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

doi:10.12307/2022.859

Abstract

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

CLC Number:

Wu Haineng, Geng Kang, Wang Jing, Xiong Aibing. Platelet-rich fibrin combined with curcumin nanoparticle hydrogel promotes wound healing in diabetic mice[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(27): 4300-4307.

Figures/Tables 7

2.3 各组糖尿病造模一般情况 A组小鼠毛色光亮，精神良好，行为活跃。B组小鼠在注射链脲佐菌素后逐渐出现多饮、多食、多尿、毛发粗糙萎黄、精神倦怠、活动减少等表现，并且体质量较A组逐渐减轻，体型消瘦。 2.4 各组小鼠体质量及血糖变化普通维持饲料适应性喂养1周后，A组与B组小鼠体质量比较差异无显著性意义(P > 0.05)。B组小鼠给予高脂高糖饲料喂养8周后，体质量比A组小鼠稍重。A组小鼠注射生理盐水1周后体质量较前稍有增长，B组小鼠注射链脲佐菌素1周后质量较前明显降低。普通维持饲料适应性喂养1周后，A组与B组小鼠血糖值比较差异无显著性意义(P > 0.05)。B组小鼠给予高脂高糖饲料喂养8周后，其血糖值较A组高(P < 0.05)。注射链脲佐菌素1周后，B组小鼠血糖值较A组明显升高(P < 0.01)，且大于16.7 mmol/L，说明2型糖尿病造模成功。 2.5 各组小鼠创面愈合情况观察期间各组小鼠创面逐渐愈合，表面覆盖大小不等的痂壳，痂下间隙及创面周围有不同程度红肿、渗出，均无感染、化脓现象。创面造模后第3天，各组创面面积均有不同程度减小，表面均有肉芽组织生长，但B组创面愈合面积不及其他各组。创面造模后第9天，各组创面面积进一步缩小，A、C、D、E组创面面积显著减小，肉芽组织逐渐纤维化，质地较韧。在各药物干预组中，以E组创面愈合情况最佳。B组创面表面仍以痂壳覆盖，肉芽组织成熟缓慢。创面造模后第12天，A、C、D、E组创面基本愈合完全，B组创面面积较前虽有明显缩小，但仍有部分创面未愈合，见图3A。创面造模后第12天，A-E组创面愈合率分别为(94.39±3.60)%，(71.17±6.32)%，(82.78±5.58)%，(88.33±6.37)%，(92.17±4.27)%。单因素方差分析显示，5组创面愈合率比较差异有显著性意义(F=54.182，P < 0.001)。LSD-t法两两比较显示，E组创面愈合率高于B、C、D组(P均< 0.05)，与A组比较差异无显著性意义(P=0.216)，见图3B。"

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