Table 3.

Morphological and physiological impairment in MSCs in the pathogenesis of autoimmune and autoinflammatory diseases. The table describes autoimmune and autoinflammatory disease with the methodological summary used to characterise associated MSC dysfunction. Amyotrophic lateral sclerosis (ALS); ankylosing spondylitis (AS); bone marrow (BM); chemokine (C-C motif) ligand-2 (CCL2); carboxyfluorescein diacetate succinimidyl ester CFSE); colony-forming unit–fibroblast (CFU-F); chronic obstructive pulmonary disease (COPD); cyclooxygenase-2 (COX2); endothelial-like MSC (EL-MSCs); indoleamin-2,3-dioxygenase (IDO); idiopathic pulmonary fibrosis (IPF); hepatocyte growth factor (HGF); mesenchymal stem cell (MSC); peripheral blood mononuclear cells (PBMCs); programmed death ligand-1 (PDL1); rheumatoid arthritis (RA); transforming growth factor-β (TGFβ); TNF-stimulated gene-6 (TSG6); synovial fluid (SF); systemic lupus erythematosus (SLE); systemic sclerosis (SSc); T-helpers (Th); T-regulatory cells (Treg); visual analogue score (VAS); vascular endothelial growth factor (VEGF); vascular endothelial growth factor receptors (VEGFRs).

Disease	Study Methodology	MSC Characteristics	References
Rheumatoid arthritis (subtype was not defined despite clinical importance and is a limitation of the study)	Synovial inflammation measured using the arthroscopic visual analogue score (VAS) and by immunohistochemistry with anti-CD3 and CD68 staining for macrophages. Expression of SOX9, p65, Galectin-3, and SUMO measured by qPCR. Synovial MSCs were analysed by population doublings, clonogenic activity, and multipotency. ELISA for IL1 β, TNFα, IL6.	The arthroscopic VAS in RA correlated significantly with synovial macrophage infiltration. RA activity negatively influences synovial MSC by decreasing their chondrogenic and clonogenic capability. CD44 in RA MSCs correlated negatively with inflammation and positively with chondrogenesis. Cytokine production and Sox9 expression was similar in RA MSCs and OA MSCs.	[126]
Rheumatoid arthritis (subtype was not defined despite clinical importance and is a limitation of the study)	Co-culture of BM-MSC, CD4+ cells, or PBMCs labelled with CFSE and measurement of Th-cells, Treg, and Th17-cells by flow cytometry. Proliferation and apoptosis assays. Migration assays. Human G-Series Cytokine Antibody Array. ELISA measured of IL17A. TGFβ1, IDO, PGE2, IL6, and CCL2 measured by qPCR.	RA MSCs showed equivalent immunophenotype, differentiation potential, cellular apoptosis, and cytokine profiles compared to controls which were OA patients who underwent knee arthroplasty. BM MSCs from RA patients did not downregulate Th17-cells proliferation. RA derived-MSCs showed impaired proliferative potential and migration capacity.	[122]
Ankylosing spondylitis	Multiple differentiation and cell viability assays. Immunomodulatory property of MSCs were analysed by two-way mixed PBMCs reactions or after stimulation with phytohemagglutinin. CCR4+CCR6+ Th/Treg cells and surface markers of BM-MSCs were analysed using flow cytometry.	AS MSCs demonstrated normal proliferation, cell viability, surface markers, and multiple differentiation characteristics. AS BM-MSCs induced an imbalance in the ratio of CCR4+CCR6+ Th/Treg cells by reducing Treg and increasing CCR4+CCR6+ Th cells. AS MSCs reduced Foxp3+ cells when co-cultured with PBMCs.	[123]
Systemic lupus erythematosus	Immunocytochemistry and flow cytometry with CD34, CD45, CD73, CD90, CD105, CD31, CD19, CD11b, HLA-ABC, CD44, CD29, and HLA-DR surface markers. qPCR with IL6, IL8, Gro1, Mcp2, Rantes, and GM-CSF. Western blotting for FNβ, MAVS, p53, p16, and 53BP1, ELISA for IL-6, IL-8, and GM-CSF. Comet assay. β-galactosidase assay.	SLE BM-MSCs were characterised by: ○ reduced proliferation rate. ○ increased production of reactive oxygen species. ○ increased expression of p53 and p16. ○ altered cytokine production, increased IL6 and IL8, increased IFNβ levels, and IFNβ-induced mRNAs.	[127]
Systemic sclerosis	Quantification of CFU-F. Osteogenic, adipogenic, and endothelial cells differentiation. Immunophenotyping by flow cytometry. Assessment of the endothelial-like MSC (EL-MSC) phenotype after culture in endothelial-specific medium and measurement of VEGFR and CXCR4 expression with flow cytometric analysis. Chemoinvasion assays of MSCs and EL-MSCs. Capillary morphogenesis assay. Telomerase activity assay.	SSc MSCs demonstrated: ○ The same phenotype (positive for CD29, CD44, CD166, CD90, CD73, HLA–A, B, and C, and CD105, low HLA–DP, DQ, and DR) and clonogenic activity as healthy MSCs. ○ A decreased percentage of VEGFR-2+, CXCR4+, VEGFR-2+/CXCR4+, and early senescence. ○ Low migration and angiogenic potential. ○ Decreased capacity to capillary morphogenesis and chemoinvasion. ○ The addition of VEGF and stromal cell-derived factor 1 to cultured SSc EL-MSCs increased their angiogenic potential less than that in controls.	[124]
Parkinson’s disease	Confocal images for identification of mitochondrial and lysosomal localisation. NADH autofluorescence. Nuclear DNA sequencing analysis with target genes: SNCA, PARK2, UCHL1, PINK1, DJ1, LRRK2, GBA, VPS35, ATP13A2, EIF4G1, HTRA2, DNAJC13, VPS13C, DNAJC6, FBXO7, PLA2G6, SYNJ1, and MAPT. Mitochondrial DNA sequencing analysis. MSC adipogenic potential.	Impaired differentiation of BM-MSCs. Mitochondrial dysfunction. Higher basal rate of mitochondrial degradation and lower levels of biogenesis. Reduction in mitochondrial mass. Increased level of oxidative stress.	[121]
Idiopathic pulmonary fibrosis	Cell senescence was determined by cell proliferation and expression of p16INK4A, p21, and β-galactosidase activity. Mitochondrial function and DNA damage were measured. Paracrine induction of senescence and profibrotic responses were analysed in human lung fibroblasts. The reparative capacity of BM-MSCs was examined in vivo using the bleomycin-induced lung fibrosis model.	BM-MSCs from patients with IPF characterised by: ○ Mitochondrial dysfunction. ○ Accumulation of DNA damage. ○ Diminished migration capacity of MSCs. ○ Less effectiveness in preventing fibrotic changes in mice after bleomycin-induced injury, increasing illness severity, and pro-inflammatory responses.	[120]
Chronic obstructive pulmonary disease	Immunophenotyping of MSCs by flow cytometry using CD73, CDw90, CD105, CD45, CD14, and CD34. Tri-lineage differentiation. The expression of migration-related chemokine receptors and their ligands in BM-MSCs: qPCR with SDF-1a, CXCR4, CCR7, CCL19, and CCL21. SDF-1α levels in MSC-conditioned media and sera evaluated by ELISA.	COPD BM-MSCs were positive for CD73, CD90, and CD105 and negative for CD45, CD14, and CD34 antigens, and were capable of differentiating towards the adipogenic, osteogenic, and chondrogenic lineages. CXCR4 mRNA expression were decreased in COPD BM-MSCs that provided the evidence that CXCR4/SDF1 is dysregulated in COPD patients. COPD affects SDF1α levels in serum and BM-MSCs.	[125]