Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysis
Introduction: Adipose tissue is widely exploited in regenerative medicine thanks to its trophic properties, mainly based on the presence of adipose-derived stromal cells. Numerous devices have been developed to promote its clinical use, leading to the introduction of one-step surgical procedures to...
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| Format: | Article |
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Frontiers Media S.A.
2023-01-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.911600/full |
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| author | Claudia Cicione Gianluca Vadalà Gianluca Vadalà Giuseppina Di Giacomo Veronica Tilotta Luca Ambrosio Luca Ambrosio Fabrizio Russo Fabrizio Russo Biagio Zampogna Biagio Zampogna Francesca Cannata Rocco Papalia Rocco Papalia Vincenzo Denaro |
| author_facet | Claudia Cicione Gianluca Vadalà Gianluca Vadalà Giuseppina Di Giacomo Veronica Tilotta Luca Ambrosio Luca Ambrosio Fabrizio Russo Fabrizio Russo Biagio Zampogna Biagio Zampogna Francesca Cannata Rocco Papalia Rocco Papalia Vincenzo Denaro |
| author_sort | Claudia Cicione |
| collection | DOAJ |
| description | Introduction: Adipose tissue is widely exploited in regenerative medicine thanks to its trophic properties, mainly based on the presence of adipose-derived stromal cells. Numerous devices have been developed to promote its clinical use, leading to the introduction of one-step surgical procedures to obtain minimally manipulated adipose tissue derivatives. However, only a few studies compared their biological properties. This study aimed to characterize micro-fragmented (MAT) and nanofat adipose tissue (NAT) obtained with two different techniques.Methods: MAT, NAT and unprocessed lipoaspirate were collected from surgical specimens. RNA extraction and collagenase isolation of stromal vascular fraction (SVF) were performed. Tissue sections were analysed by histological and immunohistochemical (collagen type I, CD31, CD34 and PCNA) staining to assess tissue morphology and cell content. qPCR was performed to evaluate the expression of stemness-related (SOX2, NANOG and OCT3/4), extracellular matrix (COL1A1) and inflammatory genes (IL1β, IL6 and iNOS). Furthermore, multilineage differentiation was assessed following culture in adipogenic and osteogenic media and staining with Oil Red O and Alizarin red. ASC immunophenotype was assessed by flow cytometric analysis of CD90, CD105, CD73 and CD45.Results: Histological and immunohistochemical results showed an increased amount of stroma and a reduction of adipocytes in MAT and NAT, with the latter displaying the highest content of collagen type I, CD31, CD34 and PCNA. From LA to MAT and NAT, an increasing expression of NANOG, SOX2, OCT3/4, COL1A1 and IL6 was noted, while no significant differences in terms of IL1β and iNOS emerged. No statistically significant differences were noted between NAT and SVF in terms of stemness-related genes, while the latter demonstrated a significantly higher expression of stress-related markers. SVF cells derived from all three samples (LA, MAT, and NAT) showed a similar ASC immunoprofile as well as osteogenic and adipogenic differentiation.Discussion: Our results showed that both MAT and NAT techniques allowed the rapid isolation of ASC-rich grafts with a high anabolic and proliferative potential. However, NAT showed the highest levels of extracellular matrix content, replicating cells, and stemness gene expression. These results may provide precious clues for the use of adipose tissue derivatives in the clinical setting. |
| first_indexed | 2024-04-10T22:30:49Z |
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| id | doaj.art-83a18a4be40b48ed999a186a4983c39b |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-10T22:30:49Z |
| publishDate | 2023-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-83a18a4be40b48ed999a186a4983c39b2023-01-17T05:36:29ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-01-011110.3389/fbioe.2023.911600911600Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysisClaudia Cicione0Gianluca Vadalà1Gianluca Vadalà2Giuseppina Di Giacomo3Veronica Tilotta4Luca Ambrosio5Luca Ambrosio6Fabrizio Russo7Fabrizio Russo8Biagio Zampogna9Biagio Zampogna10Francesca Cannata11Rocco Papalia12Rocco Papalia13Vincenzo Denaro14Laboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyLaboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyOperative Research Unit of Orthopaedic and Trauma Surgery, Fondazione Policlinico Universitario Campus Bio-Medico, Rome, ItalyLaboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyLaboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyLaboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyOperative Research Unit of Orthopaedic and Trauma Surgery, Fondazione Policlinico Universitario Campus Bio-Medico, Rome, ItalyLaboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyOperative Research Unit of Orthopaedic and Trauma Surgery, Fondazione Policlinico Universitario Campus Bio-Medico, Rome, ItalyLaboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyOperative Research Unit of Orthopaedic and Trauma Surgery, Fondazione Policlinico Universitario Campus Bio-Medico, Rome, ItalyOperative Research Unit of Endocrinology and Diabetes, Fondazione Policlinico Universitario Campus Bio-Medico, Rome, ItalyLaboratory for Regenerative Orthopaedics, Research Unit of Orthopaedic and Trauma Surgery, Department of Medicine and Surgery, Università Campus Bio-Medico di Roma, Rome, ItalyOperative Research Unit of Orthopaedic and Trauma Surgery, Fondazione Policlinico Universitario Campus Bio-Medico, Rome, ItalyOperative Research Unit of Orthopaedic and Trauma Surgery, Fondazione Policlinico Universitario Campus Bio-Medico, Rome, ItalyIntroduction: Adipose tissue is widely exploited in regenerative medicine thanks to its trophic properties, mainly based on the presence of adipose-derived stromal cells. Numerous devices have been developed to promote its clinical use, leading to the introduction of one-step surgical procedures to obtain minimally manipulated adipose tissue derivatives. However, only a few studies compared their biological properties. This study aimed to characterize micro-fragmented (MAT) and nanofat adipose tissue (NAT) obtained with two different techniques.Methods: MAT, NAT and unprocessed lipoaspirate were collected from surgical specimens. RNA extraction and collagenase isolation of stromal vascular fraction (SVF) were performed. Tissue sections were analysed by histological and immunohistochemical (collagen type I, CD31, CD34 and PCNA) staining to assess tissue morphology and cell content. qPCR was performed to evaluate the expression of stemness-related (SOX2, NANOG and OCT3/4), extracellular matrix (COL1A1) and inflammatory genes (IL1β, IL6 and iNOS). Furthermore, multilineage differentiation was assessed following culture in adipogenic and osteogenic media and staining with Oil Red O and Alizarin red. ASC immunophenotype was assessed by flow cytometric analysis of CD90, CD105, CD73 and CD45.Results: Histological and immunohistochemical results showed an increased amount of stroma and a reduction of adipocytes in MAT and NAT, with the latter displaying the highest content of collagen type I, CD31, CD34 and PCNA. From LA to MAT and NAT, an increasing expression of NANOG, SOX2, OCT3/4, COL1A1 and IL6 was noted, while no significant differences in terms of IL1β and iNOS emerged. No statistically significant differences were noted between NAT and SVF in terms of stemness-related genes, while the latter demonstrated a significantly higher expression of stress-related markers. SVF cells derived from all three samples (LA, MAT, and NAT) showed a similar ASC immunoprofile as well as osteogenic and adipogenic differentiation.Discussion: Our results showed that both MAT and NAT techniques allowed the rapid isolation of ASC-rich grafts with a high anabolic and proliferative potential. However, NAT showed the highest levels of extracellular matrix content, replicating cells, and stemness gene expression. These results may provide precious clues for the use of adipose tissue derivatives in the clinical setting.https://www.frontiersin.org/articles/10.3389/fbioe.2023.911600/fulladipose tissuemesenchymal stromal cellscell therapystromal vascular fractionregenerative medicine |
| spellingShingle | Claudia Cicione Gianluca Vadalà Gianluca Vadalà Giuseppina Di Giacomo Veronica Tilotta Luca Ambrosio Luca Ambrosio Fabrizio Russo Fabrizio Russo Biagio Zampogna Biagio Zampogna Francesca Cannata Rocco Papalia Rocco Papalia Vincenzo Denaro Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysis Frontiers in Bioengineering and Biotechnology adipose tissue mesenchymal stromal cells cell therapy stromal vascular fraction regenerative medicine |
| title | Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysis |
| title_full | Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysis |
| title_fullStr | Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysis |
| title_full_unstemmed | Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysis |
| title_short | Micro-fragmented and nanofat adipose tissue derivatives: In vitro qualitative and quantitative analysis |
| title_sort | micro fragmented and nanofat adipose tissue derivatives in vitro qualitative and quantitative analysis |
| topic | adipose tissue mesenchymal stromal cells cell therapy stromal vascular fraction regenerative medicine |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.911600/full |
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