title: Three-dimensional culture of annulus fibrosus cells within PDLLA/Bioglass composite foam scaffolds: assessment of cell attachment, proliferation and extracellular matrix production.
abstract: The objective of the present study was to assess cell attachment, proliferation and extracellular matrix (ECM) production by bovine annulus fibrosus (BAF) cells cultured in vitro in PDLLA/Bioglass composite foams. PDLLA foams incorporated with different percentages (0, 5 and 30wt%) of Bioglass particles were prepared by thermally induced phase separation (TIPS) process and characterized by scanning electron microscopy (SEM). BAF cell morphology and attachment within the PDLLA/Bioglass foams were analysed using SEM. An assessment of cell proliferation was conducted using the WST-1 assay. The amount of sulphated glycosaminoglycans (sGAG) were quantified using the 1,9-dimethylmethylene blue (DMMB) assay after 4 weeks in culture. Furthermore, the amount of collagen synthesis was determined using a hydroxyproline assay, and the presence of collagen types I and II was investigated using Western blotting. Our results reveal that PDLLA/Bioglass foam scaffolds can provide an appropriate microenvironment for BAF cell culture which enhances cell proliferation and promotes the production of sGAG, collagen type I and collagen type II. These findings provide preliminary evidence for the use of PDLLA/Bioglass composite scaffolds as cell-carrier materials for future treatments of intervertebral discs with damaged AF regions.
authors.affiliation: School of Materials, Materials Science Centre, The University of Manchester, UK.
authors.forename: Wilda
authors.initials: W
authors.lastname: Helen
authors.forename: Catherine L R
authors.initials: CL
authors.lastname: Merry
authors.forename: Jonny J
authors.initials: JJ
authors.lastname: Blaker
authors.forename: Julie E
authors.initials: JE
authors.lastname: Gough
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002417:Cattle; D002448:Cell Adhesion; D049109:Cell Proliferation; D002516:Ceramics / Q000737:chemistry*; D003094:Collagen / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D005109:Extracellular Matrix / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000737:chemistry; D006909:Hydroxyproline / Q000737:chemistry; D019344:Lactic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D008751:Methylene Blue / Q000031:analogs & derivatives / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D011091:Polyesters; D011108:Polymers / Q000737:chemistry*
chemical_list: C510584:2,2'-bis(2-oxazoline) linked poly(lactic acid); D001672:Biocompatible Materials; C022107:Bioglass; D006025:Glycosaminoglycans; D011091:Polyesters; D011108:Polymers; C435946:dimethylmethylene blue; D019344:Lactic Acid; C033616:poly(lactide); D003094:Collagen; D004247:DNA; D006909:Hydroxyproline; D008751:Methylene Blue