Optimization of primary culture, inflammatory induction, and molecular analysis of fibroblast-like synoviocytes in a rat model of osteoarthritis pain.

Ao Xiong, Zhaoxin Ma, Renping Xiong, Fei Luo

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Article in Scientific reports, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

4 authors.

Ao XiongDepartment of Orthopaedics, First Affiliated Hospital, Army Medical University, (Third Military Medical University), No. 30, Gaotanyan Zheng Street, Shapingba District, Chongqing, 400038, China.

Zhaoxin MaDepartment of Orthopaedics, First Affiliated Hospital, Army Medical University, (Third Military Medical University), No. 30, Gaotanyan Zheng Street, Shapingba District, Chongqing, 400038, China.

Renping XiongDepartment of Army Occupational Disease, State Key Laboratory of Trauma and Chemical Poisoning, Research Institute of Surgery, Daping Hospital, Army Medical University, (Third Military Medical University), Chongqing, 400042, China. xiong63@tmmu.edu.cn.

Fei LuoDepartment of Orthopaedics, First Affiliated Hospital, Army Medical University, (Third Military Medical University), No. 30, Gaotanyan Zheng Street, Shapingba District, Chongqing, 400038, China. luofly@tmmu.edu.cn.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Synovitis, driven by fibroblast-like synoviocytes (FLS), is a key early event in knee osteoarthritis (KOA). The lack of standardized protocols for FLS culture and molecular analysis hinders reproducible in vitro modeling of early KOA. We developed an integrated platform combining MIA-induced KOA pain modeling, standardized primary FLS culture, inflammation induction, and molecular analysis. We defined passages 3-7 (P3-P7) as the optimal functional passage window for FLS, during which FLS maintained > 98% purity and stable proliferation. LPS stimulation at 1000 ng/mL for 3 h robustly induced IL-1β and TNF-α expression at both mRNA and protein levels. Our optimized RT-qPCR protocols achieved high efficiency (> 91%) and linearity (r > 0.95). Using dexamethasone (DEX), we validated the platform by demonstrating suppression of NF-κB signaling and downstream inflammatory mediators (MMP3/13, VEGFA, NGF). This work provides a standardized system addressing major reproducibility challenges in FLS research.

Indexed as

ArthralgiaFibroblastsOsteoarthritis, KneePrimary Cell CultureSynoviocytesAnimalsCells, CulturedDexamethasoneDisease Models, AnimalInflammation MediatorsIodoacetic AcidLipopolysaccharidesMaleNF-kappa BRatsRats, Sprague-DawleyDexamethasoneInflammation MediatorsIodoacetic AcidLipopolysaccharidesNF-kappa BFibroblast-like synoviocytes (FLS)IL-1βOsteoarthritisPrimary cultureRT-qPCRWestern blot

Identifiers

PMID41286036

DOI10.1038/s41598-025-25494-3

PMCPMC12644989

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.