Surface Curvature Differentially Regulates Stem Cell Migration and Differentiation via Altered Attachment Morphology and Nuclear Deformation

Signals from the microenvironment around a cell are known to influence cell behavior. Material properties, such as biochemical composition and substrate stiffness, are today accepted as significant regulators of stem cell fate. The knowledge of how cell behavior is influenced by 3D geometric cues is, however, strongly limited despite its potential relevance for the understanding of tissue regenerative processes and the design of biomaterials. Here, the role of surface curvature on the migratory and differentiation behavior of human mesenchymal stem cells (hMSCs) has been investigated on 3D surfaces with well-defined geometric features produced by stereolithography. Time lapse microscopy reveals a significant increase of cell migration speed on concave spherical compared to convex spherical structures and flat surfaces resulting from an upward-lift of the cell body due to cytoskeletal forces. On convex surfaces, cytoskeletal forces lead to substantial nuclear deformation, increase lamin-A levels and promote osteogenic differentiation. The findings of this study demonstrate a so far missing link between 3D surface curvature and hMSC behavior. This will not only help to better understand the role of extracellular matrix architecture in health and disease but also give new insights in how 3D geometries can be used as a cell-instructive material parameter in the field of biomaterial-guided tissue regeneration.Peer reviewe

Development of poly(trimethylene carbonate) network implants for annulus fibrosus tissue engineering

Purpose: Intervertebral disk degeneration is the main cause of chronic back pain. Disk degeneration often leads to tearing of the annulus fibrosus (AF) and extrusion of the nucleus pulposus (NP), which compresses the nerves. Current treatment involves removing the herniated NP and suturing the damaged AF tissue. This surgical approach has several drawbacks. In this study, we designed a biodegradable AF closure system comprising a tissue engineering scaffold, a supporting membrane and an adhesive material, to not only restore the function of the herniated disc but also to promote tissue regeneration. Materials and Methods: Porous scaffolds with precisely defined architectures were built by stereolithography using resins based on poly(trimethylene carbonate) (PTMC) macromers functionalized with methacrylate endgroups. In addition, a porous photo-cross-linked PTMC membrane was developed that can be used to keep the scaffold in place in the AF tissue. Results: After synthesis and characterization, the components of the implant are glued together and to the AF tissue using a diisocyanate glue based on polyethylene glycol-PTMC triblock copolymers. The adhesion strengths of the materials to each other and to AF tissue were determined in lap-shear tests. Conclusions: This study showed that a device for AF tissue engineering can be prepared from PTMC-based scaffolds, membranes and glues

Biodegradable nanocomposite hydrogel structures with enhanced mechanical properties prepared by photo-crosslinking solutions of poly(trimethylene carbonate)-poly(ethylene glycol)-poly(trimethylene carbonate) macromonomers and nanoclay particles

Soft hydrogels with elasticity modulus values lower than 100 kPa that are tough and biodegradable are of great interest in medicine and in tissue engineering applications. We have developed a series of soft hydrogel structures from different methacrylate-functionalized triblock copolymers of poly(ethylene glycol) (PEG) with poly(trimethylene carbonate) (PTMC) by photo-crosslinking aqueous solutions of the macromonomers in 2.5 and 5 wt.% colloidal dispersions of clay nanoparticles (Laponite XLG). The length of the PTMC blocks of the macromonomers and the clay content determined the physicomechanical properties of the obtained hydrogels. While an increase in the PTMC block length in the macromonomers from 0.2 to 5 kg/mol resulted in a decrease in the gel content, the addition of 5 wt.% Laponite nanoclay to the crosslinking solution lead to very high gel contents of the hydrogels of more than 95%. The effect of PTMC block length on the mechanical properties of the hydrogels was not as pronounced, and soft gels with a compressive modulus of less than 15 kPa and toughness values of 25 kJ m(-3) were obtained. However, the addition of 5 wt.% Laponite nanoclay to the formulations considerably increased the compressive modulus and resilience of the hydrogels; swollen nanocomposite networks with compressive modulus and toughness values of up to 67 kPa and 200 kJ m(-3), respectively, could then be obtained. The prepared hydrogels were shown to be enzymatically degradable by cholesterol esterase and by the action of macrophages. With an increase in PTMC block length in the hydrogels, the rates of mass loss increased, while the incorporated Laponite nanoclay suppressed degradation. Nanocomposite hydrogel structures with a designed gyroid pore network architecture were prepared by stereolithography. Furthermore, in the swollen state the porous gyroid structures were mechanically stable and the pore network remained fully open and interconnected. (c) 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved

Differentiation of adipose stem cells seeded towards annulus fibrosus cells on a designed poly(trimethylene carbonate) scaffold prepared by stereolithography

Cell-based therapies could potentially restore the biomechanical function and enhance the self-repair capacity of annulus fibrosus (AF) tissue. However, choosing a suitable cell source and scaffold design are still key challenges. In this study, we assessed the in vitro ability of human adipose stem cells (hASCs), an easily available cell source to produce AF-like matrix in novel AF-mimetic designed scaffolds based on poly(trimethylene carbonate) and built by stereolithography. To facilitate efficient differentiation of hASCs towards AF tissue, we tested different culture medium compositions and cell seeding techniques. This is the first study to report that medium supplementation with transforming growth factor (TGF)-beta 3 is essential to support AF differentiation of hASCs while TGF-beta 1 has negligible effect after 21 days of culture. Fibrin gel seeding resulted in superior cell distribution, proliferation and AF-like matrix production of hASCs compared to direct and micromass seeding under TGF-beta 3 stimulation. Not only the production of sulphated glycosaminoglycans (sGAG) and collagen was significantly upregulated, but the formed collagen was also oriented and aligned into bundles within the designed pore channels. The differentiated hASCs seeded with fibrin gel were also found to have a comparable sGAG: collagen ratio and gene expression profile as native AF cells demonstrating the high potential of this strategy in AF repair

Surface curvature in triply-periodic minimal surface architectures as a distinct design parameter in preparing advanced tissue engineering scaffolds

Reproduction of the anatomical structures and functions of tissues using cells and designed 3D scaffolds is an ongoing challenge. For this, scaffolds with appropriate biomorphic surfaces promoting cell attachment, proliferation and differentiation are needed. In this study, eight triply-periodic minimal surface (TPMS)-based scaffolds were designed using specific trigonometric equations, providing the same porosity and the same number of unit cells, while presenting different surface curvatures. The scaffolds were fabricated by stereolithography using a photocurable resin based on the biocompatible, biodegradable and rubber-like material, poly(trimethylene carbonate) (PTMC). A numerical approach was developed to calculate the surface curvature distributions of the TPMS architectures. Moreover, the scaffolds were characterized by scanning electron microscopy, microcomputed tomography and water permeability measurements. These original scaffold architectures will be helpful to decipher the biofunctional role of the surface curvature of scaffolds intended for tissue engineering applications