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Re: Could anabolic steroids induce micronuclei in buccal cells?

Dear Editor,

I read the paper by Torres-Bugarin et al with great interest. Unfortunately, anabolic androgenic steroids (AAS) are frequently used in professional, and even in amateur sports. Hence, the aim of the study is very important. But before final decision concerning genotoxic activity of AAS, studied by means of micronucleus (MN) assay exfoliated buccal cells, some very important points of the paper must be clarified. And of course, in all cases the decision could not be final because only 5 bodybuilders were included in the study.

First of all, the design of the study is not clear, eg. when the sportsmen started to use the AAS. As I understand it they started to consume the anabolics after week 1. In this case the number of cells with MN (CMN) is two-fold higher in consumers that in the control group (1.1 vs. 0.5). Did the athletes use AAS before? If yes, the study is not valid. If no, the authors must explain why in the group of AAS consumers before the start of the study the number of CMN is significantly higher (I calculated the difference using GraphPrism, p value is equal to 0.0087, Mann-Whitney test with Gaussian approximation). If I understand the article rightly, the athletes started to consume the AAS after the 1st week when they had the level of CMN equal to 1.1‰. Just after one week the number of CMN increased to 4.8‰ – 4.3-fold. On page 592 it is written that MN in exfoliated cells “can reflect the damage occurring during the of 3 weeks” (!!). In this case after ONE week of consumption of AAS such a strike was registered which is suspicious.

The second thing is that in Table 2 some numbers are strange, e.g. 1.3, 6.3 and 6.8. How could the authors obtain such figures in case of studying 2000 cells? The numbers must be either whole numbers or half numbers, eg. 0.5, 7.5 and so on.

Figure 1 depicts CMN, according to the authors. But careful examination of the figure showed that MN in attached to main nucleus. With very high probability it mustn’t be considered as MN and it ought to be considered as a nuclear bud.

The authors wrote that “The scoring was done according to the criterion established by Torres-Bugarin et al 19(page 593, Sample analysis). But in the mentioned paper it says that the criterion was established by Page et al.

Very unsuccessful expressions are used by the authors. The example is written in the conclusions (What this study adds, page 595) – “The frequency of MN cells is increased in bodybuilders by AAS, leading to in situ or systemic loss or DNA damage”. It is completely unclear what the authors meant. Another example is “Exfoliated cells are also biomarkers of genotoxicity, with the advantage that they do not need to be kept in vivo” (page 592). I could not understand the meaning of this sentence even after reading it many times.

In conclusion, this is a very interesting paper but there are many errors and shortcomings which can confuse the readers of the journal. Hence, they must be clarified as soon as possible.