Since stem cells emerged as a new generation of medicine, there are increasing efforts to deliver the stem cells to a target tissue via intravascular injection. However, the therapeutic stem cells lack a capacity to detect and adhere to the target tissue. Therefore, this study presents synthesis of a bioactive hyper-branched polyglycerol (HPG) which can non-invasively associate with stem cells and further guide them to target sites, such as inflamed endothelium. The overall process is analogous to the way in which leukocytes are mobilized to the injured endothelium.
Extensive efforts have been made to understand the effects of extracellular microenvironments on phenotypic activities for a wide array of stem, progenitor, and precursor cells. Hydrogels have emerged as invaluable platforms for examining the effects of extracellular matrix (ECM) properties on cell activities because of their several advantageous features. Specifically, hydrogels are unique materials that enable cell studies in three-dimensional (3D) environments, similar to in vivo environments. Recently, there have been increasing efforts to assemble cell-encapsulating hydrogels; however, hydrogel design strategies for 3D cell cultures have not been systematically discussed to date. Therefore, this review article summarizes current hydrogel designs for 3D cell culture studies and further discusses current challenges and potential resolutions for enhancing the controllability of hydrogel properties and microstructures. The hydrogels discussed herein include those of natural polymers (e.g., collagen, fibrinogen, alginate, and hyaluronic acids), synthetic polymers [e.g., poly(ethylene glycol) (PEG) and its derivatives], and mixtures of natural and synthetic polymers. We envision that hydrogels that enable 3D studies will greatly assist in the understanding of emergent cell behaviors, and ultimately become important biomedical tools for enhancing the quality of in vitro drug screening and clinical treatments.
Nano-sized polymersomes functionalized with peptides or proteins are being increasingly studied for targeted delivery of diagnostic and therapeutic molecules. Earlier computational studies have suggested that ellipsoidal nanoparticles, compared to spherical ones, display enhanced binding efficiency with target cells, but this has not yet been experimentally validated. We hypothesize that hydrophilic polymer chains coupled to vesicle-forming polymers would result in ellipsoidal polymersomes. In addition, ellipsoidal polymersomes modified with cell adhesion peptides bind with target cells more actively than spherical ones. We examine this hypothesis by substituting polyaspartamide with octadecyl chains and varying numbers of poly(ethylene glycol) (PEG) chains. Increasing the degree of substitution of PEG from 0.5 to 1.0 mol% drives the polymer to self-assemble into an ellipsoidal polymersome with an aspect ratio of 2.1. Further modification of these ellipsoidal polymersomes with peptides containing an Arg-Gly-Asp sequence (RGD peptides) lead to a significant increase in the rate of association and decrease in the rate of dissociation with a substrate coated with αvβ3 integrins. In addition, in a circulation-mimicking flow, the ellipsoidal polymersomes linked with RGD peptides adhere to target tissues more favorably than their spherical equivalents do. Overall, the results of this study will greatly serve to improve the efficiency of targeted delivery of a wide array of polymersomes loaded with various biomedical modalities.
An integrated multimodal optical microscope is demonstrated for high-resolution, structural and functional imaging of engineered and natural skin. This microscope incorporates multiple imaging modalities including optical coherence (OCM), multi-photon (MPM), and fluorescence lifetime imaging microscopy (FLIM), enabling simultaneous visualization of multiple contrast sources and mechanisms from cells and tissues. Spatially co-registered OCM/MPM/FLIM images of multi-layered skin tissues are obtained, which are formed based on complementary information provided by different modalities, i.e., scattering information from OCM, molecular information from MPM, and functional cellular metabolism states from FLIM. Cellular structures in both the dermis and epidermis, especially different morphological and physiological states of keratinocytes from different epidermal layers, are revealed by mutually-validating images. In vivo imaging of human skin is also investigated, which demonstrates the potential of multimodal microscopy for in vivo investigation during engineered skin engraftment. This integrated imaging technique and microscope show the potential for investigating cellular dynamics in developing engineered skin and following in vivo grafting, which will help refine the control and culturing conditions necessary to obtain more robust and physiologically-relevant engineered skin substitutes. Multimodal microscopy images of a microporous 3D hydrogel scaffold seeded with 3T3 fibroblasts. Representative spatially co-registered images were generated based on different methodologies including optical coherence (OCM), multiphoton (MPM), and fluorescence lifetime imaging (FLIM) microscopy.
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