Injectable hydrogel microspheres that enhance autophagy can improve cartilage microenvironment and resist chondrocyte senescence

doi:10.12307/2025.439

Abstract

Abstract: BACKGROUND: Cell senescence is one of the major risk factors for osteoarthritis, but there is no widely accepted anti-osteoarthritis therapy targeting senescent cells.
OBJECTIVE: To develop a feasible treatment strategy targeting senescent cells in osteoarthritis.
METHODS: The cationic liposome containing rapamycin, RAPA@Lipo, was prepared by thin film dispersion method. Methylallylated hyaluronic acid hydrogel was synthesized, and RAPA@Lipo was added to the methylallylated hyaluronic acid hydrogel aqueous phase solution. The hydrogel microspheres were prepared by microfluidic equipment. Solid hydrogel microspheres (RAPA@Lipo@MS) were crosslinked under violet light. Primary human chondrocytes were co-cultured with RAPA@Lipo and RAPA@Lipo@MS, respectively. The biocompatibility of the materials was evaluated by CCK-8 assay and live/dead staining. Primary rat chondrocytes were cultured in four groups. Normal control group was cultured for 48 hours. The model group was stimulated with H2O2 for 24 hours to establish senescent cell model. RAPA@Lipo group and RAPA@Lipo@MS group were cultured for 24 hours after establishing senescent cell model with RAPA@Lipo and RAPA@Lipo@MS, respectively. After culture, immunofluorescence was used to observe the expression of p62 and type II collagen. RT-PCR was used to detect the mRNA expression of interleukin 6, matrix metalloproteinase 13, type II collagen, aggrecan, and ADAMTS-5.
RESULTS AND CONCLUSION: (1) The results of CCK-8 assay and live/dead staining showed that RAPA@Lipo and RAPA@Lipo@MS had good biocompatibility. (2) Compared with the normal control group, the protein expression of p62 was increased (P < 0.05); the expression of type II collagen was decreased (P < 0.05), and the mRNA expression levels of interleukin 6, matrix metalloproteinase 13, and ADAMTS-5 were increased (P < 0.05); mRNA expression levels of type II collagen and aggrecan were decreased (P < 0.05) in the model group. Compared with the model group, the expression of p62 protein was decreased (P < 0.05); the expression of type II collagen was increased (P < 0.05), and the mRNA expression levels of interleukin 6, matrix metalloproteinase 13, and ADAMTS-5 were decreased (P < 0.05); mRNA expression of type II collagen and aggrecan increased (P < 0.05) in the RAPA@Lipo@MS group. (3) These findings indicate that RAPA@Lipo@MS can control the quality of cells in vivo by enhancing autophagy, reduce senescent cells in vivo, and locally eliminate senescent cells and senescence-associated secretory phenotype factors in osteoarthritis, thereby slowing the progression of osteoarthritis and creating a cartilage microenvironment that promotes regeneration.

Key words: osteoarthritis, hydrogel microsphere, autophagy, senescent cell, senescence-associated secretory phenotype, rapamycin

CLC Number:

Li Wenming, Li Yonghang, Yan Caiping, Wang Xingkuan, Xiang Chao, Zhang Yuan, Jiang Ke, Chen Lu. Injectable hydrogel microspheres that enhance autophagy can improve cartilage microenvironment and resist chondrocyte senescence[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(22): 4710-4719.

Figures/Tables 5

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