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Calcium alginate gels as stem cell matrix-making paracrine stem cell activity available for enhanced healing after surgery.

Schmitt A, Rödel P, Anamur C, Seeliger C, Imhoff AB, Herbst E, Vogt S, van Griensven M, Winter G, Engert J - PLoS ONE (2015)

Bottom Line: Humans MSCs remained viable for the duration of 6 weeks within the gels.Human VEGF and bFGF was found in quantifiable concentrations in cell culture supernatants of gels loaded with MSCs and incubated for a period of 6 weeks.This work shows that calcium alginate gels can function as immobilization matrices for human MSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Sports Orthopedics, Technical University Munich, Ismaninger Str. 22, D-81675 Munich, Germany.

ABSTRACT
Regeneration after surgery can be improved by the administration of anabolic growth factors. However, to locally maintain these factors at the site of regeneration is problematic. The aim of this study was to develop a matrix system containing human mesenchymal stem cells (MSCs) which can be applied to the surgical site and allows the secretion of endogenous healing factors from the cells. Calcium alginate gels were prepared by a combination of internal and external gelation. The gelling behaviour, mechanical stability, surface adhesive properties and injectability of the gels were investigated. The permeability of the gels for growth factors was analysed using bovine serum albumin and lysozyme as model proteins. Human MSCs were isolated, cultivated and seeded into the alginate gels. Cell viability was determined by AlamarBlue assay and fluorescence microscopy. The release of human VEGF and bFGF from the cells was determined using an enzyme-linked immunoassay. Gels with sufficient mechanical properties were prepared which remained injectable through a syringe and solidified in a sufficient time frame after application. Surface adhesion was improved by the addition of polyethylene glycol 300,000 and hyaluronic acid. Humans MSCs remained viable for the duration of 6 weeks within the gels. Human VEGF and bFGF was found in quantifiable concentrations in cell culture supernatants of gels loaded with MSCs and incubated for a period of 6 weeks. This work shows that calcium alginate gels can function as immobilization matrices for human MSCs.

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Related in: MedlinePlus

Schematic construction of the texture analyser.(A) Compressibility and stiffness measurements using the texture analyser. The falcon tube cap was used as a constant plane surface, for stabilisation and for central positioning of the sample during the measurement. (B) Surface adhesion measurements using texture analyser. A piston reversely embedded into the gel of a syringe served as connector to the texture analyser for adhesion measurements on three different surfaces (teflon, petri dish, gelatine gel).
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pone.0118937.g001: Schematic construction of the texture analyser.(A) Compressibility and stiffness measurements using the texture analyser. The falcon tube cap was used as a constant plane surface, for stabilisation and for central positioning of the sample during the measurement. (B) Surface adhesion measurements using texture analyser. A piston reversely embedded into the gel of a syringe served as connector to the texture analyser for adhesion measurements on three different surfaces (teflon, petri dish, gelatine gel).

Mentions: A TA.XTplus Texture analyser (Stable Micro Systems, Surrey, UK) having the following technical specifications (Force Capacity: 50 kg.f (500 N); Force Resolution: 0.1 g; Loadcells: 0.5, 5, 30, 50 kg.f; Speed Range: 0.01–40 mm/s; Range Setting: 0.01–280 mm) was utilized to determine the mechanical stability of the gels. Gels were prepared in 60 ml falcon tube caps to provide a constant plane surface and to stabilize the central position of the samples during the measurement (Fig. 1a). A cylindrical probe with a circular area of 0.4 mm2 was used for all measurements. Measurements were performed at a test speed of 0.5 mm/s when a trigger force of 0.1 N was reached. Emerging forces were measured over a distance of 8.0 mm, which corresponds to half the gel height. For each formulation, three independent gels were measured. Data analysis was performed using the TextureExponent32 software (Stable Micro Systems, Surrey, UK). Gel strength was defined as the maximum force applied before rupture of the gels occurred.


Calcium alginate gels as stem cell matrix-making paracrine stem cell activity available for enhanced healing after surgery.

Schmitt A, Rödel P, Anamur C, Seeliger C, Imhoff AB, Herbst E, Vogt S, van Griensven M, Winter G, Engert J - PLoS ONE (2015)

Schematic construction of the texture analyser.(A) Compressibility and stiffness measurements using the texture analyser. The falcon tube cap was used as a constant plane surface, for stabilisation and for central positioning of the sample during the measurement. (B) Surface adhesion measurements using texture analyser. A piston reversely embedded into the gel of a syringe served as connector to the texture analyser for adhesion measurements on three different surfaces (teflon, petri dish, gelatine gel).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4368733&req=5

pone.0118937.g001: Schematic construction of the texture analyser.(A) Compressibility and stiffness measurements using the texture analyser. The falcon tube cap was used as a constant plane surface, for stabilisation and for central positioning of the sample during the measurement. (B) Surface adhesion measurements using texture analyser. A piston reversely embedded into the gel of a syringe served as connector to the texture analyser for adhesion measurements on three different surfaces (teflon, petri dish, gelatine gel).
Mentions: A TA.XTplus Texture analyser (Stable Micro Systems, Surrey, UK) having the following technical specifications (Force Capacity: 50 kg.f (500 N); Force Resolution: 0.1 g; Loadcells: 0.5, 5, 30, 50 kg.f; Speed Range: 0.01–40 mm/s; Range Setting: 0.01–280 mm) was utilized to determine the mechanical stability of the gels. Gels were prepared in 60 ml falcon tube caps to provide a constant plane surface and to stabilize the central position of the samples during the measurement (Fig. 1a). A cylindrical probe with a circular area of 0.4 mm2 was used for all measurements. Measurements were performed at a test speed of 0.5 mm/s when a trigger force of 0.1 N was reached. Emerging forces were measured over a distance of 8.0 mm, which corresponds to half the gel height. For each formulation, three independent gels were measured. Data analysis was performed using the TextureExponent32 software (Stable Micro Systems, Surrey, UK). Gel strength was defined as the maximum force applied before rupture of the gels occurred.

Bottom Line: Humans MSCs remained viable for the duration of 6 weeks within the gels.Human VEGF and bFGF was found in quantifiable concentrations in cell culture supernatants of gels loaded with MSCs and incubated for a period of 6 weeks.This work shows that calcium alginate gels can function as immobilization matrices for human MSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Sports Orthopedics, Technical University Munich, Ismaninger Str. 22, D-81675 Munich, Germany.

ABSTRACT
Regeneration after surgery can be improved by the administration of anabolic growth factors. However, to locally maintain these factors at the site of regeneration is problematic. The aim of this study was to develop a matrix system containing human mesenchymal stem cells (MSCs) which can be applied to the surgical site and allows the secretion of endogenous healing factors from the cells. Calcium alginate gels were prepared by a combination of internal and external gelation. The gelling behaviour, mechanical stability, surface adhesive properties and injectability of the gels were investigated. The permeability of the gels for growth factors was analysed using bovine serum albumin and lysozyme as model proteins. Human MSCs were isolated, cultivated and seeded into the alginate gels. Cell viability was determined by AlamarBlue assay and fluorescence microscopy. The release of human VEGF and bFGF from the cells was determined using an enzyme-linked immunoassay. Gels with sufficient mechanical properties were prepared which remained injectable through a syringe and solidified in a sufficient time frame after application. Surface adhesion was improved by the addition of polyethylene glycol 300,000 and hyaluronic acid. Humans MSCs remained viable for the duration of 6 weeks within the gels. Human VEGF and bFGF was found in quantifiable concentrations in cell culture supernatants of gels loaded with MSCs and incubated for a period of 6 weeks. This work shows that calcium alginate gels can function as immobilization matrices for human MSCs.

Show MeSH
Related in: MedlinePlus