Prevascularization of collagen-glycosaminoglycan scaffolds: stromal vascular fraction versus adipose tissue-derived microvascular fragments.

Autor: Später T; 1Institute for Clinical & Experimental Surgery, Saarland University, 66421 Homburg/Saar, Germany., Frueh FS; Division of Plastic Surgery and Hand Surgery, University Hospital Zürich, University of Zürich, 8091 Zürich, Switzerland., Nickels RM; 1Institute for Clinical & Experimental Surgery, Saarland University, 66421 Homburg/Saar, Germany., Menger MD; 1Institute for Clinical & Experimental Surgery, Saarland University, 66421 Homburg/Saar, Germany., Laschke MW; 1Institute for Clinical & Experimental Surgery, Saarland University, 66421 Homburg/Saar, Germany.
Jazyk: angličtina
Zdroj: Journal of biological engineering [J Biol Eng] 2018 Nov 13; Vol. 12, pp. 24. Date of Electronic Publication: 2018 Nov 13 (Print Publication: 2018).
DOI: 10.1186/s13036-018-0118-3
Abstrakt: Background: The seeding of scaffolds with the stromal vascular fraction (SVF) of adipose tissue is a common prevascularization strategy in tissue engineering. Alternatively, adipose tissue-derived microvascular fragments (ad-MVF) may serve as vascularization units. In contrast to SVF single cells, they represent a mixture of intact arteriolar, capillary and venular vessel segments. Therefore, we herein hypothesized that the ad-MVF-based prevascularization of scaffolds is superior to the conventional SVF single cells-based approach.
Results: SVF single cells and ad-MVF were enzymatically isolated from epididymal fat pads of green fluorescent protein (GFP) + donor mice to assess their viability and cellular composition using fluorescence microscopy and flow cytometry. Moreover, collagen-glycosaminoglycan matrices (Integra®) were seeded with identical amounts of the isolates and implanted into full-thickness skin defects within dorsal skinfold chambers of GFP - recipient mice for the intravital fluorescent microscopic, histological and immunohistochemical analysis of implant vascularization and incorporation throughout an observation period of 2 weeks. Non-seeded matrices served as controls. While both isolates contained a comparable fraction of endothelial cells, perivascular cells, adipocytes and stem cells, ad-MVF exhibited a significantly higher viability. After in vivo implantation, the vascularization of ad-MVF-seeded scaffolds was improved when compared to SVF-seeded ones, as indicated by a significantly higher functional microvessel density. This was associated with an enhanced cellular infiltration, collagen content and density of CD31 + /GFP + microvessels particularly in the center of the implants, demonstrating a better incorporation into the surrounding host tissue. In contrast, non-seeded matrices exhibited a poor vascularization, incorporation and epithelialization over time.
Conclusions: The present study demonstrates that ad-MVF are highly potent vascularization units that markedly accelerate and improve scaffold vascularization when compared to the SVF.
Competing Interests: All animal experiments were approved by the local governmental animal protection committee (permit number: 33/2016) and conducted in accordance with the European legislation on the protection of animals (Directive 2010/63/EU) and the NIH guidelines on the care and use of laboratory animals (NIH publication #85–23 Rev. 1985).Not applicable.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Databáze: MEDLINE
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