Impact of the Different Chemical-Based Decellularization Protocols on the Properties of the Caprine Pericardium.

PURPOSE: This study aims to decellularized caprine pericardium tissue with varied non-ionic surfactant and anionic detergent concentrations. METHODS: Protocol A consists of 1%, 0.5%, and 0.25% (w/v) sodium dodecyl sulphate (SDS). Protocol B uses 1%, 0.5%, and 0.25% (w/v) Triton X-100. Protocol C comprised 0.5% SDS + 0.5% Triton X-100, 0.5% + 0.25%, and 0.25% SDS + 0.5% Triton X-100. RESULTS: Protocol B left a few countable cells in the pericardium tissue, but treatments A and C removed all cells. DNA quantification also demonstrated that protocol B had the most leftover DNA after decellularization. The pericardium tissue treated with an equal combination of anionic detergent and non-ionic surfactant preserves the matrix. However, changing the anionic detergent-non-ionic surfactant ratio disrupted the microstructure. Protocol A decreased pericardium tissue secant modulus (p < 0.05). Protocol B-treated pericardium tissue matched native tissue secant modulus and ultimate tensile stress. Protocol C strengthened pericardium tissue. CONCLUSION: The intact extracellular matrix and biomechanical properties like native tissues require optimal chemical doses and combinations.

Biodegradable AZ91 magnesium alloy/sirolimus/poly D, L-lactic-co-glycolic acid-based substrate for cardiovascular device application.

Biodegradable drug-eluting stents (DESs) are gaining importance owing to their attractive features, such as complete drug release to the target site. Magnesium (Mg) alloys are promising materials for future biodegradable DESs. However, there are few explorations using biodegradable Mg for cardiovascular stent application. In this present study, sirolimus-loaded poly D, L-lactic-co-glycolic acid (PLGA)-coated/ sirolimus-fixed/AZ91 Mg alloy-based substrate was developed via a layer-by-layer approach for cardiovascular stent application. The AZ91 Mg alloy was prepared through the squeeze casting technique. The casted AZ91 Mg alloy (Mg) was alkali-treated to provide macroporous networks to hold the sirolimus and PLGA layers. The systematic characterization was investigated via electrochemical, optical, physicochemical, and in-vitro biological characteristics. The presence of the Mg17 Al12 phase in the Mg sample was found in the x-ray diffraction system (XRD) spectrum which influences the corrosion behavior of the developed substrate. The alkali treatment increases the substrate's hydrophilicity which was confirmed through static contact angle measurement. The anti-corrosion characteristic of casted-AZ91 Mg alloy (Mg) was slightly less than the sirolimus-loaded PLGA-coated alkali-treated AZ91 Mg alloy (Mg/Na/S/P) substrate. However, dissolution rates for both substrates were found to be controlled at cell culture conditions. Radiographic densities of AZ91 Mg alloy substrates (Mg, Mg/Na, and Mg/Na/S/P) were measured to be 0.795 ± 0.015, 0.742 ± 0.01, and 0.712 ± 0.017, respectively. The star-shaped structure of 12% sirolimus/PLGA ensures the bioavailability of the drugs. Sirolimus release kinetic was fitted up to 80% with the "Higuchi model" for Mg samples, whereas Mg/Na/S/P showed 45% fitting with a zero-order mechanism. The Mg/Na/S/P substrate showed a 70% antithrombotic effect compared to control. Further, alkali treatment enhances the antibacterial characteristic of AZ91 Mg alloy. Also, the alkali-treated sirolimus-loaded substrates (Mg/Na/S and Mg/Na/S/P) inhibit the valvular interstitial cell's growth significantly in in-vitro. Hence, the results imply that sirolimus-loaded PLGA-coated AZ91 Mg alloy-based substrate can be a potential candidate for cardiovascular stent application.

Evaluation of physicochemical properties of graphene oxide-decellularized pericardium biohybrid scaffold.

The decellularized pericardium has been widely used in cardiac tissue engineering, whereas its clinical applications are limited due to weak mechanical performance, high collagen exposure, and being prone to microbial contamination. In this study, a biohybrid scaffold of the decellularized caprine pericardium (DCP) and graphene oxide (GO) was fabricated by an immersion coating technique. The antimicrobial activity of GO was evaluated against Escherichia coli and showed minimum inhibitory concentration at 125 µg/mL and minimum bactericidal concentration at 250 µg/mL. The presence of GO on the surface of the biohybrid GO-DCP was confirmed through SEM analysis. The existence of glycosaminoglycan, elastin, and collagen in the DCP and GO-DCP was inferred from the FTIR spectra. The biocompatibility of GO-DCP was studied by seeding valvular interstitial cells, and the results show GO coating supports cell adhesion on the serous and fibrous sides of the DCP. Further, the biomechanical response of DCP is unaltered by the presence of GO. In conclusion, GO enhances the biological performance of decellularized pericardium, which can be used in cardiac tissue engineering applications.

Metallic Nanocarriers for Therapeutic Peptides: Emerging Solutions Addressing the Delivery Challenges in Brain Ailments.

Peptides and proteins have recently emerged as efficient therapeutic alternatives to conventional therapies. Although they emerged a few decades back, extensive exploration of various ailments or disorders began recently. The drawbacks of current chemotherapies and irradiation treatments, such as drug resistance and damage to healthy tissues, have enabled the rise of peptides in the quest for better prospects. The chemical tunability and smaller size make them easy to design selectively for target tissues. Other remarkable properties include antifungal, antiviral, anti-inflammatory, protection from hemorrhage stroke, and as therapeutic agents for gastric disorders and Alzheimer and Parkinson diseases. Despite these unmatched properties, their practical applicability is often hindered due to their weak susceptibility to enzymatic digestion, serum degradation, liver metabolism, kidney clearance, and immunogenic reactions. Several methods are adapted to increase the half-life of peptides, such as chemical modifications, fusing with Fc fragment, change in amino acid composition, and carrier-based delivery. Among these, nanocarrier-mediated encapsulation not only increases the half-life of the peptides in vivo but also aids in the targeted delivery. Despite its structural complexity, they also efficiently deliver therapeutic molecules across the blood-brain barrier. Here, in this review, we tried to emphasize the possible potentiality of metallic nanoparticles to be used as an efficient peptide delivery system against brain tumors and neurodegenerative disorders. SIGNIFICANCE STATEMENT: In this review, we have emphasized the various therapeutic applications of peptides/proteins, including antimicrobial, anticancer, anti-inflammatory, and neurodegenerative diseases. We also focused on these peptides' challenges under physiological conditions after administration. We highlighted the importance and potentiality of metallic nanocarriers in the ability to cross the blood-brain barrier, increasing the stability and half-life of peptides, their efficiency in targeting the delivery, and their diagnostic applications.

Exploring the bioactivity of reduced graphene oxide and TiO2 nanocomposite for the regenerative medicinal applications.

Millions of people globally suffer from issues related to chronic wounds due to infection, burn, obesity, and diabetes. Nanocomposite with antibacterial and anti-inflammatory properties is a promising material to promote wound healing. This investigation primarily aims to synthesize reduced graphene oxide and titanium dioxide (rGO@TiO2) nanocomposite for wound healing applications. The rGO@TiO2 nanocomposite was synthesized by the one-step hydrothermal technique, and the physicochemical characterization of synthesized nanocomposite was performed by X-ray diffraction, Fourier transforms infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy, and dynamic light scattering. Further, the nanocomposite antibacterial, cytotoxicity, and wound-healing properties were analyzed by disc diffusion method, MTT assay, and in vitro scratch assay, respectively. Based on the TEM images, the average particle size of TiO2 nanoparticles was around 9.26 ± 1.83 nm. The characteristics peak of Ti-O-Ti bonds was observed between 500 and 850 cm-1 in the Fourier transforms infrared spectrum. The Raman spectrum of graphene oxide (GO) was obtained for bands D and G at 1354 cm-1 and at 1593 cm-1, respectively. This GO peak intensity was reduced in rGO, revealing the oxygen functional group reduction. Moreover, the rGO@TiO2 nanocomposite exhibited dose-dependent antibacterial properties against the positive and negative bacterium. The cytotoxicity for 5-100 µg/mL of rGO@TiO2 nanocomposite was above the half-maximal inhibitory concentration value. The in vitro scratch assay for rGO@TiO2 indicates that the nanocomposite promotes cell proliferation and migration. The nanocomposite recovered the wound within 48 h. The rGO@TiO2 nanocomposite shows potential materials for wound healing applications.

Mixed-ligand copper(II) complex of quercetin regulate osteogenesis and angiogenesis.

Copper(II) complex of quercetin Cu+Q, mixed ligand complexes, quercetin-Cu(II)-phenanthroline [Cu+Q(PHt)] and quercetin-Cu(II)-neocuproine [Cu+Q(Neo)] have been synthesized and characterized. From the FT-IR spectroscopic studies, it was evident that C-ring of quercetin is involved in the metal chelation in all the three copper complexes. C-ring chelation was further proven by UV-Visible spectra and the presence of Cu(II) from EPR spectroscopic investigations. These complexes were found to have osteogenic and angiogenic properties, observed through in vitro osteoblast differentiation and chick embryo angiogenesis assay. In osteoblast differentiation, quercetin-Cu(II) complexes treatment increased calcium deposition and alkaline phosphatase activity (ALP) activity at the cellular level and stimulated Runx2 mRNA and protein, ALP mRNA and type 1 collagen mRNA expression at the molecular level. Among the complexes, Q+Cu(PHt) showed more effects on osteoblast differentiation when compared to that of other two copper complexes. Additionally, Q+Cu(Neo) showed more effect compared to Q+Cu. Furthermore, the effect of these complexes on osteoblast differentiation was confirmed by the expression of osteoblast specific microRNA, pre-mir-15b. The chick embryo angiogenesis assay showed that angiogenic parameters such as blood vessel length, size and junctions were stimulated by these complexes. Thus, the present study demonstrated that quercetin copper(II) complexes exhibit as a pharmacological agent for the orthopedic application.