中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (27): 4300-4307.doi: 10.12307/2022.859

• 纳米生物材料 nanobiomaterials • 上一篇    下一篇

富血小板纤维蛋白联合姜黄素纳米颗粒水凝胶促糖尿病小鼠创面愈合

吴海能1，耿  康1，王  静2，熊爱兵1   

  1. 西南医科大学附属医院，1整形烧伤外科，国家临床重点建设专科，2肿瘤科，四川省泸州市  646000
  • 收稿日期:2021-10-14 接受日期:2021-12-10 出版日期:2022-09-28 发布日期:2022-03-12
  • 通讯作者: 熊爱兵，硕士，主任医师，教授，西南医科大学附属医院整形烧伤外科，国家临床重点建设专科，四川省泸州市 646000
  • 作者简介:吴海能，男，1995年生，汉族，广东省东莞市人，西南医科大学在读硕士，主要从事整形外科临床与基础研究。
  • 基金资助:
    西南医科大学自然科学青年基金项目(2020ZRQNA008)，项目负责人：耿康

Platelet-rich fibrin combined with curcumin nanoparticle hydrogel promotes wound healing in diabetic mice

Wu Haineng1, Geng Kang1, Wang Jing2, Xiong Aibing1   

  1. 1Department of Plastic and Burn Surgery, Affiliated Hospital of Southwest Medical University, National Clinical Key Construction Specialty, Luzhou 646000, Sichuan Province, China; 2Department of Oncology, Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2021-10-14 Accepted:2021-12-10 Online:2022-09-28 Published:2022-03-12
  • Contact: Xiong Aibing, Master, Chief physician, Professor, Department of Plastic and Burn Surgery, Affiliated Hospital of Southwest Medical University, National Clinical Key Construction Specialty, Luzhou 646000, Sichuan Province, China
  • About author:Wu Haineng, Master candidate, Department of Plastic and Burn Surgery, Affiliated Hospital of Southwest Medical University, National Clinical Key Construction Specialty, Luzhou 646000, Sichuan Province, China
  • Supported by:
    Natural Science Youth Fund Project of Southwest Medical University, No. 2020ZRQNA008 (to GK)

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摘要：

文题释义：
姜黄素纳米颗粒：姜黄素是一种药理作用广泛的中药，具有良好的抗氧化活性和抗炎作用，但在实际应用中发现，姜黄素存在溶解度不高、稳定性差、吸收率低、代谢快、半衰期短等缺陷，这些问题导致其生物利用度较低，限制了其临床应用。将姜黄素制备成纳米颗粒可改善其理化性质，提高生物利用度。 
水凝胶：是一种三维网络结构的生物材料，具有高亲水高保水的特性及良好的组织相容性，将其作为创面敷料符合湿性愈合理论的要求，能为创面提供类似生理状态的愈合环境，有利于创面修复和组织再生。

背景：糖尿病难愈性创面目前仍缺乏十分理想的治疗手段，探索促进糖尿病创面愈合的方法意义重大。
目的：观察富血小板纤维蛋白联合姜黄素纳米颗粒水凝胶对糖尿病小鼠创面愈合的影响。
方法：分别制备富血小板纤维蛋白与姜黄素纳米颗粒水凝胶。从100只成年C57BL/6J小鼠中随机取20只作为正常对照(A组)，另外80只建立糖尿病模型。在建模成功的72只小鼠背部制作直径1 cm的圆形全层皮肤损伤创面，随机分4组处理：B组涂抹生理盐水，C组涂抹富血小板纤维蛋白与游离姜黄素溶液，D组涂抹富血小板纤维蛋白与姜黄素纳米颗粒溶液，E组涂抹富血小板纤维蛋白与姜黄素纳米颗粒水凝胶，每组18只。A组小鼠背部制作直径1 cm的圆形全层皮肤损伤创面，涂抹生理盐水。于创面造模当日即开始给药，之后隔日分别给予上述对应药物，直至伤后12 d。处理后第3，9，12天，观察创面愈合及创缘组织病理组织学变化。
结果与结论：①处理后第12天，A、C、D、E组创面基本愈合完全，B组创面未完全愈合，E组创面愈合率高于B、C、D组(P < 0.05)；②处理后第3天苏木精-伊红染色显示，E组创面愈合情况好于B、C、D组，炎细胞浸润及肉芽成熟度最接近A组；③免疫组化染色显示，E组处理后第3天的创面Ly6G+细胞平均数量低于B、C组(P < 0.001)，E组处理后第9天的创面F4/80+细胞平均数量低于B、C、D组(P < 0.001)；E组处理后第12天的创面CD31+细胞数量高于B、C、D组(P < 0.01)；④处理后第9天的免疫荧光染色显示，E组创面M2型巨噬细胞数量高于B、C、D组(P < 0.01)，M1型巨噬细胞数量低于B、C、D组(P < 0.001)；⑤处理后第12天的Masson染色显示，A、C、D、E组可见大量排列整齐、分布均匀致密的胶原纤维，其中A、E组胶原数量更多、染色更深；B组胶原纤维排列疏松，分布不均匀，颜色淡染，数量较少；⑥结果说明，富血小板纤维蛋白联合姜黄素纳米颗粒水凝胶可以改善糖尿病创面炎症环境，促进糖尿病小鼠创面愈合。

https://orcid.org/0000-0002-6798-0950 (吴海能) 

关键词: 富血小板纤维蛋白, 姜黄素, 纳米颗粒, 水凝胶, 糖尿病创面, 炎症

Abstract: BACKGROUND:  At present, there is still a lack of ideal treatment methods for refractory diabetic wounds. It is of great significance to explore methods to promote diabetic wounds healing.
OBJECTIVE: To observe the effect of platelet-rich fibrin combined with curcumin nanoparticles hydrogel on wound healing in diabetic mice. 
METHODS: Platelet-rich fibrin combined with curcumin nanoparticles hydrogel was prepared. 100 healthy adult C57BL/6J mice were randomly divided into normal control group (A group, n=20) and diabetic group (n=80). After model establishment, round full-thickness skin injury wounds were made in 72 mice by skin punch with a diameter of 1 cm. Seventy-two diabetic mice were randomly divided into diabetic control group (B group; physiological saline; n=18), platelet-rich fibrin + free curcumin group (C group; n=18), platelet-rich fibrin + curcumin nanoparticles group (D group; n=18), and platelet-rich fibrin + curcumin nanoparticles hydrogel group (E group; n=18). Round full-thickness skin injury wounds were made with a diameter of 1 cm in mice of A group and treated with physiological saline. The drug administration was started on the day of wound modeling, and the above-mentioned corresponding drugs were administered every other day until post-injury 12 days. On days 3, 9, and 12 after treatment, wound healing and histopathological changes of wound margin were observed. 
RESULTS AND CONCLUSION: (1) On day 12 after treatment, the wounds in A, C, D, and E groups were basically healed completely, but the wounds in B group were not completely healed. The wound healing rate of E group was higher than that of B, C, and D groups (P < 0.05). (2) On day 3 after treatment, hematoxylin-eosin staining showed that the wound healing in E group was better than that in B, C, and D groups, and the inflammatory cell infiltration and granulation maturity were the closest to the A group. (3) Immunohistochemical staining showed that the mean number of Ly6G+ cells in the wound on day 3 after treatment in E group was lower than that in B and C groups (P < 0.001), and the mean number of F4/80+ cells in E group was lower than that in B, C and D groups on day 9 after treatment (P < 0.001). The number of CD31+ cells in the wound on day 12 after treatment in group E was higher than that in B, C and D groups (P < 0.01). (4) Immunofluorescence staining on day 9 after treatment showed that the number of M2-type macrophages in the wounds of E group was higher than that of B, C, and D groups (P < 0.01), and the number of M1-type macrophages was lower than that of B, C, and D groups (P < 0.001). (5) Masson staining on day 12 after treatment showed that there were a large number of neatly arranged, evenly distributed and dense collagen fibers in A, C, D, and E groups, among which A and E groups demonstrated more collagen and deep staining; group B presented loose collagen fibers, uneven distribution, pale color, and small quantity. (6) Platelet-rich fibrin combined with curcumin nanoparticles hydrogel can improve the inflammatory environment of diabetic wound and promote wound healing in diabetic mice.

Key words: platelet-rich fibrin, curcumin, nanoparticles, hydrogels, diabetic wound, inflammation

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