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在钛植入材料上培养的MC3T3-E1细胞矿化过程中,胶原结合型富含亮氨酸小分子蛋白聚糖和赖氨酰羟化酶的差异基因表达

Differential gene expression of collagen-binding small leucine-rich proteoglycans and lysyl hydroxylases, during mineralization by MC3T3-E1 cells cultured on titanium implant material.

作者信息

Takashi Matsuura, Tsubaki Satoshi, Tsuzuki Takashi, Duarte Wagner R, Yamauchi Mitsuo, Sato Hironobu

机构信息

Department of Oral Rehabilitation, Fukuoka Dental College, Tamura 2-15-1, Sawara-ku, Fukuoka 814-0193, Japan.

出版信息

Eur J Oral Sci. 2005 Jun;113(3):225-31. doi: 10.1111/j.1600-0722.2005.00208.x.

Abstract

Titanium implants create a unique ultrastructure (composed of a collagenous zone with relatively disorganized fibril morphology) at the bone-implant interface. The objective of this study was to investigate the temporal mRNA expression patterns, using real-time polymerase chain reaction, of type I collagen (COLI) and regulators for collagen fibrillogenesis, collagen-binding small leucine-rich proteoglycans (SLRPs) and lysyl hydroxylases (LHs), during mineralization, by MC3T3-E1 cells cultured on titanium (Ti). Lysates of the cultures on Ti and on plastic wells (Pl) for 10-50 d were used for the quantification of calcium and mRNA. Although the onset of calcium accumulation in the cultures on Ti (30-40 d) was slower than that of cultures on Pl (20-30 d), the gene expression patterns during mineralization were similar in cells cultured on either material. COLI and fibromodulin were up-regulated just before the onset of mineralization and then down-regulated. Lumican and LH1 were up-regulated just before the onset of mineralization and then returned to the baseline level. Decorin and LH2 were up-regulated at the late mineralization stage. Biglycan was down-regulated once at the early mineralization stage and then returned to the original level. LH3 was maintained at a steady level throughout. This study suggests actual but distinct roles of SLRPs and LHs in the formation of a unique ultrastructure at the bone-implant interface.

摘要

钛植入物在骨-植入物界面处形成一种独特的超微结构(由具有相对无序原纤维形态的胶原区组成)。本研究的目的是通过在钛(Ti)上培养的MC3T3-E1细胞,利用实时聚合酶链反应研究矿化过程中I型胶原蛋白(COLI)以及胶原纤维生成调节剂、胶原结合型富含亮氨酸小分子蛋白聚糖(SLRPs)和赖氨酰羟化酶(LHs)的mRNA表达随时间的变化模式。将在Ti和塑料孔(Pl)上培养10 - 50天的细胞裂解物用于钙和mRNA的定量分析。虽然在Ti上培养的细胞中钙积累的起始时间(30 - 40天)比在Pl上培养的细胞慢(20 - 30天),但在这两种材料上培养的细胞在矿化过程中的基因表达模式相似。COLI和纤调蛋白在矿化开始前上调,然后下调。核心蛋白聚糖和LH1在矿化开始前上调,然后恢复到基线水平。饰胶蛋白和LH2在矿化后期上调。双糖链蛋白聚糖在矿化早期有一次下调,然后恢复到原始水平。LH3在整个过程中保持稳定水平。本研究表明SLRPs和LHs在骨-植入物界面独特超微结构的形成中发挥了实际但不同的作用。

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