Browsing by Author "Bhatt, A"

Now showing 1 - 19 of 19
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    Analysis of Whole Blood Prothrombin Time/ International Normalized Ratio Using Image Processing
    (Research in Medical and Engineering Sciences, 2021-03) Nair, SS; Rakhi, MR; Sadanandan, L; Harikrishnan, S; Bhatt, A
    Purpose: Patients who had undergone mechanical heart valve replacements, who have atrial fibrillation or deep vein thrombosis, need drugs called oral anticoagulants to prevent blood clotting and need regular testing of Prothrombin Time/International Normalized Ratio. Conventional laboratory approaches are time consuming, need blood component separation and a regular visit to clinical labs. The burden of PT measurement on the clinical laboratory is huge globally, which raise need for point of care, quick and user-friendly device. Methods: In this study we have proposed a handheld device based on the image processing for the PT/INR detection. Cost effective disposable strips were fabricated using thromboplastin as reagent. Device and strips were tested for 100 samples in clinical set up as per the ISO standard 17593 “Clinical laboratory testing and in vitro medical devices - Requirements for in vitro monitoring systems for self-testing of oral anticoagulant therapy”. Results: Data was compared with the values obtained from clinical laboratory using automated coagulometer T Coag DT-100 (Trinity), and commercially available Point of Care (POC) device from Roche, Diagnostics. A correlation coefficient (r) of 0.87 & 0.77 was observed between lab vs Chitra device and Chitra device vs commercially available device, respectively. Conclusion: Clinically accepted correlation may be obtained after automation of the strip fabrication technique. The proposed device is cost effective and easy to operate and works on the novel approach of image processing. To best of our knowledge this is the first report on the image processing-based PT/INR monitoring device.
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    Aqueous based Synthesis of CdSe/ZnS Q-dots: Study on Luminescence Properties and Cytotoxicity
    (PROCEEDING OF INTERNATIONAL CONFERENCE ON RECENT TRENDS IN APPLIED PHYSICS & MATERIAL SCIENCE (RAM 2013), 2013) Painuly, D; Bhatt, A; Krishnan, VK
    Present study aims to modify the thioacid capped CdSe Quantum-dots (Q-dots) surface by ZnS coating by direct synthesis in aqueous medium. CS formation was confirmed by red shift as well as enhancement in the luminescence peak compared to bare Q-dots. Effects of processing parameters during the shell preparation such as core concentration and sulphur concentration on the luminescence properties of CS have been studied. Processing parameters have been optimized at maximum luminescence efficiency. Cytocompatibility behavior was found to be better for CS compared to their bare Q-dots counterpart after evaluation. Cytotoxicity of CS has been further evaluated by changing the sulphur concentration and after aging for 8 days.
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    Biofabrication of skin tissue constructs using alginate, gelatin and diethylaminoethyl cellulose bioink.
    (International J of Biological Macromolecules, 2021-10) Lakshmi, TS; Riya, Raju; Suvanish, Kumar; Geevarghese, R; Renjith P., Nair; Kasoju, N; Bhatt, A
    Introduction: Biofabrication of skin tissue equivalents using 3D bioprinting technology has gained much attention in recent times due to the simplicity, the versatility of the technology and its ability in bioengineering biomimetic tissue histology. The key component being the bioink, several groups are actively working on the development of various bioink formulations for optimal skin tissue construction. Methods: Here, we present alginate (ALG), gelatin (GEL) and diethylaminoethyl cellulose (DCEL) based bioink formulation and its application in bioprinting and biofabrication of skin tissue equivalents. Briefly, DEAE cellulose powder was dispersed in alginate solution with constant stirring at 60 °C to obtain a uniform distribution of cellulose fibers; this was then mixed with GEL solution to prepare the bioink. The formulation was systematically characterized for its morphological, physical, chemical, rheological, biodegradation and biocompatibility properties. The printability, shape fidelity and cell-laden printing were assessed using the CellInk bioprinter. Results: The bioink proved to be a good printable, non-cytotoxic and stable hydrogel formulation. The primary human fibroblast and keratinocyte-loaded 3D bioprinted constructs showed excellent cell viability, collagen synthesis, skin-specific marker and biomimetic tissue histology. Conclusion: The results demonstrated the successful formulation of ALG-GEL-DCEL bioink and its application in the development of human skin tissue equivalents with distinct epidermal-dermal histological features.
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    Cytocompatibility Evaluation of the Mercaptoethanol capped CdSe Quantum Dots and CdSe/ZnS Core/Shell
    (J Biomedical Nanotechnology, 2013-03) Painuly, D; Bhatt, A; Krishnan, VK
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    DEAE- Cellulose-based composite hydrogel for 3D printing application: Physicochemical, mechanical, and biological optimization
    (Materials Today Communications, 2022-12) Rathina, Vel; Bhatt, A; Priyanka, A; Gauthaman, A; Anilkumar, V; Safeena, AS; Kartha, RS
    3D bioprinting is a layer-by-layer additive manufacturing process that requires the incorporation of biomaterials, cells, growth factors, etc. The biomaterial-ink used in bioprinting should comprise essential properties like shear thinning, proper viscosity and reduced shear stress on cells, structural integrity, porosity, biocompatible and degradable, etc., Especially in extrusion-based bioprinting, optimization of biomaterial ink is critical. Even though single-aspect biomaterials have been used for establishing a biomaterial ink, however, they often fail to meet all properties needed to be used as a biomaterial ink. Carrying this point in view, we have formulated hydrogels using Diethylaminoethyl Cellulose (DEAE-Cellulose), Alginate (ALG), and Gelatin (GEL) as biomaterial inks. Initially, six different hydrogel formulations (F1-F6) were prepared with varying concentrations of DEAE- Cellulose (0.45%−2%), alginate (1%−2%), and keeping gelatine concentration constant at 3.33%. These formulations were then assayed by swelling and degradation tests. Out of six, three hydrogels (F3, F4, and F5) were eliminated after initial studies due to the rapid degradation rate. The other three hydrogels ( F1, F2, and F6) were further thoroughly analyzed by the rheological study, mechanical study, printability assay, morphological analysis, and biocompatibility assays. Here, We have demonstrated the successful formulation of three biomaterial inks utilizing three different biopolymers for the field of tissue engineering with adequate swelling, degradation, rheological and printability properties. It was observed that the incorporation of DEAE-Cellulose significantly improved the shear thinning and viscosity recovery of hydrogels. Also, it improves mechanical integrity and printing accuracy. Moreover, all three hydrogels have shown excellent hemocompatibility and cytocompatibility. To conclude, this study proposes the optimization of composite hydrogel for 3D printing applications.
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    Differentiation of Human Adipose Tissue Derived Mesenchymal Stem Cells to Functional Hepatocytes like Cells: Exploring the Niche and Admscs Conditioned Media
    (Archives of Clinical and Biomedical Research., 2021-03) Sultana, R; Bhatt, A
    Tissue engineered approaches have been used for hepatic regeneration and growing hepatic cells in-vitro, however it remains challenging due to the lesser proliferative capacity of the hepatic cells as well as cell source availability due to damaged tissues. Stem cells could be an alternative strategy if differentiated to functional hepatic like cells by providing suitable niche or culture conditions. In the present study we tried to explore differentiation of mesodermal origin cells to the endodermal origin hepatic cells under different culture conditions including fibrin niche and adipocytes derived mesenchymal stem cells conditioned medium. Fibrin provides better cell adhesion and proliferation. Mesenchymal stem cells were isolated and characterized. Two step induction was used for differentiating into hepatic like cells. Functionality of the differentiated hepatic like cells was analyzed using Cyto P 450, glycogen storage and urea/albumin assay at different time points. It was observed the fibrin niche supports the hepatic differentiation and differentiated cells shows hepatic functions. This approach may be used to generate alternative cell source for hepatic tissue engineering.
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    Effect of photografting 2-hydroxyethyl acrylate on the hemocompatibility of electrospun poly(ethylene-co-vinyl alcohol) fibroporous mats
    (MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS, 2016) Mayuri, PV; Bhatt, A; Joseph, R; Ramesh, P
    Poly(ethylene-co-vinyl alcohol) (EVAL) has been recommended as a material suitable for blood contacting applications. Effect of ethylene content and associated hydrophobicity of EVAL on the blood-material interactions have been documented in the literature. In this work, surface chemistry of EVAL substrate was altered by photografting a hydrophilic monomer, 2-hydroxyethyl acrylate (HEA) with the aid of a photoinitiator, benzophenone (BP), and the effect of surface modification on the blood-material interactions was investigated. Since the modified material was intended to be used as leukodepletion filters, a solution containing EVAL, HEA and BP was electrospun into fibroporous mats and UV treated to induce grafting. Degree of grafting, bonding between fibers and fiber diameter increased with increase in UV exposure time whereas mechanical properties showed a decreasing trend. Decreased water contact angle indicated improved wetting characteristics. In vitro hemocompatibility tests revealed that the modified EVAL surface exhibited significantly lower hemolytic activity, protein adsorption and platelet adhesion than neat EVAL. The modification did not have any substantial effect on the activation of the complement system and coagulation parameters. Photografting led to significant reduction in the adhesion of red blood cells (RBC) whereas white blood cell (WBC) consumption remained above 90%. The results implied that photografting HEA on EVAL substantially improves hemocompatibility of EVAL and when it is used as a filter, it selectively removes leukocytes and allows easy passage of other blood components. (C) 2015 Elsevier B.V. All rights reserved.
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    Effect of photografting 2-hydroxyethylacrylate on the hemocompatibility of electrospun poly(ethylene-co-vinyl alcohol) fibroporous mats
    (Materials Science and Engineering, 2015-11) Mayuri, PV; Bhatt, A; Joseph, R; Ramesh, P
    Poly(ethylene-co-vinyl alcohol) (EVAL) has been recommended as a material suitable for blood contacting applications. Effect of ethylene content and associated hydrophobicity of EVAL on the blood–material interactions have been documented in the literature. In this work, surface chemistry of EVAL substrate was altered by photografting a hydrophilic monomer, 2-hydroxyethyl acrylate (HEA) with the aid of a photoinitiator, benzophenone (BP), and the effect of surface modification on the blood–material interactions was investigated. Since the modified material was intended to be used as leukodepletion filters, a solution containing EVAL, HEA and BP was electrospun into fibroporous mats and UV treated to induce grafting. Degree of grafting, bonding between fibers and fiber diameter increased with increase in UV exposure time whereas mechanical properties showed a decreasing trend. Decreased water contact angle indicated improved wetting characteristics. In vitro hemocompatibility tests revealed that the modified EVAL surface exhibited significantly lower hemolytic activity, protein adsorption and platelet adhesion than neat EVAL. The modification did not have any substantial effect on the activation of the complement system and coagulation parameters. Photografting led to significant reduction in the adhesion of red blood cells (RBC) whereas white blood cell (WBC) consumption remained above 90%. The results implied that photografting HEA on EVAL substantially improves hemocompatibility of EVAL and when it is used as a filter, it selectively removes leukocytes and allows easy passage of other blood components
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    Enhanced P-selectin expression on platelet-a marker of platelet activation, in young patients with angiographically proven coronary artery disease
    (MOLECULAR AND CELLULAR BIOCHEMISTRY, 2016) George, R; Bhatt, A; Narayani, J; Thulaseedharan, JV; Sivadasanpillai, H; Tharakan, JA
    P-selectin (CD62p) exposure is an established marker for platelet activation. P-selectin exposure can trigger variety of thrombotic and inflammatory reactions. In patients with coronary artery disease (CAD), platelets are activated, and hence, there is increased P-selectin exposure. The role of P-selectin exposure in patients on treatment with statins and anti-platelets is conflicting. A case-control study was performed to determine P-selectin exposure in consecutively recruited 142 patients (age aecurrency sign 55 years) with angiographically proven CAD on treatment and 92 asymptomatic controls. P-selectin exposure was determined by flow cytometry. Data on conventional risk factors were obtained along with estimation of levels of thrombotic [fibrinogen, lipoprotein (a), tissue plasminogen activator, plasminogen activator inhibitor-1, homocysteine and von Willebrand factor] and anti-thrombotic factors (antithrombin III). The P-selectin exposure was compared among patient groups who had different modes of presentation of CAD and categories of CAD disease severity. The patients were followed up for a period of 26 months. The results indicate that P-selectin exposure was significantly elevated in patients (mean +/- SD 9.24 +/- 11.81) compared to controls (mean +/- SD 1.48 +/- 2.85) with p < 0.0001. Similarly, conventional risk factors were significantly elevated in patients. P-selectin exposure showed significant negative correlation with antithrombin III levels. P-selectin exposure was higher in patients who presented with acute coronary syndromes than those who presented with effort angina. Cardiovascular event rate was 6 % on follow-up. The study establishes that thrombotic-inflammatory pathways enhancing P-selectin exposure unrelated to treatment might be activated in patients, while the event rate remained lowered, and hence, treatment strategies should be inclusive to control these factors.
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    . Exploring the Potential of Alginate-Gelatin-Diethylaminoethyl Cellulose Fibrinogen based Bioink for 3D Bioprinting of Skin
    (Carbohydrate Polymer Technologies and Applications, 2022-06) Ramakrishnan, R; Kasoju, N; Raju, R; Geevarghese, R; Gauthaman, A; Bhatt, A
    Designing printable bioinks for 3D bioprinting capable of supporting cellular viability with post-printing functionality remains challenging. Native ECM offers several physical, chemical, and biological cues that are difficult to restore using only a single component. Herein, we have optimized a multicomponent-based bioink formulation comprising alginate (ALG), gelatin (GEL), diethylaminoethyl cellulose (DCEL) and fibrinogen (FIB), termed as ALG-GEL-DCEL-FIB bioink for potential application in bioprinting and biofabrication of skin tissue equivalents. The designed formulation was extensively studied for its printability, physico-chemical, rheological, and biocompatibility properties. Excellent printability, shape fidelity and cell-laden tissue equivalent printing were established using the RegenHu 3D Discovery Bioprinter. The human primary fibroblast and keratinocyte-laden bioprinted constructs exhibited good cell viability. Long term culture of 4 weeks comprising 5 days of air-liquid-interphase followed by 21 days of submerged culture produced biomimetic tissue histology in the ALG-GEL-DCEL-FIB bioink printed constructs. Specific epidermal-dermal marker expressions proving functionality were evident in immunohistochemical, biochemical and gene expression analysis. The ALG-GEL-DCEL-FIB bioink may be explored further for potential biofabrication and therapeutic applications.
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    Fabrication and Characterization of Soy Protein/Polyvinyl Alcohol (PVA) Composite Membrane for Guided Tissue Regeneration
    (Regenerative Engineering and Translational Medicine, 2024-10) Saranya, CV; Bridget Jeyatha, W; Deepu, DR; Bhatt, A; Lizymol, PP
    Purpose Periodontitis is an inflammatory disease that damages the periodontal tissue and leads to tooth loss. Guided tissue regeneration (GTR) is a membrane-based method that prevents the down growth of epithelial and fibroblast cells and gradually restores the periodontal tissues. Currently, collagen membranes exist as the top choice in the field of GTR membranes. However, disease transmission, poor mechanical strength and unpredictable degradation limit its use. The main aim of the study is to fabricate a soy protein–based GTR membrane with good mechanical properties, cell barrier function, and cytocompatibility. Methods Soy protein isolate (SPI) was extracted from the seeds of Glycine max, and the membranes (SPG-1, SPG-2, and SPG-3) were fabricated using SPI, polyvinyl alcohol (PVA), and glycerol (Gly) by aqueous solution casting method. Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), contact angle, swelling analysis, and degradation studies of the membranes were carried out. Human periodontal ligament (hPDL) cells were used for the direct contact test, MTT assay, live-dead, cell adhesion, and membrane barrier function experiments. Results SPG-1 membrane exhibited a rough surface and significantly (p ≤ 0.05) lower contact angle (68°) than SPG-3. SPG-1 showed a lower swelling (74.03%) and weight loss percentage (42.13%) (p ≤ 0.001) than SPG-2 and SPG-3. SPG-1 membrane exhibited significantly (p ≤ 0.05) higher tensile strength of 5.7 MPa and suture pull-out strength of 9.04 N when compared with SPG-2 and SPG-3. SPG membranes were non-cytotoxic, cyto-compatible, and prevented the down growth of fibroblast cells. Conclusion SPG-1 membranes with 50% SPI stand out as a best candidate than other SPG membranes with better physiochemical properties. It favoured the growth and proliferation of hPDL cells and exhibited barrier properties. Lay Summary Periodontitis is a disease that affects the structure and function of the periodontal tissues, leading to teeth loss. Guided tissue regeneration (GTR) is a widely accepted treatment using a barrier membrane. Three different composite GTR membranes of soy protein, polyvinyl alcohol, and glycerol were fabricated by the solvent casting method by varying the amount of soy protein isolate. Physiochemical characterization and in vitro studies with human periodontal ligament cells and fibroblast cells demonstrated the suitability of the material for periodontal defect management.
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    Mercaptoethanol Capped CdSe Quantum Dots and CdSe/ZnS Core/Shell: Synthesis, Characterization and Cytotoxicity Evaluation
    (JOURNAL OF BIOMEDICAL NANOTECHNOLOGY, 2013) Painuly, D; Bhatt, A; Krishnan, VK
    CdSe Quantum dots (Q-dots) and CdSe/ZnS core/shell have been synthesized by wet chemical route using mercaptoethanol (ME) as cappant. The synthesized Q-dots and core/shell were characterized using X-ray diffraction (XRD), Transmission electron microscopy (TEM), Energy dispersive X-ray analysis (EDS), Dynamic Light Scattering (DLS), Optical absorption and luminescence spectroscopy. The core/shell formation was confirmed by both XRD and TEM analysis. The luminescence was shown to be considerably enhanced in the core/shell sample. Effect of dialysis process on the optical properties of the Q-dots and core/shell has also been discussed. Cytotoxicity studies have been carried out for Q-dots and core/shell. CdSe/ZnS core/shell was found to be non-cytotoxic as compared to CdSe Q-dots up to a certain concentration range. Polyethylene glycol (PEG) coating enhances the non-cytotoxic nature of CdSe/ZnS core/shell when compared with bare core/shell.
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    Newly Exposed Membrane Glycoprotein II β (CD41) on Activated Platelets has Higher Affinity for Plasma Fibrinogen that Blocks Anti-CD41 Binding
    (International Blood Research Reviews., 2015-04) Bhatt, A; Krishnan, LK
    The density of glycoprotein receptors on platelet membrane is dynamic and increase upon platelet activation by various stimulants to facilitate more ligand binding and functional outcome. Since CD41/61 receptor complex is responsible for platelet to platelet cross-linking, infusion of antagonists/antibodies against these molecules is considered an effective method for inhibiting platelet aggregation in certain clinical conditions. However, binding of anti-CD41 to the platelets stored in bags for transfusion was found to be lower. Probably, upon stimulation the newly exposed receptors may immediately bind fibrinogen and prevent access to the antibody. To prove this hypothesis, binding intensity of FITC-conjugated human fibrinogen to resting and stimulated platelets were compared using flow cytometry. In the presence of plasma fibrinogen, monoclonal antibody binding to the CD41 receptor decreased significantly, both onto the stored and agonistactivated platelets as compared to the resting cells. As compared to ADP-activated platelets, binding of anti-CD41 to thrombin-activated platelets was significantly lower. Variable binding intensity of anti-CD41 to platelets activated by different stimulants observed in this study allowed us to suggest that therapeutic antibodies also may not bind to platelets that are stimulated by strong agonists. Our results signify the importance of validating such antibodies under conditions of stronger activation of platelets. We observed that more platelet microparticles (PMP) were shredded upon in vitro platelet activation by thrombin as compared to induction by ADP. However, fibrinogen did not bind to the shredded microparticles Therefore; PMP in circulation may not take part in pathological thrombus formation.
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    A novel leukodepletion filter from electrospun poly(ethylene-vinyl alcohol) membranes and evaluation of its efficiency
    (International Journal of Polymeric Materials and Polymeric Biomaterials, 2016-01) Mayuri, PV; Bhatt, A; Sabareeswaran, A; Ramesh, P
    The authors describe the fabrication and evaluation of a novel leukodepletion filter using novel poly (ethylene-vinyl alcohol) membranes through electrospinning. The membranes were biocompatible and nonhemolytic. A prototype of the filter was developed by stacking the membranes in a poly(methyl methacrylate) case. Upon whole blood filtration, the filter could achieve 100% leukodepletion while capturing 8% red blood cells and 91% platelets. The length of filtration through the developed filter was high when compared to that of marketed filter while overall performance of the filter was comparable to that of the marketed filter
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    Physicochemical and invitro biocompatibility evaluation of water-soluble CdSe/ZnS core/shell
    (JOURNAL OF BIOMATERIALS APPLICATIONS, 2014) Painuly, D; Bhatt, A; Krishnan, VK
    Group II-VI semiconductor quantum dots (Q-dots) have found various applications in biomedical field during last decade. In this study, we have synthesized CdSe Q-dots and CdSe/ZnS core/shell (CS) by wet chemical route and characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM), dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FT-IR) and Photoluminescence (PL) spectroscopy. CS formation was confirmed by red shift as well as enhancement in the luminescence peak compared to bare Q-dots. Processing parameters such as core and sulfur concentrations were optimized at maximum luminescence efficiency during the shell preparation. Effects of dialysis, aging and cell culture medium on the properties of the Q-dots and CS were also studied by luminescence and DLS techniques. DLS data showed Q-dots and CS to be stable, and there was no effect on the integrity of the Q-dots and CS after various modifications. CS was found to be hemocompatible and cytocompatible for human umbilical vein endothelial cells even at a high concentration of 0.1 mg/ml up to 48 h indicating high potential for CS in various biomedical applications.
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    A preliminary study on monocyte-platelet interactions in diabetic subjects leading to phenotypic changes in monocytes
    (Indian Journal of Applied Research, 2020-07) Deepa, S; Bhatt, A
    Background and objectives: Platelets become hyperactive during progression of inflammatory disease conditions such as diabetes mellitus. We aimed to assess whether activated platelets (CD62+) interact with monocytes to form monocyteplatelet aggregates and such aggregates change the phenotype of monocytes into proinflammatory phenotype (CD14-CD16+) via a comparative study between diabetic and non diabetic subjects. Methods: Based on inclusion and exclusion criteria, study subjects were chosen. Monocytes isolated from buffy coat were characterised using immunostaining and flow cytometry. Phenotypic changes in monocytes were determined through flow cytometry; interaction between monocytes and platelets were determined through ow cytometry, ESEM. Proteomic studies were carried out to determine inflammatory marker expression. Results: Study revealed that there are monocyte-platelet interactions and alteration in monocyte phenotypes occurring in diabetic subjects compared to control subjects. Western blotting analysis showed the expression of inflammatory molecules IL-10 and MCP-1 in diabetic subjects, further confirming the proinflammatory nature of monocytes. Conclusion: Platelet monocytes interaction in diabetic subjects may lead to the phenotypic alteration in monocytes. Inflammatory protein analysis and cell surface marker analysis of monocytes may be considered as a preliminary tool for assessing complications during diabetes mellitus.
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    Scaffold for liver tissue engineering: Exploring the potential of fibrin incorporated alginate dialdehyde-gelatin hydrogel.
    (International J of Biological Macromolecules, 2020-11) Resmi, R; Anusree, KS; Joseph, R; Bhatt, A
    Introduction Development of a tissue-engineered construct for hepatic regeneration remains a challenging task due to the lack of an optimum environment that support the growth of hepatocytes. Hydrogel systems possess many similarities with tissues and have the potential to provide the microenvironment essential for the cells to grow, proliferate, and remain functionally active. Methods In this work, fibrin (FIB) incorporated injectable alginate dialdehyde (ADA) - gelatin (G) hydrogel was explored as a matrix for liver tissue engineering. ADA was prepared by periodate oxidation of sodium alginate. An injectable formulation of ADA-G-FIB hydrogel was prepared and characterized by FTIR spectroscopy, Scanning Electron Microscopy, and Micro-Computed Tomography. HepG2 cells were cultured on the hydrogel system; cellular growth and functions were analyzed using various functional markers. Results FTIR spectra of ADA-G-FIB depicted the formation of Schiff's base at 1608.53 cm−1 with a gelation time of 3 min. ADA-G-FIB depicted a 3D surface topography with a pore size in the range of 100–200 μm. The non-cytotoxic nature of the scaffold was demonstrated using L929 cells and more than 80 % cell viability was observed. Functional analysis of cultured HepG2 cells demonstrated ICG uptake, albumin synthesis, CYP-P450 expression, and ammonia clearance. Conclusion ADA-G-FIB hydrogel can be used as an effective 3D scaffold system for liver tissue engineering.
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    Urokinase -CdSe/ZnS core/shell conjugates for targeted fibrinolytic treatment: An in vitro evaluation
    (Journal of Biomaterials & Tissue Engineering., 2014-10) Painuly, D; Bhatt, A; Krishnan, VK
    Cardiac diseases are one of the major causes of mortality worldwide. The conventional fibrinolytic therapy using drugs causes fibrinolysis in a non-specific and uncontrolled manner limiting the therapeutic potential of this therapy. If the drugs used can be specifically targeted and released in a controlled fashion at the site of the clot, a better therapeutic potential can be expected. In this study, we have monitored in vitro clot lysis generated by a quantum dot [Q-dot] based targeted drug delivery system. Urokinase, a plasminogen activator was conjugated with CdSe/ZnS core–shell (CS) by carbodimide coupling method to form urokinase-core/shell conjugate (UK-CS conjugate). These conjugates were characterized using X-ray diffraction (XRD), Dynamic Light Scattering (DLS), Fourier-Transform Infrared Spectroscopy (FT-IR), absorption and Photoluminescence (PL) spectroscopy. The optimum entrapment efficiency (E. E) was found to be ∼96% and the conjugate was found to be stable at 4 °C for 20 days of storage. The storage stability was determined in terms of size, absorbance and encapsulation efficiency. In vitro drug release studies exhibited controlled sustained release of UK from conjugate. In vitro thrombolysis testing on the plasma clot was carried out at different dose concentrations (18, 36 and 72 μg) of UK and UK-CS conjugate and the efficacy of UK-CS was found similar to bare UK at higher concentrations. Accelerated thrombolysis was observed in UK-CS-Ab system by binding the drug carrier specific to the clot using anti—fibrin antibody (Ab). Clot lysis time was found to decrease and comparable to UK even at 18 μg level compared to UK-CS conjugate. Thus, the proposed conjugate system based upon CS drug carriers may find potential use in therapeutic applications of thrombolytic diseases.
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    Urokinase-CdSe/ZnS Core/Shell Conjugates for Targeted Fibrinolytic Treatment: An In Vitro Evaluation
    (JOURNAL OF BIOMATERIALS AND TISSUE ENGINEERING, 2014) Painuly, D; Bhatt, A; Krishnan, VK
    Cardiac diseases are one of the major causes of mortality worldwide. The conventional fibrinolytic therapy using drugs causes fibrinolysis in a non-specific and uncontrolled manner limiting the therapeutic potential of this therapy. If the drugs used can be specifically targeted and released in a controlled fashion at the site of the clot, a better therapeutic potential can be expected. In this study, we have monitored in vitro clot lysis generated by a quantum dot [Q-dot] based targeted drug delivery system. Urokinase, a plasminogen activator was conjugated with CdSe/ZnS core-shell (CS) by carbodimide coupling method to form urokinase-core/shell conjugate (UK-CS conjugate). These conjugates were characterized using X-ray diffraction (XRD), Dynamic Light Scattering (DLS), Fourier-Transform Infrared Spectroscopy (FT-IR), absorption and Photoluminescence (PL) spectroscopy. The optimum entrapment efficiency (E.E) was found to be similar to 96% and the conjugate was found to be stable at 4 degrees C for 20 days of storage. The storage stability was determined in terms of size, absorbance and encapsulation efficiency. In vitro drug release studies exhibited controlled sustained release of UK from conjugate. In vitro thrombolysis testing on the plasma clot was carried out at different dose concentrations (18, 36 and 72 mu g) of UK and UK-CS conjugate and the efficacy of UK-CS was found similar to bare UK at higher concentrations. Accelerated thrombolysis was observed in UK-CS-Ab system by binding the drug carrier specific to the clot using anti-fibrin antibody (Ab). Clot lysis time was found to decrease and comparable to UK even at 18 mu g level compared to UK-CS conjugate. Thus, the proposed conjugate system based upon CS drug carriers may find potential use in therapeutic applications of thrombolytic diseases.