| Background: Random skin flap is widely used for its advantages of flexibility and convenient among the fields of burns, wounds, plastic and so on. While there is no clear blood supply, the random skin flap would be necrosis partly or all. This is the most common complication in transplantation of flap, which influence the effect of the operation. Based on a large number of experimental and clinical experiences, the design of skin flap often follows by long: width =(1.5-2):1, to avoid distal flap be necrosis by ischemia-reperfusion injury and to improve the survival of the flap area. Therefore, to increase the blood flow of the skin flap and avoid ischemia-reperfusion injury has become an important mean and a research hot spot to improve the survival rate of skin flap.Ohsawa’s study has shown that 2% hydrogen could effectively reduce the cerebral infarct area that is caused by ischemia-reperfusion injury [2], which presents a research boom of that hydrogen molecule could be an antioxidant. A large number of reports in succession confirm that bi- hydrogen has a protective effect to decrease ischemia-reperfusion injury of many tissues and organs, for example, brain [3-4], liver [5], kidney [6], heart [7] and retina [8]. But there is still no study to confirm whether hydrogen has an active influence in random flap as an antioxidant. In the follow experiment, in which rats of the experimental group drink hydrogen-rich saline, to investigate the effect of hydrogen on the survival area and flow of rats’ random skin flap and to discussits possible mechanism.Objective: As an antioxidant, hydrogen-rich saline can selectively remove hydroxy-1 and nitrous anion, to reduce ischemia-reperfusion injury and do the protection. So far, there are so less reports about the effect of hydrogen-rich saline on ischemia-reperfusion injury of skin flap. In the follow experiment, we feed hydrogen-rich saline to experimental group, while feed common saline to control group, by the model of random skin flap at rats’ back and the pathological observation, the ratio of survival area and the blood flow for times, to explore the effect of hydrogen-rich saline on random skin flap. Methods: 1. The preparation o f hydrogen-rich saline: According to the specification, put hydrogen stick and 500 ml normal saline together into 550 ml plastic bottle to continual generate hydrogen. In order to ensure the hydrogen concentration, replace normal saline once every 24 h and clean hydrogen stick regularly according to the specification. Test the concentration of the hydrogen in hydrogen-rich saline by gas chromatography. The result is around 0.73mmol/L, reaching 0.6mmol/L which is the effective hydrogen concentration for treatment. 2. Model preparation and Grouping: Rats are randomly divided into two groups: the experimental group(A) and control group(B), 36 rats of each group. Each group is randomly divided into four small groups, named A1, A2, A3, A4 and B1, B2, B3, B4, 9 rats of each group. The experimental group drinks hydrogen-rich saline and the control group drinks normal saline from pre-operative 2 weeks to death. 10% chloral hydrate 3 ml/kg is injected into abdominal. After anesthesia, fix the rat on the surgery table, remove hairs and clean skin on the back. Design random pattern skin flap on the back of rat by methylene blue(figure 1): 9.0 cm x 1.5 cm. The flap pedicle is on the caudal part, about 1.5cm from the tail. Divide the flap into 3 parts(figure 1). Remain the capillary network under the leather flap. Stop bleeding completely, and use a 3-0 silk to manage the flap by interrupted suture in situ [9]. Operation must follow the principle of aseptic and rats are in single cage after operation. 3, Management of specimens: A1 and B1, A2 and B2, A3 and B3, A4 and B4 were killed by cervical dislocation at the time point of immediate post-operation, 1d, 3d, 7d. Tissue specimens that are cut from middle part(Ⅱ area) anddistal part(Ⅲ area), are fixed by 10% formaldehyde solution, paraffin embedding, serial sections and HE staining. Do histological observation under light microscope. Measure the flap blood flow by the La ser-Doppler measurement at the d ifferent times. Results: 1) Compared with control group, the blood flow of proximal and middle portions increase significantly in the experimental group(p<0.05); 2) There is a significant difference in the survival rate of flap between the experimental group and control group at 7d after operation(p<0.05); 3) Postoperative morphologic observation shows that the survival skin flap of the experimental group is better than the control group. There is less necrosis in the experimental group; 4) it confirms that the experimental group has less tissue inflammatory cell infiltration and edema by postoperative histopathological observation. Conclusion: Drinking hydrogen-rich saline has an obviously protective effect on ischemia-reperfusion injury of the skin flap. It can improve microcirculation blood flow and improve the survival ratio. The possible mechanism may lie in that the hydrogen can improve microcirculation blood flow, remove free radical selectively to relieve ischemia-reperfusion injury, maintain the dynamic balance between antioxidant and free radical, and inhibit or reduce generation of inflammatory factors. |
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