Controlling differentiation of stem cells <i>via</i> bioactive disordered cues

Zhang, Yujie; Rémy, Murielle; Apartsin, Evgeny; Prouvé, Emilie; Feuillie, Cécile; Labrugère, Christine; Cam, Nithavong; Durrieu, Marie-Christine

doi:10.1039/d3bm00605k

Cited by 2 publications

(12 citation statements)

References 113 publications

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“…As previously demonstrated, the peptide droplets sprayed using the spray technique come into contact with the activated PET surface, initiating covalent reactions between the peptides and the surface. 32 Our earlier study primarily focused on PET-spray conditions, which involved spraying with a single mimetic peptide of BMP-2 protein. 32 It was reported that the S1 condition (sprayed at 1 bar air pressure, 200 μL/min solution flow rate, and 3 s spray time) exhibited the highest osteoblast induction effect.…”

Section: ■ Resultsmentioning

confidence: 99%

“…28−31 Conversely, BMP-2 peptides can trigger cell membrane receptors, directing MSCs toward osteoblastic differentiation. 29,30,32 Notably, the FDA approves BMP-2 protein as an osteoinductive growth factor. 15,33,34 However, uncontrolled peptide release can lead to complications such as heterotopic bone formation and inflammation.…”

Section: ■ Introductionmentioning

confidence: 99%

“…PET was modified according to our previous protocol. 32 Briefly, the clean PET was immersed in EDC/NHS/MES activation solution (EDC (0.2 M), NHS (0.1 M), and MES (0.1 M) in ultrapure water. The carboxyl groups on the PET surface were activated and denoted as "PET-Activated" (Figure 1).…”

Section: ■ Introductionmentioning

confidence: 99%

“…We constructed a homemade spray prototype using an automatic spray gun (PILOT WA 450, WALTHER Spritz- and Lackiersysteme GmbH, Germany) mounted on a rail. 32 The spray gun was connected to an air cap (0.3−1.8 mm, V1136030050*) and regulated by a pressure regulator (BD-26−8−6) ranging from 0.5 to 4 bar for controlling air pressure. A syringe pump (R99-EB, Razel Scientific Instruments) was interposed between the spray gun and a 10 or 20 mL Terumo syringe without a needle (Terumo Europe N.V.).…”

Section: ■ Introductionmentioning

confidence: 99%

“…Surface Functionalization. As previously described, 32 all peptides were first dissolved in DMSO, and then diluted to the desired concentration in ultrapure water. To synthesize homogeneous bioactive surfaces, the activated surface "PET-Activated" was immersed for 16 h at room temperature in a solution of RGD-TAMRA or BMP-2-TAMRA, or a solution containing both RGD and BMP-2 peptides (Four different ratios of each peptide were used).…”

Section: ■ Introductionmentioning

confidence: 99%

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Manipulating Stem Cell Fate with Disordered Bioactive Cues on Surfaces: The Role of Bioactive Ligand Selection

Zhang,

Remy,

Leste-Lasserre

et al. 2024

ACS Appl. Mater. Interfaces

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The development of 2D or 3D bioactive platforms for rapidly isolating pure populations of cells from adult stem cells holds promise for advancing the understanding of cellular mechanisms, drug testing, and tissue engineering. Over the years, methods have emerged to synthesize bioactive micro- and nanostructured 2D materials capable of directing stem cell fate. We introduce a novel method for randomly micro- or nanopatterning any protein/peptide onto both 2D and 3D scaffolds via spray technology. Our goal is to investigate the impact of arranging bioactive micropatterns (ordered vs disordered) on surfaces to guide human mesenchymal stem cell (hMSC) differentiation. The spray technology efficiently coats materials with controlled, cost-effective bioactive micropatterns in various sizes and shapes. BMP-2 mimetic peptides were covalently grafted, individually or in combination with RGD peptides, onto activated polyethylene terephthalate (PET) surfaces through a spraying process, incorporating nano/microscale parameters like size, shape, and composition. The study explores different peptide distributions on surfaces and various peptide combinations. Four surfaces were homogeneously functionalized with these peptides (M1 to M4 with various densities of peptides), and six surfaces with disordered micro- and nanopatterns of peptides (S0 to S5 with different sizes of peptide patterns) were synthesized. Fluorescence microscopy assessed peptide distribution, followed by hMSC culture for 2 weeks, and evaluated osteogenic differentiation via immunocytochemistry and RT-qPCR for osteoblast and osteocyte markers. Cells on uniformly peptide-functionalized surfaces exhibited cuboidal forms, while those on surfaces with disordered patterns tended toward columnar or cuboidal shapes. Surfaces S4 and S5 showed dendrite-like formations resembling an osteocyte morphology. S5 showed significant overexpression of osteoblast (OPN) and osteocyte markers (E11, DMP1, and SOST) compared to control surfaces and other micropatterned surfaces. Notably, despite sharing an equivalent quantity of peptides with a homogeneous functionalized surface, S5 displayed a distinct distribution of peptides, resulting in enhanced osteogenic differentiation of hMSCs.

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Section: ■ Resultsmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Manipulating Stem Cell Fate with Disordered Bioactive Cues on Surfaces: The Role of Bioactive Ligand Selection

Zhang,

Remy,

Leste-Lasserre

et al. 2024

ACS Appl. Mater. Interfaces

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View full text Add to dashboard Cite

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Integrating Mechanics and Bioactivity: A Detailed Assessment of Elasticity and Viscoelasticity at Different Scales in 2D Biofunctionalized PEGDA Hydrogels for Targeted Bone Regeneration

López-Serrano,

Côté-Paradis,

Habenstein

et al. 2024

ACS Appl. Mater. Interfaces

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Methods for promoting and controlling the differentiation of human mesenchymal stem cells (hMSCs) in vitro before in vivo transplantation are crucial for the advancement of tissue engineering and regenerative medicine. In this study, we developed poly(ethylene glycol) diacrylate (PEGDA) hydrogels with tunable mechanical properties, including elasticity and viscoelasticity, coupled with bioactivity achieved through the immobilization of a mixture of RGD and a mimetic peptide of the BMP-2 protein. Despite the key relevance of hydrogel mechanical properties for cell culture, a standard for its characterization has not been proposed, and comparisons between studies are challenging due to the different techniques employed. Here, a comprehensive approach was employed to characterize the elasticity and viscoelasticity of these hydrogels, integrating compression testing, rheology, and atomic force microscopy (AFM) microindentation. Distinct mechanical behaviors were observed across different PEGDA compositions, and some consistent trends across multiple techniques were identified. Using a photoactivated cross-linker, we controlled the functionalization density independently of the mechanical properties. X-ray photoelectrin spectroscopy and fluorescence microscopy were employed to evaluate the functionalization density of the materials before the culturing of hMSCs on them. The cells cultured on all functionalized hydrogels expressed an early osteoblast marker (Runx2) after 2 weeks, even in the absence of a differentiationinducing medium compared to our controls. Additionally, after only 1 week of culture with osteogenic differentiation medium, cells showed accelerated differentiation, with clear morphological differences observed among cells in the different conditions. Notably, cells on stiff but stress-relaxing hydrogels exhibited an overexpression of the osteocyte marker E11. This suggests that the combination of the functionalization procedure with the mechanical properties of the hydrogel provides a potent approach to promoting the osteogenic differentiation of hMSCs.

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Controlling differentiation of stem cells via bioactive disordered cues

Abstract: Ideal bone tissue engineering is to induce bone regeneration through the synergistic integration of biomaterial scaffolds, bone progenitor cells, and bone-forming factors. Biomimetic scaffolds imitate the native extracellular matrix (ECM)...

Cited by 2 publications

References 113 publications

Manipulating Stem Cell Fate with Disordered Bioactive Cues on Surfaces: The Role of Bioactive Ligand Selection

Manipulating Stem Cell Fate with Disordered Bioactive Cues on Surfaces: The Role of Bioactive Ligand Selection

Integrating Mechanics and Bioactivity: A Detailed Assessment of Elasticity and Viscoelasticity at Different Scales in 2D Biofunctionalized PEGDA Hydrogels for Targeted Bone Regeneration

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