力度可控性滚轮微针的构建及其对大面积皮肤药物吸收影响研究

发布时间：2018-05-12 11:34

本文选题：力度可控性滚轮微针 + 增生性瘢痕　；参考：《宁夏医科大学》2014年硕士论文

【摘要】：目的构建力度可控性滚轮微针并研究其在大面积皮肤药物吸收中的影响。方法1.构建力度可控性滚轮微针，采用石蜡切片、HE染色法考察其力度可控性，Franz扩散池法考察释药重现性，Score法测定皮肤刺激性；Franz扩散池法、亚甲蓝染色、CLSM法筛选微针密度、使用力度及不同处理次数；TEWL法、激光多普勒法考察微针对健康裸鼠、健康人体及瘢痕病人皮肤的刺激性。 2.采用洗脱法、组织匀浆法比较微针及注射针对地塞米松磷酸钠在体不同时间的总吸收量、皮肤及血浆药量；Franz扩散池法研究其体外渗透特性；冰冻切片后进行组织匀浆考察微针及注射针对地塞米松磷酸钠均匀性的影响；SEM法、TEWL法、激光多普勒法考察皮肤刺激性。 3.采用激光多普勒法测定不同浓度川芎挥发油和去氧肾上腺素对健康裸鼠皮肤血流的影响，Franz扩散池法对比其对裸鼠离体皮肤的作用，洗脱法、组织匀浆法研究改变血流对地塞米松磷酸钠在体吸收及分布的影响，Score法、组织学检测皮肤刺激性。结果1.滚轮微针改装后有效的提高了力度可控性、释药重现性，且皮肤刺激性小。力度与深度实验结果为皮肤微孔深度随微针使用力度的增加而加大，微孔深度范围由65.28～106.25μm收窄为改装后的71.53～97.92μm，其RSD范围由10.45%～19.69%显著下降至改装后的2.33%～9.21%。在相同微针使用力度下，曲安奈德的累积释药量RSD范围由改装前的34.75%～55.92%，显著下降至改装后的14.29%～29.73%。改装滚轮微针仅在用药1h时出现轻度皮肤刺激性，之后消失。微针密度、使用力度筛选实验结果为540针、3.5~4.0N效果较好。540针微针组维A酸的累积渗透量是对照组的6.8倍，较192针微针组的2.5倍促渗作用更强(P＜0.05)；2.5~3.0N使用力度时，给药2h维A酸渗透深度为30μm，而3.5~4.0N与4.5~5.0N两组的深度无显著差异，渗透深度均为40μm。微针处理次数结果显示，微针使用次数越多促透作用越好，药物吸收越均匀，但8次以后趋于平衡，皮肤刺激性实验结果显示5次及以下处理次数较安全。处理10次时的48h累积渗透量和渗透速率均最高，分别为对照组的3.1倍和4.1倍（P0.05），较处理8次组的2.6倍和3.0倍增加不明显；8、10次处理组的皮肤维A酸滞留量相近，约是对照组的3.0倍。亚甲蓝染色与激光共聚焦结果显示，微针孔数量随微针使用次数的增加逐渐增多并趋于均匀，而使用8次与10次效果相似。TEWL与激光多普勒血流结果显示，微针处理后TEWL和血流均迅速增加，并随使用次数的增加对皮肤的刺激作用增大；处理10次时对TEWL和皮肤血流的影响均最大，且恢复时间也最长；1、3、5次处理组TEWL恢复时间为24h，8、10次为48h；皮肤血流恢复时间除8、10次在120min外，其余处理组均在60min内恢复基线。微针健康人体TEWL和血流影响均在90min内完全恢复，而在瘢痕患者45min完全恢复。 2.微针给药技术较皮内注射给药的优点在于皮肤促透作用高效持续、皮肤中药物分布量反而高于皮内注射组，且分布更加均匀，皮肤刺激性相对较小。皮内注射组裸鼠体外快速渗透，4h透皮累积药量高达83.76%，而后缓慢释药；而微针组则体外释药缓慢，4h透皮累积药量仅为25.75%，但相对持续。皮内注射组皮肤滞留药量较微针组少，12h皮肤滞留药量仅为微针组的36.14%。皮内注射组裸鼠体内1h皮肤滞留药量为微针组的94.52%，随后皮肤滞留药量逐渐下降，至12h仅为1h的18.96%；而微针组皮肤滞留药量随时间逐步增加，12h皮肤滞留药量是皮内注射组的6.92倍。此外，给药12h，微针组血浆药物浓度是对照组的3.16倍，是皮内注射组的2.25倍。皮内注射组注射点处皮肤滞留药量较高，注射点外药量较少，峰谷值相差286.21%；而微针组皮肤滞留药量相对均衡，峰谷值相差仅为114.61%。TEWL显示微针对皮肤表面失水量影响较注射组大，但24h后刺激消除；而激光多普勒血流量法显示皮内注射对皮肤血流的影响更大。 3.川芎挥发油和去氧肾上腺素对微针离体皮肤促透没有影响，但能通过改变在体皮肤血流达到对微针给药的调控作用。离体实验各处理组12h累积渗透量、渗透速率和12h皮肤内药物滞留量均无显著差异；在体实验中，含10%川芎挥发油组的给药部位皮肤药量最低，为1.45±0.42μg，，仅为对照组的0.5倍（P0.05），而含0.15%去氧肾上腺素组给药部位皮肤含药量为3.89±1.52μg，为对照组的1.32倍（P0.05）；血浆中药物含量规律相反，含3%川芎挥发油组和含10%川芎挥发油组分别是对照组的5.39倍和6.39倍（P0.05），而含0.15%去氧肾上腺素组仅为对照组的0.61倍（P0.05）；含10%川芎挥发油组药物总吸收量最高，为对照组的2.05倍，含0.15%去氧肾上腺素组总吸收量最小；皮肤刺激性实验各组1h时表现为轻度刺激强度，48h所有实验裸鼠均完全恢复。皮肤组织学检查结果显示，微针处理后6h，皮肤仅现轻微角质层剥离和缺失，加入川芎挥发油和去氧肾上腺素后刺激性均未增加，未现炎细胞等炎症反应。结论制备力度可控性滚轮微针后，有效提高了其力度可控性、释药重现性及给药均匀性，通过筛选滚轮微针的使用方法，增强了对增生性瘢痕大面积给药的可操作性。构建滚轮微针在解决了皮内注射给药存在的疼痛、给药不便等问题的同时，起到高效促透、皮肤药物浓集及分布均匀的作用。皮肤血流影响透皮药物在皮肤和血液中的分布，适当的去氧肾上腺素能降低局部皮肤血流有效增加给药病灶部位药量，降低药物入血。
[Abstract]:Objective to construct a dynamic controllable roller microneedle and study its effect on the absorption of large area skin drugs.
Method 1. the dynamics controllable roller microneedles were constructed. The strength controllability was examined by paraffin section and HE staining. The reproducibility of drug release was investigated by the Franz diffusion pool method. Score method was used to determine the irritation of the skin. Franz diffusion pool, methylene blue staining, and CLSM method were used to screen the density of microneedles, use intensity and different treatment times; TEWL method and laser Doppler method were used to investigate the micropin density. For healthy nude mice, skin irritation of healthy and scarred patients.
2. the method of elution and tissue homogenate were used to compare the total absorption of dexamethasone sodium phosphate and the dosage of dexamethasone sodium phosphate at different time in the body and the dosage of the skin and plasma; the Franz diffusion pool method was used to study the osmosis in vitro; the effect of microneedle and injection on the homogeneity of sodium phosphate of dexamethasone was investigated by tissue homogenate after frozen section; SEM method, TEWL Method, laser Doppler method was used to examine skin irritation.
3. the effects of different concentrations of volatile oil and epinephrine on the skin blood flow of healthy nude mice were measured by laser Doppler method. The effect of the Franz diffusion pool method on the skin of nude mice was compared. The effect of blood flow on the absorption and distribution of dexamethasone sodium phosphate in the body was studied by the elution method and tissue homogenate method. Score method and histological examination of skin were used. Skin irritation.
Results after the 1. roller microneedles were modified, the dynamic controllability, reproducibility and irritation of the skin were effectively improved. The results of strength and depth were increased with the increase of micropin intensity. The range of micropore depth was narrowed from 65.28 to 106.25 mu m to 71.53 to 97.92 mu m, and the range of RSD was from 10.45% to 19.69%. Under the use of the same micro needle, the cumulative release amount of Cu Ann Ned's RSD range from 2.33% to 9.21%. after the modification was 34.75% ~ 55.92% before the modification, which was significantly reduced to 14.29% ~ 29.73%. modified roller microneedles for mild skin irritation only at 1H.
Microneedle density, the results of the use of the strength screening test were 540 needles, and the effect of 3.5~4.0N was better than that of the control group. The cumulative permeability of the.540 needle microneedle group was 6.8 times more than that of the control group, and the infiltration effect was stronger than that of the 192 needle microneedle group (P < 0.05). The penetration depth of 2H vitamin A was 30 u m, while the depth of 3.5~4.0N and 4.5~5.0N two was not obvious when 2.5~3.0N was used. The depth of penetration is 40 u M.
The results of microneedle treatment showed that the better the use of microneedles, the better the penetration effect, the better the absorption of the drugs, but the more balanced after 8 times. The results of the skin irritation test showed that the 5 times and the following treatment times were more safe. The 48h cumulative permeability and the penetration rate of the 10 times were the highest, 3.1 times and 4.1 times of the control group (P0.05), respectively. The increase of 2.6 times and 3 times of the treatment in the 8 group was not obvious; the amount of A acid retention in the skin of the 8,10 treatment group was similar, about 3 times that of the control group. The methylene blue staining and laser confocal results showed that the number of micropinholes increased gradually with the increase of the number of microneedles, and the effect of the 8 and 10 times was similar to that of the laser Doppler. The blood flow results showed that the TEWL and blood flow increased rapidly after the microneedle treatment, and increased with the increase of use times. The effects on TEWL and skin blood flow were the largest and the recovery time was the longest when treated 10 times; the TEWL recovery time of the 1,3,5 treatment group was 24h, 8,10 was 48h, and the time of skin blood flow recovery was 1 except 8,10 times in 8,10. At 20min, the rest of the treatment groups were all restored to the baseline in 60min. The effects of TEWL and blood flow on the healthy human body were all recovered within the 90min, while the 45min in the scar patients recovered completely.
The advantage of 2. microneedle therapy is that the effect of intradermal injection is more efficient and continuous. The distribution of drugs in the skin is higher than that in the intradermal injection group, and the distribution is more uniform and the skin irritation is relatively small. The intradermal injection group is fast permeable in the nude mice in vitro, the 4H transdermal cumulative dosage is up to 83.76%, and then slowly release the drug; and the microneedle group The drug release in vitro was slow, and the cumulative dose of 4H was only 25.75%, but it was relatively sustained. The amount of skin retention in the intradermal injection group was less than that in the microneedle group. The amount of 12h retention in the 36.14%. intradermal injection group was only 94.52% in the nude mice of the group of microneedles, and the dosage of the skin stagnation gradually decreased to 12h only 18.96% of the 1H. The dosage of skin retention in the microneedle group increased gradually with time, and the amount of 12h skin retention was 6.92 times as high as that in the intradermal injection group. In addition, the drug concentration in the microneedle group was 3.16 times more than that of the control group and 2.25 times that of the intradermal injection group. The dosage of the injection point at the intradermal injection group was higher, the dosage of the injection point was less and the difference of peak and valley value was 286.21%. The amount of skin retention in the microneedle group was relatively balanced, and the difference of peak and valley value was only 114.61%.TEWL, but the effect of the skin water loss was larger than that of the injection group, but the stimulation after 24h was eliminated, while the laser Doppler blood flow method showed that intradermal injection had more effect on the skin blood flow.
3. the volatile oil of Ligusticum chuanxiong and deoxyadrenaline did not affect the penetration of microneedles in vitro, but it could regulate the effects of microneedle therapy on the blood flow in the body skin. There was no significant difference in the cumulative 12h permeability, the rate of osmosis and the drug retention in the 12h skin in the experimental group. In the body experiment, the volatile oil group of 10% Ligusticum chuanxiong was found. The dosage of the skin was the lowest, 1.45 + 0.42 mu g, only 0.5 times of the control group (P0.05), while the dosage of the 0.15% deoxyadrenaline group was 3.89 + 1.52 g, which was 1.32 times (P0.05) of the control group, and the drug content in the plasma was opposite, 3% of the volatile oil group and 10% Ligusticum chuanxiong volatile oil group were 5.3 of the control group. 9 times and 6.39 times (P0.05), and 0.15% deoxyadrenaline group was only 0.61 times of the control group (P0.05), and the total absorption of the volatile oil group containing 10% Ligusticum chuanxiong was the highest, which was 2.05 times of the control group, and the total absorption of 0.15% deoxyadrenaline group was the smallest. The 1H in the skin irritation test group was mild stimulation intensity, and all the experimental nude mice of 48h were finished. The results of skin histological examination showed that the skin was only slight cuticle dissection and deletion in 6h after microneedle treatment. After adding Ligusticum chuanxiong volatile oil and deoxyadrenaline, the irritation did not increase, and there was no inflammatory reaction.
Conclusion the controllability, reproducibility and uniformity of drug delivery are effectively improved after the preparation of the dynamics controllable roller microneedles. By screening the use of roller microneedles, the maneuverability of large area drug delivery for hypertrophic scars is enhanced. The construction of roller microneedles is a solution to the pain and inconvenience of intradermal injection. At the same time, the skin blood flow affects the distribution of the skin and blood in the skin and blood. The appropriate deoxyadrenaline can reduce the local skin blood flow effectively and reduce the dose of the focus on the focus of the focus, and reduce the entry of the drug into the blood.

【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

【参考文献】