Development of Two-Dimensional Nanomaterials Based Electrochemical Biosensors on Enhancing the Analysis of Food Toxicants.

In recent times, food safety has become a topic of debate as the foodborne diseases triggered by chemical and biological contaminants affect human health and the food industry's profits. Though conventional analytical instrumentation-based food sensors are available, the consumers did not appreciate them because of the drawbacks of complexity, greater number of analysis steps, expensive enzymes, and lack of portability. Hence, designing easy-to-use tests for the rapid analysis of food contaminants has become essential in the food industry. Under this context, electrochemical biosensors have received attention among researchers as they bear the advantages of operational simplicity, portability, stability, easy miniaturization, and low cost. Two-dimensional (2D) nanomaterials have a larger surface area to volume compared to other dimensional nanomaterials. Hence, researchers nowadays are inclined to develop 2D nanomaterials-based electrochemical biosensors to significantly improve the sensor's sensitivity, selectivity, and reproducibility while measuring the food toxicants. In the present review, we compile the contribution of 2D nanomaterials in electrochemical biosensors to test the food toxicants and discuss the future directions in the field. Further, we describe the types of food toxicity, methodologies quantifying food analytes, how the electrochemical food sensor works, and the general biomedical properties of 2D nanomaterials.

Carbon Dots-Mediated Fluorescent Scaffolds: Recent Trends in Image-Guided Tissue Engineering Applications.

Regeneration of damaged tissues or organs is one of the significant challenges in tissue engineering and regenerative medicine. Many researchers have fabricated various scaffolds to accelerate the tissue regeneration process. However, most of the scaffolds are limited in clinical trials due to scaffold inconsistency, non-biodegradability, and lack of non-invasive techniques to monitor tissue regeneration after implantation. Recently, carbon dots (CDs) mediated fluorescent scaffolds are widely explored for the application of image-guided tissue engineering due to their controlled architecture, light-emitting ability, higher chemical and photostability, excellent biocompatibility, and biodegradability. In this review, we provide an overview of the recent advancement of CDs in terms of their different synthesis methods, tunable physicochemical, mechanical, and optical properties, and their application in tissue engineering. Finally, this review concludes the further research directions that can be explored to apply CDs in tissue engineering.

Dendrimer-Functionalized Metal Oxide Nanoparticle-Mediated Self-Assembled Collagen Scaffold for Skin Regenerative Application: Function of Metal in Metal Oxides.

Functionalized metal oxide nanoparticles cross-linked collagen scaffolds are widely used in skin regenerative applications because of their enhanced physicochemical and biocompatibility properties. From the safety clinical trials point of view, there are no reports that have compared the effects of functionalized metal oxide nanoparticles mediated collagen scaffolds for in vivo skin regenerative applications. In this work, triethoxysilane-poly (amido amine) dendrimer generation 3 (TES-PAMAM-G3 or G3)-functionalized spherical shape metal oxide nanoparticles (MO NPs: ZnO, TiO2, Fe3O4, CeO2, and SiO2, size: 12-25 nm) cross-linked collagen scaffolds were prepared by using a self-assembly method. Triple helical conformation, pore size, mechanical strength, and in vitro cell viability of MO-TES-PAMAM-G3-collagen scaffolds were studied through different methods. The in vivo skin regenerative proficiency of MO-TES-PAMAM-G3-collagen scaffolds was analyzed by implanting the scaffold on wounds in Wistar albino rats. The results demonstrated that MO-TES-PAMAM-G3-collagen scaffold showed superior skin regeneration properties than other scaffolds. The skin regenerative efficiency of MO NPs followed the order ZnO > TiO2 > CeO2 > SiO2 > Fe3O4 NPs. This result can be attributed to higher mechanical strength, cell viability, and better antibacterial activity of ZnO-TES-PAMAM-G3-collagen scaffold that leads to accelerate the skin regenerative properties in comparison to other metal oxide based collagen scaffolds.

Polyphenols-loaded electrospun nanofibers in bone tissue engineering and regeneration.

Bone is a complex structure with unique cellular and molecular process in its formation. Bone tissue regeneration is a well-organized and routine process at the cellular and molecular level in humans through the activation of biochemical pathways and protein expression. Though many forms of biomaterials have been applied for bone tissue regeneration, electrospun nanofibrous scaffolds have attracted more attention among researchers with their physicochemical properties such as tensile strength, porosity, and biocompatibility. When drugs, antibiotics, or functional nanoparticles are taken as additives to the nanofiber, its efficacy towards the application gets increased. Polyphenol is a versatile green/phytochemical small molecule playing a vital role in several biomedical applications, including bone tissue regeneration. When polyphenols are incorporated as additives to the nanofibrous scaffold, their combined properties enhance cell attachment, proliferation, and differentiation in bone tissue defect. The present review describes bone biology encompassing the composition and function of bone tissue cells and exemplifies the series of biological processes associated with bone tissue regeneration. We have highlighted the molecular mechanism of bioactive polyphenols involved in bone tissue regeneration and specified the advantage of electrospun nanofiber as a wound healing scaffold. As the polyphenols contribute to wound healing with their antioxidant and antimicrobial properties, we have compiled a list of polyphenols studied, thus far, for bone tissue regeneration along with their in vitro and in vivo experimental biological results and salient observations. Finally, we have elaborated on the importance of polyphenol-loaded electrospun nanofiber in bone tissue regeneration and discussed the possible challenges and future directions in this field.

Polymethyl methacrylate (PMMA) grafted collagen scaffold reinforced by PdO-TiO2 nanocomposites.

In the past few decades, the design and fabrication of bio-scaffolds exhibiting structural stability in long-term and biocompatibility has received much attention in the field of tissue engineering application. In this direction, we have synthesized different mole ratio of PdO-TiO2 nanocomposites (1:1, 2:1 and 3:1 of Pd:Ti, size 5-11nm, 7-16 nm and 9-22 nm) through a simple single step sol-gel method. The obtained nanocomposites of different sizes were assimilated into poly (methyl methacrylate) grafted collagen biopolymer (g-PMMA-Collagen), resulting in a PdO-TiO2-g-PMMA-Collagen based scaffold. Physico-chemical properties and biocompatibility of g-PMMA-Collagen/PdO-TiO2-g-PMMA-Collagen scaffolds were analysed by using various techniques such as XRD, FT-IR, TGA, DSC, Universal Testing Machine, MTT, Alkaline phosphatase, Alizarin Red S staining assay and the obtained results were compared against pure collagen scaffold. Our results suggest that the incorporation of 1:1 mol ratio PdO-TiO2 nanocomposite (Size, 5-11 nm) offers a higher thermal stability (83.45 °C) and mechanical strength (Young's modulus 105.57 MPa) than the pure collagen scaffold (71.64 °C, 11.67 MPa). The PdO-TiO2 endowed scaffolds were not toxic to MG 63 cells (human osteosarcoma) and enhanced the ALP activity on the scaffolds during in vitro osteogenic differentiation. This work provides a new approach for mechanical reinforcing and enhanced osteogenic activity of collagen scaffolds without affecting its conformation or biocompatibility, an aspect that possibly makes them ideal for bone tissue engineering applications.

Effect of functionalized gold nanoparticle on collagen stabilization for tissue engineering application.

Functionalization of nanoparticle with specific groups is one of the most straightforward strategies to induce structural stability and specific cell responses from collagen based biomaterials. The effect of functionalised nanoparticles on triple helical conformational changes in collagen has not been understood well. For understanding the role of functionalization on collagen conformation, gold nanoparticles (Au NPs) prepared through wet chemical methods and functionalized with organic molecules (F-AuNPs) such as self-assembled monolayer (SAM), (3-aminopropyl) triethoxysilane (APTES), Polysaccharides (pectin and chitosan) and Poly(amido amine) PAMAM dendrimer (G0), were characterised and their interaction with collagen was studied. Protein conformational changes assessed by circular dichroism spectroscopy (CD) reveals that triple helical conformation of collagen was retained in presence of functionalized gold nanoparticle. The biocompatibility of functionalized gold nanoparticle was analysed against keratinocytes (HaCaT) cell by using (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) MTT assay. The result suggested that dendrimer functionalized gold nanoparticle exhibited higher cell viability when compared to other molecules functionalized gold nanoparticles studied. Based on the outcome of this study it can be envisioned that dendrimer functionalized gold nanoparticle mediated collagen materials are highly suitable for tissue engineering and cosmetic application.

Collagen-ZnO Scaffolds for Wound Healing Applications: Role of Dendrimer Functionalization and Nanoparticle Morphology.

Functionalized nanoparticle cross-linked collagen scaffolds offer improved properties to biomaterials and regenerated tissues, as influence of nanoparticle shape on collagen scaffold has received little attention. The present study evaluates the role of ZnO nanoparticle shape (sphere, rod, hexagonal, needle, flower, star, circular disk, doughnut, and cube) on collagen self-assembly. The nanoparticle was prepared by using coprecipitation method and subsequently functionalized with triethoxysilane poly(amidoamine) dendrimer generation 1 (TES-PAMAM-G1 or G1) on the nanoparticle surface. The self-assembly process of collagen, facilitated by EDC-NHS cross-linking, led to stable ZnO-TES-PAMAM-G1-collagen scaffolds. Physicochemical properties and biocompatibility of scaffolds were analyzed to determine the thermal, mechanical and pore size transformation and cell viability, etc. and obtained results compared against collagen scaffolds with/without EDC-NHS cross-linking. In vivo wound healing activity of ZnO-TES-PAMAM-G1-collagen scaffolds was tested on Albino rats that were subjected to excisional wounds and results were compared with control and collagen scaffold. Our findings suggested that the functionalized nanostructure mediated collagen scaffolds exhibited higher thermal (91.2 ± 0.3 °C) and mechanical stability (130.23-305.45 ± 0.1-2.0 MPa) than collagen scaffold (77.36 ± 0.5 °C and 7.96 ± 0.8 MPa). The result of in vivo wound healing study indicated that spherical shape of ZnO-TES-PAMAM -G1 NPs cross-linked collagen scaffold showed enhanced re-epithelization and faster collagen deposition than other scaffolds probably owing to their higher surface area, which led to higher grafting density on the surface. This work provides a new approach for designing nanoparticle mediated collagen scaffold for wound healing application.

Role of nanoparticle size in self-assemble processes of collagen for tissue engineering application.

Nanoparticle mediated extracellular matrix may offer new and improved biomaterial to wound healing and tissue engineering applications. However, influence of nanoparticle size in extracellular matrix is still unclear. In this work, we synthesized different size of silver nanoparticles (AgNPs) comprising of 10nm, 35nm and 55nm using nutraceuticals (pectin) as reducing as well as stabilization agents through microwave irradiation method. Synthesized Ag-pectin nanoparticles were assimilated in the self-assemble process of collagen leading to fabricated collagen-Ag-pectin nanoparticle based scaffolds. Physico-chemical properties and biocompatibility of scaffolds were analyzed through FT-IR, SEM, DSC, mechanical strength analyzer, antibacterial activity and MTT assay. Our results suggested that 10nm sized Ag-pectin nanoparticles significantly increased the denaturation temperature (57.83°C) and mechanical strength (0.045MPa) in comparison with native collagen (50.29°C and 0.011MPa). The in vitro biocompatibility assay reveals that, collagen-Ag-pectin nanoparticle based scaffold provided higher antibacterial activity against to Gram positive and Gram negative as well as enhanced cell viability toward keratinocytes. This work opens up a possibility of employing the pectin caged silver nanoparticles to develop collagen-based nanoconstructs for biomedical applications.

Nanorod mediated collagen scaffolds as extra cellular matrix mimics.

Creating collagen scaffolds that mimic extracellular matrices without using toxic exogenous materials remains a big challenge. A new strategy to create scaffolds through end-to-end crosslinking through functionalized nanorods leading to well-designed architecture is presented here. Self-assembled scaffolds with a denaturation temperature of 110 °C, porosity of 70%, pore size of 0.32 µm and Young's modulus of 231 MPa were developed largely driven by imine bonding between 3-mercapto-1-propanal (MPA) functionalized ZnO nanorods and collagen. The mechanical properties obtained were much higher than that of native collagen, collagen-MPA, collagen-3-mercapto-1-propanol (3MPOH) or collagen- 3-MPOH-ZnO, clearly bringing out the relevance of nanorod mediated assembly of fibrous networks. This new strategy has led to scaffolds with mechanical properties much higher than earlier reports and can provide support for cell growth and facilitation of cell attachment.

Fluorescent nanonetworks: a novel bioalley for collagen scaffolds and tissue engineering.

Native collagen is arranged in bundles of aligned fibrils to withstand in vivo mechanical loads. Reproducing such a process under in vitro conditions has not met with major success. Our approach has been to induce nanolinks, during the self-assembly process, leading to delayed rather than inhibited fibrillogenesis. For this, a designed synthesis of nanoparticles - using starch as a template and a reflux process, which would provide a highly anisotropic (star shaped) nanoparticle, with large surface area was adopted. Anisotropy associated decrease in Morin temperature and superparamagnetic behavior was observed. Polysaccharide on the nanoparticle surface provided aqueous stability and low cytotoxicity. Starch coated nanoparticles was utilized to build polysaccharide - collagen crosslinks, which supplemented natural crosslinks in collagen, without disturbing the conformation of collagen. The resulting fibrillar lamellae showed a striking resemblance to native lamellae, but had a melting and denaturation temperature higher than native collagen. The biocompatibility and superparamagnetism of the nanoparticles also come handy in the development of stable collagen constructs for various biomedical applications, including that of MRI contrast agents.