【作者】 王晖；

【导师】 罗成群；

【作者基本信息】 中南大学 ， 外科学， 2010， 博士

【摘要】 应用皮瓣修复创面是整形外科最常用的修复手段。为预防皮瓣的缺血坏死,确保皮瓣的成活率,对皮瓣预先进行延迟仍然被公认为是提高皮瓣成活率最有效的方法。外科延迟皮瓣成活率高,需要二次手术,住院时间长,患者负担加重。目前对于皮瓣的药物延迟法研究已经有了一定的进展,Karacaoglu等研究表明通过肾上腺素微球局部注射,可以使皮瓣产生延迟作用。本研究使用肾上腺素药物简单注射,在确定可以产生良好的皮瓣延迟效应的基础上,根据纳米柔性脂质体可以作为透皮给药的载体,并且有缓控释放的特点,配制了肾上腺素纳米柔性脂质体,并经过一系列的检测后,证实了肾上腺素纳米柔性脂质体可以缓释透皮给药,且制备工艺可行,质量控制方法简便、可靠。本研究以大鼠为实验动物,在背部设计跨区域的超长皮瓣,比较肾上腺素纳米柔性脂质体透皮给药组、其他方式的肾上腺素给药组、手术组及对照组的皮瓣存活率,结果显示：肾上腺素纳米柔性脂质体透皮给药可极大的提高延迟效果,同时通过透皮给药法,可避免手术延迟给患者带来的痛苦,降低医疗费用,具有广阔的临床应用价值。肾上腺素纳米柔性脂质体透皮给药这种新颖的无创给药思路可成为取代传统手术延迟法的一种新方法。目的制备肾上腺素纳米柔性脂质体,并对其进行表征。方法采用逆向蒸发法制备水溶性盐酸肾上腺素纳米柔性脂质体,研究柔性脂质体的包封率及流动性。测定不同超声条件下柔性脂质体的粒径变化,确定实验的最佳超声功率和时间值。透射电镜下观察标准实验状态下配制的肾上腺素纳米柔性脂质体的形态。结果肾上腺素柔性脂质体包封率为61.83%,柔性脂质体胶体溶液的相对透过速率值结果平均为70.49%。在透射电镜下观察,肾上腺素纳米柔性脂质体在超声条件50%功率,超声14分钟时能得到稳定的纳米粒径在80nm左右。结论肾上腺素纳米柔性脂质体制备工艺可行,质量控制方法简便、可靠。制剂的包封率,粒径大小均符合常规药典要求。目的探讨利用肾上腺素纳米柔性脂质体进行透皮给药的可行性。方法建立了肾上腺素高效液相色谱分析方法,采用Franz扩散池法进行肾上腺素纳米柔性脂质体离体经皮扩散研究,测定其24 h时累积透皮吸收百分率。结果色谱条件为流动相：甲醇：0.02mol／L磷酸二氢钾溶液(V／V)=25：75,流速：1.0 mL·min-1,检测波长：280nm,24 h肾上腺素纳米柔性脂质体累积透皮吸收百分率为47.6%结论肾上腺素纳米柔性脂质体透皮速率和吸收百分率均符合常规药典要求。目的探索皮瓣延迟新方法,提高皮瓣延迟效率,减少手术创伤和患者所承受的痛苦,利用肾上腺素纳米柔性脂质体的缓释效应,探索无创的皮外给药方法的可行性。方法健康雄性Wister大鼠252只。取大鼠下背部皮瓣,5cm×1 cm大小,跨越中线区2.5 cm,双侧主干血管在中线区有着恒定的吻合支。将252只大鼠随机分为6组：手术延迟组(A)、肾上腺素注射延迟组(B)、生理盐水对照组(C)、肾上腺素脂质体注射延迟组(D组),肾上腺素脂质体透皮给药延迟组(E组)和肾上腺素脂质体透皮加注射给药联合延迟组(F组),每组皮瓣分别再分成未经延迟组(A1、B1、C1、D1、E1、F1),延迟3天皮瓣组(A2、B2、C2、D2、E2、F2),延迟7天皮瓣组(A3、B3、C3、D3、E3、F3),延迟14天皮瓣组(A4、B4、C4、D4、E4、F4)。观测各组皮瓣毛细血管密度、乳酸含量、VEGF表达及明胶氧化铅灌注大体标本,比较各组的皮瓣成活率。结果A组皮瓣自延迟3天开始,皮瓣中线区血管密度开始增加,随着延迟时间的延长,乳酸浓度依次下降,皮瓣成活率依次升高。B组D组乳酸含量在注射7天和14天后并不如A组逐渐下降反而处于一个比较稳定的值,而F组和E组却持续性上升。A组、B组、D组、E组和F组皮瓣成活率按观测时间点的长短依次升高,A组、B组和D组组间差异无统计学意义；毛细血管密度、乳酸含量差异三组亦无统计学意义。而采取了透皮给药方式的E组和F组在各项指标上都优于上述三组,统计学上有显著性差别。其中联合用药的F组在各项指标上又优于单纯透皮给药的E组,在统计学上有显著性差别。五组数据都和生理盐水对照组有统计学上的显著性差别。结论利用脂质体的缓释效应,配制肾上腺素纳米柔性脂质体,通过透皮给药,提高皮瓣存活率,创造了一种新型的无创延迟方法,具有较高的临床实用价值。更多还原

【Abstract】 The skin flap is the most commonly used means for wound repair in plastic surgery. For the prevention of flap necrosis and ensuring flap survival rate, advance delay of the flap is still recognized as the most effective way in raising the survival rate. Surgical delay can get a high survival rate, but need for secondary surgery, much more long hospital stay, the patients’burden is increased.At present, some progress has made in drug flap delay, Karacaoglu et al have shown that local injection of adrenaline microspheres can make the role of delayed in flap. In this study, we use simple injection of epinephrine drugs, and make sure a good delayed effect of the flap. We prepare the adrenaline flexible nano-liposome according to the flexible nano-liposomes could serve as a transdermal drug delivery carrier and it’s slow controlled release characteristics. A series test show that adrenaline flexible nano-liposome could be slow-release in transdermal drug delivery, and the preparation technology is feasible, and the quality control method is simple and reliable.In this study, ultra-long cross-regional flap was designed in the back of the rats, comparing survival rate of adrenaline flexible nano-liposome transdermal delivery groups, other forms of epinephrine treatment group, surgery group and control group, the results showed that:delay effect can be greatly improved by adrenaline flexible nano-liposome transdermal delivery.At the same time, we can avoid the painful delay surgery in patients, cut medical costs and get a broad clinical application. Adrenaline flexible nano-liposome transdermal delivery can replace the traditional surgical method as a novel idea of non-invasive drug delivery way.Objective:The purpose of the present study was to prepare epinephrine nano-flexible liposomes and to exosyndrome it.Method:Epinephrine nano-Flexible liposomes were prepared by reverse evaporation method, it’s encapsulation efficiency and liquidity were observed. Flexible liposome size was measured under the conditions of different ultrasonic to determine the best experimental values of ultrasonic power and time. The morphous of Flexible nano-liposome particle was precisely measured under the transmission electron microscope under the standard experiments state.Results:The encapsulation efficiency of water-soluble hydrochloride Epinephrine flexible liposomes was 61.83%, the relative permeation rate of flexible liposome colloidal solution was 70.49%. Observed by the transmission electron microscope, epinephrine flexible nano-liposomes can get a stable nano-particle size of about 80nm under the ultrasonic conditions of 50% power and 14 minutes.Conclusion:The preparation technology of adrenaline flexible liposomes is feasible, the quality control method is simple and reliable. The adrenaline flexible liposomes’encapsulation efficiency and particle size are in line with general requirements of the pharmacopoeia.Objective:The purpose of the present study was to research the feasibility of adrenaline flexible nano-liposome transdermal delivery.Methods:The methods of analysis adrenaline by high performance liquid chromatography was established and using amended Franz diffusion cells to transdermal delivery of adrenaline flexible nano-liposome across rat skin in vitro. The cumulative transdermal absorption percentages were observed after 24h.Results:Chromatographic conditions were mobile phase:methanol: 0.02mol/L potassium dihydrogen phosphate solution (V/V)=25:75, flow rate:1.0 mL·min-1, detection wavelength:280nm, the cumulative transdermal absorption percentage in 24h was 47.6%. Conclusion:adrenaline flexible nano-liposome penetration rate and absorption rate are in line with general requirements of the pharmacopoeia.Objective:The purpose of the present study was to explore new ways in flap delay, enhance the efficiency of flap delay, reduce surgical trauma and the suffering of patients, explore the feasibility of non-invasive transdermal method.Method:252 healthy male Wister rats were taken back-flap about 5 cm×1 cm, across the midline area 2.5 cm, bilateral main vessel in the middle zone had a constant anastomosis. The 252 rats were randomly divided into 6 groups:surgical delay group (A), epinephrine injection delayed group (B), normal saline control group (C), epinephrine injection liposome delayed group (D group), epinephrine liposome transdermal delivery delay group (E group),epinephrine liposome transdermal and injected delayed group (F group), each flap were then divided into groups, without delay (A1, B1, C1, D1, E1, F1), delayed three days (A2, B2, C2, D2, E2, F2), delayed 7 days (A3, B3, C3, D3, E3, F3), delayed 14 days (A4, B4, C4, D4, E4, F4). Capillary density, lactic acid content, VEGF expression and oxide gelatin infusion in general samples were observed, comparing flap survival rate of each group.Results:Since delayed three days, A group flaps’blood vessel density were increased in the midline area, lactic acid concentration were descending with the extended delay time, following by increased flap survival rate. In B and D group delay 7 days and 14 days after the injection,lactic acid in a relatively stable value, but F and E group had sustained rise. The survival rate of A group, B group, D group, E group and F group were increasing according to the observation time. A group, B group and D group had no significant statistically difference between groups (p>0.01); capillary density, lactic acid content of the three groups had no significant statistically difference (p> 0.01). E group and the F group are better than ABD groups in all indicators, there was a significant statistical difference (p<0.01). Group F was better than E group, there was a significant statistical difference (p<0.01). Compared A, B, D, E, and F group with C group, there was a significant statistical difference (p <0.01).Conclusion:We made use of the slow-release effect of liposomes, and prepared nano-flexible liposomes epinephrine, through the transdermal delivery, improved skin flap survival rate, created a new type of non-invasive method of delays with a high clinical value.更多还原