Optimization of leukocyte concentration in platelet-rich plasma for the treatment of tendinopathy.

McCarrel, TM
·
Minas, T
·
Fortier, LA

Abstract: 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. Conclusions: 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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Publication Date: 2012-10-04 PubMed ID: 23032594DOI: 10.2106/JBJS.L.00019Google Scholar: Lookup

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MeSH
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Comparative Study
Journal Article
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Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research study focused on optimizing the concentration of leukocytes in platelet-rich plasma (PRP) for the treatment of tendinopathy. The research was carried out on young adult horses and suggested that PRP with reduced amounts of leukocytes would result in lower inflammatory responses and potentially help in more effective treatment of tendinopathy.

Research Procedure

Blood and tendons (flexor digitorum superficialis) were collected from young adult horses.
Three different PRP groups were generated varying in terms of white blood cell (WBC) concentrations but maintaining the same platelet concentration. These consisted of standard PRP, leukocyte-reduced PRP, and concentrated-leukocyte PRP.
Another 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. A 10% plasma in Dulbecco modified Eagle medium (DMEM) served as a control group.

Results Analysis

The study successfully generated the desired PRP groups.
The PRP groups showed an increase in the expression of COMP and the COL1A1:COL3A1 ratio, which are markers of tissue remodelling, compared with the control group. In contrast, MMP-13, a matrix metalloproteinase associated with tissue damage, was decreased in the PRP groups compared with the control group.
The expression of inflammatory cytokines, IL-1β and TNF-α, varied among the PRP groups. IL-1β expression was lowest in the leukocyte-reduced PRP and highest in the concentrated-leukocyte PRP. TNF-α expression was lowest in the leukocyte-reduced PRP and control groups, and highest in the high-concentration PRP and concentrated-leukocyte PRP groups.

Conclusions

The researchers found that a high WBC concentration in PRP contributes to the increased production of inflammatory cytokines in the examined tendons. This effect was not counteracted by simply maintaining the platelet:WBC ratio.
According to these findings from an animal-model study, the presence of WBC in the PRP can contribute to inflammation and scar tissue formation. As a result, for therapies involving PRP preparations, leukocyte-reduced PRP could provide the most effective healing without leading to scar tissue formation.

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APA

McCarrel TM, Minas T, Fortier LA. (2012). Optimization of leukocyte concentration in platelet-rich plasma for the treatment of tendinopathy. J Bone Joint Surg Am, 94(19), e143(1-e143(8. https://doi.org/10.2106/JBJS.L.00019

Publication

The Journal of bone and joint surgery. American volume

ISSN: 1535-1386

NlmUniqueID: 0014030

Country: United States

Language: English

Volume: 94

Issue: 19

Pages: e143(1-e143(8

Researcher Affiliations

McCarrel, Taralyn M

Rood and Riddle Equine Hospital, 2150 Georgetown Road, Lexington, KY, 40511, USA.

Minas, Tom

Fortier, Lisa A

MeSH Terms

Animals
Blood Component Transfusion
Cytokines / metabolism
Disease Models, Animal
Extracellular Matrix Proteins / metabolism
Glycoproteins / metabolism
Horses
Interleukin-1beta / metabolism
Leukocyte Count
Leukocytes
Matrilin Proteins
Matrix Metalloproteinase 13 / metabolism
Platelet-Rich Plasma
Random Allocation
Reference Values
Regeneration / physiology
Risk Assessment
Tendinopathy / pathology
Tendinopathy / therapy
Treatment Outcome
Tumor Necrosis Factor-alpha / metabolism

Citations

This article has been cited 117 times.

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