Study On The Sustained-Release Scaffold Loaded With Andrographolide Collagen Promotes The Maintenance Of Chondrocyte Phenotype In Inflammatory Environment In Vitro
Keywords
Andrographolide, Collagen Scaffold, Interleukin 1¦¢, Rabbit Chondrocytes, Phenotype Maintenance
Abstract
This article mainly studies the effect of andrographolide collagen sustained-release scaffold on maintaining the phenotype of chondrocytes in an inflammatory environment. The andrographolide collagen sustained-release scaffold was prepared by physical blending and vacuum freeze-drying. Environmental scanning electron microscopy (ESEM), true densitometer and UV-visible spectrophotometer were used to characterize the morphology, porosity and porosity of the drug-loaded scaffold. Drug release. Isolated, expanded and cultured rabbit articular chondrocytes were seeded on andrographolide collagen sustained-release scaffolds and cultured for 3 days under conventional culture medium conditions and in an inflammatory environment simulated by interleukin 1¦Â (IL-1¦Â). During 7 days, Alamar Blue, fluorescein diacetate (FDA) staining, real-time quantitative polymerase chain reaction (RT-qPCR) and other methods were used to study the effects of the prepared scaffolds on the proliferation and phenotype of chondrocytes. . Experimental results show that the collagen scaffold prepared by vacuum freeze-drying method has good pore connectivity, the average pore diameter is (120.7 ¡À 17.8) ¦Ìm, and the opening rate can reach 96%. The drug-loaded scaffold can achieve sustained release of drugs within two weeks. Alamar Blue experimental results show that the collagen sustained-release scaffold loaded with andrographolide with a mass fraction of 2.22% can significantly inhibit the proliferation of chondrocytes, while collagen sustained-release scaffolds with other drug loading concentrations have no significant effect on the proliferation of chondrocytes in an inflammatory environment. FDA staining results show that the andrographolide collagen-loaded sustained-release scaffold with a mass fraction of 0.44% can reduce the changes in chondrocyte morphology caused by IL-1¦Â, and the drug-loaded scaffold at this concentration can significantly inhibit matrix metalloproteinase-1 (MMP-1 ) and matrix metalloproteinase-13 (MMP-13) transcription, promoting matrix metalloproteinase inhibitor-1 (TIMP-1), cartilage extracellular matrix-related genes type II collagen (COL II) and aggrecan (Aggrecan) Transcription, increasing the COL II/collagen type I (COL I) ratio. Based on the above research results, it is shown that the andrographolide collagen-loaded sustained-release scaffold with a mass fraction of 0.44% can inhibit genes in chondrocytes that are not conducive to maintaining the phenotype (such as: MMP-1, MMP-13) in an inflammatory environment simulated by IL-1¦Â. ), promotes the transcription of genes that are beneficial to maintaining phenotype (such as COL II, Aggrecan, TIMP-1), thereby enhancing the ability of chondrocytes to maintain their phenotype in an inflammatory environment, and is expected to be used to repair cartilage damage in arthritis
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Original research was done by Liu Hairong, Dai Yao, Li Yongsheng, Chen Wei
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