Ginsenoside Rg3-loaded liposome hydrogel promotes chondrogenic differentiation of stem cells

doi:10.12307/2026.766

Chinese Journal of Tissue Engineering Research ›› 2026, Vol. 30 ›› Issue (26): 6760-6767.doi: 10.12307/2026.766

Ginsenoside Rg3-loaded liposome hydrogel promotes chondrogenic differentiation of stem cells

Zhao Qinglan1, 2, 3, Zhou Xinting2, Wang Huajun3, Zhong Jiaxuan2, 3, Zheng Liheng4, Tan Wencheng5, Yang Xinchun6, Huang Shusen3, #br# Wu Tingting2, Zheng Xiaofei3, Hong Jinsong1, 3

1Guangzhou University of Chinese Medicine, Guangzhou Orthopaedic Hospital, Guangzhou 510006, Guangdong Province, China; 2National Engineering Research Center for Healthcare Devices, Guangdong Provincial Key Laboratory of Medical Electronic Instruments and Materials, Institute of Biomedical and Health Engineering, Guangdong Academy of Sciences, Guangzhou 510316, Guangdong Province, China; 3Department of Sports Medicine, The First Affiliated Hospital of Jinan University, Guangdong Key Laboratory for Orthopaedic Precision and Regenerative Medicine, Guangdong Key Laboratory of Speed and Agility, Guangzhou 510630, Guangdong Province, China

Accepted:2025-10-20 Online:2026-09-18 Published:2026-03-11
Contact: Wu Tingting, Senior engineer, National Engineering Research Center for Healthcare Devices, Guangdong Provincial Key Laboratory of Medical Electronic Instruments and Materials, Institute of Biomedical and Health Engineering, Guangdong Academy of Sciences, Guangzhou 510316, Guangdong Province, China Zheng Xiaofei, Chief physician, Department of Sports Medicine, The First Affiliated Hospital of Jinan University, Guangdong Key Laboratory for Orthopaedic Precision and Regenerative Medicine, Guangdong Key Laboratory of Speed and Agility, Guangzhou 510630, Guangdong Province, China Hong Jinsong, Associate professor, Master’s supervisor, Guangzhou University of Chinese Medicine, Guangzhou Orthopaedic Hospital, Guangzhou 510006, Guangdong Province, China; Department of Sports Medicine, The First Affiliated Hospital of Jinan University, Guangdong Key Laboratory for Orthopaedic Precision and Regenerative Medicine, Guangdong Key Laboratory of Speed and Agility, Guangzhou 510630, Guangdong Province, China
About author:Zhao Qinglan, MS candidate, Guangzhou University of Chinese Medicine, Guangzhou Orthopaedic Hospital, Guangzhou 510006, Guangdong Province, China; National Engineering Research Center for Healthcare Devices, Guangdong Provincial Key Laboratory of Medical Electronic Instruments and Materials, Institute of Biomedical and Health Engineering, Guangdong Academy of Sciences, Guangzhou 510316, Guangdong Province, China; Department of Sports Medicine, The First Affiliated Hospital of Jinan University, Guangdong Key Laboratory for Orthopaedic Precision and Regenerative Medicine, Guangdong Key Laboratory of Speed and Agility, Guangzhou 510630, Guangdong Province, China Zhou Xinting, MS, Intermediate engineer, National Engineering Research Center for Healthcare Devices, Guangdong Provincial Key Laboratory of Medical Electronic Instruments and Materials, Institute of Biomedical and Health Engineering, Guangdong Academy of Sciences, Guangzhou 510316, Guangdong Province, China Wang Huajun, MD, Associate professor, Chief physician, Department of Sports Medicine, The First Affiliated Hospital of Jinan University, Guangdong Key Laboratory for Orthopaedic Precision and Regenerative Medicine, Guangdong Key Laboratory of Speed and Agility, Guangzhou 510630, Guangdong Province, China
Supported by:
Guangdong Provincial Science and Technology Project - Guangdong-Macao Joint Innovation Project, No. 2023A0505020008 (to ZXF); National Key Research & Development Program of China, No. 2022YFE0206200 (to ZXF); Macao Science and Technology Development Fund Project, No. FDCT 0032/2022/AGJ (to YXC); Macao Science and Technology Development Fund Project, No. FDCT 0009/2021/AMMJ (to ZLH); GDAS Science and Technology Development Project, No. 2023GDASZH-2023010102 (to WTT); Guangdong Provincial Medical Research Foundation Project, No. A2024428 (to HSS)

Abstract

Abstract: BACKGROUND: Ginsenoside Rg3 has potential for cartilage protection and repair, but its application suffers from drawbacks such as poor water solubility, short half-life, and low bioavailability. To improve drug pharmacokinetic properties, embedding drug-loaded liposomes into methylated gelatin, polysaccharide, or silk fibroin-based hydrogels has become a hot topic in cartilage tissue engineering research.
OBJECTIVE: To fabricate liposome-methacryloylated silk fibroin hydrogels loaded with ginsenoside Rg3 and further analyze the effects of these hydrogels on the chondrogenic differentiation of mouse bone marrow mesenchymal stem cells.
METHODS: (1) Ginsenoside Rg3 liposomes were prepared by a thin film dispersion method. Dil-labeled liposomes were co-cultured with mouse bone marrow mesenchymal stem cells, and cellular uptake of the liposomes was assessed by phalloidin staining. Methacryloylated silk fibroin (SilMA) was prepared. Ginsenoside Rg3 liposomes were mixed with SilMA and photocrosslinked to form a composite hydrogel (SilMA@Lipo-Rg3). The micromorphology, mechanical properties, rheological properties, swelling properties, and drug release performance of the hydrogel were evaluated. (2) Mouse bone marrow mesenchymal stem cells were cultured with different concentrations of SilMA hydrogel extract or SilMA@Lipo-Rg3 hydrogel extract. The cytocompatibility of the materials was evaluated by CCK-8 assay and live-dead cell staining. Mouse bone marrow mesenchymal stem cells were cultured with 1/8 concentration of SilMA hydrogel extract or SilMA@Lipo-Rg3 hydrogel extract, respectively. After chondrogenic induction, the expression of type II collagen, SOX9, and aggrecan mRNA was measured by qPCR. The expression of intracellular proteoglycans and glycosaminoglycans was detected by Alcian blue and safranin O staining.
RESULTS AND CONCLUSION: (1) Phalloidin staining revealed that Dil-labeled liposomes were successfully taken up by mouse bone marrow mesenchymal stem cells. Scanning electron microscopy revealed that the SilMA@Lipo-Rg3 hydrogel exhibited a loose and porous lattice structure. Compression tests and rheological tests revealed that the compressive stiffness of the SilMA@Lipo-Rg3 hydrogel was slightly lower than that of the SilMA hydrogel; there was no significant difference in the swelling properties of the two hydrogels. The SilMA@Lipo-Rg3 hydrogel exhibited excellent sustained-release properties, releasing ginsenoside Rg3 for more than 14 days. (2) CCK-8 assay and live-dead cell staining revealed that the SilMA and SilMA@Lipo-Rg3 hydrogels exhibited good cytocompatibility. qPCR analysis showed that the mRNA expression of type II collagen, SOX9, and aggrecan in the SilMA@Lipo-Rg3 group was higher than that in the SilMA group (P < 0.05). Alcian blue and safranin O staining revealed that the expression of proteoglycans and glycosaminoglycans in the SilMA@Lipo-Rg3 group was higher than that in the SilMA group. These results indicate that the SilMA@Lipo-Rg3 hydrogel promotes the chondrogenic differentiation of mouse bone marrow mesenchymal stem cells.

Key words: ginsenoside Rg3, silk fibroin hydrogel, stem cells, chondrogenic differentiation, methacrylation modification, liposome

CLC Number:

Zhao Qinglan, Zhou Xinting, Wang Huajun, Zhong Jiaxuan, Zheng Liheng, Tan Wencheng, Yang Xinchun, Huang Shusen, Wu Tingting, Zheng Xiaofei, Hong Jinsong. Ginsenoside Rg3-loaded liposome hydrogel promotes chondrogenic differentiation of stem cells[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(26): 6760-6767.

Figures/Tables 6

2.1 材料的表征结果使用透射电镜观察人参皂苷Rg3脂质体的外观形貌，观察到负染后人参皂苷Rg3脂质体呈现出经典的囊泡结构(图1A)，粒径约为39.71 nm(图1B)，证明纳米脂质体的成功构建。将人参皂苷Rg3脂质体与甲基丙烯酰化丝素蛋白混合，通过光交联构建SilMA@Lipo-Rg3水凝胶，扫描电镜下观察到SilMA、SilMA@Lipo-Rg3水凝胶均呈现疏松多孔的网格结构(图1C，D)。通过傅里叶红外光谱评估材料改性前后的结构变化，发现丝素蛋白、SilMA水凝胶、SilMA@Lipo-Rg3水凝胶均在1 637，1 507，1 230 cm-1附近存在酰胺Ⅰ、Ⅱ、Ⅲ区的红外吸收峰；SilMA和SilMA@Lipo-Rg3水凝胶在1 408，1 377 cm-1出现新峰，可能与甲基(-CH3)基团引入有关；此外，1 165，1 125 cm-1处的红外峰可能是甲基丙烯酸缩水甘油酯分子中酯或醚类的C-O-C振动峰，证明甲基丙烯酰化反应成功；SilMA@Lipo-Rg3水凝胶虽然引入了人参皂苷Rg3脂质体，但由于是物理掺入，没有新的化学键合成，总体吸收峰情况与SilMA水凝胶相似(图2A)。水凝胶的压缩应变-应力曲线显示(图2B)，与SilMA水凝胶相比，SilMA@Lipo-Rg3水凝胶的在相同应变下的应力略低，表明脂质体包载后的复合体系刚度略有下降，可能是因为脂质体的引入削弱了丝素蛋白交联网络的均匀性。如图2C，D所示，两种水凝胶的流变性能测试结果与压缩应力-应变实验结果基本一致，进一步证实脂质体的引入对交联网络强度产生了轻微的削弱作用。图2E显示，两组水凝胶的溶胀性能基本一致，在前10 min内，水凝胶迅速溶胀至原始质量的200%以上，此后溶胀速率逐渐减缓；在12 h内，最大溶胀倍率可达约500%，良好的溶胀能力有助于细胞黏附与营养物质交换。 SilMA@Lipo-Rg3水凝胶的体外累积释放结果显示，人参皂苷Rg3在24 h内迅速释放近20%，之后释放速率减缓，可实现缓释效果，药物持续释放时间可延长至14 d以上(图2F)，有望在早期释放高浓度药物抑制局部炎症反应，为后期软骨再生提供持续的药效支持。 "

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Ginsenoside Rg3-loaded liposome hydrogel promotes chondrogenic differentiation of stem cells

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