Mineral trioxide aggregate (MTA) would influence healing of periapical tissues by modulating the production of growth factors and cytokines from PDL fibroblasts, however, the studies are insufficient. Therefore, the purpose of this study was to monitor the expression of transforming growth factor-beta1 (TGF- beta1), fibroblast growth factor-2 (FGF-2), and interleukin-6 (IL-6) from PDL fibroblasts in the presence of MTA. The human PDL fibroblasts were seeded onto the set MTA or IRM at a level of 1 x 10(5) cells per unit well, and further incubated for 6, 12, 24, and 48 hours. The levels of TGF-beta1, FGF-2, and IL-6 from the supernatant were measured by enzyme-linked immunosorbent assay (ELISA). The data were analyzed using one-way ANOVA. The level of TGF-beta1 was down-regulated when the cells were grown in the presence of MTA except at 6 hours. The levels of FGF-2 release were significantly suppressed when PDL fibroblasts were grown in the presence of MTA or IRM at all time intervals (p < 0.05). The expressions of IL-6 from MTA treated cells were comparable to those of untreated control cells throughout the observation periods. We presume that this material inhibits the stimulatory function of growth factors on granulation tissue formation and in turn, it promotes the healing process modulated by other bone-remodeling cells.
Cytokines ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factor 2 ; Fibroblasts* ; Granulation Tissue ; Humans* ; Intercellular Signaling Peptides and Proteins* ; Interleukin-6 ; Periapical Tissue ; Periodontal Ligament* ; Transforming Growth Factor beta1 ; Pemetrexed

Mineral trioxide aggregate (MTA) would influence healing of periapical tissues by modulating the production of growth factors and cytokines from PDL fibroblasts, however, the studies are insufficient. Therefore, the purpose of this study was to monitor the expression of transforming growth factor-beta1 (TGF- beta1), fibroblast growth factor-2 (FGF-2), and interleukin-6 (IL-6) from PDL fibroblasts in the presence of MTA. The human PDL fibroblasts were seeded onto the set MTA or IRM at a level of 1 x 10(5) cells per unit well, and further incubated for 6, 12, 24, and 48 hours. The levels of TGF-beta1, FGF-2, and IL-6 from the supernatant were measured by enzyme-linked immunosorbent assay (ELISA). The data were analyzed using one-way ANOVA. The level of TGF-beta1 was down-regulated when the cells were grown in the presence of MTA except at 6 hours. The levels of FGF-2 release were significantly suppressed when PDL fibroblasts were grown in the presence of MTA or IRM at all time intervals (p < 0.05). The expressions of IL-6 from MTA treated cells were comparable to those of untreated control cells throughout the observation periods. We presume that this material inhibits the stimulatory function of growth factors on granulation tissue formation and in turn, it promotes the healing process modulated by other bone-remodeling cells.
Cytokines ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factor 2 ; Fibroblasts* ; Granulation Tissue ; Humans* ; Intercellular Signaling Peptides and Proteins* ; Interleukin-6 ; Periapical Tissue ; Periodontal Ligament* ; Transforming Growth Factor beta1 ; Pemetrexed

PURPOSE: This study analyzed the role of plasma biomarkers for TSU-68 in a previous phase II trial comparing TSU-68 plus docetaxel and docetaxel alone in patients with metastatic breast cancer. MATERIALS AND METHODS: A total of 77 patients were eligible for this study (38 in the TSU-68 plus docetaxel arm and 39 in the docetaxel alone arm). Blood samples were collected prior to the start of each cycle, and vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF)-AA, -AB, -BB, fibroblast growth factor, M30, C-reactive protein (CRP), and interleukin 6 (IL-6) levels were measured using enzyme linked immunosorbent assay. The primary endpoint was progression-free survival (PFS). RESULTS: In patients with baseline PDGF-AA ≥ median, median PFS was significantly worse in the TSU-68 plus docetaxel group than in the docetaxel alone group (5.4 months vs. 13.7 months, p=0.049), while a trend toward a PFS benefit was observed in those with baseline PDGF-AA < median (9.7 months vs. 4.0 months, p=0.18; p for interaction=0.03). In the TSU-68 plus docetaxel group, PFS showed significant association with fold changes in CRP (p=0.001), IL-6 (p < .001), PDGF-BB (p=0.02), and VEGF (p=0.047) following the first treatment cycle. CONCLUSION: Baseline PDGF-AA levels and dynamics of VEGF, PDGF-BB, CRP, and IL-6 levels were predictive for the efficacy of TSU-68.
Arm ; Biological Markers* ; Breast Neoplasms* ; Breast* ; C-Reactive Protein ; Disease-Free Survival ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factors ; Humans ; Interleukin-6 ; Pharmacology ; Plasma* ; Platelet-Derived Growth Factor ; Vascular Endothelial Growth Factor A

3-Deoxysappanchalcone (3-DSC) has been reported to possess anti-allergic, antiviral, anti-inflammatory and antioxidant activities. In the present study, we investigated the effects of 3-DSC on the proliferation of human hair follicle dermal papilla cells (HDPCs) and mouse hair growth in vivo. A real-time cell analyzer system, luciferase assay, Western blot and real-time polymerase chain reaction (PCR) were employed to measure the biochemical changes occurring in HDPCs in response to 3-DSC treatment. The effect of 3-DSC on hair growth in C57BL/6 mice was also examined. 3-DSC promoted the proliferation of HDPCs, similar to Tofacitinib, an inhibitor of janus-activated kinase (JAK). 3-DSC promoted phosphorylation of β-catenin and transcriptional activation of the T-cell factor. In addition, 3-DSC potentiated interleukin-6 (IL-6)-induced phosphorylation and subsequent transactivation of signal transducer and activator of transcription-3 (STAT3), thereby increasing the expression of cyclin-dependent kinase-4 (Cdk4), fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF). On the contrary, 3-DSC attenuated STAT6 mRNA expression and IL4-induced STAT6 phosphorylation in HDPCs. Finally, we observed that topical application of 3-DSC promoted the anagen phase of hair growth in C57BL/6 mice. 3-DSC stimulates hair growth possibly by inducing proliferation of follicular dermal papilla cells via modulation of WNT/β-catenin and STAT signaling.
Animals ; Blotting, Western ; Fibroblast Growth Factors ; Hair Follicle* ; Hair* ; Humans* ; Interleukin-6 ; Luciferases ; Mice* ; Phosphorylation ; Phosphotransferases ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; T-Lymphocytes ; Transcriptional Activation ; Transducers ; Vascular Endothelial Growth Factor A

OBJECTIVETo observe the temporal changes of basic fibroblast growth factor (FGF-2) expression in the spinal cord of a rat model of spared nerve injury (SNI) of the sciatic nerve.

METHODSA total of 100 male SD rats were randomly divided into sham-operated group and SNI group. The paw withdrawal threshold to mechanical stimulation was recorded at 1 day before and at 1, 4, 7, 14 and 28 days after the operation. The expressions of FGF-2, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the L4-6 spinal cord segments of the rats were measured at the specified time points.

RESULTSThe paw withdrawal threshold began to decline at 1 day after SNI, reached the lowest level at 7 days, and maintained a low level till 28 days (P<0.05). The expression of FGF-2 began to increase significantly on postoperative day 4, reached the peak level on day 14 and maintained the high level till day 28 (P<0.05). The rats with SNI showed significantly higher expressions of TNF-α and IL-6A in the spinal cord than those in the sham-operated group at each time point of measurement (P<0.05).

CONCLUSIONSSNI of the sciatic nerve can induce neuropathic tactile allodynia and causes up-regulation of FGF-2 and inflammatory cytokines in the spinal cord.

Animals ; Disease Models, Animal ; Fibroblast Growth Factor 2 ; metabolism ; Hyperalgesia ; Interleukin-6 ; metabolism ; Male ; Pain Threshold ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; metabolism ; Sciatica ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVES: The purpose of this study was to investigate the wound healing effect of primary cultured oral mucosal keratinocytes (OMKs) and to assess their roles in skin wounds. MATERIALS AND METHODS: OMK labeled with BromodeoxyUridine were scattered onto 1.5x1.5 cm skin defects of adult female nude mice (OMK group, n=15). For the control, culture media were placed on the wound (control group, n=15). Mice in both groups were sacrificed at three days (n=5), one week (n=5), and two weeks (n=5), and histomorphometric and immunoblot analyses with keratinocyte growth factor (KGF), interleukin (IL)-6, and IL-1alpha antibody were performed for the biopsied wound specimen. To verify the effect of the cytokine, rhIL-1alpha was applied instead of OMK transplantation, and the OMK and control groups were compared with regard to re-epithelialization. RESULTS: Histomorphometric analyses demonstrated faster re-epithelialization in the graft group than in the control group at the third day, first week, and second week. Newly forming epithelium showed maintenance of the histological character of the skin epithelium. The graft group showed superior expression of KGF, IL-6, and IL-1alpha protein, compared with the control group. Similar faster re-epithelialization was observed after treatment with rhIL-1alpha instead of OMK transplantation. CONCLUSION: We successfully confirmed that the graft of primary cultured OMKs promoted regeneration of skin defects. The mechanism of accelerated wound healing by primary cultured OMKs was attributed to inducement of cytokine expression as required for re-epithelialization.
Adult ; Animals ; Bromodeoxyuridine ; Culture Media ; Epithelium ; Female ; Fibroblast Growth Factor 7 ; Humans ; Interleukin-6 ; Interleukins ; Keratinocytes ; Mice ; Mice, Nude ; Primary Cell Culture ; Re-Epithelialization ; Regeneration ; Skin ; Tissue Engineering ; Transplants ; Wound Healing

OBJECTIVETo investigate the expressions of the aging gene P16(ink4a) and anti-aging gene HST2 in benign prostatic hyperplasia (BPH).

METHODSTwenty-three BPH and eighteen normal prostate specimens were collected and total RNA was extracted, followed by the reverse transcriptase polymerase chain reaction (RT-PCR). The expressions of P16(ink4a) was detected by semi-quantitative analysis in BPH and normal prostate tissues.

RESULTSP16(ink4a) mRNA, rather than HST2, was expressed in the BPH and normal prostate tissues. Semi-quantitative analysis showed that the P16(ink4a) mRNA expression in the normal prostate tissues (0.4868 +/- 0.545 was significantly higher than in the BPH tissues (0.2783 +/- 0.0268, with a statistical difference in between (P < 0. 05).

CONCLUSIONP16(ink4a) might play an important role in the pathogenesis of BPH and is probably one of the factors of cell aging escape.

Adult ; Aged ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Fibroblast Growth Factor 6 ; genetics ; Gene Expression Profiling ; Humans ; Male ; Pilot Projects ; Prostatic Hyperplasia ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: A loss of bone mass is usually detected after a bone marrow transplantation (BMT), especially during the early post-transplant period. We recently reported that enhanced bone resorption following a BMT was related to both the steroid dose and the increase in IL-6. We also suggested damage to the marrow stromal microenvironment, by myoablation, partly explains the impaired bone formation following a BMT. It is well known that some growth factors play important role in bone growth and osteogenesis. However, the pathogenetic role of bone growth factors in post-BMT bone loss is unknown and data on the changes in the growth factors, in accordance with bone turnover markers and bone mineral density (BMD) changes are scarce. We investigated changes in bone growth factors such as IGF-I (Insulin-like growth factor-I), fibroblast growth factor-2 (FGF-2) and Macrophage colony stimulating factor (M-CSF), during the post-BMT period, and assessed whether the growth factor changes influenced the bone turnover and post-BMT bone loss. The present study is the first prospective study to describe the changes in bone growth factors following a BMT. METHODS: We prospectively investigated 110 patients undergoing a BMT, and analyzed 36 patients (32.4+/-1.3 years, 17 men and 19 women) whose BMDs were measured before, and 1 year after, the BMT. The serum biochemical markers of bone turnover were measured before, 1, 2, 3 and 4 weeks, 3 and 6 months, and 1 year, after the BMT. The serum FGF-2, IGF-I and M-CSF levels were measured before and 1 and 3 weeks, and 3 months after the BMT. The correlation between the changes of growth factors and various bone parameters was analyzed. RESULTS: The mean bone losses in the lumbar spine and total proximal femur, calculated as the percentage change from the baseline to the level at 1 year, were 5.2 (p<0.05) and 11.6% (p<0.01), respectively. The serum type I carboxyterminal telopeptide (ICTP), a bone resorption marker, increased progressively until 6 months after the BMT, but thereafter decreased, to the base value after 1 year. Serum osteocalcin, a bone formation marker, decreased progressively, until 3 weeks after the BMT but then increased transiently, and finally returned to the base level at 1 year. The serum IGF-I and FGF-2 also decreased progressively until 3 weeks and 1 week after the BMT, respectively, then increased to the base values at 3 months. The serum M-CSF increased briskly at 1 week post-BMT, then decreased to the base level. There were positive correlations between the percentage changes from the baseline proximal femur BMD and the IGF-I levels 3 weeks and 3 months (r=0.52, p<0.01, r=0.41, p<0.05) post BMT. A Significant correlation was found between the IGF-I and osteocalcin levels pre-BMT, and 3 weeks after the BMT. Another positive correlation was found between the M-CSF and the ICTP levels at 3 weeks post BMT (r=0.54, p<0.05). CONCLUSION: In conclusion, there were significant changes in the serum IGF-I, FGF-2 and M-CSF levels in the immediate post-BMT period, which were related to a decrease in bone formation and loss in the proximal femoral BMD during the year following the BMT
Biomarkers ; Bone Density ; Bone Development ; Bone Marrow ; Bone Marrow Transplantation ; Bone Resorption ; Colony-Stimulating Factors ; Femur ; Fibroblast Growth Factor 2 ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Humans ; Insulin-Like Growth Factor I ; Intercellular Signaling Peptides and Proteins* ; Interleukin-6 ; Macrophage Colony-Stimulating Factor ; Macrophages ; Male ; Metabolism* ; Osteocalcin ; Osteogenesis ; Osteoporosis ; Prospective Studies ; Spine

The present study identified the favorable environment conditions for Spermatogomial stem cells in vitro according to their unique biological properties. Three growth factors, stem cell factor (SCF), leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF) were all found to independently contribute to the proliferation of mouse spermatogonial stem cell. The percentage of cell proliferation significantly enhanced by SCF at 30 ng/mL but decreased with heightening its combination after cultured 120 hours. The mice spermatogonial stem cells were significantly proliferated after 120 hours' culture with 10 ng/mL and 20 ng/mL (P < 0.01) of LIF, between 20 ng/mL and 50 ng/mL (P < 0.01) for bFGF. SCF and bFGF were significantly enhanced mice spermatogonial stem cells proliferation after these three factors combination. For LIF, no obvious effect was observed.
Animals ; Cell Division ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Fibroblast Growth Factor 2 ; pharmacology ; Growth Inhibitors ; pharmacology ; Interleukin-6 ; Leukemia Inhibitory Factor ; Lymphokines ; pharmacology ; Male ; Mice ; Spermatogonia ; drug effects ; physiology ; Stem Cell Factor ; pharmacology ; Stem Cells ; drug effects ; physiology

OBJECTIVETo explore an optional condition to induce mouse embryonic stem (ES) cells to differentiate into endothelial cells and to establish in vitro models of vasculogenesis and angiogenesis.

METHODSMouse ES cells were cultured in differentiation medium containing a cocktail of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), interleukin-6 (IL-6) and erythropoietin (EPO) in 1% methylcellulose to induce formation of embryoid bodies (EBs). At day 11, EBs were harvested and suspended in rat-tail collagen type I with the same cocktail of cytokines cultured for three additional days. The differentiation of ES cells into endothelial cells, processes of vasculogenesis and angiogenesis were examined using immunostaining of EBs slices and whole-mount immunocytochemistry of EBs with monoclonal antibodies (mAbs) against platelet endothelial cell adhesion molecule-1 (PECAM-1) and alpha-smooth muscle actin (SMA).

RESULTSUnder appropriate culture conditions; ES cells spontaneously differentiated and formed EBs containing vascular structures and tubular channels, which were positive for PECAM-1 co-differentiated with smooth muscle. When not treated with angiogenic growth factors, PECAM-1-positive cells could not organize into vascular structures of 11-day-old EBs. In the presence of angiogenic factors 11-day old EBs embedded into type I collagen, and rapidly developed an endothelial networks. Whole-mount immunocytochemistry of collagen gel with anti-PECAM-1 antibody showed the formation of primary vascular structures sprouting from EBs. Quantitative analysis revealed that 100 microg/ml thalidomide significantly reduced the number and length of EBs endothelial sprouting.

CONCLUSIONSMouse ES cells can differentiate into endothelial cells combined with smooth muscle differentiation during EBs formation and further develop endothelial outgrowths after EBs embedded into collagen, which respectively recapitulate vasculogenesis, angiogenesis, and arteriogenesis processes in vivo. The models provide a useful tool to investigate vasculogenesis, angiogenesis, and arteriogenesis mechanisms and evaluate the effects of angiogenic and angiostatic agents.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Collagen ; pharmacology ; Culture Media ; Embryo, Mammalian ; cytology ; physiology ; Endothelial Cells ; cytology ; physiology ; Erythropoietin ; pharmacology ; Fibroblast Growth Factor 2 ; pharmacology ; Interleukin-6 ; pharmacology ; Mice ; Neovascularization, Physiologic ; physiology ; Stem Cells ; cytology ; physiology ; Vascular Endothelial Growth Factor A ; pharmacology