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Volume 12
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10.3390/cells12162025
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Article

In Vitro Study of a Novel Vibrio alginolyticus-Based Collagenase for Future Medical Application

by
Lindsey Alejandra Quintero Sierra
1,
Reetuparna Biswas
1,
Alice Busato
1,‡,
Anita Conti
1,
Riccardo Ossanna
1,
Giamaica Conti
1,
Nicola Zingaretti
2,
Michele Caputo
3,
Christian Cuppari
3,
Pier Camillo Parodi
2,4,
Andrea Sbarbati
1,4,
Michele Riccio
4,5,*,† and
Francesco De Francesco
5,*,†
1
Department of Neuroscience, Biomedicine, and Movement Sciences, Human Anatomy and Histology Section, University of Verona, 37134 Verona, Italy
2
Clinic of Plastic and Reconstructive Surgery, Academic Hospital of Udine, Department of Medical Area (DAME), University of Udine, 33100 Udine, Italy
3
Fidia Farmaceutici S.p.A., R&D Local Unit Fidia Research Sud, Contrada Pizzuta, 96017 Noto, Italy
4
Research and Training Center in Regenerative Surgery, Accademia del Lipofilling, 61025 Montelabbate (PU), Italy
5
Department of Reconstructive Surgery and Hand Surgery, AOU “Ospedali Riuniti”, 60126 Ancona, Italy
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
‡
Current Affiliation: Safety Assessment Department, Aptuit (Verona) S.r.l., an Evotec Company, 37135 Verona, Italy.
Cells 2023, 12(16), 2025; https://doi.org/10.3390/cells12162025
Submission received: 7 July 2023 / Revised: 28 July 2023 / Accepted: 5 August 2023 / Published: 8 August 2023
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Abstract

Mesenchymal stem cells extracted from adipose tissue are particularly promising given the ease of harvest by standard liposuction and reduced donor site morbidity. This study proposes a novel enzymatic method for isolating stem cells using Vibrio alginolyticus collagenase, obtaining a high-quality product in a reduced time. Initially, the enzyme concentration and incubation time were studied by comparing cellular yield, proliferation, and clonogenic capacities. The optimized protocol was phenotypically characterized, and its ability to differentiate in the mesodermal lineages was evaluated. Subsequently, that protocol was compared with two Clostridium histolyticum-based collagenases, and other tests for cellular integrity were performed to evaluate the enzyme’s effect on expanded cells. The best results showed that using a concentration of 3.6 mg/mL Vibrio alginolyticus collagenase allows extracting stem cells from adipose tissue after 20 min of enzymatic reaction like those obtained with Clostridium histolyticum-based collagenases after 45 min. Moreover, the extracted cells with Vibrio alginolyticus collagenase presented the phenotypic characteristics of stem cells that remain after culture conditions. Finally, it was seen that Vibrio alginolyticus collagenase does not reduce the vitality of expanded cells as Clostridium histolyticum-based collagenase does. These findings suggest that Vibrio alginolyticus collagenase has great potential in regenerative medicine, given its degradation selectivity by protecting vital structures for tissue restructuration.
Keywords: collagenase; Vibrio alginolyticus; regenerative medicine; adipose tissue; disaggregation collagenase; Vibrio alginolyticus; regenerative medicine; adipose tissue; disaggregation

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MDPI and ACS Style

Quintero Sierra, L.A.; Biswas, R.; Busato, A.; Conti, A.; Ossanna, R.; Conti, G.; Zingaretti, N.; Caputo, M.; Cuppari, C.; Parodi, P.C.; et al. In Vitro Study of a Novel Vibrio alginolyticus-Based Collagenase for Future Medical Application. Cells 2023, 12, 2025. https://doi.org/10.3390/cells12162025

AMA Style

Quintero Sierra LA, Biswas R, Busato A, Conti A, Ossanna R, Conti G, Zingaretti N, Caputo M, Cuppari C, Parodi PC, et al. In Vitro Study of a Novel Vibrio alginolyticus-Based Collagenase for Future Medical Application. Cells. 2023; 12(16):2025. https://doi.org/10.3390/cells12162025

Chicago/Turabian Style

Quintero Sierra, Lindsey Alejandra, Reetuparna Biswas, Alice Busato, Anita Conti, Riccardo Ossanna, Giamaica Conti, Nicola Zingaretti, Michele Caputo, Christian Cuppari, Pier Camillo Parodi, and et al. 2023. "In Vitro Study of a Novel Vibrio alginolyticus-Based Collagenase for Future Medical Application" Cells 12, no. 16: 2025. https://doi.org/10.3390/cells12162025

APA Style

Quintero Sierra, L. A., Biswas, R., Busato, A., Conti, A., Ossanna, R., Conti, G., Zingaretti, N., Caputo, M., Cuppari, C., Parodi, P. C., Sbarbati, A., Riccio, M., & De Francesco, F. (2023). In Vitro Study of a Novel Vibrio alginolyticus-Based Collagenase for Future Medical Application. Cells, 12(16), 2025. https://doi.org/10.3390/cells12162025

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