本文選題：鈦顆粒　切入點：成骨細胞　出處：《中國修復重建外科雜志》2014年05期

【摘要】：目的通過觀察鈦顆粒對成骨細胞增殖分化和細胞形態(tài)的影響,探討其可能的內在聯(lián)系和作用機制。方法取10只新生24 h SD大鼠顱骨,采用多次酶消化法體外分離培養(yǎng)成骨細胞,ALP和茜素紅染色進行細胞鑒定。取生長良好的第3代細胞,分別采用濃度為0.01、0.05、0.1、0.5、1 mg/mL的鈦顆粒培養(yǎng)基(0.01、0.05、0.1、0.5、1 mg/mL組)培養(yǎng),7 d后用細胞計數(shù)試劑盒8檢測吸光度(A)值,比較不同濃度鈦顆粒對成骨細胞增殖能力的影響,并篩選半數(shù)致死濃度;ELISA法檢測細胞Ⅰ型膠原表達,比較不同濃度鈦顆粒對成骨細胞分化能力的影響。另取以半數(shù)致死濃度鈦顆粒培養(yǎng)7 d的成骨細胞(實驗組)進行FITC-鬼筆環(huán)肽和碘化丙啶雙重熒光染色,激光共聚焦顯微鏡下觀察吞噬鈦顆粒后細胞形態(tài)學的改變。以上觀察指標均以正常第3代成骨細胞作為對照組。結果 ALP及茜素紅染色鑒定提示培養(yǎng)的細胞為成骨細胞。培養(yǎng)7 d,各濃度組成骨細胞增殖、分化能力與對照組比較均有不同程度下降,差異均有統(tǒng)計學意義(P0.05)。同時隨濃度增加,各濃度組對成骨細胞增殖的抑制作用逐漸增強,成骨細胞Ⅰ型膠原表達逐漸降低;除0.01、0.05 mg/mL組間差異無統(tǒng)計學意義(P0.05)外,其余各濃度組間比較差異均有統(tǒng)計學意義(P0.05)。鈦顆粒的半數(shù)致死濃度為0.5 mg/mL。激光共聚焦顯微鏡下示,與對照組比較,實驗組吞噬了鈦顆粒的成骨細胞皺縮變形,偽足收縮變短,微絲排列紊亂。結論鈦顆�？梢种瞥晒羌毎脑鲋澈头只芰�,這種作用可能與吞噬鈦顆粒后細胞形態(tài)學的改變有關。
[Abstract]:Objective to observe the effect of titanium granule on proliferation, differentiation and cell morphology of osteoblasts, and to explore its possible internal relationship and mechanism. Methods the cranial bones of 10 newborn 24 h SD rats were taken out. Osteoblasts were isolated and cultured with ALP and alizarin red staining by multiple enzyme digestion in vitro. The cell count kit 8 was used to detect the absorbance of osteoblasts in 0. 01% 0. 05 and 0. 5 mg/mL titanium granule culture medium containing 0. 01 0. 05 and 0. 5 mg/mL for 7 days. The effects of different concentrations of titanium particles on the proliferation of osteoblasts were compared, and the effects of different concentrations of titanium particles on the proliferation of osteoblasts were compared. The expression of type I collagen was detected by Elisa. The effects of titanium particles at different concentrations on the differentiation of osteoblasts were compared. The osteoblasts cultured with titanium particles with LD50 for 7 days (experimental group) were stained with FITC- phloropylinin and promadime iodide double fluorescence staining. The morphologic changes of the cells after phagocytosis of titanium granules were observed by laser confocal microscopy. All the above indexes were normal third passage osteoblasts as control group. Results ALP and alizarin red staining indicated that the cultured cells were formed. Osteoblasts were cultured for 7 days. Compared with the control group, the differentiation ability decreased to some extent, and the difference was statistically significant (P 0.05). At the same time, with the increase of concentration, the inhibition of osteoblast proliferation was gradually enhanced, and the expression of type I collagen in osteoblasts was gradually decreased. There was no significant difference between 0. 01 mg/mL and 0. 01 mg/mL groups (P 0. 05), but there were significant differences among the other groups. The LD50 of titanium particles was 0. 5 mg / mL. Laser confocal microscope was used to compare the results with that of the control group, and compared with the control group, the LD50 of titanium particles was 0. 5 mg 路mL ~ (-1) 路ml ~ (-1) 路L ~ (-1). In the experimental group, the osteoblasts of the experimental group were engulfed with titanium particles, the contraction of pseudopodia became shorter and the arrangement of microfilaments was disordered. Conclusion Titanium granules can inhibit the proliferation and differentiation of osteoblasts. This effect may be related to the changes of cell morphology after phagocytosis of titanium particles.
【作者單位】： 深圳市龍崗中心醫(yī)院骨關節(jié)科;
【基金】：深圳市龍崗區(qū)科技發(fā)展基金資助項目(YS2012163)~~
【分類號】：R318.08

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本文編號：1681510