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

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

doi:10.3389/fvets.2019.00443

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

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

Authors

Affiliations

¹ Department of Veterinary Medicine (DIMEVET), Università degli Studi di Milano, Milan, Italy.
² Reproduction Unit, Centro Clinico-Veterinario e Zootecnico-Sperimentale di Ateneo, Università degli Studi di Milano, Milan, Italy.
³ Private Practitioner, Milan, Italy.
⁴ Institute of Biochemistry and Clinical Biochemistry, Università del Sacro Cuore Roma, Rome, Italy.
⁵ Fondazione Policlinico Universitario Agostino Gemelli IRCCS, Rome, Italy.

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), in vitro and in vivo, 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 Staphylococcus aureus (S. aureus) 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 in vivo study was performed using 48 dairy cows with acute and chronic mastitis, treated with CM (treated group) or antibiotics (control group). In vitro results showed that CM can attenuate bacterial growth, as evaluated by the number of CFU. After 24 h of culture with S. aureus, 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 in vivo 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 in vitro 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.

Keywords: bacteria; bovine; conditioned medium; mastitis; regenerative medicine.