Cytoprotective Self-assembled RGD Peptide Nanofilms for Surface Modification of Viable Mesenchymal Stem Cells

Abstract

Intravenous administration of mesenchymal stem cells (MSCs) has served as a clinical intervention for inflammatory diseases. Once entered to blood circulation, MSCs are exposed to a harsh environment which sharply decreases cell viability due to the fact that injected cells, being susceptible to shear stress, are subjected to the high velocities of the bloodstream and lack of proper mechanical support that keeping them in an attachment-deprived state. Here, we coated the nanofilm onto viable MSCs by depositing poly-l-lysine and hyaluronic acid molecules along with arginine-glycine-aspartic acid (RGD peptide) as building blocks to protect cells from shear stress and stabilize them in a single cell, suspension state. In this article, we found that nanofilm-coated cells showed significantly increased cell survival in vitro and in vivo, which was also supported by the activation of survival-related protein, Akt. The coated nanofilm did not interfere with the stemness of MSCs which was determined based on the colony forming unit-fibroblast (CFU-F) assay and in vitro differentiation potential. Because of the characteristics of films showing light molecular deposition density, flexibility, and looseness, application of nanofilms did not block cell migration. When the cells were administrated intravenously, the nanofilm coated MSCs not only prolonged blood circulation lifetime but also showed increased stem cell recruitment to injured tissues in the muscle injury in vivo model, due to prolonged survival. Surface modification of MSCs using nanofilms successfully modulated cell activity enabling them to survive the anoikis-inducing state, and this can provide a valuable tool to potentiate the efficacy of MSCs for in vivo cell therapy.

Cell‐Surface Engineering for Advanced Cell Therapy

“…However,e ven for nanothin films with at hickness of approximately 7-29 nm (Figure 6a), anoikis, at ype of programmed cell death induced by the attachment-depriveds tate, is significantly decreased (Figure 6b). [218][219][220] The mechanism by which nanothin film coated on cell plasma membrane inhibitsa noikis involves the interaction of matrix components with extracellularr eceptors such as integrin and CD44. [218] In cases where the thickness of the matrix is on ananometer scale, the cytoprotective effect of cell encapsulation is greatly Spatial control of cellinteraction with hydrogelb ym atrix metalloproteinase-degradable and 3d imensional RGD photopatterned encapsulation.…”

“…[67][68][69] Encapsulation in softer hydrogels, composed of biodegradable material, permits cell proliferation as long as cell viability is maintained. [218,219] Upon mass deliveryo fm ulticellular encapsulated constructs, tissue maturation and regeneration are considered complete when the transplanted cells fulfill the structuraland physiological requirementso fa ll tissues at the transplant site. Ideally,t he native matrix produced by the encapsulated cells and neighboring tissues at the transplant site completely replaces the matrix used for cell encapsulation,r esulting in total integration between transplants and natural tissues.…”

“…One study reportedt hat MSCs encapsulatedb yL bL assembly through alternating reactions with polycationic and polyanionic polymers containing RGD peptide showedr esistance to anoikis in vitro and enhanced cell survival in the circulation after systemic intravenous injection (Figure 14). [218,219] Even in cases where encapsulation thickness does not permit systemici ntravenous injection, encapsulatedM SCs can still be directly administered to the desired area by local injection. Additionally,i ncorporation of activity-modulating agents can endow encapsulated cells with ap redisposition toward the desired cellular lineage, even without differentiation induction.…”

“…MSCse ncapsulated by LbL assembly showed significantly increased survival in circulationa nd betterr ecruitmenttot he muscle injury site compared with non-encapsulated,bare MSCs. Reprinted with permission from Ref [218]. Copyright 2017 American Chemical Society.…”

Transmission and regulation of biochemical stimulus via a nanoshell directly adsorbed on the cell membrane to enhance chondrogenic differentiation of mesenchymal stem cell

“…The self‐assembled multilayer nanofilm (nanoshell) directly adsorbed on cell surface is one of the facile method for controlling the interfacial structure between cellular membrane and extracellular matrix. These artificial nano‐extracellular matrices generally shelter each cell from external stress without interfering in cell‐to‐cell communication, which results in enhanced cell viability, cell migration, and cell proliferation (Choi, Lee, et al, ; Matsuzawa, Matsusaki, & Akashi, ). In comparison with conventional tool for control of stem cell fates, the nanoshell has several unique features.…”