[Application effect of a dual release system of androgen and its antagonist in the repair of full-thickness burn wounds in mice].

R Z Huang, Y W Wang, H Y Huang, R H Jiang, N N Xue, S P Yin, H Y Zhao
{"title":"[Application effect of a dual release system of androgen and its antagonist in the repair of full-thickness burn wounds in mice].","authors":"R Z Huang, Y W Wang, H Y Huang, R H Jiang, N N Xue, S P Yin, H Y Zhao","doi":"10.3760/cma.j.cn501225-20230802-00033","DOIUrl":null,"url":null,"abstract":"<p><p><b>Objective:</b> To explore the optimal ratio of dihydrotestosterone and hydroxyflutamide (hereinafter referred to as DH), construct a dual release system of androgen and its antagonist, and analyze the application effect of this system in the repair of full-thickness burn wounds in mice. <b>Methods:</b> This study was an experimental study. The HaCaT cells were divided into blank group (without drug culture), low baseline group, medium baseline group, and high baseline group according to the random number table (the same grouping method below), and the last three groups of cells were cultured by adding three different ratios of DH. Under a medium ratio, the mass of dihydrotestosterone in the three baseline groups from low to high was 1.4, 2.8, and 4.0 µg, respectively, and the mass of hydroxyflutamide was 1.2, 1.6, and 2.0 µg, respectively. On this basis, under a small ratio, the mass of dihydrotestosterone was reduced by half and the mass of hydroxyflutamide was increased by half; under a large ratio, the mass of dihydrotestosterone was increased by half and the mass of hydroxyflutamide was reduced by half. After culture of 2 days, the cell proliferation level was detected by cell counting kit 8 (<i>n</i>=4). Sixteen 6-8-week-old male BALB/c mice were used to establish a full-thickness burn wound on the back and divided into blank group, small ratio group, medium ratio group, and large ratio group, with 4 mice in each group. On post injury day (PID) 7, normal saline containing different ratios of DH was locally dropped to the wounds of mice in the last three groups of mice (the total mass of DH in the three ratio groups from small to large was 127.5, 165.0, and 202.5 µg, respectively, and the mass ratios of dihydrotestosterone to hydroxyflutamide (hereinafter referred to as drug mass ratio) were 8∶9, 8∶3, and 8∶1, respectively), afterwards, the administration was repeated every 48 hours until PID 27; normal saline was dropped to the wound of mice in blank group at the aforementioned time points. The wound healing status on PID 0 (immediately), 7, 14, 21, and 28 was observed, and the wound healing rates on PID 7, 14, 21, and 28 were calculated (<i>n</i>=4). On PID 28, the wound tissue was taken, which was stained with hematoxylin and eosin for observing re-epithelialization and with Masson for observing collagen fibers, and the proportion of collagen fibers was analyzed (<i>n</i>=3). Twenty 6-8-week-old male BALB/c mice were used to establish a full-thickness burn wound on the back and divided into ordinary scaffold group, small proportion scaffold group, medium proportion scaffold group, and large proportion scaffold group (with 5 mice in each group). On PID 7, the wound was continuously dressed with a polycaprolactone scaffold without drug and a polycaprolactone scaffold containing DH with a drug mass ratio of 1∶3, 1∶1, or 3∶1 (i.e. the dual release system of androgen and its antagonist, with total mass of DH being about 1.7 mg) prepared by using electrospinning technology until the end of the experiment. Histopathological analyses of tissue (<i>n</i>=3) at the same time points as those in the previous animal experiment were performed. On PID 7 and 14, the wound exudates were collected and the relative abundance of bacterial communities was analyzed using 16S ribosomal RNA high-throughput sequencing (<i>n</i>=3). <b>Results:</b> After culture of 2 days, under a small ratio, the proliferation levels of HaCaT cells in low baseline group and high baseline group were significantly higher than the level in blank group (<i>P</i><0.05). As the time after injury prolonged, the wounds of all four groups of mice continued to shrink. On PID 14, the wound healing rate of mice in large ratio group was 72.5% (61.7%, 75.1%), which was close to 53.3% (49.5%, 64.4%) in blank group (<i>P</i>>0.05); the wound healing rates of mice in small and medium ratio groups were 74.2% (71.0%, 84.2%) and 70.4% (65.1%, 74.4%), respectively, which were significantly higher than the rate in blank group (with both <i>Z</i> values being -2.31, <i>P</i><0.05). On PID 21, the wound healing rate of mice in small ratio group was significantly higher than that in blank group (<i>Z</i>=-2.31, <i>P</i><0.05). On PID 28, the wounds of mice in the three ratio groups were completely re-epithelialized and the epidermis was thicker than that in blank group; compared with that in blank group, the collagen fiber content in the wound tissue of mice in the three ratio groups was higher and arranged more orderly, and the proportions of collagen fibers in the wound tissue of mice in small and large ratio groups were significantly increased (<i>P</i><0.05). On PID 28, the wounds of mice in ordinary scaffold group were partially epithelialized, while the wounds of mice in the three proportion scaffold groups were almost completely epithelialized. Among them, the wounds of mice in small proportion scaffold group had the thickest epidermis. The proportion of collagen fibers in the wound tissue of mice in small proportion scaffold group was significantly increased compared with that in ordinary scaffold group (<i>P</i><0.05). On PID 7, the bacterial communities with high relative abundance in the wound exudation of mice in the four groups included bacteria of <i>Corynebacterium</i>, <i>Staphylococcus</i>, and <i>Rhodococcus</i>. On PID 14, the bacterial communities with high relative abundance in the wound exudation of mice in the four groups included bacteria of <i>Stenotrophomonas</i>, <i>Rhodococcus</i>, and <i>Staphylococcus</i>, and the number of bacterial species in the wound exudation of mice in the three proportion scaffold groups was more than that in ordinary scaffold group. <b>Conclusions:</b> When the drug mass ratio is relatively small, DH has the effect of promoting the proliferation of HaCaT cells. The ratio of 8∶9 is the optimal mass ratio of dihydrotestosterone to hydroxyflutamide, and DH with this mass ratio can promote re-epithelialization and collagen deposition of full-thickness burn wounds in mice, and promote wound healing. The constructed dual release system of androgen and its antagonist with DH in a 1∶3 drug mass ratio contributes to the re-epithelialization and collagen deposition of the full-thickness burn wounds in mice, and can improve the diversity of wound microbiota.</p>","PeriodicalId":516861,"journal":{"name":"Zhonghua shao shang yu chuang mian xiu fu za zhi","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Zhonghua shao shang yu chuang mian xiu fu za zhi","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3760/cma.j.cn501225-20230802-00033","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Objective: To explore the optimal ratio of dihydrotestosterone and hydroxyflutamide (hereinafter referred to as DH), construct a dual release system of androgen and its antagonist, and analyze the application effect of this system in the repair of full-thickness burn wounds in mice. Methods: This study was an experimental study. The HaCaT cells were divided into blank group (without drug culture), low baseline group, medium baseline group, and high baseline group according to the random number table (the same grouping method below), and the last three groups of cells were cultured by adding three different ratios of DH. Under a medium ratio, the mass of dihydrotestosterone in the three baseline groups from low to high was 1.4, 2.8, and 4.0 µg, respectively, and the mass of hydroxyflutamide was 1.2, 1.6, and 2.0 µg, respectively. On this basis, under a small ratio, the mass of dihydrotestosterone was reduced by half and the mass of hydroxyflutamide was increased by half; under a large ratio, the mass of dihydrotestosterone was increased by half and the mass of hydroxyflutamide was reduced by half. After culture of 2 days, the cell proliferation level was detected by cell counting kit 8 (n=4). Sixteen 6-8-week-old male BALB/c mice were used to establish a full-thickness burn wound on the back and divided into blank group, small ratio group, medium ratio group, and large ratio group, with 4 mice in each group. On post injury day (PID) 7, normal saline containing different ratios of DH was locally dropped to the wounds of mice in the last three groups of mice (the total mass of DH in the three ratio groups from small to large was 127.5, 165.0, and 202.5 µg, respectively, and the mass ratios of dihydrotestosterone to hydroxyflutamide (hereinafter referred to as drug mass ratio) were 8∶9, 8∶3, and 8∶1, respectively), afterwards, the administration was repeated every 48 hours until PID 27; normal saline was dropped to the wound of mice in blank group at the aforementioned time points. The wound healing status on PID 0 (immediately), 7, 14, 21, and 28 was observed, and the wound healing rates on PID 7, 14, 21, and 28 were calculated (n=4). On PID 28, the wound tissue was taken, which was stained with hematoxylin and eosin for observing re-epithelialization and with Masson for observing collagen fibers, and the proportion of collagen fibers was analyzed (n=3). Twenty 6-8-week-old male BALB/c mice were used to establish a full-thickness burn wound on the back and divided into ordinary scaffold group, small proportion scaffold group, medium proportion scaffold group, and large proportion scaffold group (with 5 mice in each group). On PID 7, the wound was continuously dressed with a polycaprolactone scaffold without drug and a polycaprolactone scaffold containing DH with a drug mass ratio of 1∶3, 1∶1, or 3∶1 (i.e. the dual release system of androgen and its antagonist, with total mass of DH being about 1.7 mg) prepared by using electrospinning technology until the end of the experiment. Histopathological analyses of tissue (n=3) at the same time points as those in the previous animal experiment were performed. On PID 7 and 14, the wound exudates were collected and the relative abundance of bacterial communities was analyzed using 16S ribosomal RNA high-throughput sequencing (n=3). Results: After culture of 2 days, under a small ratio, the proliferation levels of HaCaT cells in low baseline group and high baseline group were significantly higher than the level in blank group (P<0.05). As the time after injury prolonged, the wounds of all four groups of mice continued to shrink. On PID 14, the wound healing rate of mice in large ratio group was 72.5% (61.7%, 75.1%), which was close to 53.3% (49.5%, 64.4%) in blank group (P>0.05); the wound healing rates of mice in small and medium ratio groups were 74.2% (71.0%, 84.2%) and 70.4% (65.1%, 74.4%), respectively, which were significantly higher than the rate in blank group (with both Z values being -2.31, P<0.05). On PID 21, the wound healing rate of mice in small ratio group was significantly higher than that in blank group (Z=-2.31, P<0.05). On PID 28, the wounds of mice in the three ratio groups were completely re-epithelialized and the epidermis was thicker than that in blank group; compared with that in blank group, the collagen fiber content in the wound tissue of mice in the three ratio groups was higher and arranged more orderly, and the proportions of collagen fibers in the wound tissue of mice in small and large ratio groups were significantly increased (P<0.05). On PID 28, the wounds of mice in ordinary scaffold group were partially epithelialized, while the wounds of mice in the three proportion scaffold groups were almost completely epithelialized. Among them, the wounds of mice in small proportion scaffold group had the thickest epidermis. The proportion of collagen fibers in the wound tissue of mice in small proportion scaffold group was significantly increased compared with that in ordinary scaffold group (P<0.05). On PID 7, the bacterial communities with high relative abundance in the wound exudation of mice in the four groups included bacteria of Corynebacterium, Staphylococcus, and Rhodococcus. On PID 14, the bacterial communities with high relative abundance in the wound exudation of mice in the four groups included bacteria of Stenotrophomonas, Rhodococcus, and Staphylococcus, and the number of bacterial species in the wound exudation of mice in the three proportion scaffold groups was more than that in ordinary scaffold group. Conclusions: When the drug mass ratio is relatively small, DH has the effect of promoting the proliferation of HaCaT cells. The ratio of 8∶9 is the optimal mass ratio of dihydrotestosterone to hydroxyflutamide, and DH with this mass ratio can promote re-epithelialization and collagen deposition of full-thickness burn wounds in mice, and promote wound healing. The constructed dual release system of androgen and its antagonist with DH in a 1∶3 drug mass ratio contributes to the re-epithelialization and collagen deposition of the full-thickness burn wounds in mice, and can improve the diversity of wound microbiota.

[雄激素及其拮抗剂双释放系统在小鼠全厚烧伤创面修复中的应用效果]。
目的探索双氢睾酮与羟基氟他胺(以下简称DH)的最佳配比,构建雄激素及其拮抗剂的双释放系统,并分析该系统在小鼠全厚烧伤创面修复中的应用效果。研究方法本研究为实验研究。将 HaCaT 细胞按随机数字表(下同)分为空白组(无药物培养)、低基线组、中基线组和高基线组,后三组细胞分别加入三种不同比例的 DH 进行培养。在中等比例下,从低到高的三个基线组中双氢睾酮的质量分别为 1.4、2.8 和 4.0 µg,羟基氟酰胺的质量分别为 1.2、1.6 和 2.0 µg。在此基础上,在小比例下,双氢睾酮的质量减少一半,羟基氟他胺的质量增加一半;在大比例下,双氢睾酮的质量增加一半,羟基氟他胺的质量减少一半。培养 2 天后,用细胞计数试剂盒 8 检测细胞增殖水平(n=4)。16 只 6-8 周大的雄性 BALB/c 小鼠背部全厚烧伤创面,分为空白组、小比例组、中比例组和大比例组,每组 4 只。在损伤后第 7 天(PID),在后三组小鼠的伤口局部滴入含有不同比例 DH 的生理盐水(从小到大三个比例组的 DH 总质量分别为 127.5、165.0 和 202.5 µg,双氢睾酮与羟基氟他胺的质量比(以下简称药物质量比)分别为 8∶9、8∶3 和 8∶1),之后每 48 小时重复给药一次,直至 PID 27;空白组小鼠在上述时间点的伤口处滴入生理盐水。观察 PID 0(即刻)、7、14、21 和 28 的伤口愈合情况,并计算 PID 7、14、21 和 28 的伤口愈合率(n=4)。PID 28时,取伤口组织,用苏木精和伊红染色观察再上皮化,用Masson染色观察胶原纤维,分析胶原纤维的比例(n=3)。用 20 只 6-8 周大的雄性 BALB/c 小鼠在背部建立全厚烧伤创面,分为普通支架组、小比例支架组、中比例支架组和大比例支架组(每组 5 只)。PID 7时,用电纺丝技术制备的不含药物的聚己内酯支架和含药物质量比为1∶3、1∶1或3∶1的DH(即雄激素及其拮抗剂的双释放系统,DH总质量约为1.7毫克)的聚己内酯支架持续包扎伤口,直至实验结束。在与之前动物实验相同的时间点对组织(n=3)进行组织病理学分析。在 PID 7 和 14,收集伤口渗出物,并使用 16S 核糖体 RNA 高通量测序分析细菌群落的相对丰度(n=3)。结果培养 2 天后,在小比例条件下,低基线组和高基线组的 HaCaT 细胞增殖水平显著高于空白组(PP>0.05);小比例组和中比例组小鼠的伤口愈合率分别为 74.2%(71.0%,84.2%)和70.4%(65.1%,74.4%),分别明显高于空白组(两组Z值均为-2.31,PZ=-2.31,PPP冠状杆菌、葡萄球菌和罗氏球菌。在 PID 14 日,四组小鼠伤口渗出液中相对丰度较高的细菌群落包括链球菌、罗氏球菌和葡萄球菌,且三组比例支架组小鼠伤口渗出液中细菌种类的数量多于普通支架组。结论当药物质量比相对较小时,DH 具有促进 HaCaT 细胞增殖的作用。8∶9是双氢睾酮与羟基氟酰胺的最佳质量比,该质量比的DH能促进小鼠全厚烧伤创面的再上皮化和胶原沉积,促进创面愈合。以 1∶3 的药物质量比构建的雄激素及其拮抗剂与 DH 双释放系统有助于小鼠全厚烧伤创面的再上皮化和胶原沉积,并能改善创面微生物群的多样性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信