title: Diffusion chamber system for testing of collagen-based cell migration barriers for separation of ligament enthesis zones in tissue-engineered ACL constructs.
abstract: A temporary barrier separating scaffold zones seeded with different cell types prevents faster growing cells from overgrowing co-cultured cells within the same construct. This barrier should allow sufficient nutrient diffusion through the scaffold. The aim of this study was to test the effect of two variants of collagen-based barriers on macromolecule diffusion, viability, and the spreading efficiency of primary ligament cells on embroidered scaffolds. Two collagen barriers, a thread consisting of a twisted film tape and a sponge, were integrated into embroidered poly(lactic-co-caprolactone) and polypropylene scaffolds, which had the dimension of lapine anterior cruciate ligaments (ACL). A diffusion chamber system was designed and established to monitor nutrient diffusion using fluorescein isothiocyanate-labeled dextran of different molecular weights (20, 40, 150, 500 kDa). Vitality of primary lapine ACL cells was tested at days 7 and 14 after seeding using fluorescein diacetate and ethidium bromide staining. Cell spreading on the scaffold surface was measured using histomorphometry. Nuclei staining of the cross-sectioned scaffolds revealed the penetration of ligament cells through both barrier types. The diffusion chamber was suitable to characterize the diffusivity of dextran molecules through embroidered scaffolds with or without integrated collagen barriers. The diffusion coefficients were generally significantly lower in scaffolds with barriers compared to those without barriers. No significant differences between diffusion coefficients of both barrier types were detected. Both barriers were cyto-compatible and prevented most of the ACL cells from crossing the barrier, whereby the collagen thread was easier to handle and allowed a higher rate of cell spreading.
authors: a Leibniz-Institut für Polymerforschung, Institute of Polymer Materials , Hohe Straße 6, 01069 Dresden , Germany.
authors: J
authors: J
authors: Hahner
authors: b Central Laboratory , DRK Manniske-Hospital , An der Wipper 2, 06567 Bad Frankenhausen , Germany.|c Department of Bioanalytics , Technical University , Seestraße 13, 13353 Berlin , Germany.
authors: M
authors: M
authors: Hoyer
authors: a Leibniz-Institut für Polymerforschung, Institute of Polymer Materials , Hohe Straße 6, 01069 Dresden , Germany.
authors: S
authors: S
authors: Hillig
authors: d Institute of Anatomy , Paracelsus Medical University , Salzburg and Nuremberg, Prof. Ernst Nathan Straße 1, 90340 Nuremberg , Germany.|e Department of Trauma and Reconstructive Surgery , Charité-Universitätsmedizin Berlin , Campus Benjamin Franklin, Garystrasse 5, 14195 Berlin , Germany.
authors: G
authors: G
authors: Schulze-Tanzil
authors: f Research Institute of Leather and Plastic Sheeting - FILK , Meißner Ring 1-5, 09599 Freiberg , Germany.
authors: M
authors: M
authors: Meyer
authors: f Research Institute of Leather and Plastic Sheeting - FILK , Meißner Ring 1-5, 09599 Freiberg , Germany.
authors: M
authors: M
authors: Schröpfer
authors: g Forschungseinrichtungen für Experimentelle Medizin, Charité-Universitätsmedizin Berlin , Campus Benjamin Franklin, Kramerstr.6-10, 12207 Berlin , Germany.
authors: A
authors: A
authors: Lohan
authors: c Department of Bioanalytics , Technical University , Seestraße 13, 13353 Berlin , Germany.
authors: L-A
authors: LA
authors: Garbe
authors: a Leibniz-Institut für Polymerforschung, Institute of Polymer Materials , Hohe Straße 6, 01069 Dresden , Germany.|h Institute of Materials Science , Technische Universität Dresden , Helmholtzstraße 10, 01062 Dresden , Germany.
authors: G
authors: G
authors: Heinrich
authors: a Leibniz-Institut für Polymerforschung, Institute of Polymer Materials , Hohe Straße 6, 01069 Dresden , Germany.
authors: A
authors: A
authors: Breier
mesh_terms: D000818:Animals; D016118:Anterior Cruciate Ligament / Q000166:cytology* / Q000378:metabolism / Q000601:surgery; D059549:Anterior Cruciate Ligament Reconstruction / Q000379:methods*; D002417:Cattle; D002448:Cell Adhesion; D054443:Cell Migration Assays / Q000379:methods; D002465:Cell Movement; D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry*; D003239:Connective Tissue Cells / Q000166:cytology* / Q000378:metabolism; D015194:Diffusion Chambers, Culture; D005260:Female; D006801:Humans; D008422:Materials Testing / Q000295:instrumentation / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D011091:Polyesters / Q000737:chemistry; D011126:Polypropylenes / Q000737:chemistry; D011817:Rabbits; D013499:Surface Properties; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D011091:Polyesters; D011126:Polypropylenes; C062968:lactide-caprolactone copolymer; D003094:Collagen
keywords: anterior cruciate ligament; collagen barrier; embroidered scaffold; enthesis; ligament tissue engineering; poly(lactic-co-caprolactone)