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Effect of repeated arthrocentesis on cytologic analysis of synovial fluid in healthy horse

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Effect of repeated arthrocentesis on cytologic analysis of synovial fluid in healthy horses Sgorbini M1, Bonelli F1, Gallo C2, Fusar Bassini R1, Marchetti V1, Corazza M1

1Dipartimento Scienze Veterinarie, via Livornese snc, San Pero a Grado, Pisa. 2Libero professionista, Napoli.

Introduction. Arthrocentesis is a common practice used for the diagnosis and therapy of many articular pathologies. The aim of the present work was to evaluate the influence of repeated arthrocentesis on synovial fluid composition in healthy horses.

Materials and methods. Approval to conduct this study was obtained from the Ethics Committee on Animal Experimentation of the University of Pisa n. 14875 – 20/11/12). Four horses not affected by muscoloskeletal diseases were submitted to repeated arthrocentesis from both the intercarpal (IC) joints. In particular, the right IC joint was sampled at Time 0 (T0), at 2 (T2), 7 (T7) days and then every week for 3 times (T14, T21, T28, respectively) after T0, while the left IC joint was sampled at T0 and then every 10 days for two times (T10 and T20, respectively) after T0. An arthrocentesis was also performed on both IC joints 60 days (T60) after T0. Arthrocentesis were always performed by the same operator (R.F.B.). The synovial fluid samples were collected in EDTA and processed within 1 hours to evaluate: 1) total protein (TP) concentration by a rephractometer (Mahaffey, 2002); 2) total WBC count by an automatic hematology analyzer (Lasercyte®, Idexx, USA) with hyaluronidase pretreatment (Moreno et al., 2000; Eckmann et al., 2010) to reduce the viscosity; 3) differential leukocyte count after cytospin preparation (1500 gpm, 5’) (Cytofuge 2, StatSpin, USA) to improve smear’s quality (Moreno et al., 2000), a modified Romanowsky staining (Diff Quik®, Dade Spa, Milano, Italia) and microscope evaluation at 100X. Data distribution was performed with KS test; Anova for repeated measures and Bonferroni test as post hoc were applied to verify differences related to sampling times. Significative level was set at p<0.05.

Results. Data were expressed as mean±standard deviation. The total WBC count did not change over time as well as the percentage of lymphocytes, eosinophils and neutrophils, while a variation over time was observed for monocytes and total protein concentration. Both decreased from T0 to T14 (right IC joint) and T10 (left IC joint), then rised again over time till T60 with values similar to T0.

Conclusions. Our results are in line to what reported in dogs and cows (White et al., 1989; Dykgraaf et al., 2007; Francoz et al., 2007; Berg et al., 2009; Sanchez Teran et al., 2012) and suggest that repeated arthrocentesis has no effects on cytologic analysis of synovial fluid.

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Berg RIM, Sykes JE, Kass PH, Vernau W (2009). Effect of repeated arthrocentesis on cytologic analysis of synovial fluid in dogs. J Vet Intern Med, 23: 814-7.

Dykgraaf S, Dechant JE, Johns JL, Christopher MM, Bolt DM, Snyder JR (2007). Effect of Intrathecal Amikacin Administration and Repeated Centesis on Digital Flexor Tendon Sheath Synovial Fluid in Horses. Vet Surg, 36(1): 57-63.

Ekmann A, Rigdal ML, Gröndahl G (2009). Automated counting of nucleated cells in equine synovial fluid without and with hyaluronidase pretreatment. Vet Clin Pathol, 39(1): 83-9.

Francoz D, Desrochers A, Latouche JS (2007). Effect of repeated arthrocentesis and single joint lavage on cytologic evaluation of synovial fluid in 5 young calves. Can J Vet Res, 71(2): 129-34. Mahaffey EA (2002). Synovial fluid. In: Cowell RL, Tyler RD (eds): Diagnostic Cytology nd Hematology of the horse. 2nd ed, Mosby Inc., USA, pp. 163-70.

Moreno MJ, Clayburne G, Schumacher HR (2000). Processing of noninflammatory synovial fluids with hyaluronidase for cytospin preparations improves the accuracy of differential counts. Diagn Cytopathol,, 22(4): 256-8.

Palmer DG (1968). Total leukocyte enumeration in pathologic synovial fluids. Am J Clin Pathol, 49(6): 812-4.

Sanchez Teran AF, Rubio-Martinez LM, Villarino NF, Sanz MG (2012). Effects of repeated intra-articular administration of amikacin on serum amyloid A, total protein and nucleated cell count in synovial fluid from healthy horses. Equine vet J, 44(43): 12-6.

White KK, Hodgson DR, Hancock D, Parry BW, Cordell C (1989). Changes in equine carpal joint synovial fluid in response to the injection of two local anesthetic agents. Cornell Vet, 79(1): 25-3.

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