Cellular and Molecular Research Center, Research Institute for Non-communicable Diseases, Qazvin University of Medical Sciences, Qazvin, Iran.
School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
Wound Manag Prev. 2020 Apr;66(4):38-46. doi: 10.25270/wmp.2020.4.3846.
Research to identify and develop compounds that facilitate wound healing is important, especially for hard-to-heal chronic wounds.
This study was conducted to investigate the effects of orally administered propolis (a resinous substance found in beehives), alone and in combination with silver nanoparticles (SNPs), on the wound healing process in male rats.
Forty (40) male Wistar rats were randomly divided into 4 groups of 10 each: 1 control group received no treatment, and 3 study groups received a daily dose of 1) propolis (100 mg/kg), 2) propolis + 30 ppm SNPs, or 3) propolis + 60 ppm SNPs. Healing rate was determined by wound surface area reduction on days 4, 6, 8, and 10 post-surgery. On day 12 after wound creation, histological changes of wound healing, including number of new vessels, inflammatory cells (neutrophils, eosinophils, and mast cells) and fibroblasts were counted based on morphology using a 400x objective lens, and collagen deposition density was determined using hematoxylin and eosin and trichrome staining, respectively. The histological scores were based on a 0 to 4 scale from lowest to highest amount of improving tissue status and were analyzed using one-way analysis of variance, Tukey test, Kruskal-Wallis test, t test, and Mann-Whitney U test to examine differences among the groups. Significance was set at P <.05.
The rate of wound healing was significantly different between the control and the treated groups on days 4, 6, 8, and 10 (percent change was not assessed on day 12) post-surgery, especially in the propolis + 30 ppm SNPs group compared to the control group. This difference was more significant on days 6 (wound healing percentage [WHP]: 75% and 45%) and 8 (WHP: 88% and 65% ) post-surgery (P <.001). Mean neutrophil count on day 12 was highest in the control (34.8 ± 2.97) and lowest in the propolis + 30 ppm SNPs group (16.55 ± 2.12). The number of eosinophils on day 12 was considerably higher in the control group (1.05 ± 4) compared to those in the propolis group (3 ± 0.70), propolis + 30 ppm SNPs group (60/0 ± 1/1), and propolis + 60 ppm SNPs group (0.5 ± 0.52) (P <.001). Mean propolis + 30 ppm SNPs scores for epithelialization and granulation tissue formation were 3 and 4, respectively; in the propolis + 60 ppm SNPs, scores were 2 and 3, respectively; in the propolis alone group scores were 2 and 3, respectively (statistical significance not computed for semiquantitative values). The highest fibroblast count was in the propolis + 30 ppm SNPs group (114.44 ± 3.90) compared to control group (73.2 ± 2.8); P <.001). The difference in collagen fiber density scores was also significant: 1.2 ± 0.42 in the control and 3.66 ± 0.50 in the propolis + 30 ppm SNPs group; (P <.001). The mean of collagen fiber density in the propolis + 60 ppm SNPs group was 2.63 ± 0.51.
Oral propolis alone and in combination with 30 ppm SNPs appears to provide anti-inflammatory effects and increase fibroblast proliferation and collagen deposition in experimental wounds, which may explain the observed differences in healing. Propolis + 60 ppm SNPs appears to have a cytotoxic effect. Research confirming these results and that examines toxicity levels in animals and humans is needed.
本研究旨在探讨口服蜂胶(一种在蜂巢中发现的树脂状物质)单独使用和与 30ppm 银纳米粒子(SNPs)联合使用对雄性大鼠伤口愈合过程的影响。
40 只雄性 Wistar 大鼠随机分为 4 组,每组 10 只:1 组对照组不接受任何治疗,3 组研究组每天接受 1)蜂胶(100mg/kg),2)蜂胶+30ppm SNPs,或 3)蜂胶+60ppm SNPs。通过术后第 4、6、8 和 10 天的伤口表面积减少来确定愈合率。在伤口形成后第 12 天,根据形态学使用 400x 物镜计数伤口愈合的组织学变化,包括新血管数量、炎症细胞(中性粒细胞、嗜酸性粒细胞和肥大细胞)和成纤维细胞,并使用苏木精和伊红以及三色染色分别确定胶原沉积密度。组织学评分基于从最低到最高改善组织状态的 0 到 4 分,使用单因素方差分析、Tukey 检验、Kruskal-Wallis 检验、t 检验和 Mann-Whitney U 检验来分析各组之间的差异。显著性水平设置为 P<.05。
与对照组相比,在术后第 4、6、8 和 10 天(术后第 12 天未评估百分比变化),治疗组的伤口愈合率差异显著,尤其是在蜂胶+30ppm SNPs 组与对照组之间。在术后第 6 天(伤口愈合百分比[WHP]:75%和 45%)和第 8 天(WHP:88%和 65%)差异更为显著(P<.001)。第 12 天对照组中性粒细胞计数最高(34.8±2.97),而蜂胶+30ppm SNPs 组最低(16.55±2.12)。第 12 天对照组嗜酸性粒细胞数量明显高于蜂胶组(3±0.70)、蜂胶+30ppm SNPs 组(60/0±1/1)和蜂胶+60ppm SNPs 组(0.5±0.52)(P<.001)。上皮化和肉芽组织形成的平均蜂胶+30ppm SNPs 评分分别为 3 和 4;在蜂胶+60ppm SNPs 组中,评分分别为 2 和 3;在单独使用蜂胶的组中,评分分别为 2 和 3(未计算半定量值的统计学意义)。在蜂胶+30ppm SNPs 组中,成纤维细胞计数最高(114.44±3.90),与对照组(73.2±2.8)相比差异显著(P<.001)。胶原纤维密度评分差异也有统计学意义:对照组为 1.2±0.42,蜂胶+30ppm SNPs 组为 3.66±0.50(P<.001)。蜂胶+60ppm SNPs 组胶原纤维密度的平均值为 2.63±0.51。
单独使用蜂胶和与 30ppm SNPs 联合使用似乎具有抗炎作用,并增加实验性伤口中的成纤维细胞增殖和胶原沉积,这可能解释了观察到的愈合差异。蜂胶+60ppm SNPs 似乎具有细胞毒性作用。需要进行证实这些结果并检查动物和人类毒性水平的研究。