De novo construction of T cell compartment in humanized mice engrafted with iPSC-derived thymus organoids.

Hematopoietic humanized (hu) mice are powerful tools for modeling the action of human immune system and are widely used for preclinical studies and drug discovery. However, generating a functional human T cell compartment in hu mice remains challenging, primarily due to the species-related differences between human and mouse thymus. While engrafting human fetal thymic tissues can support robust T cell development in hu mice, tissue scarcity and ethical concerns limit their wide use. Here, we describe the tissue engineering of human thymus organoids from inducible pluripotent stem cells (iPSC-thymus) that can support the de novo generation of a diverse population of functional human T cells. T cells of iPSC-thymus-engrafted hu mice could mediate both cellular and humoral immune responses, including mounting robust proinflammatory responses on T cell receptor engagement, inhibiting allogeneic tumor graft growth and facilitating efficient Ig class switching. Our findings indicate that hu mice engrafted with iPSC-thymus can serve as a new animal model to study human T cell-mediated immunity and accelerate the translation of findings from animal studies into the clinic.

On the origin of the biological effects of time varying magnetic fields: quantitative insights.

In a number of recently published experimental studies from our research group, the positive impact of magnetic stimuli (static/pulsed) on cell functionality modulation or bactericidal effects, in vitro, has been established. In order to develop a theoretical understanding of such magnetobiological effects, the present study aimed to present two quantitative models to determine magnetic Maxwell stresses as well as pressure acting on the cell membrane, under the influence of a time varying magnetic field. The model predicts that magnetic field-induced stress on the cell/bacteria is dependent on the conductivity properties of the extracellular region, which is determined to be too low to cause any significant effect. However, the force on the cell/bacteria due to the induced electric field is more influential than that of the magnetic field, which has been used to determine the membrane tension that can cause membrane poration. With a known critical membrane tension for cells, the field parameters necessary to cause membrane rupture have been estimated. Based on the experimental results and theoretically predicted values, the field parameters can be classified into three regimes, wherein the magnetic fields cause no effect or result in biophysical stimulation or induce cell death due to membrane damage. Taken together, this work provides some quantitative insights into the impact of magnetic fields on biological systems.

Evaluation of Outbreak Preparedness to Prevent the Reintroduction of Malaria in Ramanagara District: A Descriptive Epidemiological Study.

Background Malaria is a vector-borne disease transmitted by female anopheline mosquitoes. It is also a multidimensional disease influenced by social factors such as poor environmental conditions and awareness gaps. India has witnessed a substantial reduction in malaria cases and has declared three regions as malaria-free, with Karnataka being one of the states. However, Karnataka witnesses significant population movement and migration, which influences the spread of malaria. Ramanagara, a district in Karnataka, reported zero indigenous cases over the past three years. Hence, we selected this district to evaluate outbreak preparedness to prevent the reintroduction of malaria. This choice underscores the district's significance as a valuable model for preventing the reintroduction of malaria. Methodology Baseline survey data on malaria cases and vector survey data were evaluated for the period spanning 2018 to 2022. The data were gathered from both the regional office and the Ramanagara district health office. In addition to the documenting system, because there was no regular submission of Form-P and Form-L of the Integrated Disease Surveillance Project and Integrated Health Information Platform from the private sector, to complete the missing data, a cross-sectional study was conducted among private sector practitioners and pharmacies in Ramanagara from April 2023 to June 2023. Data was collected via interviews using a malaria surveillance assessment toolkit sourced from the National Centre for Vector Borne Diseases Control and World Health Organization protocols to assess six core areas of malaria elimination. Data collected via interviews were compiled in MS Excel and analyzed using SPSS Statistics for Windows, version 26.0 (IBM Corp., Armonk, NY, USA). Results Malaria control measures in the selected district achieved >80% coverage with notable improvements in the National Health Mission-National Vector Borne Disease Control Programme fund utilization, logistics availability, and physical performance over the past five years. The Annual Parasite Index was <1 at 0.0061 in 2018 and 0.0017 in 2022. The annual blood examination rate was consistently >10 from 22.05 in 2018 to 24.36 in 2022. The primary vectors identified were Anopheles culicifacies and Anopheles stephensi. In 2018, there were six cases of Plasmodium vivax and one case of Plasmodium falciparum reported as imported cases. In 2021 and 2022, two cases of P. vivax were reported. Notably, there were no instances of mixed infections or indigenous cases documented from 2018 to 2022 Conclusions Although the level of outbreak preparedness in the region is satisfactory, the effectiveness of vector control measures appears to be lacking. Increased government funding is needed along with comprehensive training and workshops for healthcare workers. Adequate financial resources and enhanced skills among healthcare workers are crucial to reinforce the existing efforts to control vectors and prevent potential outbreaks effectively.

Tunable Substrate Functionalities Direct Stem Cell Fate toward Electrophysiologically Distinguishable Neuron-like and Glial-like Cells.

Engineering cellular microenvironment on a functional platform using various biophysical cues to modulate stem cell fate has been the central theme in regenerative engineering. Among the various biophysical cues to direct stem cell differentiation, the critical role of physiologically relevant electric field (EF) stimulation was established in the recent past. The present study is the first to report the strategy to switch EF-mediated differentiation of human mesenchymal stem cells (hMSCs) between neuronal and glial pathways, using tailored functional properties of the biomaterial substrate. We have examined the combinatorial effect of substrate functionalities (conductivity, electroactivity, and topography) on the EF-mediated stem cell differentiation on polyvinylidene-difluoride (PVDF) nanocomposites in vitro, without any biochemical inducers. The functionalities of PVDF have been tailored using conducting nanofiller (multiwall-carbon nanotube, MWNT) and piezoceramic (BaTiO3, BT) by an optimized processing approach (melt mixing-compression molding-rolling). The DC conductivity of PVDF nanocomposites was tuned from ∼10-11 to ∼10-4 S/cm and the dielectric constant from ∼10 to ∼300. The phenotypical changes and genotypical expression of hMSCs revealed the signatures of early differentiation toward neuronal pathway on rolled-PVDF/MWNT and late differentiation toward glial lineage on rolled-PVDF/BT/MWNT. Moreover, we were able to distinguish the physiological properties of differentiated neuron-like and glial-like cells using membrane depolarization and mechanical stimulation. The excitability of the EF-stimulated hMSCs was also determined using whole-cell patch-clamp recordings. Mechanistically, the roles of intracellular reactive oxygen species (ROS), Ca2+ oscillations, and synaptic and gap junction proteins in directing the cellular fate have been established. Therefore, the present work critically unveils complex yet synergistic interaction of substrate functional properties to direct EF-mediated differentiation toward neuron-like and glial-like cells, with distinguishable electrophysiological responses.

Zirconia toughened mica glass ceramics for dental restorations: Wear, thermal, optical and cytocompatibility properties.

BACKGROUND: In an effort to design novel zirconia reinforced mica glass ceramics for dental restorations, clinically relevant properties such as wear, coefficient of thermal expansion, optical transmittance, and cytocompatibility with human gingival fibroblast cell lines were investigated in the present study. MATERIALS & METHODS: Microstructure analysis of two body wear of heat treated mica glass ceramic ceramics (47.2 SiO2-16.7 Al2O3-9.5 K2O-14.5 MgO-8.5 B2O3-6.3F wt.%) reinforced with 20wt.% YSZ, were evaluated against a steatite antagonist in a chewing simulator following Willytec Munich method. In addition, Coefficient of thermal expansion (CTE), total transmittance, scattering coefficient and cytocompatibility on human gingival fibroblast cell lines were performed and compared to the commercially available dental ceramic systems. RESULTS: The experimental mica glass ceramic demonstrate micro-ploughing, pull out and debris formation along the cutting surface, indicating abrasive wear mechanism. Thermal expansion of mica glass ceramic composite was recorded as 5×10-6/°C, which is lower than the thermal expansion of commercially available core and veneering ceramics. Further, significant differences of transmittance and scattering coefficient of mica glass ceramics with 20wt.% YSZ with commercial dental ceramics was found and extensive fibroblast cell spreading with filopodial extension, cell-to-cell bridges and proliferation with human gingival fibroblast cell lines. CONCLUSION: With acceptable cytocompatibility with human gingival fibroblast cells and better wear properties with respect to commercial IPS emax Press, the mica glass ceramic composites (47.2 SiO2-16.7Al2O3-9.5 K2O-14.5 MgO-8.5 B2O3-6.3F wt.%) with 20wt.% YSZ have the potential for dental restorative applications as machinable veneering ceramics.

The Foreign Body Response Demystified.

The human body is endowed with an uncanny ability to distinguish self from foreign. The implantation of a foreign object inside a mammalian host activates complex signaling cascades, which lead to biological encapsulation of the implant. This reaction by the host system to a foreign object is known as foreign body response (FBR). Over the last few decades, it has been increasingly important to have a deeper insight into the mechanisms of FBR is needed to develop biomaterials for better integration with living systems. In the light of recent advances in tissue engineering and regenerative medicine, particularly in the field of biosensors and biodegradable tissue engineering scaffolds, the classical concepts related to the FBR have acquired new dimensions. The aim of this review is to provide a holistic view of the FBR, while critically analyzing the challenges, which need to be addressed in the future to overcome this innate response. In particular, this review discusses the relevant experimental methodology to assess the host response. The role of erosion and degradation behavior on FBR with biodegradable polymers is largely explored. Apart from the discussion on temporal progression of FBR, an emphasis has been given to the design of next-generation biomaterials with favorable host response.

Zirconia toughened mica glass ceramics for dental restorations.

OBJECTIVE: The objective of the present study is to understand the role of yttria stabilized zirconia (YSZ) in achieving the desired spectrum of clinically relevant mechanical properties (hardness, elastic modulus, fracture toughness and brittleness index) and chemical solubility of mica glass ceramics. METHODS: The glass-zirconia mixtures with varying amounts of YSZ (0, 5, 10, 15 and 20wt.%) were ball milled, compacted and sintered to obtain pellets of glass ceramic-YSZ composites. Phase analysis was carried out using X-ray diffraction and microstructural characterization with SEM revealed the crystal morphology of the composites. Mechanical properties such as Vickers hardness, elastic modulus, indentation fracture toughness and chemical solubility were assessed. RESULTS: Phase analysis of sintered pellets of glass ceramic-YSZ composites revealed the characteristic peaks of fluorophlogopite (FPP) and tetragonal zirconia. Microstructural investigation showed plate and lath-like interlocking mica crystals with embedded zirconia. Vickers hardness of 9.2GPa, elastic modulus of 125GPa, indentation toughness of 3.6MPa·m1/2, and chemical solubility of 30µg/cm2 (well below the permissible limit) were recorded with mica glass ceramics containing 20wt.% YSZ. SIGNIFICANCE: An increase in hardness and toughness of the glass ceramic-YSZ composites with no compromise on their brittleness index and chemical solubility has been observed. Such spectrum of properties can be utilised for developing a machinable ceramic for low stress bearing inlays, onlays and veneers.