| Objective:This study aimed to investigate the effect of vacuum sealing drainage (VSD) on partial micro-environment, cytokine expression, immune cell function, inflammatory status and wound healing of rabbit wound model by different negative pressure, and prevention and treatment of crush syndrome and multiple organ dysfunction syndrome, and treatment of Gustilo III fracture.Methods:Animal experiment:12 cases of rabbit's wound models were undergoing the VSD (vaccum group)by different negative pressure(-75 mmHg, -125 mmHg,-225 mmHg and-350 mmHg)(1) Variation of oxygen partial pressure (PtO2) was measured by oxygen partial pressure admeasuring apparatus, area of VSD dressing and surface of wound were measured by vernier caliper, growth of anaerobic bacteria was detected by bacterial culture, progress of wound repair was examined by HE slice of tissue.(2) Expression of hypoxia inducible factor-la (HIF-1α) mRNA was measured by real-time fluorescent quantitative PCR, content of vascular endothelial growth factor (VEGF), tumor necrosis factor-α(TNF-a) and interleukin-6 (IL-6) were measured by ELISA after tissuehomogenate.(3) Vascular endothelial cells (VEC), new blood capillary (NBC), macrophages (Mφ), polymorphonuclear neutrophils(PMN)and lymphocytes (LC) of HE slice of tissue were counted by using light microscope.(4) Number of peripheral white blood cells (WBC) was counted by automatic rlood cell analyzer, content of C-reactive protein (CRP) was measured by BNⅡspecial protein analyzer, content of peripheral TNF-a and IL-6 were measured by ELISA, and vaccum group were compared with conventional group.Clinical research:(1) 102 cases of crush injury patients were random sampled and divided into study group (emergency VSD group 50 cases and delayed VSD group 22 cases) and control group (non-VSD group 30 cases), then analyzed the variance of index of crush syndrome after VSD between and in each group.(2) 90 patients admitted to Wuhan Tongji hospital between Oct 2005 and Feb 2009 were included in this retrospective study and were classified according to treatment method. Overall,53 cases underwent emergency VSD operations and 28 cases underwent delay VSD operations and 9 cases without VSD operations. The groups of patients were compared in terms of parameters such as repair deficiency of raw surface, promote union of fracture, prevent of post traumatic secondary strike and treatment outcomes after operation.(3) 94 cases of multiple traumatic patients with serious skin and soft tissue injury were random sampled and divided into study group (56 cases) and control group (38 cases), then analyzed the variance of "primary strike" and "secondary strike" related to the MODS and prognosis between two groups.Results:Animal experiment:(1) Average normal value of PtO2 was 5.79±0.50 Kpa, PtO2 value of wound was 3.06±1.20 Kpa, and negative pressure group was significant lower than conventional group and normal group (P<0.01). PtO2 was drop with increasing of negative pressure at the same time-point. PtO2 was reducing within 7 days, it falle to 80.94% of basic value in 5 seconds. Area of VSD dressing was significantly decreased after drainage (P＜0.01). Area was reduced to 65.36% of basic area after 5 minutes, then 62.82% after 7 days, and negative pressure group was significant lower than conventional group and normal group(P<0.01). There was no evidence of growth of anaerobic bacteria.(2) Expression of HIF-la and content of VEGF of negative group were increased after 30 minutes and significant lower than conventional group(P＜0.01), and both of them were reduced after 24 hours (P<0.05). Counting numbers of VEC and NBC of negative group were increased at the same time-point and significant higher than conventional group (P<0.05)(3) Contents of TNF-a and IL-6 of two groups were increased after 30 minutes. Counting numbers of Mφand PMN of two groups were increased after 24 hours. Counting number of Mφof two groups were reduced after 6 days, and negative pressure group was significant higher than conventional group (P＜0.05). Counting number of PMN of negative pressure group was reduced after 3 days, and it significant lower than conventional group which was reduced after 4 days (P<0.01). Counting numbers of LC of negative pressure group reached the peak value at 24 hours, then it started to decrease and significant lower than conventional group (P＜0.05). Contents of TNF-a of two groups reached the peak value at 24 hours, then they started to decrease. Negative pressure group was significant lower than conventional group after 1 hour (P<0.05). Contents of IL-6 of two groups reached the peak value at 24 hours, then they started to decrease. Negative pressure group was significant lower than conventional group after 12 hours (P＜0.05)(4) Number of WBC, content of CRP and IL-6 of negative pressure group were significant lower than conventional group after 6 hours (P<0.05). Content of TNF-a of negative pressure group was significant lower than conventional group after 24 hours (P< 0.01)Clinical research:(1) The evaluate index of emergency VSD group and delayed VSD group after operation, such as vital sign, urine volume, laboratory examination results, the incidence rate of crush syndrome, complications, fatality rate and so on were better than delayed VSD group and non-VSD group (P<0.05), the evaluate index of crush syndrome patients after operation in study group, such as complications, length of stay, fatality rate and so on were better than control group (P<0.05)(2) The speed of repair deficiency of raw surface, speed of union of fracture, systemic inflammatory response syndrome, infection of raw surface, sepsis, osteofascial compartment syndrome, ostemyelitis, ncidence rate of MODS and mortality rate in the both VSD groups were significant different from non-VSD group respectively (P<0.05)(3) The evaluate index of study group after operation, such as body temperature, shock, acidosis, blood clotting function, systemic inflammatory response syndrome, infection, sepsis and so on were significant better than control group (P<0.05), incidence rate of MODS and mortality rate were significant lower than control group (P<0.01)Conclusion:Animal experiment:(1) PtO2 could be reduced significantly by VSD without growth of anaerobic bacteria, and area of wound was reduced for promoting the wound healing.(2) Expression of HIF-la and content of VEGF were increased by VSD for enhanceing differentiated state of VEC and construction of NBC, which were better for vascularization and wound healing.(3) Microenvironment of negative pressure caused by VSD could increase the number of Mφ, and reduced the numbs of PMN and LC, decreased the contents of TNF-a and IL-6. It compartmentalized inflammation and avoided aggregation of inflammatory cell to manage uncontrolled inflammatory response.(4) Number of WBC, content of CRP, TNF-αand IL-6 were reduced by suction of nflammatory fluids, then reduce inflammatory response to avoid the systemic inflammatory response.Clinical research:VSD can promote the vascularization and granulation tissues of wound, improve to close raw surface of Gustilo III fracture early and wound healing, reduce the inflammatory response and was helpful to prevent and treat the infection, crush syndrome, MODS and reduce the mortality rate.
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