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Cytokine-Rich Adipose Tissue Extract Production from Water-Assisted Lipoaspirate: Methodology for Clinical Use

View Article: PubMed Central - PubMed

ABSTRACT

Proper functioning wound healing strategies are sparse. Adequate vascular formation to the injured area, as well as replacement of the volume loss, is fundamental in soft tissue repair. Tissue engineering strategies have been proposed for the treatment of these injury sites. Novel cell-free substance, human adipose tissue extract (ATE), has been previously shown to induce in vitro angiogenesis and adipogenesis and in vivo soft tissue formation. This study reports the translation of ATE preparation from laboratory to the operating room (OR). ATE samples for this study were derived from adipose tissue obtained with the water-jet assisted liposuction technique from 27 healthy patients. The variables studied included incubation time (15, 30, and 45 min), temperature (room temperature vs. 37°C), and filter type to determine the optimal method yielding the most consistent total protein content, as well as consistent and high expression of adipose-derived growth factors and cytokines, including: vascular endothelial growth factor, basic fibroblast growth factor, interleukin-6, adiponectin, leptin, and insulin-like growth factor. Following the optimization, samples were produced in the OR and tested for their sterility. No significant differences were observed when comparing extract incubation time points or incubation temperature. Nonetheless, when studying the different filter types used, a syringe filter with PES membrane with larger filter area showed significantly higher protein concentration (p ≤ 0.018). When studying the different growth factor concentrations, ELISA results showed less variation in cytokine concentrations in the OR samples with the optimized protocol. All of the OR samples were tested sterile. The devised protocol is an easy and reproducible OR-ready method for ATE generation. As an attractive source of growth factors, ATE is a promising alternative in the vast field of tissue engineering. Its clinical applications include volume replacement as a complement to fillers and improvement of the permanence of fat grafts and wound healing, among other bioactive functions.

No MeSH data available.


Specific protein concentrations in OR vs. laboratory. Analysis of 16 different adipokines in samples produced in laboratory in optimization phase versus samples produced in OR after optimization. Adipokines analyzed were VEGF, IL-6, adiponectin, leptin, rantes, FGFβ, resistin, IL-8, MIP-1, TNFα, IFNγ, G-CSF, GM-CSF, IGF-1 and IL-1α. Results depicted as mean ± SD, n ≥ 6.
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f5: Specific protein concentrations in OR vs. laboratory. Analysis of 16 different adipokines in samples produced in laboratory in optimization phase versus samples produced in OR after optimization. Adipokines analyzed were VEGF, IL-6, adiponectin, leptin, rantes, FGFβ, resistin, IL-8, MIP-1, TNFα, IFNγ, G-CSF, GM-CSF, IGF-1 and IL-1α. Results depicted as mean ± SD, n ≥ 6.

Mentions: To control the quality and assure the bioactivity of the samples, growth factor measurements of both laboratory and OR samples were studied. The results show that the growth factor yields were comparable between the samples produced in the laboratory and those from the OR (Fig. 5) although the correlations varied between the growth factors (Fig. 5). The samples had less variation when performed in the OR with the optimized protocol (Fig. 5). Most of the growth factors correlated well with laboratory samples and in addition, VEGF (also higher CV) and G-CSF produced higher mean growth factor concentrations in the OR with the optimized protocol.


Cytokine-Rich Adipose Tissue Extract Production from Water-Assisted Lipoaspirate: Methodology for Clinical Use
Specific protein concentrations in OR vs. laboratory. Analysis of 16 different adipokines in samples produced in laboratory in optimization phase versus samples produced in OR after optimization. Adipokines analyzed were VEGF, IL-6, adiponectin, leptin, rantes, FGFβ, resistin, IL-8, MIP-1, TNFα, IFNγ, G-CSF, GM-CSF, IGF-1 and IL-1α. Results depicted as mean ± SD, n ≥ 6.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5035828&req=5

f5: Specific protein concentrations in OR vs. laboratory. Analysis of 16 different adipokines in samples produced in laboratory in optimization phase versus samples produced in OR after optimization. Adipokines analyzed were VEGF, IL-6, adiponectin, leptin, rantes, FGFβ, resistin, IL-8, MIP-1, TNFα, IFNγ, G-CSF, GM-CSF, IGF-1 and IL-1α. Results depicted as mean ± SD, n ≥ 6.
Mentions: To control the quality and assure the bioactivity of the samples, growth factor measurements of both laboratory and OR samples were studied. The results show that the growth factor yields were comparable between the samples produced in the laboratory and those from the OR (Fig. 5) although the correlations varied between the growth factors (Fig. 5). The samples had less variation when performed in the OR with the optimized protocol (Fig. 5). Most of the growth factors correlated well with laboratory samples and in addition, VEGF (also higher CV) and G-CSF produced higher mean growth factor concentrations in the OR with the optimized protocol.

View Article: PubMed Central - PubMed

ABSTRACT

Proper functioning wound healing strategies are sparse. Adequate vascular formation to the injured area, as well as replacement of the volume loss, is fundamental in soft tissue repair. Tissue engineering strategies have been proposed for the treatment of these injury sites. Novel cell-free substance, human adipose tissue extract (ATE), has been previously shown to induce in vitro angiogenesis and adipogenesis and in vivo soft tissue formation. This study reports the translation of ATE preparation from laboratory to the operating room (OR). ATE samples for this study were derived from adipose tissue obtained with the water-jet assisted liposuction technique from 27 healthy patients. The variables studied included incubation time (15, 30, and 45 min), temperature (room temperature vs. 37°C), and filter type to determine the optimal method yielding the most consistent total protein content, as well as consistent and high expression of adipose-derived growth factors and cytokines, including: vascular endothelial growth factor, basic fibroblast growth factor, interleukin-6, adiponectin, leptin, and insulin-like growth factor. Following the optimization, samples were produced in the OR and tested for their sterility. No significant differences were observed when comparing extract incubation time points or incubation temperature. Nonetheless, when studying the different filter types used, a syringe filter with PES membrane with larger filter area showed significantly higher protein concentration (p ≤ 0.018). When studying the different growth factor concentrations, ELISA results showed less variation in cytokine concentrations in the OR samples with the optimized protocol. All of the OR samples were tested sterile. The devised protocol is an easy and reproducible OR-ready method for ATE generation. As an attractive source of growth factors, ATE is a promising alternative in the vast field of tissue engineering. Its clinical applications include volume replacement as a complement to fillers and improvement of the permanence of fat grafts and wound healing, among other bioactive functions.

No MeSH data available.