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.

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.

Melamine-Based Polymeric Crosslinker for Cleaner Leather Production.

To augment sustainable tanning, less chrome input, high functional quality leather processed via no restricted substance in processing, and ease to treat the inevitable protein waste generated are the key challenge, and currently, they have become the active part of leather research. Our work covers the synthesis of a formaldehyde-free chromium-incorporated polymeric tanning agent (FF-CIPTA) and its application in a reformed leather processing route which ensures near zero discharge of chromium containing solid waste. The preliminary characterization of FF-CIPTA reveals that the developed product is stable up to pH 5.2, and the particle size distribution ranges from 955 to 1450 nm with 12% Cr2O3 content. The present work significantly reduces the tanning agent input without compromising the thermal stability (103 °C) of the leather because of its multicrosslinking nature. Since the product exhibits a polymeric character, it provides tanning-cum-filling action which in turn reduces the retanning agent consumption in subsequent processes. Scanning electron microscopic study, porosity analysis, and hand assessment results clearly indicate the significant improvement in organoleptic properties. In addition, the process also enjoys the benefits of zero chromium containing solid waste generation, 71.4% reduction in chromium input, and high chromium transfer efficiency (92%) than the conventional process (36%), and 74.4% reduction in total dissolved solids generation. Furthermore, the water consumption and chemical input are reduced by 51.6 and 17%, respectively. Reduction in wastewater treatment cost and a high economic value of chromium-free leather scraps leads to a cumulative gain of US$ 39.84 per ton of raw material processing. Overall, a potential and practical applicability for cleaner and sustainable tanning is well established.

Nano-biocatalyst: Bi-functionalization of protease and amylase on copper oxide nanoparticles.

There are reports that co-immobilization of enzymes on solid supports can circumvent the problem of loss of enzyme activity in a soup of enzymes. To understand the mechanistic pattern by which solid support can ensure enzyme stability. Copper oxide nanoparticles (CuO Nps) were employed to immobilize protease and amylase. These are enzymes widely used together and at different stages in various industrial activities such as laundry, leather processing, etc., the immobilization of enzymes was confirmed through FTIR, TGA, and zeta potential analysis. Enzyme activity assays were carried out to understand enzyme activity with and without immobilization. The interaction between the nanoparticle surface and enzyme was studied through molecular docking studies. Thus, the study provides an insight into how the immobilization of enzymes on nanoparticles could be beneficial for industrial applications.

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.

A logical and sustainable approach towards bamboo pulp bleaching using xylanase from Aspergillus nidulans.

Driven by the environmental benefits that bio-bleaching could bring, the interest in xylanase has received enormous attention and hence, the search of xylanase with properties like no cellulase activity, function at elevated temperatures and pH continues. The present study reports the production of extracellular xylanase from Aspergillus nidulans using waste agro-residues as substrate. The optimum temperature (60 °C) and pH (9.0), classified the xylanase as thermo and alkali tolerant. The addition of salt of Mn2+ increased the xylanase activity to almost double; however, these ions were unable to protect the enzyme from thermal inactivation. The FTIR spectra of bamboo pulp treated with this xylanase, revealed reduction in lignin as evident from reduced peak intensity coupled with the reduction in kappa number. The SEM image of enzyme treated pulp, exhibited dissociation in fibers exposing the internal structure with slight roughness. Swelling was also observed there by increasing its thickness which eventually helped in improving its physical properties. The bleaching efficacy of indigenous xylanase as indicated in this study, has established its competence as a promising candidate for pre-treating the bamboo pulp.

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.

Dual utility of a novel, copper enhanced laccase from Trichoderma aureoviridae.

Ever since the ability of laccase to oxidize non-phenolic lignin models was described, the oxidative degradation reactions catalyzed by laccase have been widely studied for paper pulp production or detoxification of aromatic pollutants. The viability of developing eco-friendly, laccase aided industrial processes has been explored. Here, we report the isolation and screening of fungi to explore their lignolytic ability on solid media using various substrates as indicators. The promising fungus was cultivated in submerged and solid state conditions. The crude enzyme obtained yielded elevated activity at 75°C and pH 9.0. Addition of CuSO4 increased the activity by almost 25% proving that Cu(2+) catalytically enhances the action of laccases. Decolorization studies were carried out using industrial dye, Remazol Brilliant Blue R (CI 61200) on solid and liquid medium. Visual decolorization was observed within 2 days of inoculation on solid media whereas, liquid medium incorporated with varying concentrations of dye solution showed a final level of decolorization of up to 76%. Bamboo degradation studies revealed a decrease in lignin content by 51 and 43% within a month. To the best of our knowledge, this study for the first time reports that Trichoderma aureoviridae can produce lignolytic enzyme and degrade lignin.

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.

Enhancing collagen stability through nanostructures containing chromium(III) oxide.

Stabilization of collagen for various applications employs chemicals such as aldehydes, metal ions, polyphenols, etc. Stability against enzymatic, thermal and mechanical degradation is required for a range of biomedical applications. The premise of this research is to explore the use of nanoparticles with suitable functionalization/encapsulation to crosslink with collagen, such that the three dimensional architecture had the desired stability. Collagen solution prepared as per standard protocols is treated with chromium(III) oxide nanoparticules encapsulated within a polymeric matrix (polystyrene-block-polyacrylic acid copolymer). Selectivity towards encapsulation was ensured by the reaction in dimethyl sulfoxide, where the PS groups popped out and encapsulated the Cr(2)O(3). Subsequently when immersed in aqueous solution, PAA units popped up to react with functional groups of collagen. The interaction with collagen was monitored through techniques such as CD, FTIR, viscosity measurements, stress analysis. CD studies and FTIR showed no degradation of collagen. Thermal stability was enhanced upon interaction of nanostructures with collagen. Self-assembly of collagen was delayed but not inhibited, indicating a compete binding of the metal oxide encapsulated polymer to collagen. Metal oxide nanoparticles encapsulated within a polymeric matrix could provide thermal and mechanical stability to collagen. The formed fibrils of collagen could serve as ideal material for various smart applications such as slow/sustained drug release. The study is also relevant to the leather industry in that the nanostructures can diffuse through the highly networked collagen fibre bundles in skin matrix easily, thus overcoming the rate limiting step of diffusion.

Synthesis of aligned hematite nanoparticles on chitosan-alginate films.

Iron oxide nanoparticles are being viewed with interest owing to the great potential they have in the biomedical applications like MRI contrast enhancement, targeted drug delivery, hyperthermia and recently in magnetic separation of cancer cells from the body. Templated synthesis has been considered ideal for synthesis of iron oxide nanoparticles as particles are attracted magnetically, in addition to usual flocculation through van der Waals attraction. Biological templates are attractive owing to their biocompatibility and the attractive porosity and surface chemistry that nature provides. Polysaccharides like chitosan and alginate have been employed in the synthesis of a polyion complex, which provided the active-binding sites for iron(II) ions in solution to bind. The natural organization of chitosan and alginate into a porous film has been exploited to synthesize spherical iron oxide nanoparticles through careful calcination of the iron(II) conjugate film. Our experiments indicate that the formed nanoparticles are highly crystalline, confirm to the hematite structure and have a superparamagnetic response with a low coercivity of 116Oe. Particles thus synthesized were highly monodisperse with hydrodynamic diameter of 1.8 nm. The symmetric porosity of the film translates into the synthesis of well-aligned nanoparticles of iron oxide. Compared to synthesis in solution, the film-assisted synthesis offered a greater degree of control over the particle size distribution pattern, with the chitosan-alginate template providing the needed spatial separation to prevent the aggregation due to magnetostatic coupling. Such hematite nanoparticles can either be used directly or converted to paramagnetic magnetite by reduction. Zeta potential measurements indicate highly stable nanoparticles, which can therefore be conjugated to cationic liposomes carrying drugs and magnetically guided to target sites.

Biological removal of carcinogenic chromium(VI) using mixed Pseudomonas strains.

The contamination of soil and wastewaters with Cr(VI) is a major problem. It has been suggested that microbial methods for Cr(VI) reduction are better than chemical methods, as they do not add other ions or toxic chemicals to the environment. In this study an aerobic reduction of Cr(VI) to Cr(III) by employing mixed Pseudomonas cultures isolated from a marshy land has been reported. The role of chromium concentration, temperature, pH and additives on the microbial reduction of Cr(VI) has been investigated. NADH was found to enhance the rate of reduction of Cr(VI). Complete reduction of chromium(VI) has been possible even at chromium(VI) concentrations of 300 ppm. Ions like SO(4)(2-) and poly-phenols inhibited the metabolic activity relating to Cr(VI) reduction. Under optimal conditions 100 mg/L of Cr(VI) was completely reduced within 180 min.

Studies on the nature of interaction of iron(III) with alginates.

The interactions between the polysaccharide alginate and iron(III) were investigated. The solution properties were studied through pH-metry, viscometry, zeta potential and particle size measurements. In the presence of alginate, iron(III) was stabilized and no precipitation was observed. Studies indicate that iron(III)-alginate system was more stable than iron(III) or alginate alone. The binding constant is of the order of 10(4) M(-1). A case for 'site binding model' for the interaction between alginate and Fe(III) has been made based on the studies using circular dichroism and zeta potential experiments. The number of binding sites per molecule of alginate has been estimated to be 66. This indicates that the alginate can bind more number of Fe(III) ions and thus provide a stable complex which can find wide industrial applications.

Green route for the utilization of chrome shavings (chromium-containing solid waste) in tanning industry.

Chromium-containing wastes from various industrial sectors are under critical review. Leather processing is one such industrial activity that generates chromium-bearing wastes in different forms. One of them is chrome shavings, and this contributes to an extent of 10% of the quantum of raw skins/hides processed, amounting to 0.8 million ton globally. In this study, the high protein content of chrome shavings has been utilized for reduction of chromium(VI) in the preparation of chrome tanning agent. This approach has been exploited for the development of two products: one with chrome shavings alone as reducing agent and the other with equal proportion of chrome shavings and molasses. The developed products exhibit more masking due to the formation of intermediate organic oligopeptides. This has been corroborated through the spectral, hydrolysis, and species-wise distribution studies. The formation of these organic masking agents helps in chrome tanning by shifting the precipitation point of chromium to relatively higher pH levels. Hence, the developed products find use as chrome tanning agents for leather processing, thus providing a means for better utilization of chrome shaving wastes.