Scaffolds For Cardiac Tissue Engineering Research Articles

Comparative Analysis of Decellularization Methods on Bovine Pericardium Scaffolds for Cardiac Tissue Engineering

Background: Congenital heart disease (CHD) involves structural heart abnormalities present since fetal development. Corrective surgery is often the ideal treatment, utilizing implants or scaffolds for tissue repair. However, repeated surgeries may be required due to declining function of grafts, leading to increased healthcare costs. Bovine pericardium is a promising scaffold material for tissue engineering due to its composition of collagen, glycosaminoglycans (GAGs), and proteoglycans. This study aimed to compare the effectiveness of bovine pericardium scaffolds using different decellularization methods: sodium dodecyl sulfate (SDS) and hydrogen peroxide (H2O2). Methods: An experimental study was conducted with bovine pericardium scaffolds divided into control and treatment groups. The treatment groups used decellularization with SDS (0.5%, 1%) and H2O2 (3%). Scaffolds were evaluated for tensile strength, strain, Young's modulus, and histological properties, including nuclear density, collagen density, and GAG content. Data were analyzed using statistical methods to compare the effectiveness of each decellularization technique. Results: Histological analysis revealed that all decellularized samples showed no nuclear density, while control samples displayed light nuclear density. Collagen density was light in scaffolds treated with SDS 0.5% and H2O2, while SDS 1% resulted in no collagen presence. GAG density was absent in SDS-treated samples and minimal in H2O2-treated samples. Tensile strength was highest in H2O2-treated scaffolds (24.01 N) and lowest in SDS 0.5%. SDS 1% showed the greatest tensile strain, while H2O2-treated scaffolds had the highest stiffness. Conclusion: The decellularization methods effectively removed cellular components from the bovine pericardium scaffolds, with varying impacts on the extracellular matrix. SDS 1% provided the most elastic scaffold, while H2O2 preserved tensile strength and stiffness. These findings suggest that the choice of decellularization method can be optimized based on the desired mechanical properties of the scaffold for cardiac tissue engineering applications. Further research is needed to evaluate the long-term performance of these scaffolds in clinical settings.

Graphene-enhanced PCL electrospun nanofiber scaffolds for cardiac tissue engineering.

Cardiovascular diseases, particularly myocardial infarction, have significant healthcare challenges due to the limited regenerative capacity of injured heart tissue. Cardiac tissue engineering (CTE) offers a promising approach to repairing myocardial damage using biomaterials that mimic the heart's extracellular matrix. This study investigates the potential of graphene nanopowder (Gnp)-enhanced polycaprolactone (PCL) scaffolds fabricated via electrospinning to improve the properties necessary for effective cardiac repair. This work aimed to analyze scaffolds with varying graphene concentrations (0.5%, 1%, 1.5%, and 2% by weight) to determine their morphological, chemical, mechanical, and biocompatibility characteristics. The results presented that incorporating graphene improves PCL scaffolds' mechanical properties and cellular interactions. The optimal concentration of 1% graphene significantly enhanced mechanical properties and biocompatibility, promoting cell adhesion and proliferation. These findings suggest that Gnp-enhanced PCL scaffolds at this concentration can serve as a potent substrate for CTE providing insights into designing more effective biomaterials for myocardial restoration.

Development of a bacterial cellulose-gelatin composite as a suitable scaffold for cardiac tissue engineering.

Cardiac tissue engineering is suggested as a promising approach to overcome problems associated with impaired myocardium. This is the first study to investigate the use of BC and gelatin for cardiomyocyte adhesion and growth. Bacterial cellulose (BC) membranes were produced by Komagataeibacter xylinus and coated or mixed with gelatin to make gelatin-coated BC (BCG) or gelatin-mixed BC (mBCG) scaffolds, respectively. BC based-scaffolds were characterized via SEM, FTIR, XRD, and AFM. Neonatal rat-ventricular cardiomyocytes (nr-vCMCs) were cultured on the scaffolds to check the capability of the composites for cardiomyocyte attachment, growth and expansion. The average nanofibrils diameter in all scaffolds was suitable (~ 30-65nm) for nr-vCMCs culture. Pore diameter (≥ 10µm), surface roughness (~ 182nm), elastic modulus (0.075 ± 0.015MPa) in mBCG were in accordance with cardiomyocyte requirements, so that mBCG could better support attachment of nr-vCMCs with high concentration of gelatin, and appropriate surface roughness. Also, it could better support growth and expansion of nr-vCMCs due to submicron scale of nanofibrils and proper elasticity (~ 0.075MPa). The viability of nr-vCMCs on BC and BCG scaffolds was very low even at day 2 of culture (~ ≤ 40%), but, mBCG could promote a metabolic active state of nr-vCMCs until day 7 (~ ≥ 50%). According to our results, mBCG scaffold was the most suitable composite for cardiomyocyte culture, regarding its physicochemical and cell characteristics. It is suggested that improvement in mBCG stability and cell attachment features may provide a convenient scaffold for cardiac tissue engineering.

Conductive GelMA/alginate/polypyrrole/graphene hydrogel as a potential scaffold for cardiac tissue engineering; Physiochemical, mechanical, and biological evaluations

Cardiac failure can be a life-threatening condition that, if left untreated, can have significant consequences. Functional hydrogel has emerged as a promising platform for cardiac tissue engineering. In the proposed study, gelatin methacrylate (GelMA) and alginate, as a primary matrix to maintain cell viability and proliferation, and polypyrrole and carboxyl-graphene, to improve mechanical and electrical properties, are thoroughly evaluated. Initially, a polymer blend of GelMA/Alginate (1:1) was prepared, and then the addition of 2–5 wt% of polypyrrole was evaluated. Next, the effect of incorporating graphene-carboxyl nanosheets (1, 2, and 3 wt%) within the optimized scaffold with 2 wt% polypyrrole was thoroughly studied. The electrical conductivity of the hydrogels was significantly enhanced from 0.0615 ± 0.007 S/cm in GelMA/alginate to 0.124 ± 0.04 S/cm with the addition of 5 wt% polypyrrole. Also, 3 wt% carboxyl graphene improved the electrical conductivity to 0.27 ± 0.09 S/cm. The compressive strength of carboxyl-graphene-containing hydrogel was in the range of 175 to 520 kPa, and tensile strength was 61 and 133 kPa. The simplicity and low-cost fabrication, tunable mechanical properties, optimal electrical conductivity, blood compatibility, and non-cytotoxicity of GelMA/alginate/polypyrrole/graphene biocomposite hydrogel is a promising construct for cardiac tissue engineering.

Fabrication and Characterization of Piezoelectric PEO/SF/BaTiO3 Scaffolds for Cardiac Tissue Engineering

The existence of an intrinsic electrical platform responsible for the formation and transmission of impulses is essential, especially in cardiac tissue. However, most cardiac tissue made from biodegradable polymeric materials lacks conductive characteristics; this delays regional conduction, potentially causing arrhythmias. This study proposes a conductive polyethylene oxide (PEO)/silk fibroin (SF)-based material conjugated with conductive nanoparticles as a cardiac patch to fix any infarcted heart part. A new composite of PEO/15 wt%SF/0.2 wt%BaTiO3 was prepared and characterized in vitro. The obtained patches were characterized by conventional Bragg-platinum-conductive action (XRD), FTIR spectroscopy, Raman spectra, and thermogravimetric analysis. A PiezoTester device was used to evaluate the piezoelectric properties. The produced samples of 500 μm thickness were assessed in tapping mode. The applied load was selected to be as low as possible, and the frequencies were adjusted to simulate the heartbeats, ranging from 10 to 100 Hz. The results showed that a maximum of around 1100 mV was obtained at a load of 20 N. A maximum of about 80 mV was received at an applied force of 1 N and a frequency of 100 Hz, which matches the electricity generated by the human heart. The cytotoxicity effect of prepared films was tested against AC16 cells using microculture tetrazolium assay (MTT). The pristine PEO cell viability either was not affected by adding SF or slightly decreased. However, the cell viability dramatically increased by adding BaTiO3 to the PEO/SF composites. The confocal microscope images proved that the cells showed a spread morphology. The cells adhered to the PEO membranes and demonstrated a well-spread morphology. Overall, our study suggests that the PEO/SF/BaTiO3 composite can be a promising cardiac patch material for repairing infarcted heart tissue, as it is conductive, has good mechanical properties, and is biocompatible.

3D Graphene Oxide-Polyethylenimine Scaffolds for Cardiac Tissue Engineering.

The development of novel three-dimensional (3D) nanomaterials combining high biocompatibility, precise mechanical characteristics, electrical conductivity, and controlled pore size to enable cell and nutrient permeation is highly sought after for cardiac tissue engineering applications including repair of damaged heart tissues following myocardial infarction and heart failure. Such unique characteristics can collectively be found in hybrid, highly porous tridimensional scaffolds based on chemically functionalized graphene oxide (GO). By exploiting the rich reactivity of the GO's basal epoxydic and edge carboxylate moieties when interacting, respectively, with NH2 and NH3+ groups of linear polyethylenimines (PEIs), 3D architectures with variable thickness and porosity can be manufactured, making use of the layer-by-layer technique through the subsequent dipping in GO and PEI aqueous solutions, thereby attaining enhanced compositional and structural control. The elasticity modulus of the hybrid material is found to depend on scaffold's thickness, with the lowest value of 13 GPa obtained in samples containing the highest number of alternating layers. Thanks to the amino-rich composition of the hybrid and the established biocompatibility of GO, the scaffolds do not exhibit cytotoxicity; they promote cardiac muscle HL-1 cell adhesion and growth without interfering with the cell morphology and increasing cardiac markers such as Connexin-43 and Nkx 2.5. Our novel strategy for scaffold preparation thus overcomes the drawbacks associated with the limited processability of pristine graphene and low GO conductivity, and it enables the production of biocompatible 3D GO scaffolds covalently functionalized with amino-based spacers, which is advantageous for cardiac tissue engineering applications. In particular, they displayed a significant increase in the number of gap junctions compared to HL-1 cultured on CTRL substrates, which render them key components for repairing damaged heart tissues as well as being used for 3D in vitro cardiac modeling investigations.

Recent advances in tailoring stimuli-responsive hybrid scaffolds for cardiac tissue engineering and allied applications.

To recapitulate bio-physical properties and functional behaviour of native heart tissues, recent tissue engineering-based approaches are focused on developing smart/stimuli-responsive materials for interfacing cardiac cells. Overcoming the drawbacks of the traditionally used biomaterials, these smart materials portray outstanding mechanical and conductive properties while promoting cell-cell interaction and cell-matrix transduction cues in such excitable tissues. To date, a large number of stimuli-responsive materials have been employed for interfacing cardiac tissues alone or in combination with natural/synthetic materials for cardiac tissue engineering. However, their comprehensive classification and a comparative analysis of the role played by these materials in regulating cardiac cell behaviour and in vivo metabolism are much less discussed. In an attempt to cover the recent advances in fabricating stimuli-responsive biomaterials for engineering cardiac tissues, this review details the role of these materials in modulating cardiomyocyte behaviour, functionality and surrounding matrix properties. Furthermore, concerns and challenges regarding the clinical translation of these materials and the possibility of using such materials for the fabrication of bio-actuators and bioelectronic devices are discussed.

Design and fabrication of polyvinylidene fluoride-graphene oxide/gelatine nanofibrous scaffold for cardiac tissue engineering

Polyvinylidene fluoride (PVDF) electrospun scaffolds have recently been developed for cardiac tissue engineering applications thanks to their piezoelectricity. However, PVDFs’ hydrophobic nature requires modifications by incorporating natural polymers. In this study, we focussed on the hybrid electrospinning of PVDF and gelatine and the further introduction of graphene oxide nanoparticles to investigate either hydrophilicity or piezoelectricity enhancement and its impact on mouse embryonic cardiomyocytes. The results revealed a nanofibre diameter of 379 ± 73 nm for the PVDF/gelatine/graphene oxide (PVDF-GO-CG) platform, providing excellent tensile strength. Additionally, hydrophilicity was improved by gelatine and GO incorporation compared with pure PVDF. Cellular studies also showed an elongated morphology of cardiomyocytes, similar to the myocardial tissue, as well as high viability and non-toxicity in the PVDF-GO-CG scaffold according to the average survival rate. Furthermore, the expression of connexin 43 and troponin T genes underwent an increment of 41 and 35% in the PVDF-GO-CG compared with the PVDF-CG sample. This study proves the applicability of the PVDF-GO-CG scaffold as an alternative substrate for developing engineered cardiac tissues by providing an environment to re-establish their synchronised communications.

Cyto- and bio-compatibility assessment of plasma-treated polyvinylidene fluoride scaffolds for cardiac tissue engineering.

As part of applications dealing with cardiovascular tissue engineering, drop-cast polyvinylidene fluoride (PVDF) scaffolds have been treated by cold plasma to enhance their adherence to cardiac cells. The scaffolds were treated in a dielectric barrier device where cold plasma was generated in a gaseous environment combining a carrier gas (helium or argon) with/without a reactive gas (molecular nitrogen). We show that an Ar-N2 plasma treatment of 10min results in significant hydrophilization of the scaffolds, with contact angles as low as 52.4° instead of 132.2° for native PVDF scaffolds. Correlation between optical emission spectroscopy and X-ray photoelectron spectroscopy shows that OH radicals from the plasma phase can functionalize the surface scaffolds, resulting in improved wettability. For all plasma-treated PVDF scaffolds, the adhesion and maturation of primary cardiomyocytes is increased, showing a well-organized sarcomeric structure (α-actinin immunostaining). The efficacy of plasma treatment was also supported by real-time PCR analysis to demonstrate an increased expression of the genes related to adhesion and cardiomyocyte function. Finally, the biocompatibility of the PVDF scaffolds was studied in a cardiac environment, after implantation of acellular scaffolds on the surface of the heart of healthy mice. Seven and 28days after implantation, no exuberant fibrosis and no multinucleated giant cells were visible in the grafted area, hence demonstrating the absence of foreign body reaction and the biocompatibility of these scaffolds.

Sodium Alginate/Chitosan Scaffolds for Cardiac Tissue Engineering: The Influence of Its Three-Dimensional Material Preparation and the Use of Gold Nanoparticles.

Natural biopolymer scaffolds and conductive nanomaterials have been widely used in cardiac tissue engineering; however, there are still challenges in the scaffold fabrication, which include enhancing nutrient delivery, biocompatibility and properties that favor the growth, maturation and functionality of the generated tissue for therapeutic application. In the present work, different scaffolds prepared with sodium alginate and chitosan (alginate/chitosan) were fabricated with and without the addition of metal nanoparticles and how their fabrication affects cardiomyocyte growth was evaluated. The scaffolds (hydrogels) were dried by freeze drying using calcium gluconate as a crosslinking agent, and two types of metal nanoparticles were incorporated, gold (AuNp) and gold plus sodium alginate (AuNp+Alg). A physicochemical characterization of the scaffolds was carried out by swelling, degradation, permeability and infrared spectroscopy studies. The results show that the scaffolds obtained were highly porous (>90%) and hydrophilic, with swelling percentages of around 3000% and permeability of the order of 1 × 10−8 m2. In addition, the scaffolds proposed favored adhesion and spheroid formation, with cardiac markers expression such as tropomyosin, troponin I and cardiac myosin. The incorporation of AuNp+Alg increased cardiac protein expression and cell proliferation, thus demonstrating their potential use in cardiac tissue engineering.

Artificial Scaffolds in Cardiac Tissue Engineering.

Cardiovascular diseases are a leading cause of death worldwide. Current treatments directed at heart repair have several disadvantages, such as a lack of donors for heart transplantation or non-bioactive inert materials for replacing damaged tissue. Because of the natural lack of regeneration of cardiomyocytes, new treatment strategies involve stimulating heart tissue regeneration. The basic three elements of cardiac tissue engineering (cells, growth factors, and scaffolds) are described in this review, with a highlight on the role of artificial scaffolds. Scaffolds for cardiac tissue engineering are tridimensional porous structures that imitate the extracellular heart matrix, with the ability to promote cell adhesion, migration, differentiation, and proliferation. In the heart, there is an important requirement to provide scaffold cellular attachment, but scaffolds also need to permit mechanical contractility and electrical conductivity. For researchers working in cardiac tissue engineering, there is an important need to choose an adequate artificial scaffold biofabrication technique, as well as the ideal biocompatible biodegradable biomaterial for scaffold construction. Finally, there are many suitable options for researchers to obtain scaffolds that promote cell–electrical interactions and tissue repair, reaching the goal of cardiac tissue engineering.

Electrospun piezoelectric scaffolds for cardiac tissue engineering.

The use of smart materials in tissue engineering is becoming increasingly appealing to provide additional functionalities and control over cell fate. The stages of tissue development and regeneration often require various electrical and electromechanical cues supported by the extracellular matrix, which is often neglected in most tissue engineering approaches. Particularly, in cardiac cells, electrical signals modulate cell activity and are responsible for the maintenance of the excitation-contraction coupling. Addition of electroconductive and topographical cues improves the biomimicry of cardiac tissues and plays an important role in driving cells towards the desired phenotype. Current platforms used to apply electrical stimulation to cells in vitro often require large external equipment and wires and electrodes immersed in the culture media, limiting the scalability and applicability of this process. Piezoelectric materials represent a shift in paradigm in materials and methods aimed at providing electrical stimulation to cardiac cells since they can produce and deliver electrical signals to cells and tissues by mechanoelectrical transduction. Despite the ability of piezoelectric materials to mimic the mechanoelectrical transduction of the heart, the use of these materials is limited in cardiac tissue engineering and methods to characterise piezoelectricity are often built in-house, which poses an additional difficulty when comparing results from the literature. In this work, we aim at providing an overview of the main challenges in cardiac tissue engineering and how piezoelectric materials could offer a solution to them. A revision on the existing literature in electrospun piezoelectric materials applied to cardiac tissue engineering is performed for the first time, as electrospinning plays an important role in the manufacturing of scaffolds with enhanced piezoelectricity and extracellular matrix native-like morphology. Finally, an overview of the current techniques used to evaluate piezoelectricity and their limitations is provided.

Development and Characterization of Furfuryl-Gelatin Electrospun Scaffolds for Cardiac Tissue Engineering.

In this study, three types of electrospun scaffolds, including furfuryl-gelatin (f-gelatin) alone, f-gelatin with polycaprolactone (PCL) in a 1:1 ratio, and coaxial scaffolds with PCL (core) and f-gelatin (sheath), were developed for tissue engineering applications. Scaffolds were developed through single nozzle electrospinning and coaxial electrospinning, respectively, to serve as scaffolds for cardiac tissue engineering. Uniform fibrous structures were revealed in the scaffolds with significantly varying average fiber diameters of 760 ± 80 nm (f-gelatin), 420 ± 110 nm [f-gelatin and PCL (1:1)], and 810 ± 60 nm (coaxial f-gelatin > PCL) via scanning electron microscopy. The distinction between the core and the sheath of the fibers of the coaxial f-gelatin > PCL electrospun fibrous scaffolds was revealed by transmission electron microscopy. Thermal analysis and Fourier transformed infrared (FTIR) spectroscopy revealed no interactions between the polymers in the blended electrospun scaffolds. The varied blending methods led to significant differences in the elastic moduli of the electrospun scaffolds with the coaxial f-gelatin > PCL revealing the highest elastic modulus of all scaffolds (164 ± 3.85 kPa). All scaffolds exhibited excellent biocompatibility by supporting the adhesion and proliferation of human AC16 cardiomyocytes cells. The biocompatibility of the coaxial f-gelatin > PCL scaffolds with superior elastic modulus was assessed further through adhesion and functionality of human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes, thereby demonstrating the potential of the coaxially spun scaffolds as an ideal platform for developing cardiac tissue-on-a-chip models. Our results demonstrate a facile approach to produce visible light cross-linkable, hybrid, biodegradable nanofibrous scaffold biomaterials, which can serve as platforms for cardiac tissue engineered models.

Biodegradable functional macromolecules as promising scaffolds for cardiac tissue engineering

AbstractCardiovascular diseases, the major international health problem and the leading cause of death worldwide have been increasing in recent years due to population aging and lifestyle changes. Loss of cardiac muscle function after myocardial damage is one of the most critical challenges in cardiovascular medicine that has not yet been overcome. Tissue engineering (TE) has emerged as a promising therapeutic approach in modern medicine, targeting the substitution of damaged tissue with functional tissue grown inside an artificial scaffold. Great efforts have been made toward the construction of tissue engineering scaffolds that paved the way for extracellular matrix (ECM)‐like biomaterial. In cardiac tissue engineering, key parameters must be determined to select the ideal biomaterial, such as biocompatibility, conductivity, mechanical features, degradation and swelling rate, surface properties, and cell viability, growth and proliferation. Among different scaffolding materials, a wide range of natural biological macromolecules and synthetic macromolecules have been utilized to produce scaffolds with multifunctionality for cardiac tissue engineering (CTE). In this review, we have focused on recent achievements in the field of synthetic biodegradable macromolecules (such as aliphatic polyesters, polyurethane, poly (glycerol sebacate)) and the significant strategies to construct electrically conductive scaffolds to regenerate the function of native cardiac tissue. These biodegradable macromolecules have several attractive properties, including biocompatibility, elasticity, good mechanical properties, compatibility with native cardiac tissue, and proper surface biochemistry to increase cardiac cell adhesion, making them appropriate candidates for CTE. Recently, a growing trend in the use of conductive scaffolds for cardiac regeneration has been witnessed. Different materials ranging from metals, ceramics, and polymers have been used as parts of conductive scaffolds for CTE, possessing conductivity assortments from a range of semiconductive to conductive. Moreover, this review paper also focuses on the main strategies to create electroconductive scaffolds for in vitro cardiac muscle regeneration.

A porcine cholecystic extracellular matrix conductive scaffold for cardiac tissue repair.

Cardiac tissue engineering using cells, scaffolds or signaling molecules is a promising approach for replacement or repair of damaged myocardium. This study addressed the contemporary need for a conductive biomimetic nanocomposite scaffold for cardiac tissue engineering by examining the use of a gold nanoparticle-incorporated porcine cholecystic extracellular matrix for the same. The scaffold had an electrical conductivity (0.74±0.03S/m) within the range of native myocardium. It was a suitable substrate for the growth and differentiation of cardiomyoblast (H9c2) as well as rat mesenchymal stem cells to cardiomyocyte-like cells. Moreover, as an epicardial patch, the scaffold promoted neovascularisation and cell proliferation in infarcted myocardium of rats. It was concluded that the gold nanoparticle coated cholecystic extracellular matrix is a prospective biomaterial for cardiac tissue engineering.

Tri‐layered alginate/poly(𝜀‐caprolactone) electrospun scaffold for cardiac tissue engineering

AbstractNovel tri‐layered electrospun alginate–graphene oxide (GO)/poly(ε‐caprolactone) (PCL) conductive scaffolds were fabricated with a sequential electrospinning method for the first time, in which the middle PCL layer acts as a mechanical support of the scaffold. The top and bottom alginate–GO nanofibrous layers provide cell viability and attachment, and also electroconductivity for the scaffold. The fabrication of this nanofibrous scaffold aims to simulate cardiac extracellular matrix. Scanning electron microscopy and infrared spectroscopy were used to characterize the nanofibrous scaffolds, confirming the presence of GO in nanofibers and also crosslinking reaction of the alginate nanofibrous layer. The electroconductivity of the tri‐layered electrospun scaffolds reached 7.29 ± 0.91 μS with the incorporation of 1 wt% GO into the top and bottom alginate nanofibrous layers. The ultimate tensile strength and Young’s modulus of the alginate–0.5 wt% GO/PCL scaffold reached 22.46 ± 0.07 and 4.35 ± 0.56 MPa, respectively. The biocompatibility of the nanofibrous scaffolds was evaluated using cardiac progenitor cells. The results showed that the presence of GO accelerated the degradation of the scaffolds in phosphate‐buffered saline solution. The cell studies revealed that GO incorporation into alginate nanofibers on the surface of the tri‐layer scaffolds could enhance cell viability, adhesion and proliferation. Overall, the tri‐layered electroconductive alginate–PCL nanofibrous scaffolds could be considered as potential candidates for cardiac tissue regeneration. © 2022 Society of Industrial Chemistry.

Decellularized human amniotic membrane reinforced by MoS2-Polycaprolactone nanofibers, a novel conductive scaffold for cardiac tissue engineering.

In order to regenerate myocardial tissues with functional characteristics, we need to copy some properties of the myocardium, such as its extracellular matrix and electrical conductivity. In this study, we synthesized nanosheets of Molybdenum disulfide (MoS2), and integrated them into polycaprolactone (PCL) and electrospun on the surface of decellularized human amniotic membrane (DHAM) with the purpose of improving the scaffolds mechanical properties and electrical conductivity. For in vitro studies, we seeded the mouse embryonic cardiac cells, mouse Embryonic Cardiac Cells (mECCs), on the scaffolds and then studied the MoS2 nanocomposites by scanning electron microscopy and Raman spectroscopy. In addition, we characterized the DHAM/PCL and DHAM/PCL-MoS2 by SEM, transmission electron microscopy, water contact angle measurement, electrical conductivity, and tensile test. Besides, we confirmed the scaffolds are biocompatible by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTT assay. Furthermore, by means of SEM images, it was shown that mECCs attached to the DHAM/PCL-MoS2 scaffold have more cell aggregations and elongated morphology. Furthermore, through the Real-Time PCR and immunostaining studies, we found out cardiac genes were maturated and upregulated, and they also included GATA-4, c-TnT, NKX 2.5, and alpha-myosin heavy chain in cells cultured on DHAM/PCL-MoS2 scaffold in comparison to DHAM/PCL and DHAM. Therefore, in terms of cardiac tissue engineering, DHAM nanofibrous scaffolds reinforced by PCL-MoS2 can be suggested as a proper candidate.

Application potential of three-dimensional silk fibroin scaffold using mesenchymal stem cells for cardiac regeneration

Cardiac tissue engineering focusing on biomaterial scaffolds incorporating cells from different sources has been explored to regenerate or repair damaged area as a lifesaving approach.The aim of this study was to evaluate the cardiomyocyte differentiation potential of human adipose mesenchymal stem cells (hAD-MSCs) as an alternative cell source on silk fibroin (SF) scaffolds for cardiac tissue engineering. The change in surface morphology of SF scaffolds depending on SF concentration (1-6%, w/v) and increase in their porosity upon application of unidirectional freezing were visualized by scanning electron microscopy (SEM). Swelling ratio was found to increase 2.4 fold when SF amount was decreased from 4% to 2%. To avoid excessive swelling, 4% SF scaffold with swelling ratio of 10% (w/w) was chosen for further studies.Biodegradation rate of SF scaffolds depended on enzymatic activity was found to be 75% weight loss of SF scaffolds at the day 14. The phenotype of hAD-MSCs and their multi-linage potential into chondrocytes, osteocytes, and adipocytes were shown by flow cytometry and immunohistochemical staining, respectively.The viability of hAD-MSCs on 3D SF scaffolds was determined as 90%, 118%, and 138% after 1, 7, and 14 days, respectively. The use of 3D SF scaffolds was associated with increased production of cardiomyogenic biomarkers: α-actinin, troponin I, connexin 43, and myosin heavy chain. The fabricated 3D SF scaffolds were proved to sustain hAD-MSCs proliferation and cardiomyogenic differentiation therefore, hAD-MSCs on 3D SF scaffolds may useful tool to regenerate or repair damaged area using cardiac tissue engineering techniques.

Preparation and characterization of polyurethane/chitosan/CNT nanofibrous scaffold for cardiac tissue engineering

Myocardial infarction of cardiomyocytes is a leading cause of heart failure (HF) worldwide. Since heart has very limited regeneration capacity, cardiac tissue engineering (TE) to produce a bioactive scaffold is considered. In this study, a series of polyurethane solutions (5–7%wt) in aqueous acetic acid were prepared using electrospinning. A variety of Polyurethane (PU)/Chitosan (Cs)/carbon nanotubes (CNT) composite nanofibrous scaffolds with random and aligned orientation were fabricated to structurally mimic the extracellular matrix (ECM). Electrospun nanofibers were then characterized using field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), water contact angle, degradation studies, tensile tests, electrical resistance measurement and cell viability assay. The biocompatibility of electrospun random and aligned nanofibrous scaffolds with H9C2 Cells was confirmed. The results revealed that fabricated PU/Cs/CNT composite nanofibrous scaffolds were electro-conductive and aligned nanofibers could be considered as promising scaffolds with nano-scale features for regeneration of infarcted myocardium.

Adipose-derived mesenchymal stem cell seeded Atelocollagen scaffolds for cardiac tissue engineering

ADMSCs were isolated from subcutaneous adipose tissue, characterized and cultured in vitro. GFP-labeled ADMSCs can grow and proliferate well on the Atelocollagen scaffolds, and induced by 5-aza the cells can differentiate into cardio-like cells. 3D cultured ADMSCs on Atelocollagen scaffolds were transplanted into mice ischemia myocardium, and have good biocompatibility with host cardio tissue.