Fig 1.
Graphical presentation of the study design. Sixteen fracture patients with tinzaparin treatment and 14 non-fracture controls without anticoagulation therapy. To control for the effect of tinzaparin on miRNA analysis an identical concentration of synthetic miRNAs was added to plasma, isolated RNA and prepared cDNA from all samples in both groups prior to qPCR.
Table 1.
Fig 2.
Effect of prophylactic tinzaparin treatment on cDNA synthesis and qPCR.
Cq values for two synthetic spike-in RNAs, A) UniSp6 added to RNA prior to cDNA synthesis and B) UniSp3 added to cDNA prior to qPCR; UniSp6, ETG n = 71, LTG n = 58, CON n = 14; UniSp3, ETG n = 71, LTG n = 58, CON n = 14. ETG = early tinzaparin group, LTG = late tinzaparin group, CON = control. The horizontal line shows the median, while the box encompasses the 50th percentile and the whiskers encompass the minimum and maximum values. Outliers are displayed as points. Outliers are defined according to Tukey, as values exceeding more than the 75 percentile plus 1.5 IQR.
Fig 3.
Effect of prophylactic tinzaparin treatment on RNA extraction from plasma.
Cq values for three synthetic spike-in RNAs added to plasma prior to RNA extraction; UniSp2 ETG n = 71, LTG n = 57, CON n = 14; UniSp4, ETG n = 71, LTG n = 57, CON n = 14 & UniSp5, ETG N = 69, LTG n = 56, CON n = 14. ETG = early tinzaparin group, LTG = late tinzaparin group, CON = control. The horizontal line shows the median, while the box encompasses the 50th percentile and the whiskers encompass the minimum and maximum values. Outliers are displayed as points. Outliers are defined according to Tukey, as values exceeding more than the 75 percentile plus 1.5 IQR. Multiple comparisons showed a significant difference between ETG and CON, p = 0.017, denoted by *.
Fig 4.
Effect of prophylactic tinzaparin treatment on quantification of different endogenous circulating miRNAs in fracture patients.
Effect of a dose of tinzaparin (<6 hours) on levels of hsa-let-7i-5p (A), hsa-miR-222-3p (B), hsa-miR-30e-5p (C), hsa-miR-1-3p (D), hsa-miR-133a-3p (E) & hsa-miR-133b (F) in plasma. Data expressed as fold change from PRE (left graph) and delta CT pre vs post (right graph), mean±SD, n = 12.