Good manufacturing practice–grade fibrin gel is useful as a scaffold for human mesenchymal stromal cells and supports in vitro osteogenic differentiation

Trombi, Luisa; D’Alessandro, Delfo; Pacini, Stefania; Fiorentino, Bruno; Scarpellini, M.; Fazzi, Rita; Galimberti, Sara; Guazzini, S.; Petrini, Mario

doi:10.1111/j.1537-2995.2008.01829.x

Cited by 23 publications

(19 citation statements)

References 22 publications

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“…This was particularly evident 2 d and 1 week after seeding and is in line with previous reports showing that fibrin is supportive of proliferation of mesenchymal stem cells in tissue engineering applications (Galler et al, 2011;Nair et al, 2009;Trombi et al, 2008). The concentration of fibrinogen appears to be influential for the degree of proliferation as it affects the microstructure of the resulting fibrin matrix.…”

Section: Discussionsupporting

confidence: 76%

“…The concentration of fibrinogen appears to be influential for the degree of proliferation as it affects the microstructure of the resulting fibrin matrix. Trombi et al (2008) have shown the architecture of a fibrin matrix containing 3 % fibrinogen has shown the most favourable results. The concentration used in the present study is well within the range of 3 % and therefore suitable …”

Section: Discussionmentioning

confidence: 99%

“…While some studies have reported a clearly enhancing effect on the expression of osteogenic markers that occurs in fibrin clots (Nair et al, 2009;Trombi et al, 2008;Chen et al, 2009) or fibrin-modified scaffolds (Schantz et al, 2005), other authors question the value of fibrin (Demol et al, 2011). The composition of the fibrin-containing matrix tested appears to play an important role for the expression of bone specific markers.…”

Section: N Lohse Et Al Differentiation and Vegf Production Of Bone Cmentioning

confidence: 95%

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Effect of fibrin on osteogenic differentiation and VEGF expression of bone marrow stromal cells in mineralised scaffolds: a three-dimensional analysis

Lohse

Schulz²,

Schliephake³

2012

eCM

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The aim of the present study was to test the hypothesis that a fibrin matrix enhances the osteogenic differentiation and expression of vascular endothelial growth factor (VEGF) by human bone marrow stromal cells (hBMSCs) seeded into mineralised scaffolds. Porous calcium carbonate scaffolds were droplet seeded with hBMSCs using a matrix containing 3 % fibrinogen and cultured for 3 weeks. Seeded scaffolds without the fibrin matrix served as controls. The scaffolds were evaluated, using undecalcified thick sections, for fluorescence staining for nuclei, osteocalcin (OC) and VEGF. The sections were systematically scanned using optical sectioning and three dimensional distributions of cells and positive staining indicating expression of OC and VEGF were reconstructed from the z-stacks. The fibrin matrix maintained a significantly higher level of cell numbers after 2 d and 1 week and delayed the onset of osteogenic differentiation while sustaining a significantly higher level of OC and VEGF expression after 2 and 3 weeks, starting from the periphery of the scaffolds. There was a decrease in cell density from the periphery to the centre of the scaffolds in both groups The percentage of cells expressing OC and VEGF was significantly different between the centre and the periphery of the scaffolds in the fibrin(+) group but not in the controls. It is concluded that the fibrin matrix used appears to be a useful adjunct for supporting and sustaining osteogenic and angiogenic activity of hBMSCs in tissue engineered constructs. This could help to improve their performance in a clinical setting.

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Section: Discussionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: N Lohse Et Al Differentiation and Vegf Production Of Bone Cmentioning

confidence: 95%

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Effect of fibrin on osteogenic differentiation and VEGF expression of bone marrow stromal cells in mineralised scaffolds: a three-dimensional analysis

Lohse

Schulz²,

Schliephake³

2012

eCM

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“…In this study, the gel in addition to being the delivery vehicle also doubled up as the scaffolding material. According to studies reported in literature, a pre-formed solid matrix helps with improved retention of cells at the defect site, provides the cells with a substrate for bone deposition, acts as filler at the defect site and in many cases, the osteoconductive and osteoinductive properties of the scaffold promote faster bone healing [18][19][20][21]. In fact, we have preliminary results which indicate that the calcium phosphate ceramics are necessary to generate bone in our system (data not shown).…”

Section: Page 19 Of 35mentioning

confidence: 57%

“…vitro that fibrin based gels supported hMSC proliferation and differentiation and could be used as a delivery vehicle for hMSCs [18].…”

Section: Page 4 Of 35mentioning

confidence: 99%

Engineering New Bone via a Minimally Invasive Route Using Human Bone Marrow-Derived Stromal Cell Aggregates, Microceramic Particles, and Human Platelet-Rich Plasma Gel

Chatterjea

Yuan

Chatterjea

et al. 2013

Tissue Engineering Part A

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Tissue Engineering Part A eering new bone via a minimally invasive route using human bone marrow derived stromal cell aggregates, micro ceramic particles and human platelet rich plasma gel (doi: 10.1089(doi: 10. /ten.TEA.2012 This article has been peer-reviewed and accepted for publication, but has yet to undergo copyediting and proof correction. The final published version may differ from this proof. 2 AbstractThere is a rise in the popularity of arthroscopic procedures in orthopedics. However, the majority of cell based bone tissue engineered constructs rely on solid pre-formed scaffolding materials, which require large incisions and extensive dissections for placement at the defect site. Thus, they are not suitable for minimally invasive techniques.The aim of this study was to develop a clinically relevant, easily moldable, bone tissue engineered construct (TEC), amenable to minimally invasive techniques, using human mesenchymal stromal cells (hMSC) and calcium phosphate micro particles in combination with an in-situ forming platelet rich plasma (PRP) gel obtained from human platelets. Most conventional TECs rely on seeding and culturing single cell suspensions of hMSCs on scaffolds. However, for generating TECs amenable to the minimally invasive approach, it was essential to aggregate the hMSCs in vitro prior to seeding them on the scaffolds as unaggregated MSCs did not generate any bone. 24 hours of in vitro aggregation was determined to be optimal for maintaining cell viability in vitro and bone formation in vivo. Moreover, no statistically significant difference was observed in the amount of bone formed when the TECs were implanted via an open approach or a minimally invasive route. TECs generated using MSCs from three different human donors generated new bone through the minimally invasive route in a reproducible manner, suggesting that these TECs could be a viable alternative to pre-formed scaffoldsemployed through an open surgery for treating bone defects. Page 2 of 35Tissue Engineering Part A eering new bone via a minimally invasive route using human bone marrow derived stromal cell aggregates, micro ceramic particles and human platelet rich plasma gel

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A rapid method for obtaining mesenchymal stem cells and platelets from bone marrow aspirate

Dozza

Gobbi

Lucarelli

et al. 2012

J Tissue Eng Regen Med

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Mesenchymal stem cells (MSCs) and platelet‐rich plasma (PRP) are currently used alone or in combination for therapeutic applications especially for bone repair. We tested whether MSCs can be isolated from bone marrow (BM) aspirate using a commercially available kit commonly used to obtain PRP from peripheral blood (PB). Results revealed that mononuclear cells and platelets from both PB and BM could be efficiently isolated by obtaining a mononuclear and platelet rich fraction (PB‐MPRF and BM‐MPRF, respectively). Starting with comparable volumes, the number of platelets increased 1.5‐fold in BM‐MPRF compared to PB‐MPRF. The number of clonogenic cells in BM‐MPRF samples was significantly higher than whole BM samples as revealed by CFU‐F assay (54.92 ± 8.55 CFU‐F/1.5 x 105 nucleated cells and 32.50 ± 12.43 CFU‐F/1.5 x 105 nucleated cells, respectively). Cells isolated from BM‐MPRF after in vitro expansion fulfilled the definition of MSCs by phenotypic criteria, and differentiated along osteogenic, adipogenic and chondrogenic lineages following induction. Results showed that the kit isolated MSCs and platelets from BM aspirate. Isolated MSCs were further expanded in a laboratory and BM‐MPRF was used clinically following BM withdrawal for rapid intra‐operative cell therapy for the treatment of bone defects. Copyright © 2012 John Wiley & Sons, Ltd.

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Good manufacturing practice–grade fibrin gel is useful as a scaffold for human mesenchymal stromal cells and supports in vitro osteogenic differentiation

Abstract: These results suggest that fibrin gel could be useful as a delivery system for hMSCs.

Cited by 23 publications

References 22 publications

Effect of fibrin on osteogenic differentiation and VEGF expression of bone marrow stromal cells in mineralised scaffolds: a three-dimensional analysis

Effect of fibrin on osteogenic differentiation and VEGF expression of bone marrow stromal cells in mineralised scaffolds: a three-dimensional analysis

Engineering New Bone via a Minimally Invasive Route Using Human Bone Marrow-Derived Stromal Cell Aggregates, Microceramic Particles, and Human Platelet-Rich Plasma Gel

A rapid method for obtaining mesenchymal stem cells and platelets from bone marrow aspirate

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