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Photopolymerizable Hydrogel-Encapsulated Fibromodulin-Reprogrammed Cells for Muscle Regeneration.

ABSTRACT: A central challenge in tissue engineering is obtaining a suitable cell type with a capable delivery vehicle to replace or repair damaged or diseased tissues with tissue mimics. Notably, for skeletal muscle tissue engineering, given the inadequate availability and regenerative capability of endogenous myogenic progenitor cells as well as the tumorigenic risks presented by the currently available pluri- and multipotent stem cells, seeking a safe regenerative cell source is urgently demanded. To conquer this problem, we previously established a novel reprogramming technology that can generate multipotent cells from dermal fibroblasts using a single protein, fibromodulin (FMOD). The yield FMOD-reprogrammed (FReP) cells exhibit exceeding myogenic capability without tumorigenic risk, making them a promising and safe cell source for skeletal muscle establishment. In addition to using the optimal cell for implantation, it is equally essential to maintain cellular localization and retention in the recipient tissue environment for critical-sized muscle tissue establishment. In this study, we demonstrate that the photopolymerizable methacrylated glycol chitosan (MeGC)/type I collagen (ColI)-hydrogel provides a desirable microenvironment for encapsulated FReP cell survival, spreading, extension, and formation of myotubes in the hydrogel three-dimensionally in vitro, without undesired osteogenic, chondrogenic, or tenogenic differentiation. Furthermore, gene profiling revealed a paired box 7 (PAX7) → myogenic factor 5 (MYF5) → myogenic determination 1 (MYOD1) → myogenin (MYOG) → myosin cassette elevation in the encapsulated FReP cells during myogenic differentiation, which is similar to that of the predominant driver of endogenous skeletal muscle regeneration, satellite cells. These findings constitute the evidence that the FReP cell-MeGC/ColI-hydrogel construct is a promising tissue engineering mimic for skeletal muscle generation in vitro, and thus possesses the extraordinary potential for further in vivo validation.

SUBMITTER: Yang P

PROVIDER: S-EPMC7580647 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Many attempts have been made to repair articular cartilage defects, including mesenchymal stem cell (MSC)-based tissue engineering strategies. Although this approach shows promise, optimizing MSC sources and their delivery is challenging. This study was designed to test the feasibility of using MSCs found in the human arthroscopic flushing fluid (AFF) for cartilage regeneration, by incorporating them into a newly developed one-step rapid cross-linking hyper-branched polyPEGDA/HA hydrogel. AFF-MSCs were isolated from the original intra-articular flushing fluid of 10 patients prior to arthroscopic procedures. The hydrogel was fabricated with hyper-branched polyPEGDA and thiolated hyaluronic acid (HA). In vitro assays demonstrated that AFF-MSCs possessed the typical MSC morphology and phenotype, and maintained chondrogenic differentiation properties when encapsulated within the hydrogel. The AFF-MSC/hydrogel composite could significantly repair full-thickness cartilage defects generated in a rat model after 8 weeks of implantation; smooth cartilage was formed with evidence of hyaline cartilage formation. These data suggest that human AFF-MSCs are a novel and abundant MSC source that have high therapeutic value for cartilage regeneration.Statement of significanceMany attempts have been made to repair the defects of articular cartilage, including mesenchymal stem cell (MSC)-based tissue engineering strategies. Optimizing MSC sources and their delivery approaches still remain clinically challenging. Recent studies determined that MSCs derived from synovium and synovial fluid exhibited superior chondrogenic potential. However, no feasible methods to harvest these human tissues and cells have been impeding them for clinical application. Hereby, we explored a simple and easy accessible approach to obtain a new stem cell source from arthroscopic flushing fluid (AFF-MSCs), which probably contains plenty of MSCs from synovium and synovial fluid. Further experiments demonstrated that encapsulation of these stem cells with one-step rapid cross-linked polyPEGDA/HA hydrogel held very encouraging potential for cartilage regeneration.

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Photopolymerizable Hydrogel-Encapsulated Fibromodulin-Reprogrammed Cells for Muscle Regeneration.

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