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James M. Anderson - One of the best experts on this subject based on the ideXlab platform.
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Proteomic analysis and quantification of cytokines and chemokines from biomaterial surface-adherent macrophages and foreign body giant cells
Journal of Biomedical Materials Research Part A, 2007Co-Authors: Jacqueline A Jones, Takehisa Matsuda, Erica Colton, David T Chang, Howard J Meyerson, Il Keun Kwon, James M. AndersonAbstract:Implantation of biomaterial devices results in the well-known foreign body reaction consisting of Monocytes, macrophages, and foreign body giant cells (FBGCs) at the material/tissue interface. We continue to address the hypothesis that material surface chemistry modulates the phenotypic expression of these cells. Utilizing our human Monocyte Culture system, we have used surface-modified polymers displaying hydrophobic, hydrophilic, and/or ionic chemistries to determine the cytokines/chemokines released from biomaterial-adherent macrophages/FBGCs. This study broadens our approach by using proteomic analysis to identify important factors expressed by these cells and further quantifies these molecules with ELISAs. Proteomic profiles changed over time suggesting that the adherent macrophages underwent a phenotypic switch. Macrophage/FBGC-derived proinflammatory cytokines, IL-1β and IL-6, decreased with time, while the anti-inflammatory cytokine, IL-10, gradually increased with time. Resolution of the inflammatory response was also demonstrated by a decrease in chemoattractant IL-8 and MIP-1β production with time. Material-dependent macrophage/FBGC activation was analyzed using cytokine/chemokine production and cellular adhesion. Monocyte/macrophage adhesion was similar on all surfaces, except for the hydrophilic/neutral surfaces that showed a significant decrease in cellular density and minimal FBGC formation. Normalizing the ELISA data based on the adherent cell population provided cytokine/chemokine concentrations produced per cell. This analysis showed that although there were fewer cells on the hydrophilic/neutral surface, these adherent cells were further activated to produce significantly greater amounts of each cytokine/chemokine tested than the other surfaces. This study clearly presents evidence that material surface chemistry can differentially affect Monocyte/macrophage/FBGC adhesion and cytokine/chemokine profiles derived from activated macrophages/FBGCs adherent to biomaterial surfaces. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res 2007
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effects of adsorbed heat labile serum proteins and fibrinogen on adhesion and apoptosis of Monocytes macrophages on biomaterials
Journal of Materials Science: Materials in Medicine, 2003Co-Authors: William G. Brodbeck, Erica Colton, James M. AndersonAbstract:A previously established human Monocyte Culture protocol was used to determine the effects of varying adsorbed proteins on Monocyte/macrophage adhesion and survival on dimethyl-silane (DM) or RGD modified glass coverslips. Cells were allowed to adhere for 2 h in the absence of protein or in the presence of serum, fibrinogen (Fg), heat inactivated serum (HIS), serum supplemented with Fg or HIS with Fg. Cell adhesion and apoptosis rates were determined on days 0 (2 h), 3, 7 and 10 of Culture. The presence of serum alone in the initial Culture was sufficient to optimize Monocyte/macrophage adhesion and survival rates. Adding Fg to serum did not increase adhesion nor decrease apoptotic rates. No protein or the addition of HIS during the initial incubation period significantly decreased Monocyte/macrophage adhesion and survival on both surfaces, however, the addition of Fg to HIS restored adhesion and survival rates to those seen with in the presence of serum alone on RGD surfaces. These studies demonstrate that Monocyte/macrophage adhesion and survival on biomaterial surfaces are optimized by adsorbed heat labile serum proteins while adsorbed Fg plays a surface property-dependent role.
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biomaterial surface chemistry dictates adherent Monocyte macrophage cytokine expression in vitro
Cytokine, 2002Co-Authors: William G. Brodbeck, Takehisa Matsuda, Yasuhide Nakayama, Erica Colton, Nicholas P Ziats, James M. AndersonAbstract:Abstract An in vitro human Monocyte Culture system was used to determine whether adherent Monocyte/macrophage cytokine production was influenced by material surface chemistry. A polyethylene terephthalate (PET) base surface was modified by photograft copolymerization to yield hydrophobic, hydrophilic, anionic and cationic surfaces. Freshly isolated human Monocytes were Cultured onto the surfaces for periods up to 10 days in the presence or absence of interleukin-4 (IL-4). Semi-quantitative RT-PCR analysis on days 3, 7 and 10 of cell Culture revealed that interleukin-10 (IL-10) expression significantly increased in cells adherent to the hydrophilic and anionic surfaces but significantly decreased in the cationic surface adherent Monocytes/macrophages. Conversely, interleukin-8 (IL-8) expression was significantly decreased in cells adherent to the hydrophilic and anionic surfaces. Further analysis revealed that the hydrophilic and anionic surfaces inhibited Monocyte adhesion and IL-4–mediated macrophage fusion into foreign body giant cells (FBGCs). Therefore, hydrophilic and anionic surfaces promote an anti-inflammatory type of response by dictating selective cytokine production by biomaterial adherent Monocytes and macrophages. These studies contribute information necessary to enhance our understanding of biocompatibility to be used to improve the in vivo lifetime of implanted medical devices and prostheses.
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Alkylsilane-modified surfaces: inhibition of human macrophage adhesion and foreign body giant cell formation.
Journal of Biomedical Materials Research, 1999Co-Authors: Christopher R. Jenney, James M. AndersonAbstract:A homologous set of alkylsilane-modified glass surfaces with chain lengths ranging from methyl to octadecyl was prepared in order to examine the influence of alkyl surface chemistry on macrophage adhesion and foreign body giant cell (FBGC) formation. Contact angle and X-ray photoelectron spectroscopy analysis confirmed our silanation technique and indicated a consistent alkyl chain density independent of chain length. Human peripheral blood Monocytes were isolated and Cultured on these alkylsilane surfaces for a period of 10 days. The initial density of human Monocytes was similar on all surfaces. Beyond day 0 the clean glass, methyl (DM and C1), propyl (C3), and hexyl (C6) surfaces maintained a high cell density and supported macrophage development. In contrast, long-term macrophage density was extremely low on the tetradecyl (C14) and octadecyl (C18) surfaces. When interleukin-4 was added to induce FBGC formation in vitro, the DM, C1, C3, and C6 surfaces supported high levels of macrophage fusion while clean glass strongly inhibited fusion. The C14 and C18 surfaces did not contain sufficient macrophages to support FBGC formation. Cage implant studies revealed that in vivo macrophage density and FBGC formation on clean glass and C6 surfaces was similar to in vitro data. In contrast to the Monocyte Culture results, the C18 cage implant samples supported significant FBGC formation, possibly as a result of different conditions within each experimental system. Radiotracer adsorption studies of eight human serum proteins identified the high concentration and tenacious hold of adsorbed von Willebrand factor as being possibly involved in the poor long-term macrophage density observed on C14 and C18.
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Effects of surface-coupled polyethylene oxide on human macrophage adhesion and foreign body giant cell formation in vitro.
Journal of Biomedical Materials Research, 1999Co-Authors: Christopher R. Jenney, James M. AndersonAbstract:Surface immobilized polyethylene oxide (PEO) has been shown to efficiently reduce protein adsorption and cellular adhesion, resulting in a biologically passive surface. To explore the in vitro effects of surface immobilized PEO on the human inflammatory cells, macrophages, and foreign body giant cells (FBGCs), we developed a diisocyanate-based method for coupling PEO to amine-modified glass, a surface previously shown to enhance macrophage adhesion and FBGC formation. Contact angle analysis and X-ray photoelectron spectroscopy confirmed the presence of PEO molecules bound to the surface and revealed that PEO molecular weight significantly influenced the efficiency of PEO coupling. We used a 10-day human Monocyte Culture protocol to demonstrate that the presence of surface coupled PEO molecules does not significantly decrease initial Monocyte density or Monocyte-derived macrophage density after 3 days. However, PEO-coupled surfaces significantly reduced long-term Monocyte-derived macrophage density and virtually eliminated interleukin-4-induced FBGC formation observed at day 10. The cellular response to these PEO-coupled surfaces was related to the molecular weight of the PEO chains, which was varied between 200 Da and 18.5 kDa. These results suggest that an optimized PEO surface treatment may be effective in reducing inflammatory cell adhesion and possible degradation during the inflammatory response to an implanted biomedical device.
Christopher R. Jenney - One of the best experts on this subject based on the ideXlab platform.
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Alkylsilane-modified surfaces: inhibition of human macrophage adhesion and foreign body giant cell formation.
Journal of Biomedical Materials Research, 1999Co-Authors: Christopher R. Jenney, James M. AndersonAbstract:A homologous set of alkylsilane-modified glass surfaces with chain lengths ranging from methyl to octadecyl was prepared in order to examine the influence of alkyl surface chemistry on macrophage adhesion and foreign body giant cell (FBGC) formation. Contact angle and X-ray photoelectron spectroscopy analysis confirmed our silanation technique and indicated a consistent alkyl chain density independent of chain length. Human peripheral blood Monocytes were isolated and Cultured on these alkylsilane surfaces for a period of 10 days. The initial density of human Monocytes was similar on all surfaces. Beyond day 0 the clean glass, methyl (DM and C1), propyl (C3), and hexyl (C6) surfaces maintained a high cell density and supported macrophage development. In contrast, long-term macrophage density was extremely low on the tetradecyl (C14) and octadecyl (C18) surfaces. When interleukin-4 was added to induce FBGC formation in vitro, the DM, C1, C3, and C6 surfaces supported high levels of macrophage fusion while clean glass strongly inhibited fusion. The C14 and C18 surfaces did not contain sufficient macrophages to support FBGC formation. Cage implant studies revealed that in vivo macrophage density and FBGC formation on clean glass and C6 surfaces was similar to in vitro data. In contrast to the Monocyte Culture results, the C18 cage implant samples supported significant FBGC formation, possibly as a result of different conditions within each experimental system. Radiotracer adsorption studies of eight human serum proteins identified the high concentration and tenacious hold of adsorbed von Willebrand factor as being possibly involved in the poor long-term macrophage density observed on C14 and C18.
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Effects of surface-coupled polyethylene oxide on human macrophage adhesion and foreign body giant cell formation in vitro.
Journal of Biomedical Materials Research, 1999Co-Authors: Christopher R. Jenney, James M. AndersonAbstract:Surface immobilized polyethylene oxide (PEO) has been shown to efficiently reduce protein adsorption and cellular adhesion, resulting in a biologically passive surface. To explore the in vitro effects of surface immobilized PEO on the human inflammatory cells, macrophages, and foreign body giant cells (FBGCs), we developed a diisocyanate-based method for coupling PEO to amine-modified glass, a surface previously shown to enhance macrophage adhesion and FBGC formation. Contact angle analysis and X-ray photoelectron spectroscopy confirmed the presence of PEO molecules bound to the surface and revealed that PEO molecular weight significantly influenced the efficiency of PEO coupling. We used a 10-day human Monocyte Culture protocol to demonstrate that the presence of surface coupled PEO molecules does not significantly decrease initial Monocyte density or Monocyte-derived macrophage density after 3 days. However, PEO-coupled surfaces significantly reduced long-term Monocyte-derived macrophage density and virtually eliminated interleukin-4-induced FBGC formation observed at day 10. The cellular response to these PEO-coupled surfaces was related to the molecular weight of the PEO chains, which was varied between 200 Da and 18.5 kDa. These results suggest that an optimized PEO surface treatment may be effective in reducing inflammatory cell adhesion and possible degradation during the inflammatory response to an implanted biomedical device.
H U Rongdang - One of the best experts on this subject based on the ideXlab platform.
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effects of lps stimulated Monocyte Culture supernatant on the osteogenic function of osteoblastic cells in vitro
Chinese Journal of Orthodontics, 2011Co-Authors: Haiyan Wang, Hui Deng, X U Chunyan, Y E Jie, H U RongdangAbstract:Objective To investigate the influence of Pg-LPS stimulated Monocyte (RAW264.7) Culture supernatant on the proliferation and alkaline phosphatase (ALP) activity of osteoblastic cells (MC3T3-E1) in vitro. Methods The Culture supernatant of Monocytes stimulated with Pg-LPS was applied to osteoblasts MC3T3-E1 with different diluted concentrations(10%、20%、30%、40% and 50%) simultaneously for 24 h and 48 h in vitro.The cellular proliferation was assessed by MTT assay, Cellular ALP activity was examined using ALP measurement kit, and the results were statistically analyzed using SPSS 12.0 software package. Results LPS stimulated RAW264.7 to release inflammatory cytokines including IL-1β, IL-6 and TNF-α, whose secretion was in a time and dose dependent manner with the concentration of LPS. After the stimulation of different concentrations of inflammatory supernatant, the proliferation and ALP activity of MC3T3-El cells significantly deceased, both in a concentration-dependent manner. Conclusions These results indicated that the inflammatory cytokine secretion of Pg-LPS stimulated RAW264.7 Culture supernatant was similar to the change in vivo with inflammation. The inflammatory supernatant could inhibit the proliferation and ALP activity of osteoblastic cells. Key words: LPS; Monocyte; Osteoblasts; Proliferation; ALP activity
M C De Vernejoul - One of the best experts on this subject based on the ideXlab platform.
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increased bone resorbing activity of peripheral Monocyte Culture supernatants in elderly women
The Journal of Clinical Endocrinology and Metabolism, 1998Co-Authors: Martine Cohensolal, F Boitte, Oana Bernardpoenaru, M A Denne, A M Graulet, Michel Brazier, M C De VernejoulAbstract:Accelerated bone loss occurs in the years after menopause, and is an ongoing phenomenon in elderly women. The role of cytokines in bone loss after estrogen deficiency has been shown in ovariectomized rat and mice models. In humans, the involvement of bone resorbing cytokines is now well established. In the early years after menopause, Monocyte activation leads to increased cytokine production. We have previously shown that the bone resorbing activity (BRA) of peripheral blood Monocyte Culture supernatants from postmenopausal women is higher than in premenopausal (Pre-M) women. This increased activity was related to interleukin (IL)-1, IL-6, and tumor necrosis factor-α levels. We here investigate whether Monocyte activation still occurs in older women and whether this relates to bone resorption. We studied 19 healthy Pre-M, and 24 early (E-Post-M, menopause 10 yr) postmenopausal women. Peripheral blood Monocytes were Cultured for 48 h with 20% autologous plasma. BR...
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peripheral Monocyte Culture supernatants of menopausal women can induce bone resorption involvement of cytokines
The Journal of Clinical Endocrinology and Metabolism, 1993Co-Authors: Martine Cohensolal, M A Denne, A M Graulet, J Gueris, D Baylink, M C De VernejoulAbstract:Increased bone resorption is a mechanism contributing to bone loss in the postmenopausal period. Cytokines are involved in osteoclastic differentiation and, therefore, may play a role in the regulation of bone resorption. Several previous works showed the implication of interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF alpha) in the modulation of bone remodeling. This study determines the concomitant production of the three cytokines and tests the bone-resorbing activity of peripheral Monocyte supernatants. Four groups of women were studied: premenopausal women (n = 13; mean age, 47 +/- 0.9 yr), untreated postmenopausal women (n = 21; mean age, 52 +/- 0.6 yr), postmenopausal women treated with estrogens (n = 14; mean age, 54.2 +/- 1.1 yr), or postmenopausal women treated with ethanehydroxydiphosphonate (n = 12; mean age, 53.2 +/- 2 yr). Assignment to clinical groups was verified by plasma FSH and estradiol determinations. Lumbar spine bone mineral density was significantly higher in the pre...
Haiyan Wang - One of the best experts on this subject based on the ideXlab platform.
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effects of lps stimulated Monocyte Culture supernatant on the opg rankl of osteoblastic cells
Shanghai journal of stomatology, 2015Co-Authors: Haiyan Wang, Huaxiang Qin, Hui Deng, Chaofan SunAbstract:PURPOSE To investigate the influence of Pg-LPS stimulated Monocyte (RAW264.7) Culture supernatant on the OPG/RANKL expression of osteoblastic cells (MC3T3-E1). METHODS The Culture supernatant of Monocytes stimulated with Pg-LPS was applied to osteoblasts MC3T3-E1 with different diluted concentrations (10%, 20%, 30%, 40% and 50%) simultaneously for 24h, then RT-PCR was used to detect the expression changes of OPG/RANKL mRNA. Western blot was used to detect the expression changes of OPG/RANKL protein. The data was analyzed by ANOVA using SPSS 17.0 software package. RESULTS After stimulation of different concentrations of inflammatory supernatant, the expressions of OPGmRNA and protein significantly decreased (P<0.05), whereas the expressions of RANKLmRNA and protein significantly increased(P<0.05), both of them were in a concentration-dependent manner. CONCLUSIONS These results indicate that Pg-LPS stimulated RAW264.7 Culture supernatant can inhibit the osteogenesis and differentiation of the osteoblasts through inhibiting the expression of OPGmRNA and protein of osteoblastic cells, while increasing the expression of RANKLmRNA and protein in a concentration-dependent manner.
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effects of lps stimulated Monocyte Culture supernatant on the osteogenic function of osteoblastic cells in vitro
Chinese Journal of Orthodontics, 2011Co-Authors: Haiyan Wang, Hui Deng, X U Chunyan, Y E Jie, H U RongdangAbstract:Objective To investigate the influence of Pg-LPS stimulated Monocyte (RAW264.7) Culture supernatant on the proliferation and alkaline phosphatase (ALP) activity of osteoblastic cells (MC3T3-E1) in vitro. Methods The Culture supernatant of Monocytes stimulated with Pg-LPS was applied to osteoblasts MC3T3-E1 with different diluted concentrations(10%、20%、30%、40% and 50%) simultaneously for 24 h and 48 h in vitro.The cellular proliferation was assessed by MTT assay, Cellular ALP activity was examined using ALP measurement kit, and the results were statistically analyzed using SPSS 12.0 software package. Results LPS stimulated RAW264.7 to release inflammatory cytokines including IL-1β, IL-6 and TNF-α, whose secretion was in a time and dose dependent manner with the concentration of LPS. After the stimulation of different concentrations of inflammatory supernatant, the proliferation and ALP activity of MC3T3-El cells significantly deceased, both in a concentration-dependent manner. Conclusions These results indicated that the inflammatory cytokine secretion of Pg-LPS stimulated RAW264.7 Culture supernatant was similar to the change in vivo with inflammation. The inflammatory supernatant could inhibit the proliferation and ALP activity of osteoblastic cells. Key words: LPS; Monocyte; Osteoblasts; Proliferation; ALP activity