The use of three-dimensional nanostructures to instruct cells to produce extracellular matrix for regenerative medicine strategies

Abstract

Synthetic polymers or naturally-derived extracellular matrix (ECM) proteins have been used to create tissue engineering scaffolds; however, the need for surface modification in order to achieve polymer biocompatibility and the lack of biomechanical strength of constructs built using proteins alone remain major limitations. To overcome these obstacles, we developed novel hybrid constructs composed of both strong biosynthetic materials and natural human ECM proteins. Taking advantage of the ability of cells to produce their own ECM, human foreskin fibroblasts were grown on silicon-based nanostructures exhibiting various surface topographies that significantly enhanced ECM protein production. After 4 weeks, cell-derived sheets were harvested and histology, immunochemistry, biochemistry and multiphoton imaging revealed the presence of collagens, tropoelastin, fibronectin and glycosaminoglycans. Following decellularization, purified sheet-derived ECM proteins were mixed with poly(ε-caprolactone) to create fibrous scaffolds using electrospinning. These hybrid scaffolds exhibited excellent biomechanical properties with fiber and pore sizes that allowed attachment and migration of adipose tissue-derived stem cells. Our study represents an innovative approach to generate strong, non-cytotoxic scaffolds that could have broad applications in tissue regeneration strategies.

Taking a deep look: modern microscopy technologies to optimize the design and functionality of biocompatible scaffolds for tissue engineering in regenerative medicine

“…The methodology is sensitive to changes that occur in rsif.royalsocietypublishing.org J R Soc Interface 10: 20130263 diseases such as sclerodermia or in skin or other epithelial cancer types [139 -141]. Other application examples are the investigation of heart valves and cardiac [63] or corneal tissue [142,143]. To image collagen I with SHG, excitation wavelengths from 720 to 960 nm [62] can be used, and for detection, a bandpass filter with centre wavelength at half the excitation wavelength is applied (e.g.…”

Hyaluronan-Based Three-Dimensional Microenvironment Potently Induces Cardiovascular Progenitor Cell Populations

“…Alignment has also been observed along the direction of grooves formed in laminin [18]. We have also shown that incorporating nanostructures into a culture can influence cellular behavior [19]. Additionally, cellular morphology is greatly different between in vitro 2D culture and in vivo 3D native tissue, and the addition of this 3D component has become more widespread in many research groups.…”

Vascular cell responses to ECM produced by smooth muscle cells on TiO2 nanotubes

“…Because ECM is a fibrillar meshwork including the growth factors (cytokines, vascular endothelial growth factor), structural proteins (collagen and elastin), adhesion proteins (fibronectin and laminin), and glycosaminoglycans (chondroitin sulfate), these components arrange in a biochemically complex and intricately three-dimensional structure, which constitutes a barrier for diffusion and convection, and provides tissues with mechanical strength and elastic properties. Katja developed novel hybrid constructs composed of poly and natural ECM, and these constructs exhibited excellent biomechanical property and allowed attachment and migration of adipose tissue-derived stem cells [16]. ECM component, mainly consist of type IV collagen, Fibronectin, laminin, and growth factors, had been extensively investigated.…”