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用三种不同方法提取的同种异体骨移植物中生长因子的定量分析。

Quantification of growth factors in allogenic bone grafts extracted with three different methods.

作者信息

Wildemann B, Kadow-Romacker A, Pruss A, Haas N P, Schmidmaier G

机构信息

Center for Musculoskeletal Surgery, Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, D-13353 Berlin, Germany.

出版信息

Cell Tissue Bank. 2007;8(2):107-14. doi: 10.1007/s10561-006-9021-0. Epub 2006 Oct 25.

DOI:10.1007/s10561-006-9021-0
PMID:17063261
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2795150/
Abstract

BACKGROUND

Bony allografts are used for defect filling. A reliable sterilization method is the peracetic acid-ethanol sterilization procedure (PES). Several studies showed the antimicrobiological efficacy of this method. Aim of this study was the quantification of growth factors necessary for bone formation in PES sterilized allografts (n = 9).

METHODS

To extract the growth factors from the tissue three different methods were used: (a) use of collagenase 1 for extraction, (b) incubation of the material in a proteinase inhibitor cocktail (Complete), and (c) extraction with guanidine HCl. The supernatants from the different methods were analyzed for the total protein concentration and different growth factors.

RESULTS

The extraction with guanidine HCl resulted in the highest amount of protein measurable in the supernatants of the samples. For comparison of the individual growth factor values the results were normalized to the protein content. The highest growth factor amount/protein was detectable for BMP-2 using the GndHCL method followed by FGFa, IGF-I, TGF-beta1, VEGF, and PDGF. Comparing the three extraction methods, significant differences were measured for the individual growth factor content.

CONCLUSION

PES sterilized bony allografts contain several growth factors. Depending on the extraction method, the quantity of the analyzed growth factors varies.

摘要

背景

骨移植异体骨用于填充骨缺损。一种可靠的灭菌方法是过氧乙酸 - 乙醇灭菌程序（PES）。多项研究表明了该方法的抗菌效果。本研究的目的是对经PES灭菌的异体骨（n = 9）中骨形成所需生长因子进行定量分析。

方法

为从组织中提取生长因子，使用了三种不同方法：（a）使用胶原酶1进行提取；（b）将材料置于蛋白酶抑制剂混合物（完全型）中孵育；（c）用盐酸胍提取。对不同方法得到的上清液分析总蛋白浓度和不同生长因子。

结果

用盐酸胍提取得到的样品上清液中可测量的蛋白量最高。为比较各生长因子的值，将结果按蛋白含量进行归一化处理。使用盐酸胍方法时，骨形态发生蛋白 - 2（BMP - 2）的生长因子量/蛋白最高,其次是酸性成纤维细胞生长因子（FGFa）、胰岛素样生长因子 - I（IGF - I）、转化生长因子 - β1（TGF - β1）、血管内皮生长因子（VEGF）和血小板衍生生长因子（PDGF）。比较三种提取方法，各生长因子含量存在显著差异。

结论

经PES灭菌的骨移植异体骨含有多种生长因子。根据提取方法不同，所分析的生长因子数量有所变化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/f0ac1860f8b9/10561_2006_Article_9021_Fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/188c4805c3d4/10561_2006_Article_9021_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/f6de12f300b5/10561_2006_Article_9021_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/d07a6a3592f0/10561_2006_Article_9021_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/1dfef0c1c942/10561_2006_Article_9021_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/d08088c605fd/10561_2006_Article_9021_Fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/f0d22c7ddafc/10561_2006_Article_9021_Fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/f0ac1860f8b9/10561_2006_Article_9021_Fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/188c4805c3d4/10561_2006_Article_9021_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/f6de12f300b5/10561_2006_Article_9021_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/d07a6a3592f0/10561_2006_Article_9021_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/1dfef0c1c942/10561_2006_Article_9021_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/d08088c605fd/10561_2006_Article_9021_Fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/f0d22c7ddafc/10561_2006_Article_9021_Fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d05/2795150/f0ac1860f8b9/10561_2006_Article_9021_Fig7.jpg

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