Research LettersMyoblast transplantation for heart failure
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Cited by (1008)
Temperature-modulated separation of vascular cells using thermoresponsive-anionic block copolymer-modified glass
2024, Regenerative TherapyVascular tissue engineering is a key technology in the field of regenerative medicine. In tissue engineering, the separation of vascular cells without cell modification is required, as cell modifications affect the intrinsic properties of the cells. In this study, we have developed an effective method for separating vascular cells without cell modification, using a thermoresponsive anionic block copolymer.
A thermoresponsive anionic block copolymer, poly(acrylic acid)-b-poly(N-isopropylacryl-amide) (PAAc-b-PNIPAAm), with various PNIPAAm segment lengths, was prepared in two steps: atom transfer radical polymerization and subsequent deprotection. Normal human umbilical vein endothelial cells (HUVECs), normal human dermal fibroblasts, and human aortic smooth muscle cells (SMCs) were seeded onto the prepared thermoresponsive anionic block copolymer brush-modified glass. The adhesion behavior of cells on the copolymer brush was observed at 37 °C and 20 °C.
A thermoresponsive anionic block copolymer, poly(acrylic acid)-b-poly(N-isopropylacrylamide) (PAAc-b-PNIPAAm), with various PNIPAAm segment lengths was prepared. The prepared copolymer-modified glass exhibited anionic properties attributed to the bottom PAAc segment of the copolymer brush. On the PAAc-b-PNIPAAm, which had a moderate PNIPAAm length, a high adhesion ratio of HUVECs and low adhesion ratio of SMCs were observed at 37 °C. By reducing temperature from 37 °C to 20 °C, the adhered HUVECs were detached, whereas the SMCs maintained adhesion, leading to the recovery of purified HUVECs by changing the temperature.
The prepared thermoresponsive anionic copolymer-modified glass could be used to separate HUVECs and SMCs by changing the temperature without modifying the cell surface. Therefore, the developed cell separation method will be useful for vascular tissue engineering.
Thermoresponsive block-copolymer brush-modified interfaces for effective fabrication of hepatocyte sheets
2024, Materials and DesignHepatic tissue engineering has been investigated for the treatment of liver disease. One promising tissue is the hepatocyte sheet, cultured in temperature-responsive cell culture dishes. However, the preparation of hepatocyte sheets requires precoating of the dish with fetal bovine serum (FBS), which is not desirable for clinical applications. To overcome this issue, we developed a thermoresponsive polymer-modified glass with an affinity for hepatocytes. Poly(N-p-vinylbenzyl-O-β-d-galactopyranosyl-(1→4)-d-guliconamide)-b-poly(N-isopropylacrylamide) (PVLA-b-PNIPAAm) was prepared through two steps of atom transfer radical polymerization (ATRP). The prepared copolymer brushes were characterized using X-ray photoelectron spectroscopy, atomic force microscopy, gel permeation chromatography, contact angle measurements, and fluorescence labeled-fibronectin and -lectin adsorption assays. These characterizations showed that the smooth block copolymer layer was successfully modified by ATRP. The feasibility of using the prepared polymer brush-grafted glass as a hepatocyte sheet fabrication dish was investigated using primary rat hepatocytes. The PVLA-b-PNIPAAm brushes exhibited hepatocyte adhesion and proliferation without FBS precoating. By reducing the temperature from 37 °C to 20 °C, the hepatocyte sheet was recovered from the copolymer brush. These results indicated that the prepared PVLA-b-PNIPAAm brushes can be utilized for effective hepatocyte sheet fabrication without the need for FBS precoating.
Ethylene glycol-based thermoresponsive block copolymer brushes with cell-affinity peptides for thermally controlled interaction with target cells
2023, Materials and DesignTissue engineering has recently attracted attention as a potential remedy for intractable diseases. To be effective, such treatments require cell separation methods that do not modify cellular surfaces. In this study, we developed cell separation materials using ethylene glycol-based thermoresponsive block copolymer brushes and cell-affinity peptides. A poly(2-hydroxyethyl methacrylate-co-propargyl acrylate) (P(HEMA-co-PgA)) brush was grafted onto a glass substrate through atom transfer radical polymerization (ATRP). Subsequently, poly(2-(2-methoxyethoxy)ethyl methacrylate) (PMEO2MA), P(MEO2MA-co-HEMA-co-poly(ethylene glycol) methacrylate [PEGMA]), or P(MEO2MA-co-PEGMA) was grafted onto the P(HEMA-co-PgA) brush-coated substrates through a second ATRP. A Gly-Gly-Gly-Arg-Glu-Asp-Val (GGGREDV) peptide was conjugated to the copolymer brush via a click reaction. The prepared copolymer brushes exhibited thermoresponsive properties. The block copolymer brushes having the P(MEO2MA-co-HEMA-co-PEGMA) and P(MEO2MA-co-PEGMA) segment exhibited effective human umbilical vein endothelial cells (HUVECs) adhesion and normal human dermal fibroblasts (NHDFs) repulsion at 37 °C. By reducing temperature to 20 °C, adherent HUVECs were successfully recovered from the copolymer brushes. Using these copolymer brushes, HUVECs were separated from contaminant NHDFs and smooth muscle cells with these simple changes in temperature. The development of thermoresponsive ethylene glycol-based copolymer brushes with affinity peptides could be a useful cell separation material for tissue engineering applications.
Cardiac regeneration – Past advancements, current challenges, and future directions
2023, Journal of Molecular and Cellular CardiologyCardiovascular disease is the leading cause of mortality and morbidity worldwide. Despite improvements in the standard of care for patients with heart diseases, including innovation in pharmacotherapy and surgical interventions, none have yet been proven effective to prevent the progression to heart failure. Cardiac transplantation is the last resort for patients with severe heart failure, but donor shortages remain a roadblock. Cardiac regenerative strategies include cell-based therapeutics, gene therapy, direct reprogramming of non-cardiac cells, acellular biologics, and tissue engineering methods to restore damaged hearts. Significant advancements have been made over the past several decades within each of these fields. This review focuses on the advancements of: 1) cell-based cardiac regenerative therapies, 2) the use of noncoding RNA to induce endogenous cell proliferation, and 3) application of bioengineering methods to promote retention and integration of engrafted cells. Different cell sources have been investigated, including adult stem cells derived from bone marrow and adipose cells, cardiosphere-derived cells, skeletal myoblasts, and pluripotent stem cells. In addition to cell-based transplantation approaches, there have been accumulating interest over the past decade in inducing endogenous CM proliferation for heart regeneration, particularly with the use of noncoding RNAs such as miRNAs and lncRNAs. Bioengineering applications have focused on combining cell-transplantation approaches with fabrication of a porous, vascularized scaffold using biomaterials and advanced bio-fabrication techniques that may offer enhanced retention of transplanted cells, with the hope that these cells would better engraft with host tissue to improve cardiac function. This review summarizes the present status and future challenges of cardiac regenerative therapies.
Thermoresponsive mixed polymer brush to effectively control the adhesion and separation of stem cells by altering temperature
2023, Materials Today BioDuring the last few decades, thermoresponsive materials for modulating cell adhesion have been investigated for the application of tissue engineering. In this study, we developed thermoresponsive mixed polymer brushes consisting of poly(N-isopropylacrylamide) (PNIPAAm) and poly(N,N-dimethylaminopropylacrylamide) (PDMAPAAm). The mixed polymer brushes were prepared on a glass substrate via the reversible addition-fragmentation chain transfer polymerization of DMAPAAm and subsequent atom transfer radical polymerization of NIPAAm. The mixed polymer brushes grafted to glass exhibited increased cationic properties by increasing the grafted PDMAPAAm length. The shrinking and extension of PNIPAAm exposed and concealed PDMAPAAm, respectively, indicating that the surface cationic properties can be controlled by changing the temperature. At 37 °C, the prepared mixed polymer brushes enhanced cell adhesion through their electrostatic interactions with cells. They also exhibited various thermoresponsive adhesion and detachment properties using various types of cells, such as mesenchymal stem cells. Temperature-controlled cell adhesion and detachment behavior differed between cell types. Using the prepared mixed polymer brush, we separated MSCs from adipocytes and HeLa cells by simply changing the temperature. Thus, the thermoresponsive mixed polymer brushes may be used to separate mesenchymal stem cells from their differentiated or contaminant cells by altering the temperature.
MicroRNAs With Mega Functions in Cardiac Remodeling and Repair: The Micromanagement of Matters of the Heart
2023, MicroRNA in Regenerative Medicine, Second EditionCardiac remodeling subsequent to pressure volume load changes and myocardial injury is characterized by altered morphological and structural features with compromised cardiac function. MicroRNAs (miRNAs) are determinants of cellular function and maintain tissue homeostasis under physiological and pathophysiological conditions. Their mechanistic participation in signaling pathways of development and progression of myocardial remodeling is being studied. Research is also underway to use specific miRNAs as biomarkers of myocardial injury, and protocols are being developed to manipulate heart-specific miRNAs to achieve reverse remodeling. This chapter reviews published data relevant to miRNA-based therapeutic intervention as a novel alternative to existing treatment options in promoting cardiac remodeling and repair of the injured myocardium.