Regulating orientation and phenotype of primary vascular smooth muscle cells by biodegradable films patterned with arrays of microchannels and discontinuous microwalls

Cao, Yilin; Poon, Yin Fun; Feng, Jie; Rayatpisheh, Shahrzad; Chan, Vincent; Chan‐Park, Mary B.

doi:10.1016/j.biomaterials.2010.04.059

Cited by 67 publications

(81 citation statements)

References 30 publications

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“…At the same time, SMCs started to align along the direction of the long axis of the microwall (figure 5b). The result as mentioned above was supported by the constraint on SMC alignment which had been reported in SMCs grown between the PDMS microchannel [52]. During the intense period of cell proliferation, cells reached confluence after 7 days of culture.…”

Section: The Morphology and Phenotype Control Of Smooth Muscle Cellsupporting

confidence: 70%

“…This functional relation between the two parameters can also be calculated using the theoretical Hertzian model from equation (2.6) if Young's modulus is provided. With the given experimental data, the Young modulus can be estimated by minimizing the errors between experimental data and theoretical calculations [52]. One example is shown in figure 4a as an appropriate Young's modulus is chosen.…”

Section: Polyacrylamide-based Hydrogel Substrate Stiffness Measured Bmentioning

confidence: 99%

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Collective cell traction force analysis on aligned smooth muscle cell sheet between three-dimensional microwalls

Zhang

Wang

et al. 2014

Interface Focus.

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During the past two decades, novel biomaterial scaffold for cell attachment and culture has been developed for applications in tissue engineering, biosensing and regeneration medicine. Tissue engineering of blood vessels remains a challenge owing to the complex three-layer histology involved. In order to engineer functional blood vessels, it is essential to recapitulate the characteristics of vascular smooth muscle cells (SMCs) inside the tunica media, which is known to be critical for vasoconstriction and vasodilation of the circulatory system. Until now, there has been a lack of understanding on the mechanotransduction of the SMC layer during the transformation from viable synthetic to quiescent contractile phenotypes. In this study, microfabricated arrays of discontinuous microwalls coated with fluorescence microbeads were developed to probe the mechanotransduction of the SMC layer. First, the system was exploited for stimulating the formation of a highly aligned orientation of SMCs in native tunica medium. Second, atomic force microscopy in combination with regression analysis was applied to measure the elastic modulus of a polyacrylamide gel layer coated on the discontinuous microwall arrays. Third, the conventional traction force assay for single cell measurement was extended for applications in three-dimensional cell aggregates. Then, the biophysical effects of discontinuous microwalls on the mechanotransduction of the SMC layer undergoing cell alignment were probed. Generally, the cooperative multiple cell–cell and cell–microwall interactions were accessed quantitatively by the newly developed assay with the aid of finite-element modelling. The results show that the traction forces of highly aligned cells lying in the middle region between two opposing microwalls were significantly lower than those lying adjacent to the microwalls. Moreover, the spatial distributions of Von Mises stress during the cell alignment process were dependent on the collective cell layer orientation. Immunostaining of the SMC sheet further demonstrated that the collective mechanotransduction induced by three-dimensional topographic cues was correlated with the reduction of actin and vinculin expression. In addition, the online two-dimensional LC–MS/MS analysis verified the modulation of focal adhesion formation under the influence of microwalls through the regulation in the expression of three key cytoskeletal proteins.

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Section: The Morphology and Phenotype Control Of Smooth Muscle Cellsupporting

confidence: 70%

Section: Polyacrylamide-based Hydrogel Substrate Stiffness Measured Bmentioning

confidence: 99%

Collective cell traction force analysis on aligned smooth muscle cell sheet between three-dimensional microwalls

Zhang

Wang

et al. 2014

Interface Focus.

Self Cite

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“…6,8 Despite these great advances in basic research, the incorporation of geometric cues into threedimensional (3D) tubular vascular TE scaffolds remains a challenging issue. Geometric features presently have oen been fabricated on stiff and non-biodegradable substrates (e.g.…”

Section: 10mentioning

confidence: 99%

“…3,5 Geometric cues serving as articial cell micro-environments could facilitate vascular TE scaffolds to replicate the complex architecture of tunica media. [6][7][8] It has been known that geometric cues could trigger cell response via contact guidance.…”

Section: Introductionmentioning

confidence: 99%

Enhancing mesenchymal stem cell response using uniaxially stretched poly(ε-caprolactone) film micropatterns for vascular tissue engineering application

et al. 2014

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Regeneration of tunica media with anisotropic architecture still remains a challenging issue for vascular tissue engineering (TE). Herein, we present the development of flexible poly(3-caprolactone) (PCL) film micropatterns to regulate mesenchymal stem cells (MSCs) function for tunica media construction.Results showed that uniaxial thermal stretching of PCL films resulted in topographical micropatterns comprising of ridges/grooves, and improved mechanical properties, including yield stress, Young's modulus, and fracture stress without sacrificing film elasticity. Culturing on such PCL film micropatterns, MSCs self-aligned along the ridges with a more elongated morphology as compared to that of the unstretched film group. Moreover, MSCs obtained a contractile SMCs-like phenotype, with ordered organization of cellular stress filaments and upregulated expression of the contractile makers, including SM-a-actin, calponin, and SM-MHC. The PCL film micropatterns could be rolled into a small-diameter 3D tubular scaffold with circumferential anisotropy of ridges/grooves, and in the incorporation of MSCs, which facilitated a hybrid sandwich-like vascular wall construction with ordered cell architecture similar to that of the tunica media. These results provide insights of how geometric cues are able to regulate stem cells with desired functions and have significant implications for the designing of a functionalized vascular TE scaffold with appropriate topographical geometries for guiding tunica media regeneration with microscale control of cell alignment and genetic expression.

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“…Furthermore, the depth of the groove can determine whether the cells within the grooves can make contact with the cells between and within the neighboring grooves. A shallow groove can produce a more continuous sheet of cells whereas the deeper grooves create more distinct lines of cells [14,25] .…”

Section: Resultsmentioning

confidence: 99%

A novel 3D printing method for cell alignment and differentiation

Bhuthalingam¹,

Lim²,

Irvine³

et al. 2024

IJB

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Abstract:The application of bioprinting allows precision deposition of biological materials for bioengineering applications. Here we propose a 2 stage methodology for bioprinting using a back pressure-driven, automated robotic dispensing system. This apparatus can prepare topographic guidance features for cell orientation and then bioprint cells directly onto them. Topographic guidance features generate cues that influence adhered cell morphology and phenotype. The robotic dispensing system was modified to include a sharpened stylus that etched on a polystyrene surface. The same computer-aided design (CAD) software was used for both precision control of etching and bioink deposition. Various etched groove patterns such as linear, concentric circles, and sinusoidal wave patterns were possible. Fibroblasts and mesenchymal stem cells (MSC) were able to sense the grooves, as shown by their elongation and orientation in the direction of the features. The orientated MSCs displayed indications of lineage commitment as detected by fluorescence-activated cell sorting (FACS) analysis. A 2% gelatin bioink was then used to dispense cells onto the etched features using identical, programmed co-ordinates. The bioink allows the cells to contact sense the pattern while containing their deposition within the printed pattern.

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Regulating orientation and phenotype of primary vascular smooth muscle cells by biodegradable films patterned with arrays of microchannels and discontinuous microwalls

Cited by 67 publications

References 30 publications

Collective cell traction force analysis on aligned smooth muscle cell sheet between three-dimensional microwalls

Collective cell traction force analysis on aligned smooth muscle cell sheet between three-dimensional microwalls

Enhancing mesenchymal stem cell response using uniaxially stretched poly(ε-caprolactone) film micropatterns for vascular tissue engineering application

A novel 3D printing method for cell alignment and differentiation

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