Adult ; Antibodies, Anticardiolipin ; blood ; Antibodies, Antiphospholipid ; blood ; Autoantibodies ; blood ; Female ; Humans ; Male ; Middle Aged ; Phosphatidic Acids ; immunology ; Phosphatidylcholines ; immunology ; Phosphatidylethanolamines ; immunology ; Phosphatidylinositols ; immunology ; Phosphatidylserines ; immunology ; Reference Values

A lipidomic study on extensive plasma lipids in bacterial peritonitis (cecal ligation and puncture, CLP)-induced sepsis in mice was done at 24 h post-CLP. The effects of administration of lysophosphatidylcholine (LPC) and lysophosphatidic acid (LPA), compounds known to have beneficial effects in CLP, on the sepsis-induced plasma lipid changes were also examined. Among the 147 plasma lipid species from 13 lipid subgroups (fatty acid [FA], LPA, LPC, lysophosphatidylethanolamine [LPE], phosphatidic acid [PA], phosphatidylcholine [PC], phosphatidylethanolamine [PE], phosphatidylinositol [PI], monoacylglyceride [MG], diacylglyceride [DG], triacylglyceride [TG], sphingomyelin [SM], and ceramide [Cer]) analyzed in this study, 40 and 70 species were increased, and decreased, respectively, in the CLP mice. Treatments with LPC and LPA affected 14 species from 7 subgroups, and 25 species from 9 subgroups, respectively. These results could contribute to finding the much needed reliable biomarkers of sepsis.
Animals ; Biomarkers ; Ligation ; Lysophosphatidylcholines ; Mice* ; Peritonitis ; Phosphatidic Acids ; Phosphatidylcholines ; Phosphatidylinositols ; Plasma* ; Punctures ; Sepsis

The purpose of this study is to investigate the response of human pulp cell on Portland cement mixed with beta-glycerophosphate. To investigate the effect of beta-glycerophosphate and/or dexamethasone on human pulp cell, ALP activity on various concentration of beta-glycerophosphate and dexamethasone was measured and mineral nodule of human pulp cell was stained with Alizarin red S. MTS assay and ALP activity of human pulp cell on Portland cement mixed with various concentration of beta-glycerophosphate (10 mM, 100mM, 1M) was measured and the specimens were examined under SEM. Addition of beta-glycerophosphate or dexamethasone alone had no effect however, the addition of 5 mM beta-glycerophosphate and 100 nM dexamethasone had the largest increasement in ALP activity. There was no toxicity in all samples and the data showed that Portland cement mixed with 10 mM beta-glycerophosphate had more increase in ALP activity compared with control. In conclusion, Portland cement mixed with beta-glycerophosphate has no toxicity and promotes differentiation and mineralization of pulp cell compared with additive-free Portland cement. This implicated that application of Portland cement mixed with beta-glycerophosphate might form more reparative dentin and in turn it would bring direct pulp capping to success.
Anthraquinones ; Dental Pulp Capping ; Dentin ; Dexamethasone ; Glycerophosphates ; Humans

Bone and Bones ; Calcium ; Glycerophosphates ; Mandible ; Osseointegration ; Sulfur ; Swine

Acetates ; Bone and Bones ; Calcium ; Calcium Compounds ; Glycerophosphates ; Mandible ; Osseointegration ; Osteogenesis ; Phosphoric Acids ; Sulfur ; Sulfuric Acids ; Titanium

OBJECTIVETo culture human dental papilla cells (HDPCs)and to study its cytobiological characters in vitro.

METHODSHDPCs were isolated and cultured with explant culture technique in vitro; Type I collagen, fibronection and laminin were detected in HDPCs and its secreted matrix with the immunocyto-chemical stain; HDPCs were incubated in mineralized promoting solution containing 10 mmol/L beta-glycerophosphate, 100 mg/L of ascorbic acid and 10 nmol/L dexamethasone supplemented with 10% FBS and the form of mineralized nodules was tested with Alizarin Red S stainning.

RESULTSCultured HDPCs in vitro were well growing in DMEM/F12. Type I collagen, fibronection and laminin staining were all positive in both HDPCs and its secreted matrix, and laminin was stained with bunchiness in matrix. Mineralized nodules formed after cultured 27 days by Alizarin Red S stainning.

CONCLUSIONHDPCs isolated and cultured are well growing in vitro, have a capability of synthesizing and secreting matrix and in mineralized promoting solution, are able to form mineralizer, so, HDPCs have a capacity of seed cell of tissue engineering regeneration tooth.

Cell Culture Techniques ; Cells, Cultured ; Collagen Type I ; Dental Papilla ; Dexamethasone ; Glycerophosphates ; Humans ; In Vitro Techniques ; Tissue Engineering ; Tooth

OBJECTIVETo investigate the potential of synovial mesenchymal stem cells (SMSC) in osteogenic differentiation.

METHODSSMSC were obtained by limited dilution method and expanded to culture in 25-milliliter flasks. The attached cells were treated with inductive medium containing dexamethasone, glycerophosphate and vitamin C at 3rd passage SMSC. The mineralized nodule was stained by Von Kossa method. The expression of ALP and osteopontin were detected by histochemical, immunohistological staining technique, respectively, while the expression of cbfa1 mRNA by RT-PCR.

RESULTSPure SMSC which were of spindle shape and star shape, uniform in size, could be induced to pleomorphism osteoblast in vitro, which were intensive positive in ALP and osteopontin. The expression of cbfa1 mRNA were also verified by RT-PCR and the polygonal cells formed nodular structure at 4 weeks. All these were coincident with the characters of osteoblast.

CONCLUSIONSMSC can be purified and induced into osteoblast in vitro.

Ascorbic Acid ; Cell Differentiation ; Dexamethasone ; Glycerophosphates ; Humans ; Mesenchymal Stromal Cells ; Osteoblasts ; Osteogenesis ; Osteopontin ; metabolism ; RNA, Messenger ; metabolism

INTRODUCTION: Skeletal homeostasis is normally maintained by the stability between bone formation by osteoblasts and bone resorption by osteoclasts. However, the correlation between the inflammatory reaction and osteoblastic differentiation of cultured osteoprogenitor cells has not been fully investigated. This study examined the effects of inflammatory cytokines on the osteoblastic differentiation of cultured human periosteal-derived cells. MATERIALS AND METHODS: Periosteal-derived cells were obtained from the mandibular periosteum and introduced into the cell culture. After passage 3, the periosteal-derived cells were further cultured in an osteogenic induction Dulbecco's modified Eagle's medium (DMEM) medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. In this culture medium, tumor necrosis factor (TNF)-alpha with different concentrations (0.1, 1, and 10 ng/mL) or interleukin (IL)-1beta with different concentrations (0.01, 0.1, and 1 ng/mL) were added. RESULTS: Both TNF-alpha and IL-1beta stimulated alkaline phosphatase (ALP) expression in the periosteal-derived cells. TNF-alpha and IL-1beta increased the level of ALP expression in a dose-dependent manner. Both TNF-alpha and IL-1beta also increased the level of alizarin red S staining in a dose-dependent manner during osteoblastic differentiation of cultured human periosteal-derived cells. CONCLUSION: These results suggest that inflammatory cytokines TNF-alpha and IL-1beta can stimulate the osteoblastic activity of cultured human periosteal-derived cells.
Alkaline Phosphatase ; Anthraquinones ; Ascorbic Acid ; Bone Resorption ; Cell Culture Techniques ; Cytokines ; Dexamethasone ; Durapatite ; Glycerophosphates ; Homeostasis ; Humans ; Interleukins ; Osteoblasts ; Osteoclasts ; Osteogenesis ; Periosteum ; Tumor Necrosis Factor-alpha

PURPOSE: To investigate the effects of irradiation on transforming growth factor beta1 (TGF-beta1) mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line. MATERIALS AND METHODS: Cells were cultured in alpha-minimum essential medium (alpha-MEM) supplemented with 10% fetal bovine serum and antibiotics. When the cells reached the level of 70-80% confluence, culture media were changed with alpha-MEM supplemented with 10% FBS, 5 mM beta-glycerol phosphate, and 50 microgram/mL ascorbic acid. Thereafter the cells were irradiated with a single dose of 2, 4, 6, 8 Gy at a dose rate of 1.5 Gy/min. The expression pattern of TGF-beta1 mRNA, calcium content and calcific nodule formation were examined on day 3, 7, 14, 21, 28, respectively, after the irradiation. RESULTS: The amount of TGF-beta1 mRNA expression decreased significantly on day 7 after irradiation of 4, 6, 8 Gy. It also decreased on day 14 after irradiation of 6, 8 Gy, and decreased on day 21 after irradiation of 8 Gy. The amount of calcium deposition decreased significantly on day 7 after irradiation of 4, 8 Gy (P.0.01) and showed a decreased tendency on day 14, 21 after irradiation of 4, 6, 8 Gy. The number of calcific nodules was decreased on day 7 after irradiation of 4, 8 Gy. CONCLUSION: Irradiation with a single dose of 4, 6, 8 Gy influences negatively the bone formation at the molecular level by affecting the TGF-beta1 mRNA expression that was associated with proliferation and the production of extracellular matrix in MC3T3-E1 osteoblastic cell line.
Anti-Bacterial Agents ; Ascorbic Acid ; Calcium ; Cell Line ; Culture Media ; Extracellular Matrix ; Glycerophosphates ; Organic Chemicals ; Osteoblasts ; Osteogenesis ; RNA, Messenger ; Transforming Growth Factor beta1

The present study aimed to investigate the osteogenic potentials of differentiated osteoblast-like cells (DOCs) induced from bone marrow-derived mesenchymal stem cells (MSCs) on beta-tricalcium phosphate (beta-TCP) with recombinant human bone morphogenetic protein (rhBMP-2) in vitro. Osteoblast differentiation was induced in confluent cultures by adding 100 nM dexamethasone, 10 mM beta-glycerophosphate, 50 mM L-ascorbic acid. The Alizarin red S staining and reverse transcriptase-polymerase chain reaction (RT-PCR) were perfomed to examine the mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN), receptor activator for nuclear factor kappaB ligand (RANKL), runt-related transcription factor 2 (RUNX2), collagen-I (COL-I). There were no significant differences in the osteogenic potentials of DOCs induced from MSCs on beta-TCP(+/-). According to the incubation period, there were significant increasing of Alizadin red S staining in the induction 3 weeks. The mRNA expression of ALP, RUNX2, and RANKL were higher in DOCs/beta-TCP(-) than DOCs/beta-TCP(+). According to rhBMP-2 concentrations, the mRNA expression of BSP was significantly increased in DOCs/beta-TCP(+) compared to that of DOCs/beta-TCP(-) on rhBMP 10 ng/ml. Our study presented the beta-TCP will have the possibility that calcium phosphate directly affect the osteoblastic differentiation of the bone marrowderived MSCs.
Alkaline Phosphatase ; Anthraquinones ; Ascorbic Acid ; Bone Morphogenetic Proteins ; Calcium ; Calcium Phosphates ; Dexamethasone ; Durapatite ; Glycerophosphates ; Humans ; Integrin-Binding Sialoprotein ; Mesenchymal Stromal Cells ; Osteoblasts ; Osteocalcin ; Osteogenesis ; RNA, Messenger ; Transcription Factors