Research

Advancement in the field of healthcare requires a collaborative effort of engineers and biologists to develop new tools for therapeutic or diagnostic applications. The Biomaterials group i.e. materials engineers and biologists at BSSE are working together to develop new materials and devices for different biomedical applications. The research in the field of biomaterials is very diverse; at BSSE several groups are working on various materials for multitude of biomedical applications. Bikramjit Basu and V Kumaran are working towards development of PMMA micro-fluidics devices to study the effect of shear forces on cells. Suryasarathi Bose and Giridhar Madras are developing membranes for purification of water using PVDF based antibacterial membranes. Kaushik Chatterjee and Annapoorni Rangarajan are studying cancer cells in 3D tissue scaffolds towards engineering tumor tissues in vitro to facilitate study of cell biology and drug screening in tissue-like environments. Shilpee Jain is developing multifunctional micro devices for various biomedical applications with external electric and magnetic field stimulations. Ashok M. Raichur and Dipshikha Chakravortty are studying the efficacy of drug loaded polyelectrolyte capsules designed for encapsulation of active molecules with various bacterial and animal cell lines. Chandan Srivastava and Hanudatta S. Atreya are actively engaged in synthesizing ferrite nanoparticle-graphene oxide composites for application as contrast agents in magnetic resonance imaging.

Although the research in the Biomaterials group at BSSE is focused on developing new materials and testing their property, however, it is not limited in its scope: we have developed a water purification system based on antibacterial membranes.

BSSE offers a course on Fundamental of Biomaterials and Living Systems to familiarize the students with basic concepts in Biomaterials.

Projects

  • Water purification system based on PVDF antibacterial membranes
  • BSSE student Paresh and team are working on Purification of water using membranes. They have synthesized PVDF based membranes tethered with surface sensitive biocides imparting excellent antimicrobial action, reversible anti-fouling action and unimpeded flow to water. This will be achieved by engineering advanced functional grafts onto the membranes rendering the membrane its antimicrobial and anti-foulant properties.

Mechanical engineers and biologists in BSSE are working together in collaborative research and in training students to become adept in both areas. The current research themes in these efforts in BSSE are better characterized as biomechanics (application of mechanics to understand living systems) rather than mechano-biology (studies to understand the effects of mechanical stimuli applied on the living entities). The difference is subtle and debatable. Current projects include biomechanics of breast cancer cells and liver cells, and mechanics of motor learning.

Prof. Ashitava Ghosal and Prof. Aditya Murthy are investigating what role redundancy in human arm influences motor learning. It is a combination of kinematics and motor-learning experiments whose results are interpreted from the neurological perspectives. Prof. G. K. Ananthasuresh and Prof. Saumitra Das have an ongoing project in which the mechanical response of hypatocytes is changed presumably because of the action of viral proteins. Prof. Namrata Gundiah and Prof. P. Kondaiah are looking into the interplay between mechanics and biological response of cancer cells.

Mechanical engineers working on cells, tissues, and general biomechanics problems with biology researchers are also developing the required tools themselves rather than limiting themselves to what is available commercially and used extensively in biology laboratories. Examples include a miniature biorector, a viscometer, and a setup for conducting neuroengineering experiments.

BSSE offers a course on Mechanics of Biomaterials to familiarize students with basics in biomechanics.

Projects

  • The role of variability in motor learning
  • BSSE student Puneet Singh and team explore whether motor variability, often unwanted characteristic of motor performance has any significance in motor learning. They propose that motor variability has two components one caused by redundancy and other is random noise. In this work, Singh et al. quantify redundancy space and investigate its significance and effect on motor learning. They propose that a larger redundancy space leads to faster learning across subjects and the redundant component of motor variability is not noise. They also tested this hypothesis in neurologically diseased conditions to get a mechanistic understanding of how reward-based learning and error-based learning interact and how such learning is affected by redundancy space.

    Publication:
    P. Singh, S. Jana, A. Ghosal, A. Murthy "Exploration of joint redundancy but not task space variability facilitates supervised motor learning", PNAS, 2016 DOI: 10.1073/pnas.1613383113

  • Incubator-compatible Perfusion Bioreactor Array
  • BSSE PhD student Sreenath Balakrishnan and others have designed and built a perfusion bioreactor array that can be used inside a standard CO2 incubator. Each bioreactor has its own custom-made perstaltic pump, whose flow-rate can be adjusted as desired with its integrated electronics. A standard coverslip can be placed inside the biorector with or without scaffold to culture cells. This allows for live imaging and other assays. More information of this can be obtained from the following publication.

    Publication: S. Balakrishnan, M. S. Suma, S. R. Raju, S. D. B. Bhargav, S. Arunima, S. Das and G. K. Ananthasuresh, "A Scalable Perfusion Culture System with Miniature Peristaltic Pumps for Live-Cell Imaging Assays with Provision for Microfabricated Scaffolds", BioRes Open Access, 4(1), 343–357. DOI: 10.1089/biores.2015.0024

Projects

  • How do pathogens escape drug treatment?
  • Pathogenic bacteria display an uncanny ability to resurface after a course of antibiotic treatment. In many cases, this might happen even when no new drug resistant strain has emerged. It has been shown that there exists a tiny fraction of so called ‘persister’ bacteria that are able to tolerate the drugs in spite of being genetically identical to the drug susceptible ones in a bacterial population.

    Several evidences now suggest connection between gene motifs called the ‘toxin-antitoxin’ systems and bacterial persistence. Toxin proteins are usually lethal to cells in absence of its tight binding partner, the antitoxin. Levels of the unbound toxin results in multiphasic behaviour ranging from normal to slow-growing to deadbacteria.

    BSSE student Ratnasri, from Prof. Rahul Roy's lab studies the coupling between the toxin-antitoxin levels and persister state of the bacteria through live cell imaging and super-resolution microscopy. In parallel, a microdroplet based approach is used to enable high throughput analysis in order to measure cellular variability leading to persistence.

  • Siddharth Jhunjhunwala

  • Providing protection from pathogenic microorganisms is the primary function of our immune system, but its dysfunction has been shown to play a central role in a large number of non-pathogenic diseases such as autoimmunity, cancer, cardiovascular conditions, and diabetes. Modulating immune function using engineering tools to treat these disorders is often referred to as immuno-engineering. Siddharth Jhunjhunwala's laboratory is pursuing research in this area through two projects: one that involves creating new drug delivery systems capable of altering immune function in diabetic ulcers in order to enable wound healing; and another in collaboration with Annapoorni Rangarajan that focuses on developing biomaterial scaffolds to recreate the immune microenvironment present in tumors.

Students in the Bioengineering programme use the laboratories of their two advisers. However, a central facility has been created for general use. Currently, the lab is equipped with an Atomic Force Microscope (AFM), a BioAFM, a growth chamber, In Cell Analyzer 6000 high-content imaging system, and a bacterial culture hood in the Bioengineering laboratory. A fume hood and wet-benches are also available. In addition, a Tissue Culture Facility is in the process of being set up. The Bioengineering Office houses a small conference room, office space for Visiting Scientists, the office of the administrative assistant, and workspace for students/researchers.

  • Atomic Force Microscope (AFM)

  • Park systems NX-10 AFM
    Atomic force microscopes (AFMs), or scanning force microscopes, provide a high-resolution 3-D surface profile using a cantilever with a sharp tip to scan the surface to be imaged. AFM measures the deflection of the lever, not the force itself, with most systems using a laser beam deflection system. Park NX-10 AFM system is designed so that the X-Y scanner scans a sample in two-dimensional space, while the Z scanner moves the tip only in the z direction. X-Y scanner has a scan range of 50x50µm and z scanner of 12µm.
    Park NX-10 AFM modes
    Standard Imaging - True Non-Contact AFM (Air and Liquid), Basic Contact AFM (Air and Liquid), Phase Imaging, Intermittent (tapping) AFM, Force Modulation Microscopy (FMM), Nano indentation, Conductive AFM, I-V Spectroscopy, Force Distance (F-D) Spectroscopy
    Cantilevers Available
    Contact Cantilevers : CONTSCR, Spring Constant - 0.2N/m, MSNL-10 and MSCT : Spring Constant - 0.01-0.6 N/m, HYDRA Probes with 5um diameter spheres attached to the lever : Spring Constant - 0.01N/m, Non-Contact Cantilevers:ACTA, Spring Constant - 40N/m
    Usage Charges
    External Industrial Users: Rs. 5,700/- per hour (incl. of taxes)
    External Educational Users: Rs. 1,140/- per hour (incl. of taxes)


  • BioAFM

  • Park Systems XE-BIO AFM
    XE-Bio is a powerful 3-in-1 nanoscience research tool that uniquely combines industry's only True Non-Contact AFM with Ion Conductance Microscopy (ICM) and inverted optical microscope on the same platform. Designed for non-invasive in-liquid imaging, the combined imaging capability of AFM, ICM, and inverted optical microscopy makes the XE-Bio ideal for imaging biological samples, such as living cells, in dynamic conditions.
    This system has an SPM system on the stage of an inverted optical Microscope. The SPM system has an X-Y flat scanner (100µm × 100µm) and a separate SICM or AFM head with a 25µm z scanner. The SICM probe consists of a glass pipette filled with electrolyte with an Ag/AgCl electrode plugged into it.
    SICM can be utilized for obtaining contact-free images of the surface topography of biological samples.
    Usage Charges
    External Industrial Users: Rs. 6,900/- per hour (incl. of taxes)
    External Educational Users: Rs. 1,400/- per hour (incl. of taxes)


  • High-content imaging

  • GE InCell Analyzer 6000
    The InCell Analyzer 6000 is a high-content, laser line scanning confocal microscope equipped with a scientific cMOS camera, specially suited for use in high content assays such as genome-wide screens.
    Specifications
    Camera : sCMOS 5.5Mp (2560x2160 pixels)
    Imaging modes : Line confocal (aperture adjustable), widefield, transmitted light (brightfield, DIC and phase contrast imaging)
    Laser lines : 405, 488, 561 and 642 nm
    Objectives : 10X/0.45 NA, 20x/0.45 NA and 60x/0.95 NA
    Plate compatibility : Compatible with 6-, 12-, 24-, 48-, 96-, 384-, and 1536-well microplates
    Other features : Temperature control, liquid handling, Slide handling
    Software : IN Cell Investigator, IN Cell Miner HCM
    Usage Charges
    External Industrial Users: Rs. 6,900/- per hour (incl. of taxes)
    External Educational Users: Rs. 1,400/- per hour (incl. of taxes)


  • Flow cytometer

  • Becton Dickinson FACSCelesta
    BD FACSCelesta is a flow cytometer analyzer with three lasers (405 nm, 488 nm and 647 nm). It can simultaneously measure fourteen parameters, 1 each of FSc and SSc along with 12 colors. The filter arrangement is such that the following dyes (or their equivalents) may be measured – BV421, BV510, BV605, BV650, BV786, BB515/FITC, PE, PE-CF594, PerCP-Cy5.5, APC, APC-700, APC-Cy7. Samples may be loaded only with a 5 ml tube.



  • Micropipette puller

  • Sutter Instrument – Laser Based Micropipette Puller P-2000/F
    The P-2000/F is a microprocessor-controlled CO2 Laser-based micropipette puller. The default configuration allows fabrication of micropipettes for intracellular recording, patch clamping, microinjection and micro perfusion. CO2 laser heat source can work with quartz glass, a much stronger and more pure glass formulation than standard glass capillary tubing. The Model P-2000/F can also be used to pull tubing and optical fibres to exceedingly small diameters for research applications such as HPLC and near-field scanning microscopy.


  • Upright Fluorescence Microscope

  • Olympus BX53F
    Olympus BX53F is an upright fluorescence microscope equipped with Q-click cooled monochrome CCD camera. It is useful for imaging fluorescence on samples that are opaque such as thick materials.
    Specifications:
    Camera: Q-click cooled monochrome CCD camera, 1.4 Megapixel, pixel pitch: 6.45x6.45 microns
    Filters: Dapi, FITC, TRITC and Cy5
    Objectives: 4X, 10X, 20X, 40X and 100X (oil)
    DIC attacment for 10X and 100X
    No plates allowed, Only samples on slides.
    Usage Charges
    IISc users: Rs. 300 per hour










  • Fast performance liquid chromatography (FPLC)

  • Biorad NGC Quest 10 Chromatography System
    Used to purify proteins based on several parameters such as size, charge or specific interaction.
    Specifications:
    10ml/min pumps
    Inlet valve module
    Column switching valve
    BioFrac Fraction Collector
    Multi-Wavelength Detector



















  • Lyophilizer

  • Taitec VD-250R
    Freeze dryer used to dry frozen samples. Only aqueous samples allowed.
    Specifications:
    Cooling temperatue: -45°C
    Chamber volume: 4L
    Vacuum pump: 150lpm
    Multi branched tube with 6 ports
    Two lyophilizer bottles 750ml
    Two lyophilizer bottles 500ml
    Two lyophilizer bottles 100ml
    Usage Charges
    IISc users: Rs. 200 per day per port; minimum usage 1 day.








  • Inverted Fluorescence Microscope with Spinning Disk Confocal and Total Internal Reflection Fluorescence (TIRF) Modules











  • Bacterial cell culture facility

  • The bacterial cell culture facility at BE comprises a hortizontal laminar flow hood (Shalom Instruments, India), with a UV sterilization system that provides an aseptic environment. In addition, there is a 37°C incubator shaker (Shalom Instruments, India) for growth and incubation of bacterial cultures, a 4°C refrigerator for storage of stock solutions, reagents etc. and a visible range absorbance spectrophotometer (Spectrum, India) for measuring absorbance/ optical density of bacterial cultures to determine growth, proliferation etc.





  • Centrifuge-1

  • Large swinging bucket rotor refrigerated centrifuge from Eppendorf (model number 5804R)











  • Centrifuge-2

  • Small fixed angle rotor refrigerated centrifuge from Eppendorf (model number 5418R)











  • Cell Culture - CO2 incubator

  • CO2 incubator from New Brunswick (model number Galaxy 170S)











  • Cell Culture - Laminar hood

  • Laminar hood for sterile cell culture from Cleanair











  • Cell Culture - Microscope

  • Viewing microscope from Labomed (model number TCM-400)








  • Homogenizer

  • Homogenizer – from IKA (model number T18 digital disperser). Used for emulsification and homogenization of fluids











  • Electrospinning and Electrospraying

  • Electrospinning and Electrospraying – from Spraybase®. Used for preparation of microfibers, micro and macro-particles.










  • Incubator

  • for reactions and drug release studies











  • Other equipment

  • In addition, there are several small laboratory instruments that have been procured at the Bioengineering Lab:
    - pH meter
    - Sonicator
    - Precision weighing balance
    - Spin coater
    - Vortexer
    - Water bath
    - Gel rocker
    - Hot plates
    - Binocular microscope
    - Magnetic stirrer

The research carried out by the PhD students at BSSE cover a wide range of topics from biological molecules, cells, and tissues to organs, organisms, and systems and includes allied topics of biodesign, healthcare, agriculture, and studies on plant systems. Details of research of parcitipating faculty members can be found in their respective web sites. Brief descriptions of research projects of current PhD students appear below.


1. Breast Cancer Stem Cells in 3D Tissue Scaffolds

Gowri Balachander advised by Dr. Kaushik Chatterjee and Dr. Anu Rangarajan

Cancer contains a very small fraction of stem like cells with tumorigenic potential. These cells are highly implicated in therapy because of their resistance to anti-cancer treatment like drugs and radiation and most importantly because of their ability of self-renewal by which they continuously supply the tumor with cancer cells and also maintain the cancer stem cell population. Currently, the study of cancer stem cells is through conventional 2D culture system, which fails to mimic the in vivo tissue environment and 3D culture systems are likely to perform better in this regard. 3D tissue engineering scaffolds provide improved cell-cell and cell-matrix interactions because of which the cell morphology and signalling is more physiologic. Also, 3D culture systems are used as surrogates for in vivo drug testing in animals because the animals may respond to drugs differently from humans. The project aims to culture cancer stem cells in 3D matrices to give them a more in vivo environment and study their molecular behaviour and drug response.


2. Investigations into the changes in biomechanics of liver cells upon Hepatitis C Virus infection

Sreenath Balakrishnan advised by Prof. G. K. Ananthasuresh and Prof. Saumitra Das

We are interested in investigating the changes to the biomechanics of liver cells upon Hepatitis C Virus infection. We plan to use these alterations in the biomechanics as a mechanical biomarker for the diagnosis of the infection. In order to study the changes in the physical properties of liver cells in its physiological context, we have built a perfusion culture system to provide an in vivo like culture environment to the cells. Using this system we are characterizing the differential response of liver cells and those infected with Hepatitis C Virus to shear stress due to flow.

The perfusion culture system consists of miniature bioreactors, miniature peristaltic pumps and a control circuit. The bioreactors were designed for perfusion, high magnification live-cell imaging studies, easy incorporation of microfabricated scaffolds, and convenience of operation in standard cell culture techniques. By combining with miniature peristaltic pumps—one for each bioreactor to avoid cross-contamination and to maintain desired flow rate in each—we have made a culture system that facilitates perfusion culture inside standard incubators. This scalable system can support multiple parallel perfusion experiments. The major components were fabricated by three-dimensional printing using VeroWhite, which we show to be amenable to ex vivo cell culture. Furthermore, the components of the system can be reused, thus making it economical. We have validated the system and illustrated its versatility by culturing primary rat hepatocytes, live imaging the growth of mouse fibroblasts (NIH 3T3) on microfabricated ring- scaffolds inserted into the bioreactor, performing perfusion culture of breast cancer cells (MCF7), and high-magnification imaging of hepatocarcinoma cells (HuH7).
Figure: Clockwise from left: Perfusion culture system, Hepatocarcinoma cells and those expressing Hepatitis C Viral proteins cultured in the system


3. Engineering Nanosized Particles for Targeting Intracellular Pathogens

Rajeev Mudakavi advised by Dr. Dipshikha Chakravortty and Prof. Ashok Raichur

Intracellular pathogens such as Salmonella, M.tuberculosis, Legionella etc. are difficult to treat and eradicate from the mammalian host cells as they have developed ingenious defenses. Conventional antimicrobial agents cannot breach the bacterial defenses to attain the concentration required for killing the bacteria. They also modulate the host cell’s antibacterial response to suit their intracellular survival. This calls for delivering higher payloads of antibiotics into the infected cells, which we have carried out by formulating nanoparticles having functionally active coating. We have synthesized mesoporous silica nanoparticles (MSN) and investigated the role of lipid and amine coating in retarding the release of antibiotic and targeting the intracellular pathogen. The lipid coat was achieved through sonication with liposomes while amine coating was based on electrostatic attraction between the positively charged amine and the negatively charged polymer coated on MSN surface. Ciprofloxacin, a fluoroquinolone antibiotic, loaded into MSN showed enhanced antibacterial activity as against free drug in in-vitro assays and in-vivo studies.


4. Pore Forming Toxins

S. Pradeep advised by Prof. Sandhya Visweswariah and Prof. Ganapathy Ayyappa

Pore forming toxins (PFTs) belong to a class of bacterial exotoxins that disrupt the biological membrane barrier by formation of nanopores which cause cellular ion imbalance. These toxins are central to the virulence of pathogens that cause diseases such as cholera, anthrax and pneumonia. Unique to this class of proteins is their ability to exist in bi-stable states i.e. they are produced in soluble forms and upon exposure to membrane, undergo conformational changes, oligomerization and form a stable membrane integrated structure. We are trying to understand the mechanism of toxin assembly by a combination of biochemical techniques, single-molecule fluorescence, fluorescence correlation spectroscopy and molecular dynamics simulation. Towards this, we have identified novel means of regulation of toxin function which involve both binding mediated stabilization of conformation as well as allosteric regulation of channel function by distal protein segments.


5. How Does the Central Nervous System Control Movements?

Puneet Singh advised by Prof. Ashitava Ghosal and Prof. Aditya Murthy

A major question in motor control is how the Central Nervous System (CNS) generates muscle activity patterns to control movements. Predominant hypothesis in the literature is that CNS controls movements through muscle synergies. Muscle synergies represent coordinated activity patterns over groups of muscles. Complex movements can be constructed by a combination of muscle synergies. They simplify the problem of movement control by constraining the degrees of freedom. This research is aimed at understanding whether muscle synergies represent a functional architecture of neural networks or if it is just a simple correlation of muscle activities imposed by the task.


6. BioMicrofluidics : A tool to revisit cell engineering

Sharmistha Naskar advised by Prof. Bikramjit Basu and Prof. V. Kumaran

One of the central themes in cell and tissue engineering applications is to develop an understanding as how biophysical cues can influence cell functionality changes. The shear flow induced stress is regarded as one such biophysical cues to influence functionality changes in muscle precursor cells. The conventional two dimensional cell culture cannot be used to probe into the above aspect. While addressing such issue, the present work demonstrates the intriguing role played by the shear stress in inducing aligned growth and differentiation of myoblast cells to myotubes, after constraining the cells within a custom-made microfluidic device. In particular, the regulation of flow parameters within a narrow window was obtained in PMMA-based Lab-on-Chip (LOC) device, wherein the murine muscle cells (C2C12), chosen for its phenotypical differentiation stages, were cultured under graded shear conditions within a single chip. Quantitatively, the mRNA expression of myogenic biomarkers i.e myogenin, MyoD and neogenin exhibited a potent increase, compared to the culture under static condition. Also, the change in nuclear morphology with aligned pattern of the cells and specially the extent of myotube formation are modulated with shear stress and are in commensurate with gene expression changes. This emerging technique not only provides a close solution to mimic the in vivo conditions, but also opens up a newer approach to understand the specific role of biophysical cue on cell fate processes.


7. Dynamic Networks of Two-component Signaling (TCS) in Mycobacterium tuberculosis

Gaurav Sankhe advised by Dr. Narendra Dixit and Dr. Deepak K. Saini

Mycobacterium tuberculosis communicates with dynamic extra cellular environment primarily with help of battery of two- component signaling proteins. These two component protein relay the signal essentially through phosphorylation reaction which act as a switch tuning the kinetics of adaptive response. This signal to response scenario of the TCS can be best described through developing models of the network. The project aims at developing models and also to obtain parameters for model through experimentation. These models will elucidate features of disease like virulence, pathogenesis and drug resistance enabling to design more potent strategies of intervention.


8. Novel Polymer-Nanocomposites for Tissue Engineering

Queeny Dasgupta advised by Prof. Giridhar Madras and Dr. Kaushik Chatterjee

The primary aim of tissue engineering is to provide a suitable microenvironment to cells that promote their growth and proper function. Polymers are used most commonly as biomaterials. However, the scope of tissue engineering is limited by the constraints that commercially available polymers pose on it. The lack of tunability of properties to suit tissue specific requirements makes it necessary to synthesize novel, biodegradable and biocompatible polymers that can substitute or replace damaged tissue function. The aim of my work is to prepare resorbable, 3D scaffolds to serve as implants. It is very important to modulate the degradation properties of the polymer since it is absolutely necessary that the rate of tissue regeneration matches that of the polymer degradation. Also the mechanical properties of the scaffold material influence the growth, differentiation and overall proliferation of the cell. I am currently working on a family of polyesters and polyanhydrides, which can be used for tissue engineering and drug delivery respectively. The influence of variable stoichiometric ratios, slight changes in precursors and curing time on the mechanical and degradation properties will be studied in detail. Subsequently, the effect of functionalization on these polymers will also be investigated in order to improve cell response and function. The primary objective of this study is therefore, to match the properties of the native tissue in order to provide physical and chemical cues to direct appropriate cell growth and differentiation.


9. Shockwaves in Biology

Akshay Datey advised by Prof. G. Jagdeesh and Dr. Dipshikha Chakravortty

Shock waves are being extensively used for lithotripsy. Shock waves are also used as a treatment for angiogenesis and osteoporosis. Recently shock waves have been used to treat different medical conditions like intestinal anastomosis, to accelerate wound healing and articular cartilage defects .There are various reports on the other aspects of shock waves as well. Shock waves are also known to cause deleterious effects like severe nerve damage and brain injury. Shock wave treatment can also produce oxidative stress in the kidney and other organs leading to a diseased condition. There are several useful effects as well as ill effects of shock waves in the biological field yet the mechanism(s) of these effects are not clearly understood. Apart from the people treated with shockwaves, other groups of people constantly exposed to shockwaves are the soldiers on the war field. The effect of shock waves on infection is not well studied. We are interested to study the effects of shock waves on the human body with respect to various aspects, immune response being the main focus of our study. We are also developing shockwave driven devices to be used for procedures of transformation in prokaryotes, mammalian cells and plants.


10. Microfluidic Approach to Study Specific Neuronal Activity in C. elegans

Anjali Gupta advised by Dr. Manoj Varma and Dr. Varsha Singh

The nervous system of C. elegans consists of 302 neurons and responds to external cues exhibiting either avoidance/attraction response. C. elegans feeding on bacteria in its natural habitat has the sense to differentiate between friendly and pathogenic bacteria. We want to investigate the neuronal involvement of such an ability to differentiate between the two. In particular, we will be targeting sensory neurons to study their response against molecules from pathogens. A microfluidic approach is taken to study calcium transients in Amphid neurons using GCaMP sensor.


11. Feedback Control of Eye and Hand Movement

Varsha Vasudevan advised by Prof. Radhakant Padhi and Prof. Aditya Murthy

Successful motor control is crucial for life in determining action capabilities, regulating balance and stability. It is indeed a wonder that intricate movements are carried out by living beings so effortlessly. Different movements like eye and hand movements show stereotypical profiles like the bell shaped velocity profile. This suggests that the brain could be following a common principle/computation to carry out these movements. However, natural movements are variable in nature. This movement variability may arise from processes being corrupted by noise at various levels of motor act. A possible manner in which brain could be solving this problem caused by inherent delays and noise is by using a continuous feedback mechanism for updating of present state of the system along with a forward model that estimates these states initially. The objective of this research is to investigate the possible role of feedback mechanism in motor control by studying eye and hand movements and thus to gain insight into the control strategy involved in motor behaviour by using control theory principles widely used in engineering.


12. Biological applications of Vibrational Microspectroscopy

Taru Verma advised by Prof. Dipankar Nandi and Prof. S. Umapathy

Vibrational spectroscopy involves techniques like Infra-red spectroscopy and Raman spectroscopy. Both these methods probe the bond vibrations of the molecules present in the sample of interest and generate unique spectral profiles based on the chemical groups in the sample. Traditionally a chemist’s tool, vibrational spectroscopy has found several applications in the field of biology and medicine in the recent past. All biological samples are made up of different biomolecules like DNA, RNA, proteins, lipids and carbohydrates, which have unique structures and consequently yield unique spectra. Exploiting these advantages of vibrational spectroscopy, we aim to understand sepsis progression using these methods to identify biomarkers for early diagnosis of sepsis. Additionally, we are trying to differentiate closely related bacterial strains as well as understand the development of antibiotic resistance in bacteria.


13. Computational Models of Biological Vision

Aakash Agarwal advised by Dr. S P Arun and Prof. K.V.S. Hari

If we can make computers play chess, why can’t we make them see? The distorted letter tasks we see on websites (right) are proof that we cannot make computers solve even the simplest vision tasks. The brain is an engineering masterpiece. Decades of research in computer vision, pattern recognition and more recently machine learning has shown that using a pixel-based representation is insufficient to solve even simple vision tasks. For example, two noise images look similar to us despite differing in every pixel, whereas two faces look distinct despite being similar in pixels. So how does the brain accomplish vision?

My research will be focused on understanding the underlying neural mechanisms involved in biological vision. This research will be useful in improving the performance of existing computer vision algorithms. It will also help us understand complex disorders of vision in humans. I plan to use a variety of experimental techniques such as behavioral testing, brain imaging (fMRI), brain stimulation (TMS) in combination with computational modeling to further our understanding of biological vision.


14. Novel Nanomaterials Based Cancer Diagnostics, Imaging and Therapies

Zinia Mohanta advised by Dr. Chandan Srivastava and Dr. Hanudatta S. Atreya

This project involves development of graphene based nanomaterials for imaging/detection of cancer cells and targeted destruction of cancerous tumors. Magnetic resonance imaging (MRI) is the most popular tool for diagnosis in medical science today. Inspite of rendering an excellent imaging spatial resolution, MRI suffers from low sensitivity which often results in several diseases such as cancers remaining undetected in their early stages of development. This PhD project work addresses this particular issue with the objective of developing and investigating nanoparticle-graphene oxide/graphene/carbon nano-tube composites that will act not only as strong image contrast enhancing agents in MRI but also help in targeting the tumor cells. Targeting will be achieved by covalent coupling of the tri-peptide: RGD (known to bind to cancer cells) to graphene oxide along with a fluorescent probe (Au nanorods). Photothermal properties of gold nanorods will then be used to kill the cancer cells.


15. Interaction, Structure and Dynamics of Artificial Lipid Membranes

Ilanila I.P. advised by Prof. K. Ganapathy Ayappa and Prof. Jaydeep Kumar Basu

The cell membrane or the plasma membrane which is made up of a lipid bilayer, acts as a barrier protecting the cellular components from the external environment. The complexity of this lipid bilayer is not only due to differences in the chemistry and composition of phospholipids, sphingomyelins, and sterols, but also due to the existence of different protein molecules which crowd the membrane. Artificial membrane mimicking lipid bilayers of varying composition can be prepared either as free standing membranes or as supported membranes on a solid substrate. Our objective is to study the interaction, structure and dynamics of proteins and nanoparticles with artificial membranes using several imaging techniques with the focus on super resolution fluorescence imaging such as stimulated emission depletion spectroscopy (STED) which can break the optical resolution barrier. The differences in physicochemical properties and concentration of the interacting molecules (proteins or synthetic polymers) are expected to alter the membrane structure and lipid dynamics. An understanding of these interactions on supported membrane platforms with specific phospholipids, proteins and synthetic polymers, can potentially be used to develop sensitive and biocompatible sensors.


16. Understanding Cardiac Failure using Engineered Biomaterials

Aditi Jain advised by Dr. Kaushik Chatterjee and Dr. N. Ravi Sundaresan

Heart failure is the leading cause of deaths in the world. It is majorly attributed to the inability of the cardiomyocytes to proliferate sufficiently to restore the function of the damaged heart muscle. This compromises cardiac function and makes the heart more susceptible to heart failure. In the past, the strategies focused on cell transplantation advanced to clinical trials but failed to show any substantial improvements in cardiac function due to poor retention and survival of transplanted cells. Therefore, we are interested in creating 3D microenvironments using engineered biomaterials that simulates the in vivo condition and capacitates the cells to have functionality close to that of the native heart tissue. Culturing cells on these engineered biomaterials could provide better insights into the molecular mechanisms governing defects in cardiac regeneration and wound healing.


17. PVDF-based Membranes for Water Filtration

Paresh Kumar Samantaray advised by Dr. Suryasarathi Bose and Prof. Giridhar Madras

When PVDF membrane for filtration purpose are exposed to the feed stream, which often contain micro-organisms like bacteria. They tend to lose their effectiveness due to bacterial clogging the surface and formation of biofilms. It increases the resistance offered by the membrane and increases the overall energy required for separation . This leads to decrease in the membrane’s shelf life. Therefore, preventing this biofilm by suitable method is required. It is done by the use of functional groups that prevents the attachment or growth of microorganisms on the surface, while maintaining a good permeation rate of water. The aim of the work is to design cost effective PVDF based membranes with antimicrobial surface that can provide unimpeded permeation of water.


18. Speech signal-based monitoring and prediction of health parameters – towards a personalized medical device

Shivani Yadav advised by Dr. Prasanta Kumar Ghosh and Dr. Dipanjan Gope

Non-invasive processing of health signals is becoming increasingly popular and effective in diagnosis and monitoring of health parameters for medical applications. In our research we collects voice samples from subjects who are suffering from diseases such as Asthma, Parkinson, Amyotrophic lateral sclerosis and essential tremor and do analysis of voice to develop algorithms for monitoring health parameters of a patient. The project would involve discovering the cues from the voice signal that can reliably estimate the health parameters. It would also involve development of novel machine-learning algorithms, which can accurately predict the levels of attack of various diseases from the speech thereby helping in diagnosis and constant monitoring during treatment. Finally, the algorithms would be implemented in a platform that offers a personalized medical device for fast and accurate monitoring of an individual’s vital health parameters and data collected can be used for telemedicine.


20. Phagocytosis of non-degradable micro-particles by Macrophages: Effect on cell proliferation

Jayashree Raghavan advised by Dr. Siddharth Jhunjhunwala

Non-degradable micro-particles are of great interest in medical field for imaging, diagnosis, etc. Entry of such micro-particles into a living system may call for unfavorable responses despite the advantages offered. An often observed response is the phagocytosis of such micro-particles by macrophages. The cell fate of these macrophages after engulfing micro-particles is seldom studied. Hence our research interest lies in exploring the cell fate of phagocytized macrophages from a cell biology perspective. The hypothesis of the study is that one of the three states or a combination of them viz. apoptosis/necrosis, cell cycle arrest or senescence, the cells can enter into after micro-particle uptake. In order to study and ascertain this hypothesis, as a first step, the system has been optimized for phagocytosis of micro-particles by macrophages. The cell line used is RAW 264 and the micro-particles used are silica and polystyrene. Favorable results for particle uptake were observed with polystyrene micro-particles of diameter 3 µm. From literature and previous experiments, the rate of cell proliferation is observed to decline with increase in micro-particle concentration as well as uptake. To validate this observation, three obvious changes in cell cycle parameters are to be confirmed viz DNA content, total protein concentration and cell counts. Upon validation, the cell state is to be determined by performing standard assays. This insight could be exploited to tweak the cell state the macrophages choose to enter after particle uptake by engineering the micro-particle used in an application dependent manner.


21. Development of artificial neural networks for motor control

Saurabh Kothari advised by Prof. Ashitava Ghosal and Prof. Aditya Murthy

In what way the central nervous system directs a large number of muscles to produce complex motor behaviors is an open question. By using Artificial Neural Networks (ANNs) this project seeks to understand how representations in the central nervous system that encode decision signals from the sensory input are transformed into representations of movement plans that are eventually executed by the spinal cord and muscles in the periphery.


22. Quantifying Mechanobiology Of Fibroblasts To Cyclic Stretching & Tgf-β.

Aritra Chatterjee advised by Dr. Namrata Gundiah and Prof. Paturu Kondaiah

My current work involves investigating cell mechanobiology in context of tissue fibrosis using a growth and remodelling framework. I am using experimental methods to quantify changes in fibroblast mechanobiology in response to combined effects of cyclic stretching and a cytokine involved in fibrosis, TGF-β, using a custom designed dynamic cell stretching device. Preliminary results show dynamic changes in the stress fiber orientation due to stretch. I hope to integrate these responses within a theoretical and computational model to understand tissue fibrosis that are implicated in pathologies like aneurysms, cancers and wound healing. .


23. Protein decoration of Nano and Micro Carrier Surfaces for Improved Radio-Imaging and Drug Delivery Applications.

Preeti Sharma advised by Dr. Siddharth Jhunjhunwala

Micro and nanoparticles are widely used for various diagnostic and therapeutic purposes including radio-imaging and drug-delivery. However, when such systems encounter the biological fluid, the serum/blood proteins begin to interact with their surface. Proteins that are abundant in serum, such as albumin and fibrinogen adsorb onto the surface of these carriers, which eventually leads to unfolding of the proteins. This acts as a nucleation event, allowing less abundant proteins to adsorb non-specifically and unfold denature on the surface.The layer of proteins on the particle surface , consisting is referred to as “corona”. Unfolding of proteins might expose previously hidden domains which may potentially generate unwanted immune reactions including phagocytosis of the particles. Here, we propose a hypothesis that covalent linkage of specific proteins on material surface will result in altered serum protein adsorption, which has the potential to lead to reduced immune cell-particle surface interaction. Understanding protein-particle surface interaction is crucial to design improved drug delivery systems. For the initial studies, Polystyrene particles (P(S/2% DivenylBenzene/V-COOH) of 3µm diameter and albumin have been chosen as model particles and protein, respectively. Flow cytometry is being used to understand protein-protein interaction, protein adsorption and desorption phenomena on particle surface and in vitro-particle uptake by macrophages.


27. Electro-Thermo-Mechanical Phenotyping of Breast Cancer: From Onset Through Disease Progression.

Anil Vishnu G K advised by Dr. Hardik J Pandya and Dr. Annapoorni Rangarajan

The objective of my study is to design and develop a portable tool consisting of a disposable biochip for measuring and characterizing the electro thermo mechanical (ETM) properties of breast cancer tissues. The biochip will be integrated with microheaters, force sensors, electrical sensors, and temperature sensors and will be fabricated and packaged using microtechnology. A portable tool capable of holding the tissue and the packaged biochip will be designed. The studies will be carried out through various stages of cancer progression to understand how the ETM properties change.


28. Estimation of Neutrophils Half Life In Vivo using mouse models.

Alakesh advised by Dr. Siddharth Jhunjhunwala

Immune response against biomaterial implants is a major challenge for biomedical industry. Innate immune cells respond to the implant, by creating an inflammatory microenvironment, which eventually leads to implant fibrosis. Among the many different cells that take part in this immune action, recently neutrophils have been shown to play an important role in establishing the inflammation. However, many questions regarding the role of neutrophils remains unanswered. One of the primary questions is the length of neutrophil survival at implant sites. We are interested to know for how long neutrophils are recruited at the implant site. To answer this question, we plan to model neutrophil kinetics in tissues mathematically with the help of rate equations and validate the equation rate parameters with the help of experimental data. Our final objective is to uncover the kinetics and half life of neutrophils in tissues for better understanding of the crosstalk that happens at the immuno-synthetic interface.


29. Cellular Senescence in 3D Scaffolds.

PARUL YADAV advised by Dr. Kaushik Chatterjee and Dr. Deepak Saini

To study the biological mechanisms of ageing, it is essential to mimic the conditions in the human body, which is the 3D environment. The project involves fabrication of 3D biomaterial scaffolds to study the process of ageing and its related side effects such as inflammation. Thus, the fate of aged cells will be determined by assessing their behaviour in the 3D microenvironment. Further, the effect of age on various biomaterials will too be examined.


30. Dry powder micro- and nano- carriers for treatment of tuberculosis

Pallavi Raj Sharma advised by Dr. Rachit Agarwal

The project aims to devise an efficient drug delivery system for treatment of tuberculosis. Tuberculosis is a major health concern worldwide and the current treatment regimens have a few shortcomings, like long treatment duration and development of antibiotic-resistant strains. The project includes synthesis and optimization of nano- or micro-particles, encapsulation of antimicrobials and their deep lung delivery via the inhalation route. The potential antimicrobials include bacteriophages along with other antibiotic or non-antibiotic drugs which may contribute synergistically towards the bactericidal activity of the system. The drug-particle system will first be optimized in vitro and then tested in vivo to study its efficiency in a pre-clinical model. This proposed treatment method is more patient compliant and can potentially treat antibiotic-resistant tuberculosis lung infections.


31. Delivery of Resolvin carrier for treatment of Osteoarthritis

Ameya Dravid advised by Dr. Rachit Agarwal

Osteoarthritis is a joint disease that is characterized by joint pain, degradation and subsequent loss of function of cartilage. Mechanistically, cells present in (e.g. Chondro-cytes) and associated with the joint (e.g. Adipocytes of fat pad) sense cellular stress, and release "stress" signals (like cytokines). This attracts several immune cells to the joint and launches a state of chronic inflammation that is associated with pain and loss of cartilage function. Resolution of inflammation is an active process of clearing up inflammation and is mediated by molecules like Resolvins (Rv). Exogenous Rv delivered intra-articularly has shown to ameliorate inflammation, with no reported side-effects. However, Rv are small lipid mediators that are cleared rapidly from joint due to efficient lymphatic clearance of joints. We plan to study the effect of Resolvin D1 on joints of mice with surgically induced OA via sustained release from nano- and micro- carriers. The objective of this project is to analyze how sustained delivery of Resolvin D1 achieved via means of biomaterial based approaches prevents and/or ameliorate OA progression. .


1. Targeting intracellular Salmonella infection using lipid coated nanoparticles

Salmonella is the causative organism of gastroenteritis, typhoid and paratyphoid diseases and is responsible for 2 million fatalities throughout the world. Salmonella has developed ingenious methods to survive even in detrimental intracellular environment of macrophages. BSSE student Rajeev Mudakavi et al. have developed lipid coated mesoporous silica nanoparticle (L-MSN) which are able to target intracellular Salmonella and eliminate them from their host cell by delivering antibiotic directly into its intracellular niche. They also observed that a lower dose of antibiotic encapsulated into L-MSN was sufficient to clear the Salmonella infection from mice. More information of this can be obtained from the following publication.

Publication: R. Mudakavi, A. M. Raichur, and D. Chakravortty , "Lipid coated mesoporous silica nanoparticle as oral delivery system for targeting and treatment of intravacuolar Salmonella infection", RSC Adv., 2014, 4, 61160-61166. DOI: 10.1039/C4RA12973C



3. Pore-forming toxins on artificial membranes

Pore-forming toxins are produced by pathogenic bacteria and punch holes in cell membranes, thereby killing the cell. These toxins are proteins that are unique in their ability to bind to the cell membrane and form pores that are comprised of multiple stands of the same protein. Research has shown which region of the ClyA toxin is important for pore formation, and molecular dynamics simulations have revealed the molecular details that underlie pore formation and membrane interaction.

Publication: M. S. Vaidyanathan, P. Sathyanarayana, P. K. Maiti, S. S. Visweswariah and K. G. Ayappa, "Lysis dynamics and membrane oligomerization pathways for Cytolysin A (ClyA) pore-forming toxin", RSC Adv., 2014, 4, 4930-4942. DOI: 10.1039/C3RA45159C


4. Synthesis of biodegradable, crosslinked polyesters

This work encompasses the synthesis of an array of biodegradable, crosslinked polyesters with independently tailored degradation, mechanical, release and bioactive properties for biomedical applications. Diacids and polyols endogenous to the body are used as precursors to minimize immune responses to degradation products. The motivation of the present work is to develop a combinatorial strategy to synthesize a library of polymers where synthesis parameters can control various polymer properties independently. Furthermore, it is important to impart bioactivity to the polymers to reduce immunogenicity and susceptibility for microbial infection. Thus, the objective is to chemically incorporate salicylates (salicylic acid and p-aminosalicylic acid) and a benzofuran derivative (usnic acid) by esterification onto the polyester backbone to ensure pharmacological activity. Unlike conventional entrapment of drugs in delivery vehicles, this chemical incorporation allows higher loading, better processability and controlled release.

Publications:

  • Q. Dasgupta, G. Madras and K. Chatterjee, "Controlled Release Kinetics of p-Aminosalicylic Acid from Biodegradable Crosslinked Polyesters for Enhanced Anti-mycobacterial Activity", Acta Biomaterialia, 2016 DOI: 10.1016/j.actbio.2015.11.032
  • Q. Dasgupta, K. Chatterjee and G. Madras, "Physical insights into salicylic acid release from poly(anhydrides)", Phys. Chem. Chem. Phys., 2015 DOI: 10.1039/C5CP06858D
  • Q. Dasgupta, K. Chatterjee, and G. Madras, "Controlled Release of Salicylic Acid from Biodegradable Cross-linked Polyesters", Molecular Pharmaceutics, 2015, 12(9), 3479-3489. DOI: 10.1021/acs.molpharmaceut.5b00515




5. Shockwaves- Consequence and Complement at the battlefield!

In the era of emergence of sophisticated weapons and war techniques,bioweapons still pose as a great threat to the war field personnel. Bacterial pathogens like Mycobacterium tuberculosis, Bacillus anthrasis and Pseudomonas aeruginosa infect through the respiratory route. It commonly spreads in public by contaminated aerosols. Once inhaled, the pathogen lodges itself in the pulmonary system mainly in the lungs. In situ, it multiplies and forms a complex structure called as biofilm, which encompasses multiple systematically architectured bacterial micro-colonies held together by extra-polymeric substance(EPS) mainly curli, fimbriae proteins and cellulose. The presence of EPS makes the biofilm impermeable to antibiotics and thus resistant to the conventional mode of treatment. The antimicrobial dose required to treat biofilms is at least 1000 fold higher than the MIC. The formation and sustenance of biofilms lead to chronic and latent infections particularly in Mycobacterium tuberculosis. Shockwaves, on the other hand, are generated when any form of energy transformation occurs in a time scale of micro seconds. Shockwaves being the most efficient form of energy dissipation also make them extremely potent for various biomedical applications which include needle free injection system, accelerated wound healing, cancerous tissue disruption to name a few. Recently, we have successfully demonstrated the use of controlled shockwaves for disruption and effective clearance of Pseudomonas aeruginosa biofilm in the lungs as well as Salmonella Typhimurium and Staphylococcus aureus biofilm in the urinary catheters and topical skin wounds respectively in the murine model. The study also highlights the increase in the efficacy of conventional antibiotics when administered in combination with controlled shockwaves. Therefore, shockwaves can be proposed to be routinely used as a therapeutic and a prophylactic approach to treat biofilm related infections.

Publications:

  • A. Datey, A. Thaha, S. R. Patil, G. Jagadeesh and D. Chakravortty, "Enhancing the efficiency of desensitizing agents with shockwave treatment – A new paradigm in dentinal hypersensitivity management", RSC Advances, 2016 DOI: 10.1039/C6RA12342B
  • D.P. Gnanadhas, M. Elango, A. Datey, S. Janardhanraj, C.S. Srinandan, R.A. Strugnell, J. Gopalan, D. Chakravortty, "Successful treatment of biofilm infections using shockwaves combined with antibiotic therapy", Scientific Reports, 2015 DOI: 10.1038/srep17440



6. In vitro model for breast cancer metastasis

Breast cancer is the most common cause of cancer and its metastasis accounts for about 90% of mortality in women. Metastasis is a multi-step process wherein the cancer cells migrate from the primary tumor into another site and establish a tumor. Understanding the molecular mechanisms that orchestrate metastasis provide useful insights into developing therapeutics that prevent or treat metastasis. While conventional two-dimensional (2D) cell culture systems fail to mimic in vivo signaling due to the planar architecture and inappropriate substrate stiffness, animal based xenograft models are expensive and time consuming. To overcome these drawbacks, three dimensional (3D) scaffold based models have been developed which help recapitulate in vivo tissue micro-environment. In this study, for the first time, we have developed a comprehensive Polycaprolactone (PCL) based 3D model to study the events associated with breast cancer metastasis. Using tissue engineering techniques we fabricated 3D scaffolds of polycaprolactone (PCL), with porous geometry and of elastic modulus that was close to that of metastatic breast tumor. Cells cultured in scaffolds grew in multiple layers forming 3D contacts and networks in contrast to cells cultured on 2D dishes, which remained flat with spread morphology. The cells over prolonged culture gave rise to tumoroids. Gene expression and functional analyses showed that the scaffold-cultured cells presented greater EMT, invasive and stemness potential which are critical for manifold events of metastasis like initiation, progression and colonization. Also the scaffold-cultured cells had improved metastatic potential in vivo. Thus these results indicate that culturing breast cancer cells in tissue-like 3D matrices improves their metastatic potential and could hence serve as a better model for metastasis and cancer related inflammation.

Publication:
G.M. Balachander, S.A. Balaji, A. Rangarajan, K. Chatterjee “Enhanced metastatic potential in a 3D tissue scaffold toward a comprehensive in vitro model for breast cancer metastasis” , ACS Applied Materials and Interfaces, 2015 DOI: 10.1021/acsami.5b09064



7. The role of variability in motor learning

BSSE student Puneet Singh and team explore whether motor variability, often unwanted characteristic of motor performance has any significance in motor learning. They propose that motor variability has two components one caused by redundancy and other is random noise. In this work, Singh et al. quantify redundancy space and investigate its significance and effect on motor learning. They propose that a larger redundancy space leads to faster learning across subjects and the redundant component of motor variability is not noise. They also tested this hypothesis in neurologically diseased conditions to get a mechanistic understanding of how reward-based learning and error-based learning interact and how such learning is affected by redundancy space.

Publication:
P. Singh, S. Jana, A. Ghosal, A. Murthy "Exploration of joint redundancy but not task space variability facilitates supervised motor learning", PNAS, 2016 DOI: 10.1073/pnas.1613383113



8. The Curious Case of Dynein: Regulation by Myosin I

Molecular motors are tiny machines that convert the energy from the breakdown of ATP to mechanical work and hence bring about changes in the organisation of cellular components, including cargo, organelles and nuclear material. There are three major classes of motor proteins that have been identified: (i) myosins, which are associated with cellular polymers called filamentous actin, and (ii) kinesins and (iii) cytoplasmic dynein, which are associated with pipe-like structures called microtubules. Cell division requires the precise orchestration of a number of proteins in the cell, primarily the microtubules and molecular motors. Cytoplasmic dynein is especially important during cell division for ensuring that the nuclear material is organised properly. In this study, we have uncovered that the localisation and activity of dynein during meiotic cell division in fission yeast is dependent on myosin I. This discovery highlights one of the few examples of interplay between actin-based and microtubule-based motors inside the cell. Studies such as these are important for understanding division of not just healthy cells, but also in cases when the cell division machinery goes rogue, such as during cancer.

Publication:
J. M. Thankachan, S. S. Nuthalapati, N. A. Thirumala, V. Ananthanarayanan "Fission yeast myosin I facilitates PI(4,5)P2–mediated anchoring of cytoplasmic dynein to the cortex", PNAS, 2017 DOI: 10.1073/pnas.pnas.1615883114



9. Beating cardiomyocytes on keratin coated substrates

In the current study, our aim was to develop a facile, efficient and cost-effective protocol for culturing the neonatal cardiomyocytes using keratin derived from human hair, which in turn could be used for investigating cardiac hypertrophy in vitro. The nanoscale coating with keratin was characterized using SEM and AFM. Our optimized protocol for culturing of cardiomy- ocytes yielded atleast ~106 cells per heart. The characterization of cardiomyocytes with specific markers revealed that they can attach, grow and show spontaneous contraction on keratin-coated substrates similar to fibronectin-coated surfaces. Cardiomyocyte differentiation was assessed by immunofluorescence. Cardiac hypertrophy was induced using Phenylephrine (PE). Effect of PE-induced hypertrophy on signalling pathways was analysed using western blotting. Signaling proteins such as p-Akt, p-ERK, p-mTOR were up-regulated along with marked increase in protein synthesis on development of hypertrophy. Up-regulation of transcript levels of genes associated with hypertrophy was observed in qRT-PCR. The transient intracellular calcium spikes in beating cardiomyocytes were clearly detected using the fluorescent dye, Fluo-4. We conclude that cardiomyocytes grown on such keratin-coated surfaces are capable of developing the characteristic features of hypertrophy and hence serve as an inexpensive alternative and candidate model for understanding heart failure. Our culture protocol can be used interchangeably for both neonatal rats and mice to study cardiac hypertrophy in vitro. .

Publication:
A. Jain, V. Ravi, J. Muhamed, K. Chatterjee, N. R. Sundaresan, "A simplified protocol for culture of murine neonatal cardiomyocytes on nanoscale keratin coated surfaces", International Journal of Cardiology, 2017 DOI: 10.1016/j.ijcard.2017.01.036


India, despite the much talked about demographic dividend, has a large aging population. Elderly care is inadequate even in urban and affluent segments. Elderly care in the rural and poor segments is much worse. Assistive devices, proper diagnosis and treatment, and regular care of the elderly are all important problems that must be addressed right now before they become unmanageable. BSSC@IISc has begun look at the healthcare of the elderly by brainstorming with geriatricians in Bengaluru.

Three projects supported by the DST-TIDE (Technology for the Disabled and the Elderly by the Department of Science and Technology, Govt. of India) have been initiated.

If you are interested in joining hands with us, please contact us.

In the last few decades, the Indian healthcare system has been gradually embracing the western procedures and protocols for diagnostics, treatment, and patient-care. As a result, the quality and efficacy of healthcare have significantly improved, but the cost of quality healthcare has escalated beyond the reach of low-income population and has become a burden for most segments of the population. The principal reason for this can be attributed to the high cost of imported biomedical instruments. Some reports say that as much as 80% of the biomedical equipment is imported in India today. The prohibitively high cost of purchase and equally high cost of maintenance of the imported equipment are transferred from hospitals to patients.

Biosensors and other diagnostic devices, including medical imaging, are used extensively in hospitals today but indigenous efforts that see the light of the day in the market are heavily lagging. The situation is not different with therapeutic devices. There is also a dearth of good indigenous assistive devices for the disabled and the elderly population. Hence, it is imperative that the development of competent and competitive indigenous biomedical devices is a need of the hour for the Indian society. A silver lining in this dark cloud is the emerging pockets of innovation in academic institutions and industry. What is lacking is the supporting infrastructure that fosters the development of biomedical devices. Bridging this gap within IISc and then extending help to researchers, inventors, and industry in the nation is a goal that BSSE has set for itself.

We have begun to work towards this. If anyone is interested in joining hands with us, please contact us.

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