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羊膜祖细胞条件培养基的抗菌作用:迈向牛乳腺炎的组织再生疗法

Antimicrobial Effects of Conditioned Medium From Amniotic Progenitor Cells and : Toward Tissue Regenerative Therapies for Bovine Mastitis.

作者信息

Lange-Consiglio Anna, Gusmara Claudia, Manfredi Emanuela, Idda Antonella, Soggiu Alessio, Greco Viviana, Bonizzi Luigi, Cremonesi Fausto, Zecconi Alfonso

机构信息

Department of Veterinary Medicine (DIMEVET), Università degli Studi di Milano, Milan, Italy.

Reproduction Unit, Centro Clinico-Veterinario e Zootecnico-Sperimentale di Ateneo, Università degli Studi di Milano, Milan, Italy.

出版信息

Front Vet Sci. 2019 Dec 19;6:443. doi: 10.3389/fvets.2019.00443. eCollection 2019.

Abstract

There is increasing evidence to suggest that, in addition to their regenerative effect, mesenchymal stromal cells (MSCs), and their secretome have an anti-inflammatory and antimicrobial role in the innate immune response in conditions such as sepsis. However, there is no published information on the effect of MSCs in bovine mastitis. Mastitis often results in extensive tissue damage due to multi-microorganism co-infection. This study investigated the ability of amniotic-derived conditioned medium (CM), and , to counteract microbial action and restore healthy tissue capable of milk production. Following determination of a dose-response curve, 10,000 colony-forming units (CFU) of () were inoculated into bovine mammary epithelial cell culture with and without 10% CM (supplemented either at the time of bacteria inoculation or after 4 h). Acridine orange staining was used to assess cell viability/apoptosis. Additionally, an study was performed using 48 dairy cows with acute and chronic mastitis, treated with CM (treated group) or antibiotics (control group). results showed that CM can attenuate bacterial growth, as evaluated by the number of CFU. After 24 h of culture with , 89.67% of mammary epithelial cells treated with CM were still alive, whereas all cells cultured without CM were dead. Rates of epithelial cell survival (60.67%) were similar when CM was added 4 h after bacteria inoculation. There was no difference in somatic cell count between cases of acute mastitis in the CM-treated or control group in the study. However, relapses in chronic mastitis were less common in the group receiving CM. Our results show that CM is able to mitigate bacterial growth and may be particularly useful in the treatment of chronic mastitis, aiding restoration of milk production in cows that would otherwise be removed from the production cycle.

摘要

越来越多的证据表明,间充质基质细胞(MSCs)及其分泌组除了具有再生作用外,在脓毒症等病症的先天免疫反应中还具有抗炎和抗菌作用。然而,关于MSCs对牛乳腺炎影响的公开信息尚无报道。乳腺炎常因多种微生物共同感染导致广泛的组织损伤。本研究调查了羊膜来源的条件培养基(CM)抵消微生物作用并恢复能够产奶的健康组织的能力。在确定剂量反应曲线后,将10,000个金黄色葡萄球菌的菌落形成单位(CFU)接种到有或无10% CM(在细菌接种时或4小时后添加)的牛乳腺上皮细胞培养物中。使用吖啶橙染色评估细胞活力/凋亡。此外,对48头患有急性和慢性乳腺炎的奶牛进行了一项研究,分别用CM(治疗组)或抗生素(对照组)进行治疗。结果表明,通过CFU数量评估,CM可减弱细菌生长。与金黄色葡萄球菌共培养24小时后,用CM处理的乳腺上皮细胞中有89.67%仍存活,而未用CM培养的所有细胞均死亡。在细菌接种4小时后添加CM时,上皮细胞存活率(60.67%)相似。在该研究中,CM治疗组或对照组的急性乳腺炎病例之间的体细胞计数没有差异。然而,接受CM治疗的慢性乳腺炎组复发情况较少。我们的结果表明,CM能够减轻细菌生长,可能对慢性乳腺炎的治疗特别有用,有助于恢复那些否则将被从生产周期中淘汰的奶牛的产奶能力。

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