Abstract:Various types of synthetic polyesters have been developed as biomaterials for tissue engineering. These materials commonly possess biodegradability, biocompatibility, and formability, which are preferable properties for bone regeneration. The major challenge of using synthetic polyesters is the result of low cell affinity due to their hydrophobic nature, which hinders efficient cell seeding and active cell dynamics. To improve wettability, plasma treatment is widely used in industry. Here, we performed surface… Show more
“… 67 In rBMSC, the consequence of the robust mRNA expression of RUNX2 may be either cytoskeletal expression/diffusion or nuclear localisation. 29 In the present study, the quantification and line investigation confirmed that the mRNA upregulation of RUNX2 by fluid flow was accompanied by a degree of nuclear localisation. Similarly, collagen formation, ALP activity and mineral deposition were found to be enhanced in the dynamic culture groups.…”
Section: Discussionsupporting
confidence: 78%
“… 68 Nevertheless, the induction of osteogenesis, particularly in terms of collagen formation and mineral deposition, by fluid flow is apparently not as robust as that achieved by the osteogenic supplements as previously tested in a study of osteogenic differentiation of rBMSC on LTMC scaffolds. 29 This could be attributable to the characteristics of each component of the common osteogenic supplement (i.e. dexamethasone, β-glycerophosphate and ascorbic acid).…”
Section: Discussionmentioning
confidence: 99%
“…The scaffolds were placed in 48 well plates and sterilised using 70% ethanol and ultraviolet exposure for 2 h. The characterisation of LTCM scaffolds was described previously. 29 …”
Section: Methodsmentioning
confidence: 99%
“…Porous properties were parameterised using data previously obtained by microCT analysis of the scaffold, including porosity 91.708% and permeability 5.80216e-09 m 2 . 29 From the inlet to the outlet, a fully developed flow was applied at a flow rate of either 0.8 ml/min (FL-L) or 1.6 ml/min (FL-H). Non-slip wall condition was applied to the boundary condition of the fluid paths (i.e.…”
Section: Methodsmentioning
confidence: 99%
“…The scaffolds were placed in 48 well plates and sterilised using 70% ethanol and ultraviolet exposure for 2 h. The characterisation of LTCM scaffolds was described previously. 29 Before cell seeding, the scaffolds were prewetted with α-MEM for 24 h, and 250,000 cells were then seeded per scaffold and incubated at 37°C in 5% CO 2 humidified atmosphere for 72 h, before being transferred into the bioreactor.…”
Section: Fabrication Of 3d Microporous Scaffolds Of Ltmc and Cell Seedingmentioning
The fatal determination of bone marrow mesenchymal stem/stromal cells (BMSC) is closely associated with mechano-environmental factors in addition to biochemical clues. The aim of this study was to induce osteogenesis in the absence of chemical stimuli using a custom-designed laminar flow bioreactor. BMSC were seeded onto synthetic microporous scaffolds and subjected to the subphysiological level of fluid flow for up to 21 days. During the perfusion, cell proliferation was significantly inhibited. There were also morphological changes, with F-actin polymerisation and upregulation of ROCK1. Notably, in BMSC subjected to flow, mRNA expression of osteogenic markers was significantly upregulated and RUNX2 was localised in the nuclei. Further, under perfusion, there was greater deposition of collagen type 1 and calcium onto the scaffolds. The results confirm that an appropriate level of fluid stimuli preconditions BMSC towards the osteoblastic lineage on 3D scaffolds in the absence of chemical stimulation, which highlights the utility of flow bioreactors in bone tissue engineering.
“… 67 In rBMSC, the consequence of the robust mRNA expression of RUNX2 may be either cytoskeletal expression/diffusion or nuclear localisation. 29 In the present study, the quantification and line investigation confirmed that the mRNA upregulation of RUNX2 by fluid flow was accompanied by a degree of nuclear localisation. Similarly, collagen formation, ALP activity and mineral deposition were found to be enhanced in the dynamic culture groups.…”
Section: Discussionsupporting
confidence: 78%
“… 68 Nevertheless, the induction of osteogenesis, particularly in terms of collagen formation and mineral deposition, by fluid flow is apparently not as robust as that achieved by the osteogenic supplements as previously tested in a study of osteogenic differentiation of rBMSC on LTMC scaffolds. 29 This could be attributable to the characteristics of each component of the common osteogenic supplement (i.e. dexamethasone, β-glycerophosphate and ascorbic acid).…”
Section: Discussionmentioning
confidence: 99%
“…The scaffolds were placed in 48 well plates and sterilised using 70% ethanol and ultraviolet exposure for 2 h. The characterisation of LTCM scaffolds was described previously. 29 …”
Section: Methodsmentioning
confidence: 99%
“…Porous properties were parameterised using data previously obtained by microCT analysis of the scaffold, including porosity 91.708% and permeability 5.80216e-09 m 2 . 29 From the inlet to the outlet, a fully developed flow was applied at a flow rate of either 0.8 ml/min (FL-L) or 1.6 ml/min (FL-H). Non-slip wall condition was applied to the boundary condition of the fluid paths (i.e.…”
Section: Methodsmentioning
confidence: 99%
“…The scaffolds were placed in 48 well plates and sterilised using 70% ethanol and ultraviolet exposure for 2 h. The characterisation of LTCM scaffolds was described previously. 29 Before cell seeding, the scaffolds were prewetted with α-MEM for 24 h, and 250,000 cells were then seeded per scaffold and incubated at 37°C in 5% CO 2 humidified atmosphere for 72 h, before being transferred into the bioreactor.…”
Section: Fabrication Of 3d Microporous Scaffolds Of Ltmc and Cell Seedingmentioning
The fatal determination of bone marrow mesenchymal stem/stromal cells (BMSC) is closely associated with mechano-environmental factors in addition to biochemical clues. The aim of this study was to induce osteogenesis in the absence of chemical stimuli using a custom-designed laminar flow bioreactor. BMSC were seeded onto synthetic microporous scaffolds and subjected to the subphysiological level of fluid flow for up to 21 days. During the perfusion, cell proliferation was significantly inhibited. There were also morphological changes, with F-actin polymerisation and upregulation of ROCK1. Notably, in BMSC subjected to flow, mRNA expression of osteogenic markers was significantly upregulated and RUNX2 was localised in the nuclei. Further, under perfusion, there was greater deposition of collagen type 1 and calcium onto the scaffolds. The results confirm that an appropriate level of fluid stimuli preconditions BMSC towards the osteoblastic lineage on 3D scaffolds in the absence of chemical stimulation, which highlights the utility of flow bioreactors in bone tissue engineering.
There is an overwhelming demand for new scaffolding materials for tissue engineering (TE) purposes. Polymeric scaffolds have been explored as TE materials; however, their high glass transition state (Tg) limits their applicability. In this study, a novel materials platform for fabricating TE scaffolds is proposed based on solvent‐free two‐component heterocyclic triazine‐trione (TATO) formulations, which cure at room temperature via thiol‐ene/yne photochemistry. Three ester‐containing thermosets, TATO‐1, TATO‐2, and TATO‐3, are used for the fabrication of TE scaffolds including rigid discs, elastic films, microporous sponges, and 3D printed objects. After 14 days’ incubation the materials covered a wide range of properties, from the soft TATO‐2 having a compression modulus of 19.3 MPa and a Tg of 30.4 °C to the hard TATO‐3 having a compression modulus of 411 MPa and a Tg of 62.5 °C. All materials exhibit micro‐ and nano‐surface morphologies suited for bone tissue engineering, and in vitro studies found them all to be cytocompatible, supporting fast cell proliferation while minimizing cell apoptosis and necrosis. Moreover, bone marrow‐derived mesenchymal stem cells on the surface of the materials are successfully differentiated into osteoblasts, adipocytes, and neuronal cells, underlining the broad potential for the biofabrication of TATO materials for TE clinical applications.
The fate determination of bone marrow mesenchymal stem/stromal cells (BMSC) is tightly regulated by mechanical cues, including fluid shear stress. Knowledge of mechanobiology in 2D culture has allowed researchers in bone tissue engineering to develop 3D dynamic culture systems with the potential for clinical translation in which the fate and growth of BMSC are mechanically controlled. However, due to the complexity of 3D dynamic cell culture compared to the 2D counterpart, the mechanisms of cell regulation in the dynamic environment remain relatively undescribed. In the present study, we analyzed the cytoskeletal modulation and osteogenic profiles of BMSC under fluid stimuli in a 3D culture condition using a perfusion bioreactor. BMSC subjected to fluid shear stress (mean 1.56 mPa) showed increased actomyosin contractility, accompanied by the upregulation of mechanoreceptors, focal adhesions, and Rho GTPase‐mediated signaling molecules. Osteogenic gene expression profiling revealed that fluid shear stress promoted the expression of osteogenic markers differently from chemically induced osteogenesis. Osteogenic marker mRNA expression, type 1 collagen formation, ALP activity, and mineralization were promoted in the dynamic condition, even in the absence of chemical supplementation. The inhibition of cell contractility under flow by Rhosin chloride, Y27632, MLCK inhibitor peptide‐18, or Blebbistatin revealed that actomyosin contractility was required for maintaining the proliferative status and mechanically induced osteogenic differentiation in the dynamic culture. The study highlights the cytoskeletal response and unique osteogenic profile of BMSC in this type of dynamic cell culture, stepping toward the clinical translation of mechanically stimulated BMCS for bone regeneration.
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