Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum

Autologous conditioned serum (ACS) is a common intra-articular treatment for osteoarthritis in horses. The objective of this study was to investigate the influence of ACS preparation method on product contamination and concentrations of relevant cytokines and the influence of multiple freeze/thaw cy...

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Autores principales:	Josephine Hale, Kristopher Hughes, Sarah Hall, Raphael Labens
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	ACS osteoarthritis IRAP horse thermal stability Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/785ca0c0104542b7923af6da104c088d
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spelling	oai:doaj.org-article:785ca0c0104542b7923af6da104c088d2021-12-01T02:27:58ZEffects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum2297-176910.3389/fvets.2021.759828https://doaj.org/article/785ca0c0104542b7923af6da104c088d2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fvets.2021.759828/fullhttps://doaj.org/toc/2297-1769Autologous conditioned serum (ACS) is a common intra-articular treatment for osteoarthritis in horses. The objective of this study was to investigate the influence of ACS preparation method on product contamination and concentrations of relevant cytokines and the influence of multiple freeze/thaw cycles. Blood was obtained from 10 healthy Thoroughbred horses and processed in parallel using a commercial and a non-commercial method to obtain ACS. Fluorescent microsphere immunoassay (FMIA) analysis was performed to quantify Interleukin 1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations in ACS obtained by both production methods. Effect of 3, 4 and 5 freeze/thaw cycles on concentrations of IL-1Ra, IL-10, IL-1β and TNF-α were assessed against baseline samples (2 cycles) in commercial ACS products. Standard aerobic and anaerobic culture methods were applied to both ACS products. Mixed effect one-way analyses of variance (ANOVA) were used to compare the two ACS production method for each cytokine. Repeated measures, mixed effect ANOVA were used to assess the effect of freeze/thaw on cytokine concentrations. Significance was set at P < 0.05. There was no difference in cytokine concentration between production methods (IL-1Ra P = 0.067, IL-1β P = 0.752, IL-10 P = 0.211 and TNF-α P = 0.25). Microbial growth was only observed in two samples obtained using the commercial production method. When compared to baseline, IL-1Ra concentration was decreased following the 5th freeze/thaw cycle (P < 0.001). These results suggest that the concentration of important cytokines are not influenced by ACS production method. When storing ACS samples for future use, freeze/thaw cycles associated with standard clinical practice are unlikely to influence cytokine concentrations. However, the lack of outcome measures associated with 1 or 2 freeze/thaw cycles represents a limitation of this study.Josephine HaleKristopher HughesSarah HallRaphael LabensFrontiers Media S.A.articleACSosteoarthritisIRAPhorsethermal stabilityVeterinary medicineSF600-1100ENFrontiers in Veterinary Science, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	ACS osteoarthritis IRAP horse thermal stability Veterinary medicine SF600-1100
spellingShingle	ACS osteoarthritis IRAP horse thermal stability Veterinary medicine SF600-1100 Josephine Hale Kristopher Hughes Sarah Hall Raphael Labens Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum
description	Autologous conditioned serum (ACS) is a common intra-articular treatment for osteoarthritis in horses. The objective of this study was to investigate the influence of ACS preparation method on product contamination and concentrations of relevant cytokines and the influence of multiple freeze/thaw cycles. Blood was obtained from 10 healthy Thoroughbred horses and processed in parallel using a commercial and a non-commercial method to obtain ACS. Fluorescent microsphere immunoassay (FMIA) analysis was performed to quantify Interleukin 1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations in ACS obtained by both production methods. Effect of 3, 4 and 5 freeze/thaw cycles on concentrations of IL-1Ra, IL-10, IL-1β and TNF-α were assessed against baseline samples (2 cycles) in commercial ACS products. Standard aerobic and anaerobic culture methods were applied to both ACS products. Mixed effect one-way analyses of variance (ANOVA) were used to compare the two ACS production method for each cytokine. Repeated measures, mixed effect ANOVA were used to assess the effect of freeze/thaw on cytokine concentrations. Significance was set at P < 0.05. There was no difference in cytokine concentration between production methods (IL-1Ra P = 0.067, IL-1β P = 0.752, IL-10 P = 0.211 and TNF-α P = 0.25). Microbial growth was only observed in two samples obtained using the commercial production method. When compared to baseline, IL-1Ra concentration was decreased following the 5th freeze/thaw cycle (P < 0.001). These results suggest that the concentration of important cytokines are not influenced by ACS production method. When storing ACS samples for future use, freeze/thaw cycles associated with standard clinical practice are unlikely to influence cytokine concentrations. However, the lack of outcome measures associated with 1 or 2 freeze/thaw cycles represents a limitation of this study.
format	article
author	Josephine Hale Kristopher Hughes Sarah Hall Raphael Labens
author_facet	Josephine Hale Kristopher Hughes Sarah Hall Raphael Labens
author_sort	Josephine Hale
title	Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum
title_short	Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum
title_full	Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum
title_fullStr	Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum
title_full_unstemmed	Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum
title_sort	effects of production method and repeated freeze thaw cycles on cytokine concentrations and microbial contamination in equine autologous conditioned serum
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/785ca0c0104542b7923af6da104c088d
work_keys_str_mv	AT josephinehale effectsofproductionmethodandrepeatedfreezethawcyclesoncytokineconcentrationsandmicrobialcontaminationinequineautologousconditionedserum AT kristopherhughes effectsofproductionmethodandrepeatedfreezethawcyclesoncytokineconcentrationsandmicrobialcontaminationinequineautologousconditionedserum AT sarahhall effectsofproductionmethodandrepeatedfreezethawcyclesoncytokineconcentrationsandmicrobialcontaminationinequineautologousconditionedserum AT raphaellabens effectsofproductionmethodandrepeatedfreezethawcyclesoncytokineconcentrationsandmicrobialcontaminationinequineautologousconditionedserum
_version_	1718405910798270464

Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum

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