Table 1.
Summary of preclinical in vitro studies.
| Cell Source | Cell Phenotype | OA Diagnosis | Culture Conditions | Analysis and Experimental Times | Main Results | References |
|---|---|---|---|---|---|---|
| Cartilage and synovial fluid from 372 late stage OA patients (64% were female) undergoing TKA | Chondrogenic progenitor cells | The patients met the ACR classification criteriafor knee OA | 50,000 cells into 3D alginate bead with human fibronectin or bovine Col IV or Col II or Matrigel with or without E2 or testosterone supplementation | Measurement in synovial fluids (E2 and testosterone). IHC and immunoblotting (ERα, ERβ, and AR). Microarray analysis and RT-qPCR (ESR1, ESR2, AR, Sox9, Col I, Col II, and Runx2). FACS analyses |
↑testosterone in male synovial fluid vs. female. Microarrays show that 4.9% genes exhibited expression activities different between sexes. ↓ESR1 and ESR2 after testosterone in females. ↑ESR1 and ESR2 after testosterone in males. ↑AR after E2 and testosterone in males. ↑Sox9 and Col I after E2 in females and after testosterone in males. ↓Col II after E2 and testosterone in males. ↓Runx2 after E2 in males |
Koelling et al. 2010 [25] |
| Osteochondral tissue, meniscus, synovial membrane, and synovial fluid from 20 patients (10 males and 10 females) undergoing TKA | Chondrocytes (from 6 males and 6 females) and osteoblasts | Radiographic diagnosis (Kellgren–Lawrence grading scale) | Chondrocytes and osteoblasts were harvested from both minimally or maximally eroded zones with and without 1α,25(OH)2D3 or E2 | Histology on meniscus, synovium, bone, and cartilage. Measurements on synovial fluid (1α,25(OH)2D3, MMPs, sGAGs, TGF-β1,TGF-β2, and TGF-β3). RT-qPCR on cultures (VDR, PDIA3, ESR1, ERα36, IL1A, IL1B, IL6, IL8, IL10, WNT3A, WNT5A, CTNNB, DKK1, DKK2, ACAN, Col II, and COMP) (12 h). ALP in chondrocyte culture and OC and OP in osteoblast culture (24 h) |
↓pain thresholds in females at baseline. In synovial fluid: ↑MMP-1, MMP-7, MMP-9, MMP-13, HGF, SCF, SCGF-β, sGAGs, TGF-β1 and TGF-β2 in males vs. females. ↑IL2α, IL3, IL12p40, IL16, IL18, TNF-β, TRAIL, LIF, M-CSF, MIF, GRO-α, MCP-3, and MIG in females vs. males. In chondrocyte culture: ↓VDR and PDIA3 and ↑ESR1 in females vs. males. ↑IL1A, IL6, and IL8ESR1 in females vs. males. ↑WNT5A, DKK1, and DKK2 in males vs. females. E2 induced ↑ALP, ACAN, COL2A1, and COMP in females vs. males. In osteoblast culture: ↑ESR1 and ERα36 in females vs. males. ↑VDR in males vs.females. E2 induced ↑ALP and OC and ↓OP in females vs. males. 1α,25(OH)2D3 induced ↑ALP in males vs. females |
Pan et al.2016 [26] |
| Articular cartilage from 8 patients (3 males and 5 females) undergoing TKA | Chondrocytes | Not reported | Chondrocyte cultures with or without proliferative stimulus (FGF-2) | DNA extraction, aCGH, FISH and qRT-PCR (Col I, Col II, ACAN, and IL1-β) at day 0, 4, 14, and 19 | Cell expansion and growth factor addition did not cause any autosomal imbalances or loss of DNA. Cell expansion from females did not cause sex chromosome abnormalities while in males a loss of Y-chromosome sequences was observed (aCGH and FISH). No changes in gene expression between males and females |
Stumm et al. 2012 [27] |
| Subchondral, trabecular, and cortical bone from femoral head of 11 dogs (7 males and 4 females) undergoing THR | Osteoblasts | Clinical and radiographic diagnosis | Osteoblast culture from subchondral, trabecular and cortical bone in mineralizing medium (DMEM plus β-glycerophosphate and ascorbate) | Cell number and viability (6 days). TNAP activity (1 day). Mineralization analysis (40 days) |
No differences in any of the analysis parameters irrespective of the bone types between males and female | Meeson et al. 2019 [28] |
| Synovial membrane of 8 male and 8 female Sprague-Dawley rats (8-weeks-old) | FLSs | TMJ-OA chemically induced by Freund’s adjuvant combined with MIA | FLSs treated with or without TNF-α for 6 h; FLSs (lower chamber) in co-culture system with macrophage cell line (NR8383, in the upper chamber) | ELISA and Western blot (MCP-1). RT-PCR (CD68, MCP-1, iNOS, and IL-1β expression). Macrophage migration assay in the co-culture system (24 h) |
↑iNOS, IL-1β, MCP-1 expression, macrophages number, and migration in female FLSs vs. males. No difference in the mRNA expression of iNOS, IL-1β, MCP-1, and in the migration assay from male and female FLSs without treatment of TNF-α. ↑iNOS, IL-1β, and MCP-1 in female FLSs treated with TNF-α vs. male FLSs. Female FLSs recruited more macrophages vs. male FLSs upon stimulation with TNF-α |
Xue et al. 2018 [29] (in vitro) |
Abbreviations: OA, Osteoarthritis; TKA, Total knee arthroplasty; ACR, American College of Rheumatology; 3D, Three-dimensional; Col, Collagen; E2, 17β-estradiol; IHC, Immunohistochemistry; ER, Estrogen receptor; AR, androgen receptor; RT-PCR, Real-time C-reactive Protein; FACS, Fluorescence-activated cell sorting; MMPs, Matrix metalloproteinases; sGAGs, Sulphated glycosaminoglycans; TGF, Transforming growth factor; IL, Interleukin; ALP, Alkaline phosphatase; VDR, Vitamin D receptor; THR, Total hip replacement; FLSs, Fibroblast-like synoviocytes; TMJ, Temporomandibular joint; MIA, Monosodium iodoacetate; FGF-2, Fibroblast growth factor-2; DMEM, Dulbecco’s modified Eagle Medium; TNF, Tumor necrosis factor; MCP, Monocyte Chemoattractant Protein; TNAP, Tissue non-specific alkaline phosphatase; ACAN, Aggrecan; COMP, Cartilage oligomeric matrix protein; OP, Osteoprotegerin; OC, Osteocalcin; DNA, Deoxyribonucleic acid; FISH, Fluorescent in situ hybridization; aCGH, Array-based comparative genomic hybridization; ELISA, Enzyme-linked immunosorbent assay; iNOS, Inducible nitric oxide synthase; HGF, Hepatocyte growth factor; M-CSF, Macrophage colony-stimulating factor; LIF, Leukemia inhibitory factor; PDIA3, Protein disulfide isomerase A3; GRO-α, Growth-regulated oncogene α; MIG, Monokine induced by gamma interferon; TRAIL, TNF-related apoptosis-inducing ligand; MIF, Macrophage migration inhibitory factor.