Hello, I am interested to preserve bone without lost osteoinductivity, I read the best method to preserve bone is fresh-frozen, but how much can I storage without lost osteoinductivity?
Thank you for your help
Luz
http://synapse.koreamed.org/Synapse/Data/PDFData/1150JPIS/jpis-42-224.pdf
Dear Luz
You can preserve the Bone in -80 for 6 months but you believe that osteo inductive related to your method and preserving it with special method for freezing.(nutrition media).
Sincerely
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For best long term results you should use demineralized freeze-dried bone (hueso desmineralizado y liofilizado) you must see: Schwartz Z, Mellonig JT, Carnes DL Jr, de la Fontaine J, Cochran DL, Dean DD, Boyan BD. Ability of commercial demineralized freeze-dried bone allograft to induce new bone formation. Periodontol. 1996 Sep;67(9):918-26.
Dears José Francisco, Ahad and Kristijan PhD, thank you so much for your recommendations and suggestions.
Best
Luz
At -80° you can store bone indefinately. No influence on osteoinductivity (which is doubtful anyway). However, there are legal restrictions to 5years because after that time sterility cannot be guaranteed.
It depends what you wish to analyze later!
For mRNA expression analysis of bone or bone marrow you need to isolate the mRNA asap.
For protein analysis you must snap freeze in N2 and latter store it at -80°C.
For histology of cellular structures you should fix PFA, dehydrate and embed the tissue. It is possible to store it in ethanol or paraffin for very long time.
For analysis of the mineralized bone structure you can freeze -80°C or better fix it with PFA,
For mechanical testing you must snap freeze in N2 and latter store it at -80°C.
I agree to prof Winkler that the time period don´t really matter (as far as we know now). One study had detected the osteoindictive protein BMP-7 to leak out from bone allograft that had been deep freezed (Ref. Board T.N, Rooney P, Kay P.R. Strain imparted during impaction grafting may contribute to bony incorporation. An in vitro study of the release of BMP-7 from allograft. J. Bone Joint Surg (Br) 2008; 90-B: 821-4).
To set up a study for analyzing if less or "inactivated" BMP-7 would leak out from long time stored bone compared to short time stored, could be interesting.
Thank you so much, all your answers are really interesting and so important to me.
Best
Dear Luz
I agree with Dr. Winkler , but in Brazil the law allows storage for a maximum of five years
MICRONIC (https://www.micronic.com/ ) makes extremely durable tubes, certainly suitable for -80C. RNAse, DNAse, pyrogen-free, available sterile, external or internal threaded, and free of leachables. Check out some of their tech notes here: (https://www.micronic.com/tech-notes) The Generation R study may be of some interest to you in particular, the study has preserved biosamples from infant to adulthood, going on 15 years.
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