOSS), which includes socioeconomic characteristics, health behavior, workrelated characteristics, and BMI, was used to access the subjects' job stress. Multiple regression analysis of the job stressors identified by the KOSS was performed on salivary CgA and cortisol levels. Results : The salivary CgA level peaked at 7:00 (time of awakening), then decreased and were maintained at a low level throughout the day, and increased slightly at 17:30. The salivary cortisol level increased steeply within the 1st hour after awakening, followed by a gradual decrease by 12:00, and was then maintained at a low level throughout the day. The salivary cortisol levels of subjects who worked ${\leq}$5 days per week and graduated from the university were significantly lower at 8:00 (p=0.006). The salivary cortisol levels of non-smokers were significantly lower at 7:00 p=0.040) and 8:00 (p=0.003) compared to smokers. There were no significant differences in salivary CgA and cortisol levels at 10:30 and 12:00 in general characteristics. The regression coefficients on salivary CgA level were significant with interpersonal conflict at 17:30 and job insecurity at 22:30. Regression coefficients on salivary cortisol level were significant with organizational system and total job stressors at 17:30. Conclusions : We suggest that the appropriate sampling times for the salivary stress markers, CgA and cortisol, are at 7:00 (time of awakening), 8:00 (1 hour after awakening), 17:30 (early evening), and 22:30 (before sleep).

Adverse Reproductive Effects on Plasma Vitellogenin and Sex Steroid Levels, and Gonadosomatic Index in Juvenile Common Carps (Cyprinus carpio) Exposed to 17$\beta$-Estradiol and D-2-Ethylhexyl Phthalate

  • Seo, Jinwon;Park, Kyung-Seo;Moon, Woon-Gi;Lee, Sung-Kyu
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2002.11a
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    • pp.141-144
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    • 2002
  • Environmental estrogens are natural or synthetic substances present in the aquatic environment, especially in effluent from sewage treatment. However, the adverse effects of these estrogenic substances on fish reproduction are unknown. Di-2-ethylhexyl phthalate (DEHP) is the most common phthalate, which Ps used as a plasticizer in polyvinylchloride (PVC), and it is widespread in the environment and has been found in aquatic organisms and sediments. Therefore, juvenile common carps (Cyprinus carpio) were exposed to nominal concentrations of 17$\beta$-estradiol (E2) (0.5, 5, 50 $\mu\textrm{g}$/L) and DEHP (10, 100, 500 $\mu\textrm{g}$/L) for 21 days, to determine the adverse reproductive effects of these compounds on plasma vitellogenin (VTG) induction, sex steroid level, and gonad weight. Electrophoresis (SDS-PAGE) revealed that much of VTG was induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Enzyme-linked immunosorbent assay (ELISA) revealed that VTG was significantly induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, and combination of 50 E$_2$ $\mu\textrm{g}$/L with 10 and 500 DEHP $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Analysis of sex steroid levels in some fish revealed that testosterone (T) was detected in both male and female fish of the control and DEHP exposures, but none of fish exposed to 22 concentrations had detectable testosterone level. On the other hand, E$_2$ exposure induced 17$\beta$-estradiol in plasma of male fish, but there was no induction of 17$\beta$-estradiol in plasma of male fish exposed to DEHP. Comparison of gonadosomatic index (GSI) revealed that maximal E$_2$ exposure inhibited ovarian growth, but maximal DEHP exposure stimulated testicular growth. The results indicated that those comparisons can be a useful bio-indicator for determining adverse reproductive effect of endocrine disrupting chemicals (EDCs).

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Evaluation of stress-induced analgesia in acupuncture analgesic effect - An Approach on diameters of acupuncture needles and acupuncture point needlings - (침진통 효과에서 스트레스에 의한 진통 효과 개입 여부 평가 - 침굵기, 침자극 부위를 중심으로 -)

  • Jeong, Byeong-Ju;Choe, Il-Hwan;Shin, Hee-Sup;Lim, Sabina
    • Korean Journal of Acupuncture
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    • v.25 no.3
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    • pp.65-80
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    • 2008
  • Objective : In recent years, many investigators have questioned whether the analgesic effect of acupuncture is simply related to the stress-induced analgesia (SIA). However, there has been lack of studies on this issue. In this study, the stress levels induced by manual acupunctures are compared with the stress in animal experiment models. The experiments have been carried out with Sprague Dawley (SD) rats. Method : For stress level evaluation, Hot plate test has been used. Maximum Possible Effect (MPE) has been measured by checking the pre-test time and post-test time. Cortisol and corticosterone concentrations in serum were measured by enzyme-linked immunosorbent assay (ELISA). Results : In the hot plate test, MPE values of post-test time were significantly decreased after 10 minutes than after 5 minutes. Therefore, optimal time interval was chosen as 10 minutes. There was significant difference of MPE values between Suspension group and all other treatment groups. However, there were no significant differences of MPE values between Sham group and all other treatment groups. However, MPE values showed tendency to decrease when acupuncture needle diameter increased. MPE values of ST040, ST040(lido), NAP040(lido) groups were markedly decreased than that of Suspension group, while that of NAP040 group was substantially increased than that of Sham group increased in acupoint and nonacupoint models. Serum cortisol concentrations of treatment groups were not significantly different from that of Suspension and Sham groups. Serum corticosterone concentration of 0.25 mm group was substantially increased than that of compared with Sham group. Serum cortisol and corticosterone concentrations of treatment groups were not significantly different from those of Suspension and Sham groups in acupoint and nonacupoint models. Conclusion : From hot plate test and serum stress hormones concentrations, it is found that manual acupuncture treatment induces negligible stress or SIA on ST36. And the stress induced by manual acupuncture is more closely related to acupuncture point needlings than diameters of acupuncture needles.

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Anti-inflammatory effect of ethanol extract from Ecklomia cava on gram-positive bacteria(Peptidoglycan)-induced macrophages (그람-양성균(Peptidoglycan)으로 유도한 대식세포에서 감태 에탄올 추출물의 항염증 효과)

  • Kang, Ok-Hwa;Kim, Sung-Bae;Keum, Joon-Ho;Mun, Su-Hyun;Kim, Yong-Sik;An, Byung-Kwan;An, Hyeon-Jin;Kwon, Dong-Yeul
    • Herbal Formula Science
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    • v.19 no.1
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    • pp.195-205
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    • 2011
  • Objectives : Ecklonia cava is brown alga(Laminariaceae) which grows is sea, it has antioxidant, diarrhea and anticoagulant effect. In this study, the effect of ethanol extract of Ecklonia cava (EC) on peptidoglycan(PGN) -induced NO production in RAW 264.7 cells was investigated. Methods : In the present study, IL-6 production was measured using the enzyme-linked immunosorbent assay(ELISA), prostaglandin $\E_2$($\PGE_2$) production was measured using the EIA kit, and inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) activation, as determined by western blot analysis and reverse transcription -polymerase chain reaction(RT-PCR). Results : EC inhibited PGN-induced NO and IL-6 production. Consistent with these observations, the protein expression of iNOS and COX-2 were inhibited by EC. Moreover, EC suppressed the phosphorylation of extracellular signal-regulated kinase(ERK) 1/2 in PGN-induced RAW 264.7. Conclusions : These results suggest that EC has inhibitory effects on PGN-induced $\PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

Controlled release of nerve growth factor from heparin-conjugated fibrin gel within the nerve growth factor-delivering implant

  • Lee, Jin-Yong;Kim, Soung-Min;Kim, Myung-Jin;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.40 no.1
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    • pp.3-10
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    • 2014
  • Objectives: Although nerve growth factor (NGF) could promote the functional regeneration of an injured peripheral nerve, it is very difficult for NGF to sustain the therapeutic dose in the defect due to its short half-life. In this study, we loaded the NGF-bound heparin-conjugated fibrin (HCF) gel in the NGF-delivering implants and analyzed the time-dependent release of NGF and its bioactivity to evaluate the clinical effectiveness. Materials and Methods: NGF solution was made of 1.0 mg of NGF and 1.0 mL of phosphate buffered saline (PBS). Experimental group A consisted of three implants, in which $0.25{\mu}L$ of NGF solution, $0.75{\mu}L$ of HCF, $1.0{\mu}L$ of fibrinogen and $2.0{\mu}L$ of thrombin was injected via apex hole with micropipette and gelated, were put into the centrifuge tube. Three implants of experimental group B were prepared with the mixture of $0.5{\mu}L$ of NGF solution, $0.5{\mu}L$ HCF, $1.0{\mu}L$ of fibrinogen and $2.0{\mu}L$ of thrombin. These six centrifuge tubes were filled with 1.0 mL of PBS and stirred in the water-filled beaker at 50 rpm. At 1, 3, 5, 7, 10, and 14 days, 1.0 mL of solution in each tubes was collected and preserved at $-20^{\circ}C$ with adding same amount of fresh PBS. Enzyme-linked immunosorbent assay (ELISA) was done to determine in vitro release profile of NGF and its bioactivity was evaluated with neural differentiation of pheochromocytoma (PC12) cells. Results: The average concentration of released NGF in the group A and B increased for the first 5 days and then gradually decreased. Almost all of NGF was released during 10 days. Released NGF from two groups could promote neural differentiation and neurite outgrowth of PC12 cells and these bioactivity was maintained over 14 days. Conclusion: Controlled release system using NGF-HCF gel via NGF-delivering implant could be an another vehicle of delivering NGF to promote the nerve regeneration of dental implant related nerve damage.

The Effect of Poria Cocos Pharmacopuncture at SP6(Sameumgyo) and ST36(Joksamni) on Chronic Stress Model(CSM) Rats (삼음교(三陰交)(SP6)·족삼리(足三里)(ST36) 백복령약침(白茯笭藥鍼)이 만성 스트레스 모델 흰쥐에 미치는 영향)

  • Kim, Seon Wook;Kim, Jae Hong;Cho, Myoung Rae;Shin, Jeong Cheol
    • Journal of Acupuncture Research
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    • v.32 no.2
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    • pp.187-195
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    • 2015
  • Objectives : The purpose of this study is evaluating the effect of Poria cocos pharmacopuncture (PCP) at $SP_6$(Sameumgyo) and $ST_{36}$(Joksamni) on chronic stress model(CSM) rats. Methods : The experimental rats were assigned to five groups: normal, CSM(control), PCP1X, PCP2.5X and PCP5X. The experimental rats in CSM(control), PCP1X, PCP2.5X and PCP5X were put through a timetable to cause CSM. PCP($20{\mu}l$) was administered at $SP_6$, $ST_{36}$ to groups PCP1X, PCP2.5X and PCP5X once every three days for a total of six times. The weight of the rats was measured by an electronic scale after PCP injection, once every three days for a total of six times. Blood samples were taken from the rats for analysis of aspartate aminotransferase(AST) and alanine aminotransferase(ALT). Serotonin and norepinephrine were measured by enzyme-linked immunosorbent assay(ELISA) kit. Results : The experimental groups(PCP1X, 2.5X, 5X) showed significant body weight increase compared to the control group. Regarding AST and ALT changes in serum, the experimental groups(PCP2.5X, 5X) showed a significant decrease compared to the control group. For Serotonin changes in serum, the experimental group(PCP5X) showed a significant increase compared to the control group. For Norepinephrine changes in serum, the experimental groups(PCP2.5X) showed a significant decrease compared to the control group. Conclusions : These data suggest that PCP at $SP_6$, $ST_{36}$ has therapeutic effects on CSM rats.

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.