Dataset Information

Proteomics Reveals Increased Periostin in Synovial Fluid from Canine and Human Anterior Cruciate Ligament Injury

ABSTRACT: Post-traumatic osteoarthritis (PTOA) is a common sequela to anterior cruciate ligament (ACL) injury in humans and dogs. Identification of conserved targets between species may hasten biomarker identification and therapeutic development. To identify differentially regulated synovial fluid (SF) proteins in ACL injury, SF samples were collected from knees with ACL injury or control knees (n=8 per species) in human and dog patients. The SF proteome was evaluated using nano-scale reverse phase chromatography and tandem mass spectroscopy. Associations between proteins and ACL injury were analyzed in both species. Immunoassays were used to validate key proteins identified via mass spectrometry, including periostin, α-2-macroglublin, and lubricin, in additional SF samples (dog: n=57 ACL, n=31 controls; human: n=20 ACL, n=15 advanced OA) and plasma samples (human: n=18 ACL, n=15 advanced OA, and n=12 controls). Periostin was the most upregulated SF protein (log2 fold-change) in both dog (2.6) and human (3.5) ACL injury, followed by complement C1q (2.3) and α-2-macroglobulin (1.8) in dogs and fibrinogen (1.1) and α-2-macroglobulin (1.1) in humans. Top downregulated proteins included serum amyloid A1 (-1.9) and aggrecan (-1.8) in dogs and carbonic anhydrase 2 (-3.2) and hemoglobin subunit β (-2.8) in humans. Approximately 60% of proteins detected were shared between species, and immunoregulatory and macromolecular transport proteins were the most common families. Clinical Relevance: Findings in both species support further investigation into periostin as a potential biomarker or therapeutic target and the use of spontaneous ACL injury in dogs as a potential translational large animal model for human ACL injury.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human) Canis Familiaris (dog) (canis Lupus Familiaris)

TISSUE(S): Femorotibial Joint

DISEASE(S): Osteoarthritis

SUBMITTER: Sydney Womack

LAB HEAD: Heidi Lynn Reesink

PROVIDER: PXD061915 | Pride | 2025-06-16

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	JinSu10438661TMT16plexCanine_Fr1.raw	Raw
	JinSu10438661TMT16plexCanine_Fr2.raw	Raw
	JinSu10438661TMT16plexCanine_Fr3.raw	Raw
	JinSu10438661TMT16plexCanine_Fr4.raw	Raw
	JinSuHeidi10438661TMT16plexCanine_Fr1to4.mgf	Mgf

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Publications

Proteomics Reveals Increased Periostin in Synovial Fluid From Canine and Human Anterior Cruciate Ligament Injury.

Womack Sydney J SJ Carballo Camila B CB Secor Erica J EJ Rodeo Scott A SA Reesink Heidi L HL

Journal of orthopaedic research : official publication of the Orthopaedic Research Society 20250417 7

Posttraumatic osteoarthritis (PTOA) is a common sequela to anterior cruciate ligament (ACL) injury in humans and dogs. Identification of conserved targets between species may hasten biomarker identification and therapeutic development. To identify differentially regulated synovial fluid (SF) proteins in ACL injury, SF samples were collected from knees with ACL injury or control knees (n = 8 per species) in human and dog patients. The SF proteome was evaluated using nano-scale reverse-phase chrom ...[more]

PMID: 40247443

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Project description:The anterior cruciate ligament (ACL) is an essential stabilizer of the tibiofemoral articulation. ACL tears often lead to functional instability and are associated with an increased risk for osteoarthritis. The healing potential of the injured ACL is poorly understood and is considered to be limited. Transcriptome-wide expression profiles of 24 human ACL remnants recovered at the time of surgical reconstruction were analyzed utilizing the Agilent human 8x60K microarray platform. Gene ontology was performed on differentially expressed transcripts based on time-from-injury (acute, <3 months; intermediate, 3-12 months; chronic, >12 months). A subset of transcripts was validated via microfluidic digital polymerase-chain-reaction. Expression of periostin, a highly differentially expressed transcript, was tested by immunohistochemistry. Numerous transcripts covering important functional classifications were differentially expressed by time-from-injury. In acute tears, processes representing angiogenesis were repressed while those representing stem-cell differentiation were elevated. In intermediate tears, processes representing stem-cell proliferation concomitant with cellular component organization/cellular localization were elevated. In chronic tears, processes denoting myosin filament organization were elevated while those representing cellular component organization/cell localization and extracellular matrix organization were repressed. Expression levels of periostin were down-regulated in chronic tears compared to acute (42-fold) and intermediate (29-fold) tears. Immunohistochemistry confirmed a decline in periostin expression in tissues from chronic tears. These findings suggest an initial attempt of the injured ACL to repair, which declines with time-from-injury. These findings have implications for efforts to repair the ACL and may be relevant for reconstruction of the ACL. The functional role of periostin in ACL injuries, and the potential implication for surgical treatment, warrants further investigation. Total RNA obtained frominjured anterior cruciate ligament (ACL) tissues from pateints undergoing ACL surgery.

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