Leveraging the Recent Advancements in GelMA Scaffolds for Bone Tissue Engineering: An Assessment of Challenges and Opportunities.

The field of bone tissue engineering has seen significant advancements in recent years. Each year, over two million bone transplants are performed globally, and conventional treatments, such as bone grafts and metallic implants, have their limitations. Tissue engineering offers a new level of treatment, allowing for the creation of living tissue within a biomaterial framework. Recent advances in biomaterials have provided innovative approaches to rebuilding bone tissue function after damage. Among them, gelatin methacryloyl (GelMA) hydrogel is emerging as a promising biomaterial for supporting cell proliferation and tissue regeneration, and GelMA has exhibited exceptional physicochemical and biological properties, making it a viable option for clinical translation. Various methods and classes of additives have been used in the application of GelMA for bone regeneration, with the incorporation of nanofillers or other polymers enhancing its resilience and functional performance. Despite promising results, the fabrication of complex structures that mimic the bone architecture and the provision of balanced physical properties for both cell and vasculature growth and proper stiffness for load bearing remain as challenges. In terms of utilizing osteogenic additives, the priority should be on versatile components that promote angiogenesis and osteogenesis while reinforcing the structure for bone tissue engineering applications. This review focuses on recent efforts and advantages of GelMA-based composite biomaterials for bone tissue engineering, covering the literature from the last five years.

Structural and biological engineering of 3D hydrogels for wound healing.

Chronic wounds have become one of the most important issues for healthcare systems and are a leading cause of death worldwide. Wound dressings are necessary to facilitate wound treatment. Engineering wound dressings may substantially reduce healing time, reduce the risk of recurrent infections, and reduce the disability and costs associated. In the path of engineering of an ideal wound dressing, hydrogels have played a leading role. Hydrogels are 3D hydrophilic polymeric structures that can provide a protective barrier, mimic the native extracellular matrix (ECM), and provide a humid environment. Due to their advantages, hydrogels (with different architectural, physical, mechanical, and biological properties) have been extensively explored as wound dressing platforms. Here we describe recent studies on hydrogels for wound healing applications with a strong focus on the interplay between the fabrication method used and the architectural, mechanical, and biological performance achieved. Moreover, we review different categories of additives which can enhance wound regeneration using 3D hydrogel dressings. Hydrogel engineering for wound healing applications promises the generation of smart solutions to solve this pressing problem, enabling key functionalities such as bacterial growth inhibition, enhanced re-epithelialization, vascularization, improved recovery of the tissue functionality, and overall, accelerated and effective wound healing.

Advanced bioengineering of female germ cells to preserve fertility.

Oogenesis and folliculogenesis are considered as complex and species-specific cellular differentiation processes, which depend on the in vivo ovarian follicular environment and endocrine cues. Considerable efforts have been devoted to driving the differentiation of female primordial germ cells toward mature oocytes outside of the body. The recent experimental attempts have laid stress on offering a suitable microenvironment to assist the in vitro folliculogenesis and oogenesis. Despite developing a variety of bioengineering techniques and generating functional mature gametes through in vitro oogenesis in earlier studies, we still lack knowledge of appropriate microenvironment conditions for building biomimetic culture systems for female fertility preservation. Therefore, this review paper can provide a source for a large body of scientists developing cutting-edge in vitro culture systems for female germ cells or setting up the next generation of reproductive medicine as feasible options for female infertility treatment. The focal point of this review outlines advanced bioengineering technologies such as 3D biofabricated hydrogels/scaffolds and microfluidic systems utilized with female germlines for fertility preservation through in vitro folliculogenesis and oogenesis.

Multifunctional Conductive Biomaterials as Promising Platforms for Cardiac Tissue Engineering.

Adult cardiomyocytes are terminally differentiated cells that result in minimal intrinsic potential for the heart to self-regenerate. The introduction of novel approaches in cardiac tissue engineering aims to repair damages from cardiovascular diseases. Recently, conductive biomaterials such as carbon- and gold-based nanomaterials, conductive polymers, and ceramics that have outstanding electrical conductivity, acceptable mechanical properties, and promoted cell-cell signaling transduction have attracted attention for use in cardiac tissue engineering. Nevertheless, comprehensive classification of conductive biomaterials from the perspective of cardiac cell function is a subject for discussion. In the present review, we classify and summarize the unique properties of conductive biomaterials considered beneficial for cardiac tissue engineering. We attempt to cover recent advances in conductive biomaterials with a particular focus on their effects on cardiac cell functions and proposed mechanisms of action. Finally, current problems, limitations, challenges, and suggested solutions for applications of these biomaterials are presented.

Reduced graphene oxide: osteogenic potential for bone tissue engineering.

Collagen (Col) type I, as the major component of the bone extracellular matrix has been broadly studied for bone tissue engineering. However,inferior mechanical properties limit its usage for load bearing applications. In this research, freeze dried Col scaffolds are coated with graphene oxide (GO) through a covalent bond of the amine Col with the graphene carboxyl groups. The prepared scaffolds were then reduced using a chemical agent. Scanning electron microscopy exhibited a porous structure for the synthesized scaffolds with an approximate pore size of 100-220 ± 12 µm, which is in the suitable range for bone tissue engineering application. Reducing the GO coating improved the compressive modulus of the Col from 250 to 970 kPa. Apatite formation was also indicated by immersing the scaffolds in simulated body fluid after five days. The cytocompatibility of the scaffolds, using human bone marrow-derived mesenchymal stem cells, was confirmed with MTT analysis. Alkaline phosphatase assay revealed that reducing the Col-GO scaffolds can effectively activate the differentiation of hBM-MSCs into osteoblasts after 14 days, even without the addition of an osteogenic differentiation medium. The results of this study highlight that GO and its reduced form have considerable potential as bone substitutes for orthopaedic and dental applications.

Electroactive cardiac patch containing reduced graphene oxide with potential antibacterial properties.

Graphene based nanomaterials are promising candidates for cardiac tissue engineering due to the excellent electrical and mechanical properties and the robust surface chemistry. This research was designed to investigate the physicochemical and biological effects of increasing concentration of reduced graphene oxide (rGO) coating on collagen (Col) scaffolds as well as their antibacterial properties. Enhanced GO coating content to 400 µg/ml and its reduction showed improvement of HUVECs viability, however, following reduction of more GO concentration, decreased cell viability was observed. Compared with the Col counterpart, electroactive containing rGO scaffolds upregulated cardiac gene expression involve in electrical coupling (Cx43), muscle contraction and relaxation (troponin-T) and cytoskeleton alignment (actinin-4) after 7 days even without external electrical stimulation. rGO coating significantly improved mechanical properties and the electroactivity of the Col scaffolds reaching to 1100 ±â€¯31 kPa and 4 × 10-4 ±â€¯1.20 S/m for GO concentration of 800 µg/ml, respectively. Also, the antibacterial properties of Col-rGO-400 scaffolds against Escherichia coli, Staphylococcus aureus and Streptococcus pyogenes were confirmed by culture and FESEM observation. Taken together, the results indicated that rGO coating presents promising properties to Col scaffolds providing a desirable micro environment for cardiomyocytes coupling and gene upregulation as well as antibacterial activities for cardiac patch application.

Effects of collagen/ß-tricalcium phosphate bone graft to regenerate bone in critically sized rabbit calvarial defects.

Bone defects remain a significant health issue and a major cause of morbidity in elderly patients. Composites based on collagen/calcium phosphate have been widely used for bone repair in clinical applications, owing to their comparability to bone extracellular matrix. This study aimed to evaluate the effects of a scaffold of collagen/calcium phosphate (COL/ß-TCP) on bone formation to assess its potential use as a bone substitute to repair bone defects. Bilateral full-thickness critically sized calvarial defects (8 mm in diameter) were created in New Zealand white rabbits and treated with COL/ß-TCP or COL scaffolds. One defect was also left unfilled as a control. Bone regeneration was assessed through histological evaluation using hematoxylin and eosin and Masson's trichrome staining after 4 and 8 weeks. Alizarin Red staining was also utilized to observe the mineralization process. Our findings indicated that COL/ß-TCP implantation could better enhance bone regeneration than COL and exhibited both new bone growth and scaffold material degradation.

Electroactive graphene oxide-incorporated collagen assisting vascularization for cardiac tissue engineering.

The application of a cardiac patch over the epicardial surface has shown positive effects in protecting cardiac function postinfarction. Electroactive patches could enhance electrical signal propagation among cardiac cells. In the present study, an electrically active composite of collagen and graphene oxide (Col-GO) was fabricated as a cardiac patch. Col scaffolds were fabricated using a freeze-drying method and coated covalently with GO. Some scaffolds were also reduced by a reduction agent to restore the high conductivity of GO. GO was shown to be a single layer with suitable lateral size for biological application. The Col-GO scaffolds contained randomly oriented interconnected pores with appropriate pore sizes of 120-138 ± 8 µm. GO flakes were also well distributed in the pore walls. By increasing the GO concentration, the tensile strength of the scaffolds was enhanced from 75 kPa for Col-GO-5 to 162 kPa for Col-GO-90. Young modulus also followed the same trend. Electrical conductivity of the scaffolds was in the range of semi-conductive materials (~10-4 S/m), which is suitable for cardiac tissue engineering applications. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay indicated no toxic effects on human umbilical vein endothelial cells (HUVECs) after 96 h. Also, a 10-days degradation product of the samples was compatible with HUVECs. Reduced scaffolds supported neonatal cardiomyocyte adhesion and upregulated the expression of the cardiac genes, including Cx43, Actin4, and Trpt-2 than their nonconductive counterparts. The obtained results confirmed the angiogenic properties of reduced Go-containing materials for cardiovascular applications where angiogenesis plays an important role, especially postinfarction. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 204-219, 2019.

Development of a bioactive porous collagen/ß-tricalcium phosphate bone graft assisting rapid vascularization for bone tissue engineering applications.

We developed collagen (COL) and collagen/beta tricalcium phosphate (COL/ß-TCP) scaffolds with a ß-TCP/collagen weight ratio of 4 by freeze-drying. Mouse bone marrow-derived mesenchymal stem cells (BMMSCs) were cultured on these scaffolds for 14 days. Samples were characterized by physicochemical analyses and their biological properties such as cell viability and alkaline phosphatase (ALP) activity was, also, examined. Additionally, the vascularization potential of the prepared scaffolds was tested subcutaneously in Wistar rats. We observed a microporous structure with large porosity (∼95-98%) and appropriate pore size (120-200 µm). The COL/ß-TCP scaffolds had a much higher compressive modulus (970 ± 1.20 KPa) than pure COL (0.8 ± 1.82 KPa). In vitro model of apatite formation was established by immersing the composite scaffold in simulated body fluid for 7 days. An ALP assay revealed that porous COL/ß-TCP can effectively activate the differentiation of BMMSCs into osteoblasts. The composite scaffolds also promoted vascularization with good integration with the surrounding tissue. Thus, introduction of ß-TCP powder into the porous collagen matrix effectively improved the mechanical and biological properties of the collagen scaffolds, making them potential bone substitutes for enhanced bone regeneration in orthopedic and dental applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 73-85, 2018.