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Biochemical analysis associated with HL-60 cells incubated with various ceramic materials
L Jones1, M Tucci, H Benghuzzi
1School of Health Related Professions, University of Mississippi Medical Center, Jackson 39216, USA.
Abstract:
The purpose of this experiment was to determine the effect of TCP, ALCAP, and HA ceramic particles on the viability and proliferation rate of HL-60 cells in culture. Previous experiments have suggested that growing cell lines such as macrophages, fibroblasts, and endothelial cells on discs made of TCP, ALCAP, and HA induced immunochemical and morphological changes. The effect of ceramic particles (< 38 um) on HL-60 cells in culture have never been investigated. A total of 1 x 10(5) (n = 5) cells were placed in culture with 2 mg of ALCAP, TCP, HA particles, or 10 micrograms/ml lipopolysaccharides (LPS). At the end of 24, 48, and 96 hours, the cells were evaluated morphologically and then counted. The supernatants were collected and biochemical analysis for LDH and immunochemical analysis for cytokines IL-1 was performed. The data from this investigation revealed the following: (1) At the end of the 24 hour phase, the cell number in wells containing HA and ALCAP particles were lower than the control cells, whereas cells in the presence of TCP were similar to the control. (2) Microscopic evaluation of cells treated with HA showed hydropic change at 24, 48 and 96 hours. (3) At 96 hours, there were no significant changes in cell number between the groups. This phenomenon could be attributed to the depletion of nutrients within the wells or possibly the cells reaching confluence. (4) The ease of ingestion of macrophages to CPB is as follows: TCP > ALCAP > HA. (5) Biochemical analysis of markers for cellular disturbances revealed that there were no significant differences between the different group at any time point (p < 0.05).