Expansion of human bone marrow stromal cells on poly-(dl-lactide-co-glycolide) (PDLLGA) hollow fibres designed for use in skeletal tissue engineering

Morgan, Sara; Tilley, Simon; Perera, Semali; Ellis, Marianne; Kanczler, Janos M.; Chaudhuri, Julian B.; Oreffo, Richard O.C.

doi:10.1016/j.biomaterials.2007.08.029

Cited by 50 publications

(40 citation statements)

References 64 publications

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“…Various in vitro bone regeneration strategies have relied on the use of the osteogenic supplements (dexamethasone, ascorbic acid, and b-glycerophosphate) to induce mineralization in vitro, 12,13,49,51,[53][54][55][56][57][58][59][60][61][62] and each supplement has been shown to be imperative to induce mineralization of MSCs in vitro. 9,[11][12][13][14][15][16][17]22,29,[63][64][65][66][67] The main rationale behind the use of these supplements is to provide signaling proteins or enzymes to activate mineral production by MSCs.…”

Section: Discussionmentioning

confidence: 99%

Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Freeman

Stevens

Owens

et al. 2016

Tissue Engineering Part A

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In vitro bone regeneration strategies that prime mesenchymal stem cells (MSCs) with chondrogenic factors, to mimic aspects of the endochondral ossification process, have been shown to promote mineralization and vascularization by MSCs both in vitro and when implanted in vivo. However, these approaches required the use of osteogenic supplements, namely dexamethasone, ascorbic acid, and b-glycerophosphate, none of which are endogenous mediators of bone formation in vivo. Rather MSCs, endothelial progenitor cells, and chondrocytes all reside in proximity within the cartilage template and might paracrineally regulate osteogenic differentiation. Thus, this study tests the hypothesis that an in vitro bone regeneration approach that mimics the cellular niche existing during endochondral ossification, through coculture of MSCs, endothelial cells, and chondrocytes, will obviate the need for extraneous osteogenic supplements and provide an alternative strategy to elicit osteogenic differentiation of MSCs and mineral production. The specific objectives of this study were to (1) mimic the cellular niche existing during endochondral ossification and (2) investigate whether osteogenic differentiation could be induced without the use of any external growth factors. To test the hypothesis, we evaluated the mineralization and vessel formation potential of (a) a novel methodology involving both chondrogenic priming and the coculture of human umbilical vein endothelial cells (HUVECs) and MSCs compared with (b) chondrogenic priming of MSCs alone, (c) addition of HUVECs to chondrogenically primed MSC aggregates, (d-f) the same experimental groups cultured in the presence of osteogenic supplements and (g) a noncoculture group cultured in the presence of osteogenic growth factors alone. Biochemical (DNA, alkaline phosphatase [ALP], calcium, CD31 + , vascular endothelial growth factor [VEGF]), histological (alcian blue, alizarin red), and immunohistological (CD31 + ) analyses were conducted to investigate osteogenic differentiation and vascularization at various time points (1, 2, and 3 weeks). The coculture methodology enhanced both osteogenesis and vasculogenesis compared with osteogenic differentiation alone, whereas osteogenic supplements inhibited the osteogenesis and vascularization (ALP, calcium, and VEGF) induced through coculture alone. Taken together, these results suggest that chondrogenic and vascular priming can obviate the need for osteogenic supplements to induce osteogenesis of human MSCs in vitro, while allowing for the formation of rudimentary vessels.

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Section: Discussionmentioning

confidence: 99%

Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Freeman

Stevens

Owens

et al. 2016

Tissue Engineering Part A

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“…Functionalization of end-groups: the end-capped polymers having ester end (opposed by free carboxylic group) indicates longer degradation half-lives 46,47 . However, the degradation behaviour of PLGA is highly influenced by the shape of the device based on the penetrability of water.…”

Section: Properties Of Plgamentioning

confidence: 99%

“…Several processes to obtain micro and Nano-fibrous composite scaffolds have used 53 . Morgan (2007) used the wet-spinning method for obtaining hollow fibers, as scaffolds applied in combination with human bone marrow stromal cell that helps in initiating natural bone fixation and reconstruction 47 . In comparison, the nano-fibrous composites possess similar structure to natural bone extracellular matrix (ECM) and they can take secondary stimuli to the cultured cells.…”

Section: B) Fibrous Scaffoldsmentioning

confidence: 99%

Untitled

2018

IJPSR

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“…Poly(lactide-co-glycolide) (PLGA) flat sheet (FS) and hollow fibre membranes are suitable scaffolds for bone tissue engineering Chaudhuri 2007, 2008; Morgan et al 2007). Scaffolds must endure a number of chemical treatments both in preparation for cell culture, i.e.…”

Section: Introductionmentioning

confidence: 99%

Post-culture treatment protocols for PLGA membrane scaffolds

2009

Self Cite

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The interactions of post-culture treatments reagents used for fixing, lysing and cell quantification on poly(lactide-co-glycolide) (PLGA) flat sheet membrane scaffolds are presented. Lysing with Alkaline buffer solution/Triton X-100/MilliQ water (ATM) and fixing with 10% Neutral Buffered Formalin (10% NBF) had no affect on membrane structure while fixing with 95% ethanol caused smoothing of the surface, shrinkage and a reduction in surface area of 55, 48 and 33, for 100:0, 75:25 and 50:50 (PLA:PGA), respectively. PicoGreen assay was selected for cell (560pZIPv.neo) quantification since the background noise would not affect readings for cell numbers over 3,000 cells/cm(2), while the background reading was too high for MTT and Methylene Blue (MB). MB at 0.5% (w/v) was, however, deemed suitable for visualising cell morphology on the membranes. Furthermore ATM buffer was suitable for the PicoGreen assay, which allows the same samples to be used for quantification of alkaline phosphatase activity.

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Expansion of human bone marrow stromal cells on poly-(dl-lactide-co-glycolide) (PDLLGA) hollow fibres designed for use in skeletal tissue engineering

Cited by 50 publications

References 64 publications

Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Untitled

Post-culture treatment protocols for PLGA membrane scaffolds

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