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

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

B Wildemann  1 , A Kadow-Romacker, A Pruss, N P Haas, G Schmidmaier
Affiliations
Comparative Study

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

B Wildemann et al. Cell Tissue Bank. 2007.

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.

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Figures

Fig. 1
Fig. 1
Protein content per gram sample. Significantly more protein was extracted from the samples after GndHCl extraction compared to both other methods, *P < 0.001
Fig. 2
Fig. 2
Quantification of BMP-2 [pg/µg Protein] in allografts using three different extraction methods. The BMP- 2 content is correlated to the total protein content of the samples. Using the Collagenase or the Complete Method in only two of nine samples was BMP-2 detectable
Fig. 3
Fig. 3
Quantification of IGF-I [pg/µ Protein] in allografts using three different extraction methods, *P = 0.003 compared to GndHCl
Fig. 4
Fig. 4
Quantification of VEGF [pg/µ Protein] in allografts using three different extraction methods, *P = 0.014 compared to GndHCl
Fig. 5
Fig. 5
Quantification of FGFa [pg/µg Protein] in allografts using three different extraction methods, *P < 0.001 compared to both methods
Fig. 6
Fig. 6
Quantification of TGF-β1 [pg/µg Protein] in allografts using three different extraction methods
Fig. 7
Fig. 7
Quantification of PDGF [pg/µg Protein] in allografts using three different extraction methods
See this image and copyright information in PMC

References

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