Whether nano-hydroxyapatite particles can influence apoptosis of mononuclear macrophage in rat abdominal cavity?
10.3969/j.issn.1673-8225.2009.51.018
- VernacularTitle:纳米羟基磷灰石颗粒能够影响大鼠腹腔单核巨噬细胞的凋亡吗?
- Author:
Tingting DING
;
Jiao SUN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(51):10093-10096
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Because of their size effect, nanometer particles (NPs) can combine molecular within cells, which can result in cell necrosis or apoptosis. But there are no systematic mechanisms of apoptosis induced by NPs about biological safety of NPs.OBJECTIVE: To investigate the effect of nano-hydroxyapatite particles on mononuclear macrophage in rat abdominal cavity at celluar and molecular level.DESIGN, TIME AND SETTING: A materials-cytology observation was performed at Shanghai Biomaterials Research & Testing Center from January 2001 to December 2008.MATERIALS: SD rats of clean grade were provided by SINO-BRITISH SIPPR/BK LAB. ANIMAL Co., Ltd.; NPs were provided by Shanghai Institute of Ceramics.METHODS: Peritoneal fluid was extracted under a sterility environment to in vitro separate and culture mononuclear macrophage using adherence method. The concentration of cell was adjusted at 2×10~9/L. At 300 W/40 kHz ultrasound, cell suspension containing 20,100 and 200 mg/L nano-hydroxyapatite particles was prepared to induce mononuclear macrophage for 24 hours, respectively. A normal control group was established.MAIN OUTCOME MEASURES: Ultrastructural phenotype was detected using transmission electron microscope; apoptotic rate was measured using AnnexinV-EGFP/PI staining; variation of apoptosis-related P53 gene expression was detected using Western Blot.RESULTS: Pseudopodia of mononuclear macrophage in the normal control group were intact, nuclear membrane was normal, and nucleoplasm was uniformed. After inducing by 20,100, 200 mg/L nano-hydroxyapatite particles for 24 hours, apoptotic morphological characteristics were typical in mononuclear macrophage. Compared with normal control group, apoptotic rate was significantly increased following the induction of 20 and 100 mg/L nano-hydroxyapatite (P< 0.01), and the increasing in the 100 mg/L nano-hydroxyapatite particle group was greater than 20 mg/L nano-hydroxyapatite particle group (P < 0.05). P53 protein was not observed in mononuclear macrophage in the normal control group. Following induction of varying concentrations of nano-hydroxyapatite particles, P53 protein expression was increased in the mononuclear macrophage, and the expression was positively related to concentration of nano-hydroxyapatite particles.CONCLUSION: Nano-hydroxyapatite particles can induce p53 expression through phosphorylation, which promotes downstream genes and finally results in cell apoptosis.
