Moellerberndt Julia, Hagen Alina, Niebert Sabine, Büttner Kathrin, Burk Janina
Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
Unit for Biomathematics and Data Processing, Faculty of Veterinary Medicine, Justus-Liebig-University Giessen, Giessen, Germany.
Front Vet Sci. 2023 Mar 9;10:1117829. doi: 10.3389/fvets.2023.1117829. eCollection 2023.
In equine medicine, the use of regenerative therapeutics has gained growing attention, but is still a new and complex field with room for improvement. Platelet lysate (PL) can be used as therapeutic agent but is also a promising supplement for the culture of multipotent mesenchymal stromal cells. To enable a targeted use of PL both in clinic and laboratory, it is crucial to learn more details on its effective ingredients. While so far, mainly growth factor components have been analyzed in platelet-based products such as PL, the current study focuses on the content of cytokines in serum, plasma, platelet concentrate and PL. Blood was harvested from 20 clinically healthy horses and subjected to blood count and chemistry analysis, as well as to further processing to PL. Plasma and platelet concentrate were produced by a buffy-coat-based method and PL was produced from the concentrate by freeze-thawing. Samples from each horse were analyzed regarding interleukin (IL)-1β, -4, -6 and -10, interferon-γ and tumor necrosis factor-α concentrations using sandwich ELISAs. Cytokine concentrations in serum, plasma, concentrate and PL were similar and correlated significantly. However, there was a large inter-individual variability in cytokine concentrations between the different donor horses. The samples from some donor animals had overall very high cytokine concentrations, while samples from other donors had no measurable cytokine ingredient. This pattern was observed for all cytokines. There was a noticeable link between high cytokine concentrations in the blood products and abnormal findings in blood chemistry. Cytokine concentrations in samples from horses with abnormal findings were significantly higher than in samples from the remaining horses. The interindividual differences in cytokine concentrations could be highly relevant when using PL for therapy and cell culture, as the mode of action of the PL is likely changed depending on the presence of pro- and anti-inflammatory cytokines. Blood chemistry might be useful to predict cytokine concentrations in blood products.
在马医学中，再生疗法的应用越来越受到关注，但它仍是一个新兴且复杂的领域，仍有改进空间。血小板裂解物（PL）可作为治疗剂，也是多能间充质基质细胞培养的一种有前景的补充剂。为了在临床和实验室中能够有针对性地使用PL，了解其有效成分的更多细节至关重要。虽然到目前为止，主要在基于血小板的产品（如PL）中分析了生长因子成分，但本研究重点关注血清、血浆、血小板浓缩物和PL中细胞因子的含量。从20匹临床健康的马采集血液，进行血细胞计数和化学分析，并进一步加工成PL。通过基于血沉棕黄层的方法制备血浆和血小板浓缩物，通过冻融从浓缩物中制备PL。使用夹心酶联免疫吸附测定法分析每匹马的样本中白细胞介素（IL）-1β、-4、-6和-10、干扰素-γ和肿瘤坏死因子-α的浓度。血清、血浆、浓缩物和PL中的细胞因子浓度相似且显著相关。然而，不同供体马之间细胞因子浓度存在很大的个体间差异。一些供体动物的样本总体细胞因子浓度非常高，而其他供体的样本则没有可测量的细胞因子成分。所有细胞因子均观察到这种模式。血液制品中高细胞因子浓度与血液化学异常结果之间存在明显联系。有异常结果的马的样本中的细胞因子浓度显著高于其余马的样本。当使用PL进行治疗和细胞培养时，细胞因子浓度的个体间差异可能高度相关，因为PL的作用方式可能会根据促炎和抗炎细胞因子的存在而改变。血液化学可能有助于预测血液制品中的细胞因子浓度。
