Functionalized self-assembled micelles enhance effect of tranexamic acid in treatment of cutaneous hyperpigmentation

doi:10.12307/2025.488

Abstract

Abstract: BACKGROUND: Topical administration of tranexamic acid can be used for anti-skin pigmentation, but its large polarity makes it difficult to break through the cuticle barrier and cell membrane when administered topically, and the subcutaneous accumulation concentration is not easy to reach the effective therapeutic concentration.
OBJECTIVE: To design functionalized self-assembled micelles to enhance the anti-pigmentation effect of tranexamic acid.
METHODS: The plant polyphenol derivative epigallocatechin gallate palmitate and metformin were used as carrier materials. The self-assembled micelles with synergistic anti-pigging activity and enhanced drug permeability were prepared by hydrogen bonding and electrostatic interaction. The nanoscale properties and stability of self-assembled drug-loaded micelles were tested, and their transdermal permeability was evaluated, and their biocompatibility and cellular effects were investigated.
RESULTS AND CONCLUSION: (1) Functional self-assembled drug-carrying micelles with average particle size of (176.27±5.23) nm, polydispersity coefficient of 0.23±0.02, and the Zeta potential of (-25.67±0.98) mV had good stability. The mass concentrations of tranexamic acid and metformin in the self-assembled drug-carrying micelles were (20.03±0.12) and (6.67±0.08) mg/mL, respectively. The drug loadings of tranexamic acid and metformin in the self-assembled drug-carrying micelles were (19.97±0.12)% and (6.65±0.08)%, respectively. (2) In vitro transdermal results showed that the self-assembled drug-carrying micelles had higher penetration and intradermal retention per unit skin area, and could penetrate and diffuse to deeper parts of the skin. (3) MTT assay results demonstrated that undrug-loaded self-assembled micelles containing tranexamic acid 50-250 μg/mL had no toxic effects on mouse fibroblasts and mouse skin melanoma cells. The self-assembled drug-carrying micelles containing tranexamic acid 500 μg/mL had a slight toxic effect on mouse skin melanoma cells. The self-assembled drug-carrying micelles containing 50-500 μg/mL of tranexamic acid did not cause hemolysis and had good biological safety. (4) In vitro cell culture results showed that self-assembled drug-carrying micelles containing 500 μg/mL tranexamic acid could significantly inhibit the tyrosinase activity and melanin release of mouse skin melanoma cells, and the inhibitory effect was stronger than that of tranexamic acid solution or metformin solution alone. These results indicated that functionalized self-assembled micelles could synergize with tranexamic acid to inhibit tyrosinase activity, reduce melanin synthesis, and enhance the anti-skin pigmentation effect of tranexamic acid.

Key words: tranexamic acid, functionalized self-assembled micelles, skin pigmentation, in vitro transdermal, engineered skin material

CLC Number:

Qi Junlong, Liu Junyi, He Yuzhou, Qiang Wei, Zhang Shiying, Liu Qiao, Zhu Hongda. Functionalized self-assembled micelles enhance effect of tranexamic acid in treatment of cutaneous hyperpigmentation[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(28): 6061-6069.

Figures/Tables 4

2.4 载药自组装胶束的生物安全性评价结果通过MTT检测得出，使用不同质量浓度空白自组装胶束作用于B16F10细胞和L929细胞后，两种细胞存活率最低分别为(91.8±6.6)%和(90.3±0.8)%，见图4A，B。使用不同质量浓度载药自组装胶束、二甲双胍溶液和氨甲环酸溶液分别作用于B16F10细胞后，50-250 μg/mL载药自组装胶束溶液组细胞存活率均高于80%，500 μg/mL载药自组装胶束组呈现轻微细胞毒性，细胞存活率为75%，见图4C，说明空白自组装胶束和一定质量浓度载药自组装胶束对于L929细胞和B16F10细胞具有良好的生物安全性。溶血实验结果显示，离心后阴性对照组中红细胞在离心管内形成沉淀，阳性对照组中红细胞在纯化水中破裂，离心后未分层；在50-500 μg/mL载药自组装胶束溶液中的红细胞经离心后在血液中发生分层，上层溶液无明显血红色，各样品溶血率均小于1.1%，故可视为无溶血风险，见图4D。 2.5 载药自组装胶束对B16F10细胞黑色素含量及酪氨酸酶活性影响的检测结果 B16F10细胞在培养过程中能分泌黑色素，各组B16F10细胞孵育后黑色素含量检测结果，见图4E所示。与空白对照组比较，氨甲环酸溶液和二甲双胍溶液作用后细胞黑色素含量均显著下降(P < 0.05)，证实两种活性物质均能影响B16F10细胞黑色素的生成[19-20]；药物物理混合组对B16F10细胞黑色素分泌的抑制作用显著强于氨甲环酸组和二甲双胍组(P < 0.01)，说明两种活性物质联合使用能增强抑制B16F10细胞的黑色素生成；使用载药自组装胶束作用后B16F10细胞黑色素含量下降至71.0%，黑色素含量低于药物物理混合组(P < 0.005)，说明自组装胶束的联合输送能增强抑制B16F10细胞黑色素生成的作用。各组B16F10细胞孵育后酪氨酸酶活性检测结果，见图4F。与空白对照组比较，氨甲环酸或二甲双胍溶液作用后B16F10细胞内酪氨酸酶活性略有下降但差异无显著性意义(P > 0.05)，而两者物理混合溶液作用后B16F10细胞内酪氨酸酶活性下降显著(P < 0.05)，酶活性降至88.1%；使用载药自组装胶束作用后B16F10细胞内酪氨酸酶活性下降至64.5%(P < 0.005)，与物理混合组比较差异有显著性意义(P < 0.01)。B16F10细胞内黑色素分泌及酪氨酸酶活性检测结果说明，功能性自组装胶束包载氨甲环酸后具有协同抗黑色素分泌的作用。"

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