Recapitulating cranial osteogenesis with neural crest cells in 3-D microenvironments

The experimental systems that recapitulate the complexity of native tissues and enable precise control over the microenvironment are becoming essential for the pre-clinical tests of therapeutics and tissue engineering. Here, we described a strategy to develop an in vitro platform to study the developmental biology of craniofacial osteogenesis. In this study, we directly osteo-differentiated cranial neural crest cells (CNCCs) in a 3-D in vitro bioengineered microenvironment. Cells were encapsulated in the gelatin-based photo-crosslinkable hydrogel and cultured up to three weeks. We demonstrated that this platform allows efficient differentiation of p75 positive CNCCs to cells expressing osteogenic markers corresponding to the sequential developmental phases of intramembranous ossification. During the course of culture, we observed a decrease in the expression of early osteogenic marker Runx2, while the other mature osteoblast and osteocyte markers such as Osterix, Osteocalcin, Osteopontin and Bone sialoprotein increased. We analyzed the ossification of the secreted matrix with alkaline phosphatase and quantified the newly secreted hydroxyapatite. The Field Emission Scanning Electron Microscope (FESEM) images of the bioengineered hydrogel constructs revealed the native-like osteocytes, mature osteoblasts, and cranial bone tissue morphologies with canaliculus-like intercellular connections. This platform provides a broadly applicable model system to potentially study diseases involving primarily embryonic craniofacial bone disorders, where direct diagnosis and adequate animal disease models are limited

Fish traps in inland waters of north kerala

Result of the study on traditional traps in the inland waters of three northern districts viz, Kasargod, Kannur and Kozhikode in Kerala state during 2003-2004 is presented. Mainly six types of traps are found in operation. Chempally koode is a rectangular bamboo trap with" D" shape in cross section operated without bait in some rivers of Kannur and Kasargod. Bamboo screen barriers are almost completely replaced with durable HDPE net screen to make handling easy. Thottil vala is a unique aerial trap operated from the dam in Pazhassi reservoir during monsoon to catch big fishes jumping against flowing water.cochin university of science and technologyfishery technology,2008 vol 45(2)pp 137-14

Plug-and-actuate on demand: Multimodal individual addressability of microarray plates using modular hybrid acoustic wave technology

The microarray titre plate remains a fundamental workhorse in genomic, proteomic and cellomic analyses that underpin the drug discovery process. Nevertheless, liquid handling technologies for sample dispensing, processing and transfer have not progressed significantly beyond conventional robotic micropipetting techniques, which are not only at their fundamental sample size limit, but are also prone to mechanical failure and contamination. This is because alternative technologies to date suffer from a number of constraints, mainly their limitation to carry out only a single liquid operation such as dispensing or mixing at a given time, and their inability to address individual wells, particularly at high throughput. Here, we demonstrate the possibility for true sequential or simultaneous single- and multi-well addressability in a 96-well plate using a reconfigurable modular platform from which MHz-order hybrid surface and bulk acoustic waves can be coupled to drive a variety of microfluidic modes including mixing, sample preconcentration and droplet jetting/ejection in individual or multiple wells on demand, thus constituting a highly versatile yet simple setup capable of improving the functionality of existing laboratory protocols and processes

High frequency acoustic permeabilisation of drugs through tissue for localised mucosal delivery

The majority of infectious diseases enter the body through mucosal membranes that line the ocular, nasal, oral, vaginal and rectal surfaces. As infections can be effectively prevented by instigating a local immune response in the immunocyte-rich regions of the mucosa, an efficacious route of vaccine administration is to directly target their delivery to these surfaces. It is nevertheless challenging to provide sufficient driving force to penetrate both the mucus lining as well as the epithelial barrier of the mucosal surfaces, which are designed to effectively keep foreign entities out, but not excessively such that the therapeutic agent penetrates deeper into the vascularised submucosal regions where they are mostly taken up by the systemic circulation, thus resulting in a far weaker immune response. In this work, we demonstrate the possibility of controllably localising and hence maximising the delivery of both small and large molecule model therapeutic agents in the mucosa of a porcine buccal model using high frequency acoustics. Unlike their low (kHz order) frequency bulk ultrasonic counterpart, these high frequency (>10 MHz) surface waves do not generate cavitation, which leads to large molecular penetration depths beyond the 100 μm order thick mucosal layer, and which has been known to cause considerable cellular/tissue damage and hence scarring. Through system parameters such as the acoustic irradiation frequency, power and exposure duration, we show that it is possible to tune the penetration depth such that over 95% of the delivered drug are localised within the mucosal layer, whilst preserving their structural integrity

Not Available

Not AvailableWomen form an integral part of the workforce in fisheries sector of Gujarat, India. A brief description of the post-harvest activities carried out by fisherwomen in Gujarat is given in the paper.Not Availabl

Tweaking of Peripheral Moieties in Catalytic Amyloid for Modulating Hydrogel Strength and Hydrolase Activity

The de novo design and synthesis of peptide-based biocatalysts that can mimic the activity of natural enzymes is an exciting field with unique opportunities and challenges. In a natural enzyme, the active site is composed of an assembly of different amino acid residues, often coordinated with a metal ion. A metalloenzyme’s catalytic activity results from the dynamic and concerted interplay of various interactions among the residues and metal ions. Aiming to mimic such enzymes, simple peptide fragments, drawing structural inspiration from natural enzymes, can be utilized as a model. In our effort to mimic a metal-containing hydrolase, we designed peptide amphiphiles (PA) 1 and 2 with a terminal histidine having amide and acid functionalities, respectively, at its C-terminal, imparting differential ability to coordinate with Zn and Cu ions. The PAs demonstrate remarkable self-assembly behavior forming excellent nanofibers. Upon coordination with metal ions, depending on the coordination site the nanofibers become rigidified or weakened. Rheological studies revealed excellent mechanical properties of the hydrogels formed by the PAs and the PA–metal co-assemblies. Using such co-assemblies, we mimic hydrolase activity against a p-nitrophenyl acetate (p-NPA) substrate. Michaelis–Menten’s enzyme kinetic parameters indicated superior catalytic activity of 2 with Zn amongst all the assemblies

Acoustically-mediated intracellular delivery

Recent breakthroughs in gene editing have necessitated practical ex vivo methods to rapidly and efficiently re-engineer patient-harvested cells. Many physical membrane-disruption or pore-forming techniques for intracellular delivery, however, result in poor cell viability, while most carrier-mediated techniques suffer from suboptimal endosomal escape and hence cytoplasmic or nuclear targeting. In this work, we show that short exposure of cells to high frequency (>10 MHz) acoustic excitation facilitates temporal reorganisation of the lipid structure in the cell membrane that enhances translocation of gold nano- particles and therapeutic molecules into the cell within just ten minutes. Due to its transient nature, rapid cell self-healing is observed, leading to high cellular viabilities (>97%). Moreover, the internalised cargo appears to be uniformly distributed throughout the cytosol, circumventing the need for strategies to facilitate endosomal escape. In the case of siRNA delivery, the method is seen to enhance gene silencing by over twofold, demonstrating its potential for enhancing therapeutic delivery into cells

On‐Chip Generation of Vortical Flows for Microfluidic Centrifugation

Microcentrifugation constitutes an important part of the microfluidic toolkit in a similar way that centrifugation is crucial to many macroscopic procedures, given that micromixing, sample preconcentration, particle separation, component fractionation, and cell agglomeration are essential operations in small scale processes. Yet, the dominance of capillary and viscous effects, which typically tend to retard flow, over inertial and gravitational forces, which are often useful for actuating flows and hence centrifugation, at microscopic scales makes it difficult to generate rotational flows at these dimensions, let alone with sufficient vorticity to support efficient mixing, separation, concentration, or aggregation. Herein, the various technologiesboth passive and activethat have been developed to date for vortex generation in microfluidic devices are reviewed. Various advantages or limitations associated with each are outlined, in addition to highlighting the challenges that need to be overcome for their incorporation into integrated microfluidic devices

Continuous tuneable droplet ejection via pulsed surface acoustic wave jetting

We report a miniaturised platform for continuous production of single or multiple liquid droplets with diameters between 60 and 500 mm by interfacing a capillary-driven self-replenishing liquid feed with pulsed excitation of focussed surface acoustic waves (SAWs). The orifice-free operation circumvents the disadvantages of conventional jetting systems, which are often prone to clogging that eventuates in rapid degradation of the operational performance. Additionally, we show the possibility for flexibly tuning the ejected droplet size through the pulse width duration, thus avoiding the need for a separate device for every different droplet size required, as is the case for systems in which the droplet size is set by nozzles and orifices, as well as preceding ultrasonic jetting platforms where the droplet size is controlled by the operating frequency. Further, we demonstrate that cells can be jetted and hence printed onto substrates with control over the cell density within the droplets down to single cells. Given that the jetting does not lead to significant loss to the cell's viability or ability to proliferate, we envisage that this versatile jetting method can potentially be exploited with further development for cell encapsulation, dispensing and 3D bioprinting applications