In vitro culture and characterization of alveolar bone osteoblasts isolated from type 2 diabetics.
In vitro culture and characterization of alveolar bone osteoblasts isolated from type 2 diabetics.
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DOI：
10.1590/s0100-879x2012007500054
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发表时间：
2012-06
期刊：
影响因子：
--
通讯作者：
Song YL
中科院分区：
文献类型：
--
作者：
Sun DC;Li DH;Ji HC;Rao GZ;Liang LH;Ma AJ;Xie C;Zou GK;Song YL
为了了解2型糖尿病患者牙种植体骨结合不良的机制，研究从这些患者中分离的牙槽骨成骨细胞的生物学特性是很重要的。在无菌条件下收集牙槽骨碎片，采用组织块贴壁法进行体外培养。采用碱性磷酸酶（ALP）化学染色法检测细胞活力，茜素红染色法检测成骨特性，MTT法检测细胞增殖，酶动力学检测ALP含量，放免法检测骨钙素（BGP）含量，ELISA法检测培养上清中Ⅰ型胶原（COL-I）含量。采用非特异性免疫荧光染色和细胞计数检测两种细胞在钛片上的粘附能力。在相同的培养条件下，两种细胞形态无明显差异。然而，2型糖尿病患者的牙槽骨成骨细胞生长缓慢，细胞活性和钙结节形成低于正常人。2型糖尿病患者牙槽骨成骨细胞培养上清中ALP、BGP和COL-I浓度均低于正常人（P < 0.05）。2型糖尿病患者牙槽骨成骨细胞可成功地在体外培养，其形态和生物学特性与正常人相同，但生长速度慢，特异性分泌物浓度低，与钛的结合能力较低。
In order to understand the mechanisms of poor osseointegration following dental implants in type 2 diabetics, it is important to study the biological properties of alveolar bone osteoblasts isolated from these patients. We collected alveolar bone chips under aseptic conditions and cultured them in vitro using the tissue explants adherent method. The biological properties of these cells were characterized using the following methods: alkaline phosphatase (ALP) chemical staining for cell viability, Alizarin red staining for osteogenic characteristics, MTT test for cell proliferation, enzyme dynamics for ALP contents, radio-immunoassay for bone gla protein (BGP) concentration, and ELISA for the concentration of type I collagen (COL-I) in the supernatant. Furthermore, we detected the adhesion ability of two types of cells from titanium slices using non-specific immunofluorescence staining and cell count. The two cell forms showed no significant difference in morphology under the same culture conditions. However, the alveolar bone osteoblasts received from type 2 diabetic patients had slower growth, lower cell activity and calcium nodule formation than the normal ones. The concentration of ALP, BGP and COL-I was lower in the supernatant of alveolar bone osteoblasts received from type 2 diabetic patients than in that received from normal subjects (P < 0.05). The alveolar bone osteoblasts obtained from type 2 diabetic patients can be successfully cultured in vitro with the same morphology and biological characteristics as those from normal patients, but with slower growth and lower concentration of specific secretion and lower combining ability with titanium than normal ones.
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