Long term efficacy of absorbable sutures coated with silver nanoparticles on the suture tract of skin wounds in mice
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摘要:
目的 探讨纳米银涂层可吸收缝线对小鼠皮肤切口缝合线道的远期作用。 方法 该研究为实验研究。将18只10~12周龄雌雄各半BALB/c小鼠,按照随机数字表法分为普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组,每组6只,雌雄各半。于所有小鼠背部制备皮肤切口后,对普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组小鼠皮肤切口分别采用可吸收薇乔线、三氯生抗菌涂层可吸收薇乔线和纳米银涂层可吸收薇乔线行皮内连续缝合。术后25 d,采集切口缝合线道皮肤组织,行免疫组织化学染色观测小鼠切口缝合线道F4/80、白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）阳性面积百分比,行原位末端标记（TUNEL）染色观测小鼠切口缝合线道TUNEL染色阳性面积百分比,行Masson染色观察小鼠切口缝合线道胶原纤维形成情况,行免疫组织化学染色观测小鼠切口缝合线道组织蛋白酶K阳性面积百分比。以上实验中样本数均为6。 结果 术后25 d,纳米银薇乔线组小鼠切口缝合线道F4/80、IL-6及TNF-α阳性面积百分比[（19.2±1.6）%、（20.2±1.7）%、（16.0±1.6）%]均明显低于普通薇乔线组[（100±6.4）%、（100±7.6）%、（100±9.6）%]和抗生素薇乔线组[（47.2±3.2）%、（53.8±5.0）%、43.2%,P<0.05],抗生素薇乔线组小鼠切口缝合线道F4/80、IL-6及TNF-α阳性面积百分比均明显低于普通薇乔线组（P<0.05）。术后25 d,纳米银薇乔线组小鼠切口缝合线道TUNEL染色阳性面积百分比明显低于普通薇乔线组和抗生素薇乔线组（P值均<0.05）,抗生素薇乔线组小鼠切口缝合线道TUNEL染色阳性面积百分比明显高于普通薇乔线组（P<0.05）。术后25 d,普通薇乔线组小鼠切口缝合线道胶原纤维沉积不明显,抗生素薇乔线组小鼠切口缝合线道无明显胶原纤维,纳米银薇乔线组小鼠切口缝合线道胶原纤维致密程度最高且排列有序。术后25 d,纳米银薇乔线组小鼠切口缝合线道组织蛋白酶K阳性面积百分比明显低于普通薇乔线组和抗生素薇乔线组（P值均<0.05）,抗生素薇乔线组小鼠切口缝合线道组织蛋白酶K阳性面积百分比明显低于普通薇乔线组（P<0.05）。 结论 在小鼠切口缝合线道,纳米银薇乔线的远期抗炎疗效优于三氯生抗菌涂层可吸收薇乔线和普通可吸收薇乔线；同时,采用纳米银薇乔线缝合的小鼠切口缝合线道细胞凋亡减少,胶原纤维沉积显著,迟发型超敏反应最弱,说明纳米银薇乔线具备潜在的临床应用价值。 Abstract:Objective To explore the long term efficacy of absorbable sutures coated with silver nanoparticles on the suture tract of skin wounds in mice. Methods This study was an experimental research. Eighteen half male and half female BALB/c mice aged 10-12 weeks were divided into normal Vicryl suture group, antibiotic Vicryl suture group, and silver nanoparticle Vicryl suture group, according to the random number table, with 6 mice in each group, half male and half female. After the skin wounds were created on the back of all mice, the skin incisions of mice in normal Vicryl suture group, antibiotic Vicryl suture group, and silver nanoparticle Vicryl suture group were performed with continuous intradermal suture by absorbable Vicryl sutures, absorbable Vicryl sutures coated with triclosan antimicrobial, and absorbable Vicryl sutures coated with silver nanoparticles, respectively. On the 25th day after surgery, the skin tissue in the suture tract was collected, immunohistochemical staining was used to detect the positive area percentages of F4/80, interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) in the wound suture tract of mice, the TdT-mediated dUTP nick-end labeling (TUNEL) staining was used to detect positive area percentage of TUNEL in the wound suture tract of mice, Masson staining was used to observe the collagen fiber formation in the wound suture tract of mice, and immunohistochemical staining was used to detect the positive area percentage of tissue cathepsin K in the wound suture tract of mice. The number of the above experimental samples was 6. Results On the 25th day after surgery, the positive area percentages of F4/80, IL-6, and TNF-α in the wound suture tract of mice in silver nanoparticle Vicryl suture group ((19.2±1.6)%, (20.2±1.7)%, and (16.0±1.6)%) were significantly lower than those in normal Vicryl suture group ((100±6.4)%, (100±7.6)%, and (100±9.6)%) and antibiotic Vicryl suture group ((47.2±3.2)%, (53.8±5.0)%, and (43.2±0)%, P<0.05). The positive area percentages of F4/80, IL-6, and TNF-α in the wound suture tract of mice in antibiotic Vicryl suture group were significantly lower than those in normal Vicryl suture group (P<0.05). On the 25th day after surgery, the positive area percentage of TUNEL in the wound suture tract of mice in silver nanoparticle Vicryl suture group was significantly lower than those in normal Vicryl suture group and antibiotic Vicryl suture group (with P values all <0.05), the positive area percentage of TUNEL in the wound suture tract of mice in antibiotic Vicryl suture group was significantly higher than that in normal Vicryl suture group (P<0.05). On the 25th day after surgery, the collagen fiber deposition was not obvious in the wound suture tract of mice in normal Vicryl suture group, no obvious collagen fiber formation was observed in the wound suture tract of mice in antibiotic Vicryl suture group, while the collagen fibers were mostly densely arranged and well-ordered in the wound suture tract of mice in silver nanoparticle Vicryl suture group. On the 25th day after surgery, the positive area percentage of tissue cathepsin K in the wound suture tract of mice in silver nanoparticle Vicryl suture group was significantly lower than those in normal Vicryl suture group and antibiotic Vicryl suture group (with P values all <0.05), the positive area percentage of tissue cathepsin K in the wound suture tract of mice in antibiotic Vicryl suture group was significantly lower than that in normal Vicryl suture group (P<0.05). Conclusions In the wound suture tract of mice, silver nanoparticle Vicryl sutures exhibit superior long-term anti-inflammatory effects compared with absorbable Vicryl sutures coated with triclosan antimicrobial, and absorbable Vicryl sutures. Additionally, cell apoptosis is reduced, collagen fiber deposition is obvious, and delayed-type hypersensitivity reactions is the weakest in the wound suture tract of mice sutured by silver nanoparticle Vicryl sutures. The silver nanoparticle Vicryl sutures have potential value for clinical application. -
Key words:
- Wounds and injuries /
- Inflammation /
- Apoptosis /
- Nanoparticles /
- Wound repair
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参考文献
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图 3 3组小鼠术后25 d切口缝合线道F4/80、IL-6和TNF-α的阳性表达情况 辣根过氧化物酶-二氨基联苯胺×200。3A、3B、3C与3D、3E、3F与3G、3H、3I.分别为普通薇乔线组、抗生素薇乔线组切口缝合线道F4/80、IL-6及TNF-α阳性表达,和纳米银薇乔线组切口缝合线道F4/80、IL-6及TNF-α阳性表达,其中图3C的阳性表达低于图3A、3B,图3F的阳性表达低于图3D、3E,图3I的阳性表达低于图3G、3H
注：对普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组小鼠皮肤切口分别采用可吸收薇乔线、三氯生抗菌涂层可吸收薇乔线和纳米银涂层可吸收薇乔线缝合；IL-6为白细胞介素6,TNF-α为肿瘤坏死因子α；F4/80、IL-6和TNF-α阳性染色均为棕色,其中F4/80染色阳性反映巨噬细胞分布
图 4 3组小鼠术后25 d切口缝合线道细胞凋亡情况 辣根过氧化物酶-二氨基联苯胺×200。4A、4B、4C.分别为普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组切口缝合线道细胞凋亡情况,图4C的细胞凋亡情况轻于图4A、4B
注：对普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组小鼠皮肤切口分别采用可吸收薇乔线、三氯生抗菌涂层可吸收薇乔线和纳米银涂层可吸收薇乔线缝合；细胞原位末端标记阳性染色为棕色,反映细胞凋亡
图 5 3组小鼠术后25 d切口缝合线道胶原纤维形成情况 Masson×200。5A、5B、5C.分别为普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组小鼠切口缝合线道胶原纤维形成情况,图5C胶原纤维形成情况优于5A、5B
注：对普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组小鼠皮肤切口分别采用可吸收薇乔线、三氯生抗菌涂层可吸收薇乔线和纳米银涂层可吸收薇乔线缝合；肌细胞呈红色,胶原纤维呈蓝色
图 6 3组小鼠术后25 d切口缝合线道异物巨细胞积聚情况 辣根过氧化物酶-二氨基联苯胺×200。6A、6B、6C.分别为普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组切口缝合线道异物巨细胞积聚情况,图6C的异物巨细胞积聚少于图6A、6B
注：对普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组小鼠皮肤切口分别采用可吸收薇乔线、三氯生抗菌涂层可吸收薇乔线和纳米银涂层可吸收薇乔线缝合；组织蛋白酶K阳性染色为棕色,反映异物巨细胞表达
Table 1. 3组小鼠术后25 d切口缝合线道F4/80、IL-6和TNF-α阳性面积百分比比较（%,
组别 样本数 F4/80 IL-6 TNF-α 普通薇乔线组 6 100±6.4 100±7.6 100±9.6 抗生素薇乔线组 6 47.2±3.2 53.8±5.0 43.2 纳米银薇乔线组 6 19.2±1.6 20.2±1.7 16.0±1.6 F值 1 500.84 607.06 5 117.59 P值 <0.001 <0.001 <0.001 P1值 <0.001 <0.001 <0.001 P2值 <0.001 <0.001 <0.001 P3值 <0.001 <0.001 <0.001 注：对普通薇乔线组、抗生素薇乔线组和纳米银薇乔线组小鼠皮肤切口分别采用可吸收薇乔线、三氯生抗菌涂层可吸收薇乔线和纳米银涂层可吸收薇乔线缝合；IL-6为白细胞介素6,TNF-α为肿瘤坏死因子α；F值、P值为组间各指标总体比较所得；P1值、P2值、P3值分别为普通薇乔线组与抗生素薇乔线组、普通薇乔线组与纳米银薇乔线组、抗生素薇乔线组与纳米银薇乔线组比较所得 
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