中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (22): 3524-3531.doi: 10.12307/2024.524

• 水凝胶材料Hydrogel materials • 上一篇    下一篇

脂质体凝胶负载油酸促进慢性烧伤创面的修复

王毛毛1,张  庆2,3,吴博文1,谢  岩3   

  1. 1宁夏医科大学临床医学院,宁夏回族自治区银川市  750004;2宁夏医科大学生育力保持教育部重点实验室,宁夏回族自治区银川市  750004;3宁夏医科大学总医院人体组织与器官库,宁夏回族自治区银川市  750003
  • 收稿日期:2023-08-26 接受日期:2023-10-12 出版日期:2024-08-08 发布日期:2024-01-20
  • 通讯作者: 谢岩,博士,宁夏医科大学总医院人体组织与器官库,宁夏回族自治区银川市 750003
  • 作者简介:王毛毛,男,1985年生,内蒙古自治区巴彦淖尔市人,汉族,硕士,主治医师,主要从事慢性创面修复的研究。 张庆,女,1993年生,宁夏回族自治区银川市人,汉族,硕士,主要从事慢性创面修复的研究。
  • 基金资助:
    宁夏回族自治区重点研发项目(2019BEG03069),项目负责人:谢岩

Liposome gel loaded oleic acid promotes the repair of chronic burn wounds

Wang Maomao1, Zhang Qing2, 3, Wu Bowen1, Xie Yan3   

  1. 1College of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 2Key Laboratory of Fertility Maintenance, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3Tissue Organ Bank & Tissue Engineering Center, General Hospital of Ningxia Medical University, Yinchuan 750003, Ningxia Hui Autonomous Region, China
  • Received:2023-08-26 Accepted:2023-10-12 Online:2024-08-08 Published:2024-01-20
  • Contact: Xie Yan, PhD, Tissue Organ Bank & Tissue Engineering Center, General Hospital of Ningxia Medical University, Yinchuan 750003, Ningxia Hui Autonomous Region, China
  • About author:Wang Maomao, Master, Attending physician, College of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China Zhang Qing, Master, Key Laboratory of Fertility Maintenance, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Tissue Organ Bank & Tissue Engineering Center, General Hospital of Ningxia Medical University, Yinchuan 750003, Ningxia Hui Autonomous Region, China
  • Supported by:
    Key Research and Development Plan of Ningxia Hui Autonomous Region, No. 2019BEG03069 (to XY)

摘要:


文题释义:

慢性烧伤创面:是指严重烧伤造成的经1个月以上正规治疗仍未愈合且无明显愈合趋势的创面。慢性烧伤创面的修复是临床上的一大棘手问题,也是烧伤治疗领域研究的重点和难点之一。
脂质体凝胶:脂质体是由磷脂和其他两亲性物质分散于水中,由一层或多层同心脂质双分子膜包封而成的球状体,类似细胞结构,有生物膜的特性和功能,可以包裹水溶性或脂溶性药物,独特的结构使其成为药物递送的载体。脂质体凝胶是由脂质体和凝胶基质混合而制成的。


背景:油酸能够调节炎症和免疫反应,具有修复皮肤创面的潜力,但油酸在病灶处滞留时间短、在空气中易自氧化而变质,需要合适的药物载体才能充分发挥疗效。

目的: 探究油酸脂质体凝胶修复慢性烧伤创面的疗效。
方法:采用薄膜分散法制备油酸脂质体溶液,随后溶于泊洛沙姆凝胶基质中制备油酸脂质体凝胶。①体外实验:将不同体积油酸脂质体凝胶加入细胞培养基中制备油酸脂质体凝胶溶液(体积比分别为1∶3、1∶9、1∶27),应用阿尔玛蓝染色检测不同体积比油酸脂质体凝胶溶液对人角质形成细胞和人成纤维细胞增殖的影响,结晶紫染色观察细胞形态。②体内实验:取成年SD大鼠,采用背部全层烧伤叠加创面皮下注射表柔比星复合因素构建慢性烧伤创面模型,将造模成功的30只大鼠随机分5组,每组6只:油酸脂质体凝胶组、油酸组、脂质体凝胶组、阳性对照组、阴性对照组创面分别外敷涂抹油酸脂质体凝胶、油酸、脂质体凝胶、重组人表皮生长因子凝胶与生理盐水的纱布,隔日换药一次,总共给药16次,观察创面愈合情况。

结果与结论:①体外实验:阿尔玛蓝试剂检测与结晶紫染色显示,体积比为1∶9的油酸脂质体凝胶溶液可促进人角质形成细胞和人成纤维细胞的增殖。②体内实验:油酸脂质体凝胶组创面愈合时间短于其他4组(P < 0.01),用药第4,8,12,16,20天的创面愈合率高于其他4组(P < 0.01)。给药结束后,苏木精-伊红染色显示5组创面均上皮化愈合,其中油酸脂质体凝胶组表皮厚度最接近于正常皮肤,并且优于其他4组;免疫组化染色显示,油酸脂质体凝胶组创面中细胞角蛋白10、肿瘤蛋白63、α-平滑肌肌动蛋白、Ⅰ型胶原、肿瘤坏死因子α、白细胞介素6、丙二醛及超氧化物歧化酶的表达最接近正常皮肤,并且优于其他4组。用药第12,32天,油酸脂质体凝胶组创面匀浆上清中肿瘤坏死因子α、白细胞介素6、丙二醛及超氧化物歧化酶的表达最接近正常皮肤,并且优于其他4组。③结果表明:油酸脂质体凝胶可促进角质形成细胞和成纤维细胞的增殖,减轻炎症反应及氧化应激损伤,促进慢性烧伤创面的愈合。

https://orcid.org/0000-0002-5066-728X(王毛毛)

中国组织工程研究杂志出版内容重点:生物材料;骨生物材料口腔生物材料纳米材料缓释材料材料相容性组织工程

关键词: 慢性烧伤创面, 脂质体凝胶, 油酸, 创面愈合, 炎症反应, 氧化应激, 细胞增殖

Abstract: BACKGROUND: Oleic acid can regulate inflammation and immune responses, and has the potential to repair skin wounds. Oleic acid has a short retention time at the lesion. It is prone to self oxidation and deterioration in the air, and suitable drug carriers are needed to fully exert the therapeutic effect of oleic acid. 
OBJECTIVE: To investigate the efficacy of oleic acid-liposome gel in the treatment of chronic burn wounds.  
METHODS: Oleic acid liposome solution was prepared by thin film dispersion method, and then dissolved in Poloxamer gel matrix to prepare oleic acid-liposome gel. (1) In vitro experiment: Oleic acid-liposome gel solution was prepared by adding different volumes of oleic acid-liposome gel into cell medium (volume ratio: 1:3, 1:9, 1:27, respectively). Alma-blue reagent was used to detect the effects of different concentrations of oleic acid-liposome gel on the proliferation of human keratinocytes and human fibroblasts. Crystal violet staining was used to observe cell morphology. (2) In vivo experiment: The animal model of chronic burn wounds was established by using full-thickness burn of SD rat back skin combined with local subcutaneous injection of epirubicin. The 30 successfully modeled rats were randomly divided into five groups with six rats in each group. The wounds of oleic acid liposome gel group, oleic acid group, liposome gel group, positive control group and negative control group were applied with gauze of oleic acid liposome gel, oleic acid, liposome gel, recombinant human epidermal growth factor gel and normal saline. The dressing was changed once every other day. A total of 16 doses were administered. The wound healing was observed.
RESULTS AND CONCLUSION: (1) In vitro experiments: Alma-blue reagent detection and crystal violet staining showed that oleic acid liposome gel solution with volume ratio of 1:9 could promote the proliferation of human keratinocytes and human fibroblasts. (2) In vivo experiment: The wound healing time of the oleic acid liposome gel group was shorter than that of the other four groups (P < 0.01), and the wound healing rate at 4, 8, 12, 16, and 20 days was higher than that of the other four groups (P < 0.01). After administration, hematoxylin-eosin staining showed epithelialization and healing of wounds in all five groups, and the epidermal thickness of oleic acid liposome gel group was the closest to normal skin and better than the other four groups. Immunohistochemical staining showed that the expressions of cytokeratin 10, tumor protein 63, α-smooth muscle actin, collagen I, tumor necrosis factor α, interleukin 6, malonaldehyde, and superoxide dismutase in oleic acid liposome gel group were closest to those in normal skin, and superior to those in other four groups. On days 12 and 32 of administration, the expressions of tumor necrosis factor α, interleukin 6, malondialdehyde, and superoxide dismutase in wound homogenate supernatant in oleic acid liposome gel group were closest to those in normal skin, and superior to those in other four groups. (3) The results showed that oleic acid liposome gel could promote the proliferation of keratinocytes and fibroblasts, reduce inflammation and oxidative stress injury, and promote the healing of chronic burn wounds. 

Key words: chronic burn wound, liposome gel, oleic acid, wound healing, inflammatory reaction, oxidative stress, cell proliferation

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