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Scientific Articles   |    
Optimization of Leukocyte Concentration in Platelet-Rich Plasma for the Treatment of Tendinopathy
Taralyn M. McCarrel, DVM1; Tom Minas, MD, MS2; Lisa A. Fortier, DVM, PhD3
1 Rood and Riddle Equine Hospital, 2150 Georgetown Road, Lexington, KY, 40511
2 Brigham and Women’s Hospital, 850 Boylston Street, Suite 112, Chestnut Hill, MA 02467
3 Cornell University, VMC C3-129, Ithaca, NY, 14853. E-mail address: laf4@cornell.edu
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Investigation performed at Cornell University, Ithaca, New York



Disclosure: One or more of the authors received payments or services, either directly or indirectly (i.e., via his or her institution), from a third party in support of an aspect of this work. In addition, one or more of the authors, or his or her institution, has had a financial relationship, in the thirty-six months prior to submission of this work, with an entity in the biomedical arena that could be perceived to influence or have the potential to influence what is written in this work. No author has had any other relationships, or has engaged in any other activities, that could be perceived to influence or have the potential to influence what is written in this work. The complete Disclosures of Potential Conflicts of Interest submitted by authors are always provided with the online version of the article.

Copyright © 2012 by The Journal of Bone and Joint Surgery, Inc.
J Bone Joint Surg Am, 2012 Oct 03;94(19):e143 1-8. doi: 10.2106/JBJS.L.00019
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Abstract

Background: 

Numerous methods are available for platelet-rich plasma (PRP) generation, but evidence defining the optimum composition is lacking. We hypothesized that leukocyte-reduced PRP would result in lower inflammatory cytokine expression compared with concentrated-leukocyte PRP and that maintaining the platelet:white blood cell (WBC) ratio would compensate for the effect of increased WBC concentration.

Methods: 

Blood and flexor digitorum superficialis tendons were collected from young adult horses. Three PRP groups were generated with the same platelet concentration but different WBC concentrations: intermediate-concentration standard PRP, leukocyte-reduced PRP, and concentrated-leukocyte PRP. An additional high-concentration PRP group was generated with the same WBC concentration as the concentrated-leukocyte PRP group and the same platelet:WBC ratio as the standard PRP group. The PRP groups were used as media for flexor digitorum superficialis tendon explants in culture for seventy-two hours with 10% plasma in Dulbecco modified Eagle medium (DMEM) serving as control. Tendon gene expression for collagen types I (COL1A1) and III (COL3A1), cartilage oligomeric matrix protein (COMP), matrix metalloproteinase (MMP-13), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) was performed.

Results: 

The desired PRP groups were successfully generated. The expression of COMP, the COL1A1:COL3A1 ratio, and the expression of MMP-13 in flexor digitorum superficialis tendon explants was not different between PRP groups. The expression of COMP (p = 0.0027) and the COL1A1:COL3A1 ratio (p < 0.0001) were increased in the PRP groups as compared with the control group, and the expression of MMP-13 was decreased in the PRP groups as compared with the control group (p < 0.0001). The expression of IL-1β was lowest in leukocyte-reduced PRP and highest in concentrated-leukocyte PRP (p = 0.0001). The leukocyte-reduced PRP group and the control group had the lowest TNF-α expression, whereas the high-concentration PRP and concentrated-leukocyte PRP groups had the highest expression (p = 0.0224).

Conclusions: 

A high absolute WBC concentration in PRP contributes to the expression of inflammatory cytokines in flexor digitorum superficialis tendon explants, and maintenance of the platelet:WBC ratio is not able to counteract this effect.

Clinical Relevance: 

The optimum composition of PRP for the treatment of tendinopathy has not been directly investigated. Persistent inflammation results in inferior repair with scar tissue. The present study indicates that in an animal model, WBC in PRP contributes to inflammatory cytokine production. Therefore, leukocyte-reduced PRP may be the optimum preparation to stimulate superior healing without scar tissue formation.

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    References

    Accreditation Statement
    These activities have been planned and implemented in accordance with the Essential Areas and policies of the Accreditation Council for Continuing Medical Education (ACCME) through the joint sponsorship of the American Academy of Orthopaedic Surgeons and The Journal of Bone and Joint Surgery, Inc. The American Academy of Orthopaedic Surgeons is accredited by the ACCME to provide continuing medical education for physicians.
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