Decellularized human bone as a 3D model to study skeletal progenitor cells in a natural environment

Pereira, Ana Rita; Rudert, Maximilian; Herrmann, Marietta

doi:10.1016/bs.mcb.2019.11.018

Cited by 10 publications

(9 citation statements)

References 23 publications

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“…Furthermore, it has been reported that partially or fully demineralized bone can provide not only osteoinductive factors, but also superior mechanical, biochemical, and architectural properties supporting scaffold functionality into physiological conditions [ 42 , 43 ]. We have previously developed a protocol based on physical, chemical, and enzymatic methods to consistently achieve human-femoral head-derived decellularized bone scaffolds [ 34 ]. The effective removal of all cellular components was previously confirmed, and the cell seeding protocol was optimized to ensure sustained cell viability.…”

Section: Discussionmentioning

confidence: 99%

“…Decellularized bone scaffolds were obtained from human trabecular femoral head specimens (permission number: 187/18, University of Wuerzburg ethics committee), as previously described in [ 34 ]. Briefly, freshly thawed samples were precisely cut in 3 mm thick slides using an electric diamond band saw (300, Exakt; D64, Walter Messner GmbH, Oststeinbek, Germany) to ensure homogeneous penetration of washing solutions through the complete sample volume.…”

Section: Methodsmentioning

confidence: 99%

“…The cell count was determined, and cells were cultured for adhesion selection of hMSCs. Cells were further expanded with basal medium (DMEM/F-12 GlutaMAX, Gibco, Bleiswijk, Netherlands) supplemented with 10% fetal bovine serum (FCS, Bio&Sell, Feucht, Germany), 1% Pen/Strep (100 U/mL, Gibco), 1% HEPES (Sigma-Aldrich), and 5 ng/mL fibroblast growth factor (FGF, 100-18C, PeproTech, Hamburg, Germany) until passage 4–6 and seeded in decellularized bone scaffolds as previously described in [ 34 ]. Briefly, 50 µL containing 1.5 × 10 6 cells was distributed on top of each scaffold and incubated for 3 h at 37 °C with 5% CO 2 to allow cell attachment, followed by 21 h static incubation with the supplemented basal medium.…”

Section: Methodsmentioning

confidence: 99%

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Modeling of the Human Bone Environment: Mechanical Stimuli Guide Mesenchymal Stem Cell–Extracellular Matrix Interactions

et al. 2021

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In bone tissue engineering, the design of in vitro models able to recreate both the chemical composition, the structural architecture, and the overall mechanical environment of the native tissue is still often neglected. In this study, we apply a bioreactor system where human bone-marrow hMSCs are seeded in human femoral head-derived decellularized bone scaffolds and subjected to dynamic culture, i.e., shear stress induced by continuous cell culture medium perfusion at 1.7 mL/min flow rate and compressive stress by 10% uniaxial load at 1 Hz for 1 h per day. In silico modeling revealed that continuous medium flow generates a mean shear stress of 8.5 mPa sensed by hMSCs seeded on 3D bone scaffolds. Experimentally, both dynamic conditions improved cell repopulation within the scaffold and boosted ECM production compared with static controls. Early response of hMSCs to mechanical stimuli comprises evident cell shape changes and stronger integrin-mediated adhesion to the matrix. Stress-induced Col6 and SPP1 gene expression suggests an early hMSC commitment towards osteogenic lineage independent of Runx2 signaling. This study provides a foundation for exploring the early effects of external mechanical stimuli on hMSC behavior in a biologically meaningful in vitro environment, opening new opportunities to study bone development, remodeling, and pathologies.

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Section: Discussionmentioning

confidence: 99%

“…Decellularized bone scaffolds were obtained from human trabecular femoral head specimens (permission number: 187/18, University of Wuerzburg ethics committee), as previously described in [ 34 ]. Briefly, freshly thawed samples were precisely cut in 3 mm thick slides using an electric diamond band saw (300, Exakt; D64, Walter Messner GmbH, Oststeinbek, Germany) to ensure homogeneous penetration of washing solutions through the complete sample volume.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Modeling of the Human Bone Environment: Mechanical Stimuli Guide Mesenchymal Stem Cell–Extracellular Matrix Interactions

et al. 2021

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“…Decellularized bone scaffolds were obtained from human trabecular femoral head specimens (permission number: 187/18, University of Wuerzburg ethics committee), as previously described in. 25 , 45 Briefly, freshly thawed samples (kept at −20°C for no more than 4 months after surgery) were precisely cut in 3 mm thick slides using an electric diamond band saw (300 Exakt D64, Walter Messner, Germany) to ensure homogeneous penetration of washing solutions through the complete sample volume. Blood and residual fat material were removed by several washing cycles in water and a chloroform (288306, Sigma-Aldrich) and methanol (8388.6, Carl-Roth) mix solution.…”

Section: Methodsmentioning

confidence: 99%

“…To confirm expression of CXCL12 by BM-MSC seeded on decellularized models after 5 days in basal culture (initial seeding: 10 4 cells per cm 2 for dECM models and 4 × 10 5 cells per dBone scaffold, seeding protocol previously described in Pereira et al 45 ), immunofluorescence analysis was performed on both dECM monolayers and 12 µm cryosections of dBone scaffolds (sectioning protocol previously described in Pereira et al 45 ). Samples were fixed in 4% PFA for 15 min at 4°C and permeabilized with 0.1% (v/v) Triton-X /PBS for 30 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Preservation of the naïve features of mesenchymal stromal cells in vitro: Comparison of cell- and bone-derived decellularized extracellular matrix

et al. 2022

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The fate and behavior of bone marrow mesenchymal stem/stromal cells (BM-MSC) is bidirectionally influenced by their microenvironment, the stem cell niche, where a magnitude of biochemical and physical cues communicate in an extremely orchestrated way. It is known that simplified 2D in vitro systems for BM-MSC culture do not represent their naïve physiological environment. Here, we developed four different 2D cell-based decellularized matrices (dECM) and a 3D decellularized human trabecular-bone scaffold (dBone) to evaluate BM-MSC behavior. The obtained cell-derived matrices provided a reliable tool for cell shape-based analyses of typical features associated with osteogenic differentiation at high-throughput level. On the other hand, exploratory proteomics analysis identified native bone-specific proteins selectively expressed in dBone but not in dECM models. Together with its architectural complexity, the physico-chemical properties of dBone triggered the upregulation of stemness associated genes and niche-related protein expression, proving in vitro conservation of the naïve features of BM-MSC.

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Synthesis and biological activity of waltherione F‐derived diamide derivatives containing 4‐quinolone group

Chen

Hua

Zhu

et al. 2022

Journal of Heterocyclic Chem

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The exploitation of new fungicides and the discovery of action targets are effective ways to solve the problem of resistance, and structural optimization of natural products has become an resultful method to discover lead compounds. In this work, in conjunction with the structures of the fungicidal lead compounds h‐I‐9, X‐I‐3, and X‐II‐5 found in our previous project, waltherione F‐derived diamide compounds were synthesized and characterized by 1H NMR, 13C NMR, and HRMS. The biological assays suggested that the target compounds exhibited weak to good fungicidal activity against the tested pathogens, while most of the target compounds displayed high cellular cytotoxicity against Hela cells. The investigation of accidentally discovered fluorescence characteristics revealed that the compound I‐1 exhibited the most vigorous fluorescence intensity at 350 nm, which was significantly higher than that of I‐3 at 225 nm and was further elucidated by density functional theory (DFT) calculations. Overall, the above results would provide theoretical guidance for the design of active target molecules and the development of fluorescent probes.

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Decellularized human bone as a 3D model to study skeletal progenitor cells in a natural environment

Cited by 10 publications

References 23 publications

Modeling of the Human Bone Environment: Mechanical Stimuli Guide Mesenchymal Stem Cell–Extracellular Matrix Interactions

Modeling of the Human Bone Environment: Mechanical Stimuli Guide Mesenchymal Stem Cell–Extracellular Matrix Interactions

Preservation of the naïve features of mesenchymal stromal cells in vitro: Comparison of cell- and bone-derived decellularized extracellular matrix

Synthesis and biological activity of waltherione F‐derived diamide derivatives containing 4‐quinolone group

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