Mass Spectrometry-based Biomarkers for Knee Osteoarthritis: A Systematic Review

Figures & data

Figure 1. Frontal (left) and parasagittal (middle) view of the knee’s anatomy, as well as a visualization of the osteoarthritic knee (right) with osteoarthritic features such as synovial thickening, cartilage degradation and development of bone spurs. The bones in the human knee are covered by a layer of hyaline cartilage. This cartilage, in combination with the menisci and a layer of lubricating synovial fluid, facilitate the movement of the knee. Other tissues such as synovium and Hoffa’s fat pad are there to maintain joint homeostasis. Image was modified with permission from Servier Medical Art, licensed under a Creative Common Attribution 3.0 Generic License. http://smart.servier.com/

Table 1. Data extraction and study characteristics of selected articles: proteins analyzed in synovial fluid, cartilage, synovium or meniscus, sorted on tissue type and mass spectrometry method

Author, year (ref)	Subjects (number)	MS(/MS) method and ionization source (if stated)	Identification database
Synovial fluid
Ahmed, 2016 [12]	eOA (46) lOA (17)	LC-MS/MS	Cross references
Ahrman, 2014 [13]	Non-OA (35) OA (20)	LC-MS/MS	UniProtKB
Brophy, 2019 [14]	Non-OA (6) OA (3)	LC-MS/MS	UniProt
Gobezie, 2007 [15]	Non-OA (20) eOA (21) lOA (18)	LC-MS/MS	NCBI RefSeq Human
Kamphorst, 2007 [16]	Non-OA (1) OA (1)	LC-MS/MS	Human FASTA
Liao, 2018 [17]	Non-OA (10) lOA (10)	LC-MS/MS, SWATH	NS
Roller, 2016 [18]	Non-OA (16) OA (5)	RP-LC-MS/MS	NS
Steinbeck, 2007 [19]	Non-OA (4) eOA (11) lOA (18)	LC-MS	NS
Timur, 2021 [20]	Non-OA (10) OA (10)	ESI-LC-MS/MS	Unihuman Reviewed
Corigliano, 2017 [21]	eOA (13) lOA (12)	MALDI-MS and MS/MS	UniProtKB
Liao, 2013 [22]	Non-OA (12) eOA (5) lOA (7)	MALDI-MS	NCBInr
Liao, 2015 [23]	Non-OA (10) OA (10)	MALDI-MS and MS/MS	NCBInr
Ritter, 2013 [44]	Non-OA (10) eOA (10) lOA (10)	2D-DIGE-MS and MS/MS	UniProt
Cartilage
Cillero-Pastor, 2013 [35]	Non-OA (10) lOA (10)	MALDI-MSI and MS/MS	UniProt
Guo, 2008 [24]	Non-OA (3) OA (3)	ESI and MS/MS	UniProt
Hsueh, 2016 [62]	Non-OA (NS) OA (NS)	Nano-spray LC-MS/MS	UniProt
Rosenthal, 2011 [25]	Non-OA (10) OA (10)	Nano-spray LC-MS/MS	UniProt
Wu, 2007 [26]	Non-OA (7) OA (7)	Nano-spray LC-MS/MS	NCBInr
Synovium
Cillero-Pastor, 2015 [27]	Non-OA (3) lOA (3)	MALDI-MSI and MS/MS	UniProt
Tang, 2018 [28]	Non-OA (3) OA (4)	LC-MS/MS	UniProt
Meniscus
Folkesson, 2020 [45]	Non-OA (10) lOA (9)	LC-MS/MS	Human protein database
Roller, 2015 [29]	Non-OA (6) eOA (6) lOA (3)	LC-MS/MS	NS

2D-DIGE-MS = two-dimensional difference gel electrophoresis mass spectrometry, eOA = early osteoarthritis, ESI = electrospray ionization, (RP)-LC-MS(/MS) = (reversed phase) (tandem) liquid chromatography mass spectrometry, lOA = late osteoarthritis, MALDI-MS(I) = matrix-assisted laser desorption/ionization mass spectrometry (imaging), NS = not stated, OA = osteoarthritis, SWATH-MS = SWATH-mass spectrometry, blue = analysis done in synovial fluid, green = analysis done in cartilage, red = analysis done in synovium, yellow = analysis done in meniscus.

Table 2. Data extraction and study characteristics of selected articles analyzing metabolites and lipids in synovial fluid and cartilage

Author, year (ref)	Subjects (number)	Sample type	MS(/MS) method and ionization source (if stated)	Identification database
Metabolites
Carlson, 2018 [46]	Non-OA (5) OA (5)	Synovial fluid	LC-MS	METLIN
Carlson, 2019 [38]	Non-OA (7) eOA (55) lOA (17)	Synovial fluid	LC-MS	NS
Kim, 2017 [43]	eOA (8) lOA (7)	Synovial fluid	ESI-GC-MS	BinBase (in-house)
Mickiewicz, 2015 [37]	Non-OA (13) OA (55)	Synovial fluid	H-NMR, GC-MS	GOLM, NIST
Zheng, 2017 [30]	Non-OA (21) OA (49)	Synovial fluid	ESI-GC-MS, LC-MS/MS	NIST, LECO
Lipids
Cillero-Pastor, 2012 [39]	Non-OA (3) lOA (3)	Cartilage	SIMS	LIPIDMAPS
Kosinska, 2013 [31]	Non-OA (9) eOA (17) lOA (13)	Synovial fluid	ESI-MS/MS	Lipidomic Net
Kosinska, 2014 [32]	Non-OA (9) eOA (17) lOA (13)	Synovial fluid	ESI-MS/MS, LC-MS/MS	LIPIDMAPS
Kosinska, 2015 [33]	Non-OA (16) eOA (26) lOA (22)	Synovial fluid	ESI-MS/MS, GC-MS	LIPIDMAPS
Vyver, 2020 [42]	Non-OA (6) OA (23)	Synovial fluid	ESI-LC-MS	Authentic standards

eOA = early osteoarthritis, ESI-MS/MS = electrospray ionization tandem mass spectrometry, GC-MS = gas chromatography mass spectrometry, LC-MS(/MS) = (tandem) liquid chromatography mass spectrometry, lOA = late osteoarthritis, OA = osteoarthritis.

Figure 2. Prisma flow diagram, screened abstracts and articles. A total of 32 and 24 articles were used for qualitative and quantitative analysis respectively in this systematic review

Table 3. Pathway analysis in MetaboAnalyst 5.0 using all upregulated-highlighted proteins (n = 35), lipids and metabolites (n = 13) in OA patients, found in the article search of this review. Pathways with high impact (impact > 0.1) are highlighted in bold. These pathways include complement and coagulation cascades, extracellular matrix receptor interaction and multiple inflammatory pathways

Pathway name	P-value	FDR	Impact	Database
Complement and coagulation cascades	1.19E-20	3.95E-18	0.16	KEGG
Staphylococcus aureus infection	2.69E-7	4.45E-5	0.37	KEGG
Systemic lupus erythematosus	3.57E-5	0.0039	0.10	KEGG
Fatty acid biosynthesis	5.79E-5	0.0045	0.01	KEGG
Biosynthesis of unsaturated fatty acids	6.75E-5	0.0045	0.03	KEGG
ECM-receptor interaction	1.07E-4	0.0059	0.35	KEGG
Prion diseases	1.61E-4	0.0076	0.0	KEGG
Cholesterol metabolism	6.25E-4	0.026	0.02	KEGG
Pertussis	0.0018	0.065	0.10	KEGG
Focal adhesion	0.0023	0.077	0.03	KEGG
Platelet activation	0.0067	0.20	0.01	KEGG
Vitamin digestion and absorption	0.014	0.35	0.0	KEGG
Human papillomavirus infection	0.014	0.35	0.02	KEGG
Fatty acid elongation	0.015	0.36	0.01	KEGG
PI3K-Akt signaling pathway	0.017	0.38	0.03	KEGG
Adipocytokine signaling pathway	0.019	0.40	0.07	KEGG
Proteoglycans in cancer	0.021	0.41	0.06	KEGG
Leishmaniasis	0.022	0.41	0.07	KEGG
Fatty acid degradation	0.029	0.50	0.04	KEGG
Chagas disease (American trypanosomiasis)	0.037	0.57	0.03	KEGG
Choline metabolism in cancer	0.038	0.57	0.0	KEGG
AGE-RAGE signaling pathway in diabetic complications	0.038	0.57	0.03	KEGG
Amoebiasis	0.041	0.60	0.0	KEGG

ECM = extracellular matrix, FDR = false discovery rate.

Figure 3. Upregulated proteins in cartilage, synovial fluid, synovium and meniscus of osteoarthritis patients, identified by at least two authors, without contradiction. Recurring proteins measured in at least three tissue types are highlighted in bold. Image was modified with permission from Servier Medical Art, licensed under a Creative Common Attribution 3.0 Generic License. http://smart.servier.com/

Figure 4. Upregulated metabolites and lipids in cartilage and synovial fluid osteoarthritis patients, identified by at least two authors, without contradiction. Lipid or metabolite species that were found in cartilage and synovium are highlighted in bold. Image was modified with permission from Servier Medical Art, licensed under a Creative Common Attribution 3.0 Generic License. http://smart.servier.com/

Figure 5. The Pathway Enrichment vs. -log10 (p) with p < 0.05 cutoff for proteins, lipids and metabolites in osteoarthritis patients. P-values are subdivided on a white to red color scheme, red indicating higher significance (P < 0.05). Circle size is determined based on Enrichment Ratio (ER), with small circles representing low ER and big circles representing high ER. ECM = extracellular matrix

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