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2001 Volume No 1 -
pages 52-58
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Title: In vitro evaluation of acute cytotoxicity
of human chemically treated allografts
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Authors: D. Dufrane, O. Cornu, T. Verraes, N. Schecroun,
X. Banse, Y.J. Schneider and Ch. Delloye
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Address: Orthopaedics Research Laboratory & Cell
Biochemical Laboratory, Université Catholique de Louvain,
Bruxelles, Belgium
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E-mail: denis.dufrane at clin.ucl.ac.be
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Key Words: Allografts, virus inactivation, biocompatibility,
cytotoxicity, in vitro studies.
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Publication date: 10th January 2001
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Abstract: In order to minimize the risk of contamination
associated with tissue transplantation, tissue banks commonly
chemically treat the tissues whenever possible. As viral inactivation
uses agents lethal to microorganisms, it is imperative to
assure that chemically inactivated tissue remains biocompatible.
In vitro assays can be an effective means to assess
the acute cytotoxicity of chemically treated human allografts.
We have used different types of cells cultured in the presence
of treated tissue extract. A standard cell line, a human fibroblast
(WI38), which was the same for all the samples, was chosen.
In addition, as the banked tissues (bone and fascia lata)
were prepared to be used in bone or as a dura mater substitute,
two other cell types were also used: an osteoblastic cell
line (SaOS-2) and a neuronal cell line (Neuro 2A). Cytotoxic
assessment was performed by qualitative evaluation of cell
morphology based on confluence, granulation, vacuolization
and swelling analysis. In addition, quantitative methods based
on the use of neutral red (NR) and 3- (4,5-dimethylthiazol-2-yl)-2,5
diphenyl tetrazolium bromide (MTT) were assayed. Qualitative
and quantitative evaluation of fascia lata and bone extracts
did not show deleterious effects on cell cultures. These results
show that in vitro methods can be appropriate to select
a non-toxic procedure before it is used in the human body
and that several strong chemical treatments can result in
a tissue suitable for human implantation.
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