Fluorescent hydrogels for embryoid body formation and osteogenic differentiation of embryonic stem cells.

Substrate mechanics (e.g., stiffness and topography of the microenvironment) are likely critical for driving normal morphogenesis and tissue development.

The structural analysis of three-dimensional fibrous collagen hydrogels by Raman microspectroscopy.

To investigate molecular effects of 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), EDC/N-hydroxysuccinimide (NHS), glyceraldehyde cross-linking as well as polymerization temperature and concentration on the three-dimensional (3D) collagen hydrogels, we analyzed the structures in situ by Raman microspectroscopy. The increased intensity of the 814 and 936 cm(-1) Raman bands corresponding to the C-C stretch of a protein backbone and a shift in the amide III bands from 1241 cm(-1)/1268 cm(-1) in controls to 1247 cm(-1)/1283 cm(-1) in glyceraldehyde-treated gels indicated changes to the alignment of the collagen molecules, fibrils/fibers and/or changes to the secondary structure on glyceraldehyde treatment. The increased intensity of 1450 cm(-1) band and the appearance of a strong peak at 1468 cm(-1) reflected a change in the motion of lysine/arginine CH2 groups.

Effects of zero-length and non-zero-length cross-linking reagents on the optical spectral properties and structures of collagen hydrogels.

We compared the effects of zero-length cross-linkers 1-ethyl-3 (3dimethylaminopropyl) carbodiimide (EDC) and non-zero-length cross-linkers glycolaldehyde and glyceraldehyde on the optical and structural properties of three-dimensional (3D) collagen hydrogels. We evaluated these effects by multiphoton microscopy (MPM) that combined two-photon fluorescence (TPF) and second harmonic generation (SHG) contrasts and transmission electron microscopy (TEM). The collagen hydrogels were incubated separately with the above-mentioned reagents present at the concentration of 0.

Effect of genipin crosslinking on the optical spectral properties and structures of collagen hydrogels.

Genipin, a natural cross-linking reagent extracted from the fruits of Gardenia jasminoides, can be effectively employed in tissue engineering applications due to its low cytotoxicity and high biocompatibility. The cross-linking of collagen hydrogels with genipin was followed with one-photon fluorescence spectroscopy, second harmonic generation, fluorescence and transmission electron microscopy. The incubation with genipin induced strong auto-fluorescence within the collagen hydrogels.

Collagen hydrogel characterization: multi-scale and multi-modality approach.

The complex supramolecular architecture of collagen biopolymer plays an important role in tissue development and integrity. Developing methods to report on collagen structures assembled in vitro would accelerate the pace of utilizing it in biomedical applications. Employing imaging techniques and turbidity measurements, we mapped the light scattering properties of 3D collagen hydrogels formed at initial concentrations of 1 mg ml-1 to about 5 mg ml-1 and several incubation temperatures.

Multiphoton imaging of actin filament formation and mitochondrial energetics of human ACBT gliomas.

We studied the three-dimensional (3D) distribution of actin filaments and mitochondria in relation to ACBT glioblastoma cells migration. We embedded the cells in the spheroid form within collagen hydrogels and imaged them by in situ multiphoton microscopy (MPM). The static 3D overlay of the distribution of actin filaments and mitochondria provided a greater understanding of cell-to-cell and cell-to-substrate interactions and morphology.

Multiphoton optical image guided spectroscopy method for characterization of collagen-based materials modified by glycation.

The cross-linking with reducing sugars, known as glycation, is used to increase stiffness and strength of tissues and artificial collagen-based scaffolds. Nondestructive characterization methods that report on the structures within these materials could clarify the effects of glycation. For doing this nondestructive evaluation, we employed an in situ one-photon fluorescence as well as multiphoton microscopy method that combined two-photon fluorescence and second harmonic generation signals.