Mineralization of marrow-stromal osteoblasts MBA-15 on three-dimensional carriers

Abstract

The present study describes a new three-dimensional (3-D) culture system that enables the maintenance and phenotypic expression of bone marrow stromal osteoblasts. This culture substratum is advantageous in that it provides suitable conditions for attachment, growth, and differentiation of cells forming 3-D layers. The MBA-15 cell line was grown in unlimited quantities on 3-D Fibro-Cel carriers. These cells mineralized when exposed to ascorbic acid and beta-glycerophosphate (beta GP). Under these mineralization conditions, mRNA expressions of procollagen alpha 2(I) and [3H]-proline-labeled protein were increased. The expression of mRNA for osteonectin and to a lesser extent, for osteopontin was increased, whereas alkaline phosphatase and biglycan remained unaffected under similar conditions. Exposure of mineralizing cultures to dexamethasone reduced mRNA of procollagen alpha 2 (I) and osteonectin to control level. Scanning electron microscopy revealed that cells were grown along the fabric's fibers and produced collagen fibrils. Under appropriate conditions, extensive mineralization had taken place. The mineralization process involves the formation of calcospherites, and correlates with an increase in calcium content. The Fibro-Cel carriers enable formation of 3-D architecture and mineralized tissue in vitro.

Selective differentiation of mammalian bone marrow stromal cells cultured on three-dimensional polymer foams

“…This result is consistent with previous studies in which dex and ␤GP induced the osteogenic differentiation of BMSC in monolayers 23 and on 3D matrices. [7][8][9] In previous 3D culture systems, however, ECM mineralization was restricted to an approximately 0.2-mm-thick peripheral layer, both on microcarriers 7,9 and in cell-polymer constructs. 8,10 In the present study, the mineralized foci were distributed throughout the construct volume and represented more than 12% of the total available area in 4-week constructs.…”

“…[7][8][9] We previously reported that 3D poly(glycolic acid) (PGA) meshes supported the formation of cartilaginous and bone-like tissues by avian BMSC expanded in the presence of fibroblast growth factor-2 (FGF-2). 10 However, mammalian (bo-vine calf) BMSC cultured on PGA meshes under the same conditions failed to differentiate over 4 weeks in vitro and cell-polymer constructs collapsed.…”

The effect of surface treatment on the surface texture and contact angle of electrochemically deposited hydroxyapatite coating and on its interaction with bone-forming cells

“…We used mouse osteogenic cell line MBA-15, which expresses osteoblastic phenotype in vitro and forms bone in vivo [33,34]. On each Ti sample, either coated or uncoated, 7000 cells were seeded.…”

RGD‐modified dihydrolipoamide dehydrogenase as a molecular bridge for enhancing the adhesion of bone forming cells to titanium dioxide implant surfaces

“…In this study, DLDH RGD was prepared by adding GRGDSP residues at the N‐ and C‐termini of DLDH monomer, in order to serve as a molecular bridge between MAB‐15 osteogenic cells and TiO 2 implants . Recently, we have shown that DLDH retains its structural conformation and that its TiO 2 binding is not hindered by the RGD modification .…”