Effects of apatite particle size in two apatite/collagen composites on the osteogenic differentiation profile of osteoblastic cells.
Effects of apatite particle size in two apatite/collagen composites on the osteogenic differentiation profile of osteoblastic cells.
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DOI：
10.3892/ijmm.2013.1516
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发表时间：
2013-12
影响因子：
5.4
通讯作者：
Kondo H
中科院分区：
文献类型：
--
作者：
Hatakeyama W;Taira M;Chosa N;Kihara H;Ishisaki A;Kondo H
新型骨传导替代材料的开发在医学上是值得期待的。在这项研究中，我们试图用两种不同大小的羟基磷灰石（HAP）颗粒和猪I型胶原制备新的羟基磷灰石(HAP)/胶原（Col）复合材料。这两种HAP颗粒要么是纳米级（平均直径40 nm； n-HAP），要么是大孔径为0.5-1.0 mm，且孔隙完全连通（m-HAP）。本研究的目的是研究两种HAP/Col复合材料中磷灰石粒径对成骨细胞样细胞（SaOS-2）成骨分化谱的影响。采用冷冻干燥和脱水交联技术制备胶原蛋白对照海绵（Col）和两种HAP/Col复合海绵（n-HAP/Col和m-HAP/Col），然后打孔出直径6 mm、高度1 mm的样品。SaOS-2细胞分别在三种实验材料上培养1、2、3、4周。从培养细胞中提取总RNA，利用碱性磷酸酶（ALP）、1型胶原（COL1）、骨唾液蛋白（BSP）、骨钙素前体[骨γ -羧谷氨酸（gla）蛋白（BGLAP）]基因以及β-肌动蛋白基因的引物，采用逆转录PCR （RT-PCR）检测成骨分化相关基因的表达情况。细胞分别在Col、n-HAP/Col和m-HAP/Col样品上培养1周和4周，然后在扫描电镜下观察。实验结果如下：RT-PCR结果显示，细胞在n-HAP/Col培养基上培养的成骨分化，尤其是BSP基因的表达速度最快，其次是m-HAP/Col培养基，而在Col培养基上培养的成骨分化速度最慢。扫描电镜（SEM）显示，SaOS-2细胞在冷标本上培养4周后呈成纤维细胞；在n-HAP/Col样品上培养1周时，它们是成纤维细胞，但在4周时呈现球形，同时积极吞噬n-HAP颗粒；然而，当与颗粒分离后，在m-HAP/Col标本上培养时，它们表现为变形的成纤维细胞。尽管实验结果有限，但我们的研究表明n-HAP/Col可能作为一种新的骨传导替代材料。
The development of new osteoconductive bone substitute materials is expected in medicine. In this study, we attempted to produce new hydroxylapatite (HAP)/collagen (Col) composites using two HAP particles of different sizes and porcine type I collagen. The two HAP particles were either nano-sized (40 nm in average diameter; n-HAP) or had macro-pore sizes of 0.5–1.0 mm in length with fully interconnected pores (m-HAP). The aim of this study was to investigate the effects of apatite particle size in two HAP/Col composites on the osteogenic differentiation profile in osteoblast-like cells (SaOS-2). We created a collagen control sponge (Col) and two HAP/Col composite sponges (n-HAP/Col and m-HAP/Col) using freeze-drying and dehydrothermal cross-linking techniques, and then punched out samples of 6 mm in diameter and 1 mm in height. The SaOS-2 cells were cultured on three test materials for 1, 2, 3 and 4 weeks. Total RNA was extracted from the cultured cells and the expression of osteogenic differentiation-related genes was evaluated by reverse transcription PCR (RT-PCR) using primer sets of alkaline phosphatase (ALP), type 1 collagen (COL1), bone sialoprotein (BSP) and osteocalcin precursor [bone gamma-carboxyglutamate (gla) protein (BGLAP)] genes, as well as the β-actin gene. The cells were also cultured on Col, n-HAP/Col and m-HAP/Col specimens for 1 and 4 weeks, and were then observed under a scanning electron microscope (SEM). The experimental results were as follows: RT-PCR indicated that osteogenic differentiation, particularly the gene expression of BSP, was most accelerated when the cells were cultured on n-HAP/Col specimens, followed by m-HAP/Col, whilst the weakest accelaeration was observed when the cells were cultured on Col specimens. As shown by the SEM images, the SaOS-2 cells were fibroblastic when cultured on Col specimens for up to 4 weeks; they were fibroblastic when cultured on n-HAP/Col specimens for 1 week, but appeared as spheroids, while actively phagocytizing n-HAP particles at 4 weeks; however, they appeared as deformed fibroblasts when cultured on m-HAP/Col specimens, detached from the particles. Despite limited experimental results, our study suggests that n-HAP/Col may be employed as a new osteoconductive bone substitute material.
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影响因子：
8
作者：
Liu X;Zhao M;Lu J;Ma J;Wei J;Wei S
通讯作者：
Wei S
影响因子：
5
作者：
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影响因子：
4.3
作者：
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DOI：
10.1002/jbm.b.31318
发表时间：
2009-08-01
影响因子：
3.4
作者：
Taira, Masayuki;Nezu, Takashi;Araki, Yoshima
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DOI：
10.1016/j.ijbiomac.2012.10.002
发表时间：
2013-01-01
影响因子：
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作者：
Sionkowska, A.;Kozlowska, J.
通讯作者：
Kozlowska, J.