## 4 Discussion
\begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline
**Groups** & **Leukocytes** & **Erythrocytes** & **Hemoglobin** & **Hematocrit** & **VEM** & **Platelets** & **PDW** & **MPV** & **Segmented Neutroph** \\ \hline Control & \(15.52\pm 1.25\) & \(7.07\pm 0.15\) & \(13.01\pm 0.21\) & \(35.38\pm 0.48\) & \(50.12\pm 0.44\) & \(113.00\pm 58.25\) & \(8.57\pm 0.21\) & \(13.05\pm 5.33\) & \(5.30\pm 0.50\) \\ CAP & \(10.97\pm 0.92\) & \(7.22\pm 0.205\) & \(13.47\pm 0.39\) & \(36.65\pm 0.92\) & \(50.82\pm 0.83\) & \(734.50\pm 214.30\) & \(9.22\pm 0.36\) & \(8.03\pm 0.17\) & \(3.43\pm 0.49\) \\ CAP & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) \\ PRP & \(12.26\pm 1.39\) & \(7.01\pm 0.16\) & \(12.63\pm 0.41\) & \(35.55\pm 0.85\) & \(50.73\pm 0.21\) & \(957.80\pm 166.60\) & \(8.57\pm 0.35\) & \(7.82\pm 0.26\) & \(3.25\pm 0.62\) \\  & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) \\ L-NAME & \(13.85\pm 1.54\) & \(7.09\pm 0.185\) & \(13.18\pm 0.23\) & \(36.15\pm 0.45\) & \(51.08\pm 0.92\) & \(1141.00\pm 186.50\) & \(8.80\pm 0.32\) & \(7.92\pm 0.17\) & \(4.25\pm 0.61\) \\  & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) & (ns) \\ \hline \hline \end{tabular}
\end{table} 
Table 3: Hematological parameters of the groups (ns—non-significant). Another hypothesis in such an experimental timescale (2 h) is that NO could have totally reacted with dioxygen and superoxide, and hence transformed into nitrate and nitrite in a nM concentration, resulting in the chemistry of NO being incompatible with the observed effects under the used conditions. Another observation of this study refers to the pH solution, but as no buffer was used in the CAP activation, it is reasonable to expect a significant pH drop in the range of 3 to 5, an effect reported in many papers [29; 35; 38]. Plasma activation of biocompatible liquids was studied previously so as to quantify the chemical transformations [35; 39], the bacterial inactivation [40], and the anti-cancer capacity [31; 36; 41; 42; 43; 44; 45]. To the best of our knowledge, its injection on a rat skin flap has never been reported in previous studies. The results of this type of flap after subcutaneous injection of the CAP activated solution denote a positive potential. Furthermore, apart from having the lowest necrosis rate, the histopathological examination showed a reduced inflammatory response and a strong angiogenic effect. From the laboratory data, it could be observed that the CAP group had the lowest number of white cells. The CAP activated solution has been used in chronic human wounds, with good results regarding the stimulation of the wound skin cells, as well as with a strong antimicrobial effect. There is a possibility that the CAP activated solution stimulates angiogenesis in a strong manner and pushes the maximal vasodilation of the anastomotic skin vessels higher, as it does to keratinocytes, fibroblasts, or other cells [23; 46; 47]. Its effects on the skin microcirculation are related to the release of the nitric oxide (NO) and nitrogen dioxide (NO\({}_{2}\)). An efficient CAP activated solution should maximize the NO/NO\({}_{2}\) levels and lower the reactive oxygen species level 1. Nowadays, the devices that use CAP for skin treatment are non-invasive. The model we depicted required injections in the flap, at a fixed distance from the pedicles. A minor area of skin necrosis was also found in the L-NAME group. A single dose of this substance was injected, and as it is an inhibitor of NO synthesis, the resulted necrosis rate of 18.2% was not expected. It is known that L-NAME administration has a dose dependent effect on flap survival [20; 21], and we showed that, therefore, a single administration of a dose of L-NAME (30 mg/kg) can increase flap survival. The triggers for this may be that the ischemia and the lesions produced by the surgical manipulation of the flap, as well as the single dose that blocks the synthesis of the NO, might be able to stimulate the compensatory production of NO [21], despite its use as an NOS antagonist.