Comprehensive Ocular and Systemic Safety Evaluation of Polysialic Acid-Decorated Immune Modulating Therapeutic Nanoparticles (PolySia-NPs) to Support Entry into First-in-Human Clinical Trials.

An inflammation-resolving polysialic acid-decorated PLGA nanoparticle (PolySia-NP) has been developed to treat geographic atrophy/age-related macular degeneration and other conditions caused by macrophage and complement over-activation. While PolySia-NPs have demonstrated pre-clinical efficacy, this study evaluated its systemic and intraocular safety. PolySia-NPs were evaluated in vitro for mutagenic activity using Salmonella strains and E. coli, with and without metabolic activation; cytotoxicity was evaluated based on its interference with normal mitosis. PolySia-NPs were administered intravenously in CD-1 mice and Sprague Dawley rats and assessed for survival and toxicity. Intravitreal (IVT) administration in Dutch Belted rabbits and non-human primates was assessed for ocular or systemic toxicity. In vitro results indicate that PolySia-NPs did not induce mutagenicity or cytotoxicity. Intravenous administration did not show clastogenic activity, effects on survival, or toxicity. A single intravitreal (IVT) injection and two elevated repeat IVT doses of PolySia-NPs separated by 7 days in rabbits showed no signs of systemic or ocular toxicity. A single IVT inoculation of PolySia-NPs in non-human primates demonstrated no adverse clinical or ophthalmological effects. The demonstration of systemic and ocular safety of PolySia-NPs supports its advancement into human clinical trials as a promising therapeutic approach for systemic and retinal degenerative diseases caused by chronic immune activation.

PolySialic Acid Nanoparticles Actuate Complement-Factor-H-Mediated Inhibition of the Alternative Complement Pathway: A Safer Potential Therapy for Age-Related Macular Degeneration.

The alternative pathway of the complement system is implicated in the etiology of age-related macular degeneration (AMD). Complement depletion with pegcetacoplan and avacincaptad pegol are FDA-approved treatments for geographic atrophy in AMD that, while effective, have clinically observed risks of choroidal neovascular (CNV) conversion, optic neuritis, and retinal vasculitis, leaving room for other equally efficacious but safer therapeutics, including Poly Sialic acid (PSA) nanoparticle (PolySia-NP)-actuated complement factor H (CFH) alternative pathway inhibition. Our previous paper demonstrated that PolySia-NP inhibits pro-inflammatory polarization and cytokine release. Here, we extend these findings by investigating the therapeutic potential of PolySia-NP to attenuate the alternative complement pathway. First, we show that PolySia-NP binds CFH and enhances affinity to C3b. Next, we demonstrate that PolySia-NP treatment of human serum suppresses alternative pathway hemolytic activity and C3b deposition. Further, we show that treating human macrophages with PolySia-NP is non-toxic and reduces markers of complement activity. Finally, we describe PolySia-NP-treatment-induced decreases in neovascularization and inflammatory response in a laser-induced CNV mouse model of neovascular AMD. In conclusion, PolySia-NP suppresses alternative pathway complement activity in human serum, human macrophage, and mouse CNV without increasing neovascularization.

PolySialic acid-nanoparticles inhibit macrophage mediated inflammation through Siglec agonism: a potential treatment for age related macular degeneration.

Age-related macular degeneration (AMD) is a chronic, progressive retinal disease characterized by an inflammatory response mediated by activated macrophages and microglia infiltrating the inner layer of the retina. In this study, we demonstrate that inhibition of macrophages through Siglec binding in the AMD eye can generate therapeutically useful effects. We show that Siglecs-7, -9 and -11 are upregulated in AMD associated M0 and M1 macrophages, and that these can be selectively targeted using polysialic acid (PolySia)-nanoparticles (NPs) to control dampen AMD-associated inflammation. In vitro studies showed that PolySia-NPs bind to macrophages through human Siglecs-7, -9, -11 as well as murine ortholog Siglec-E. Following treatment with PolySia-NPs, we observed that the PolySia-NPs bound and agonized the macrophage Siglecs resulting in a significant decrease in the secretion of IL-6, IL-1ß, TNF-α and VEGF, and an increased secretion of IL-10. In vivo intravitreal (IVT) injection of PolySia-NPs was found to be well-tolerated and safe making it effective in preventing thinning of the retinal outer nuclear layer (ONL), inhibiting macrophage infiltration, and restoring electrophysiological retinal function in a model of bright light-induced retinal degeneration. In a clinically validated, laser-induced choroidal neovascularization (CNV) model of exudative AMD, PolySia-NPs reduced the size of neovascular lesions with associated reduction in macrophages. The PolySia-NPs described herein are therefore a promising therapeutic strategy for repolarizing pro-inflammatory macrophages to a more anti-inflammatory, non-angiogenic phenotype, which play a key role in the pathophysiology of non-exudative AMD.

Luminous, relativistic, directional electron bunches from an intense laser driven grating plasma.

Bright, energetic, and directional electron bunches are generated through efficient energy transfer of relativistic intense (~ 1019 W/cm2), 30 femtosecond, 800 nm high contrast laser pulses to grating targets (500 lines/mm and 1000 lines/mm), under surface plasmon resonance (SPR) conditions. Bi-directional relativistic electron bunches (at 40° and 150°) are observed exiting from the 500 lines/mm grating target at the SPR conditions. The surface plasmon excited grating target enhances the electron flux and temperature by factor of 6.0 and 3.6, respectively, compared to that of the plane substrate. Particle-in-Cell simulations indicate that fast electrons are emitted in different directions at different stages of the laser interaction, which are related to the resultant surface magnetic field evolution. This study suggests that the SPR mechanism can be used to generate multiple, bright, ultrafast relativistic electron bunches for a variety of applications.

Subpicosecond pre-plasma dynamics of a high contrast, ultraintense laser-solid target interaction.

Using the spectral interferometry technique, we measured subpicosecond time-resolved pre-plasma scale lengths and early expansion (<12 ps) of the plasma produced by a high intensity (6 × 1018 W/cm2) pulse with high contrast (109). We measured pre-plasma scale lengths in the range of 3-20 nm, before the arrival of the peak of the femtosecond pulse. This measurement plays a crucial role in understanding the mechanism of laser coupling its energy to hot electrons and is hence important for laser-driven ion acceleration and the fast ignition approach to fusion.

Efficient second-harmonic generation of a high-energy, femtosecond laser pulse in a lithium triborate crystal.

We demonstrate the highest efficiency (∼80%) second harmonic generation of joule level, 27 fs, high-contrast pulses in a type-I lithium triborate (LBO) crystal. In comparison, potassium dihydrogen phosphate gives a maximum efficiency of 26%. LBO thus offers high-intensity (>1018-19W/cm2), ultra-high contrast femtosecond pulses, which have great potential for high energy density science and applications, particularly with nanostructured targets.

Measuring the lifetime of intense-laser generated relativistic electrons in solids via gating their Cherenkov emission.

Optical Kerr gating technique has been employed to investigate the life history of relativistic electrons in solids by temporally gating their Cherenkov emission. Mega-ampere currents of relativistic electrons are created during ultra-intense (2 × 1019 W/cm2) laser-solid interactions. In order to measure the lifetime of these relativistic electrons in solids, we temporally gate their Cherenkov emission using an optical Kerr gate (OKG). The OKG is induced in a nonlinear medium, namely, carbon-di-sulphide (CS2), with a measured gate-width (FWHM) of 2 ps. The gate femtosecond laser pulse is synchronized with the intense interaction pulse generating relativistic electrons. The arrival time of the gate laser pulse on the CS2 cell is varied with the help of a delay stage. We find that Cherenkov emission from relativistic electrons created with a ultra-short laser pulse (25 fs) lives as long as 120 ps, a few thousand times that of the incident light pulse.

Towards Remote Lightning Manipulation by Meters-long Plasma Channels Generated by Ultra-Short-Pulse High-Intensity Lasers.

Remote manipulation (triggering and guiding) of lightning in atmospheric conditions of thunderstorms has been the subject of intense scientific research for decades. High power, ultrashort-pulse lasers are considered attractive in generating plasma channels in air that could serve as conductors/diverters for lightning. However, two fundamental obstacles, namely the limited length and lifetime of such plasma channels prevented their realization to this date. In this paper, we report decisive experimental results of our multi-element broken wire concept that extends the generated plasma channels to the required tens of meters range. We obtain 13-meter-long plasma wire, limited only by our current experimental setup, with plasma conditions that could be sufficient for the leader initiation. This advance, coupled with our demonstrated method of laser heating for long time sustenance of the plasma channel, is a major, significant step towards controlling lightning.

Recombination of Protons Accelerated by a High Intensity High Contrast Laser.

Short pulse, high contrast, intense laser pulses incident onto a solid target are not known to generate fast neutral atoms. Experiments carried out to study the recombination of accelerated protons show a 200 times higher neutralization than expected. Fast neutral atoms can contribute to 80% of the fast particles at 10 keV, falling rapidly for higher energy. Conventional charge transfer and electron-ion recombination in a high density plasma plume near the target is unable to explain the neutralization. We present a model based on the copropagation of electrons and ions wherein recombination far away from the target surface accounts for the experimental measurements. A novel experimental verification of the model is also presented. This study provides insights into the closely linked dynamics of ions and electrons by which neutral atom formation is enhanced.

Mapping the Damping Dynamics of Mega-Ampere Electron Pulses Inside a Solid.

We report the lifetime of intense-laser (2×10^{19} W/cm^{2}) generated relativistic electron pulses in solids by measuring the time evolution of their Cherenkov emission. Using a picosecond resolution optical Kerr gating technique, we demonstrate that the electrons remain relativistic as long as 50 picoseconds-more than 1000 times longer than the incident light pulse. Numerical simulations of the propagation of relativistic electrons and the emitted Cherenkov radiation with Monte Carlo geant4 package reproduce the striking experimental findings.

Highly efficient broadband terahertz generation from ultrashort laser filamentation in liquids.

Generation and application of energetic, broadband terahertz pulses (bandwidth ~0.1-50 THz) is an active and contemporary area of research. The main thrust is toward the development of efficient sources with minimum complexities-a true table-top setup. In this work, we demonstrate the generation of terahertz radiation via ultrashort pulse induced filamentation in liquids-a counterintuitive observation due to their large absorption coefficient in the terahertz regime. The generated terahertz energy is more than an order of magnitude higher than that obtained from the two-color filamentation of air (the most standard table-top technique). Such high terahertz energies would generate electric fields of the order of MV cm-1, which opens the doors for various nonlinear terahertz spectroscopic applications. The counterintuitive phenomenon has been explained via the solution of nonlinear pulse propagation equation in the liquid medium.

A gated Thomson parabola spectrometer for improved ion and neutral atom measurements in intense laser produced plasmas.

Ions of high energy and high charge are accelerated from compact intense laser produced plasmas and are routinely analysed either by time of flight or Thomson parabola spectrometry. At the highest intensities where ion energies can be substantially large, both these techniques have limitations. Strong electromagnetic pulse noise jeopardises the arrival time measurement, and a bright central spot in the Thomson parabola spectrometer affects the signal to noise ratio of ion traces that approach close to the central spot. We present a gated Thomson parabola spectrometer that addresses these issues and provides an elegant method to improvise ion spectrometry. In addition, we demonstrate that this method provides the ability to detect and measure high energy neutral atoms that are invariably present in most intense laser plasma acceleration experiments.

Micron-scale mapping of megagauss magnetic fields using optical polarimetry to probe hot electron transport in petawatt-class laser-solid interactions.

The transport of hot, relativistic electrons produced by the interaction of an intense petawatt laser pulse with a solid has garnered interest due to its potential application in the development of innovative x-ray sources and ion-acceleration schemes. We report on spatially and temporally resolved measurements of megagauss magnetic fields at the rear of a 50-µm thick plastic target, irradiated by a multi-picosecond petawatt laser pulse at an incident intensity of ~1020 W/cm2. The pump-probe polarimetric measurements with micron-scale spatial resolution reveal the dynamics of the magnetic fields generated by the hot electron distribution at the target rear. An annular magnetic field profile was observed ~5 ps after the interaction, indicating a relatively smooth hot electron distribution at the rear-side of the plastic target. This is contrary to previous time-integrated measurements, which infer that such targets will produce highly structured hot electron transport. We measured large-scale filamentation of the hot electron distribution at the target rear only at later time-scales of ~10 ps, resulting in a commensurate large-scale filamentation of the magnetic field profile. Three-dimensional hybrid simulations corroborate our experimental observations and demonstrate a beam-like hot electron transport at initial time-scales that may be attributed to the local resistivity profile at the target rear.

Magnetic turbulence in a table-top laser-plasma relevant to astrophysical scenarios.

Turbulent magnetic fields abound in nature, pervading astrophysical, solar, terrestrial and laboratory plasmas. Understanding the ubiquity of magnetic turbulence and its role in the universe is an outstanding scientific challenge. Here, we report on the transition of magnetic turbulence from an initially electron-driven regime to one dominated by ion-magnetization in a laboratory plasma produced by an intense, table-top laser. Our observations at the magnetized ion scale of the saturated turbulent spectrum bear a striking resemblance with spacecraft measurements of the solar wind magnetic-field spectrum, including the emergence of a spectral kink. Despite originating from diverse energy injection sources (namely, electrons in the laboratory experiment and ion free-energy sources in the solar wind), the turbulent spectra exhibit remarkable parallels. This demonstrates the independence of turbulent spectral properties from the driving source of the turbulence and highlights the potential of small-scale, table-top laboratory experiments for investigating turbulence in astrophysical environments.

Transition from Coherent to Stochastic electron heating in ultrashort relativistic laser interaction with structured targets.

Relativistic laser interaction with micro- and nano-scale surface structures enhances energy transfer to solid targets and yields matter in extreme conditions. We report on the comparative study of laser-target interaction mechanisms with wire-structures of different size, revealing a transition from a coherent particle heating to a stochastic plasma heating regime which occurs when migrating from micro-scale to nano-scale wires. Experiments and kinetic simulations show that large gaps between the wires favour the generation of high-energy electrons via laser acceleration into the channels while gaps smaller than the amplitude of electron quivering in the laser field lead to less energetic electrons and multi-keV plasma generation, in agreement with previously published experiments. Plasma filling of nano-sized gaps due to picosecond pedestal typical of ultrashort pulses strongly affects the interaction with this class of targets reducing the laser penetration depth to approximately one hundred nanometers. The two heating regimes appear potentially suitable for laser-driven ion/electron acceleration schemes and warm dense matter investigation respectively.

Intense femtosecond laser driven collimated fast electron transport in a dielectric medium-role of intensity contrast.

Ultra-high intensity (> 1018 W/cm2), femtosecond (~30 fs) laser induced fast electron transport in a transparent dielectric has been studied for two laser systems having three orders of magnitude different peak to pedestal intensity contrast, using ultrafast time-resolved shadowgraphy. Use of a 400 nm femtosecond pulse as a probe enables the exclusive visualization of the dynamics of highest density electrons (> 7 × 1021 cm-3) observed so far. High picosecond contrast (~109) results in greater coupling of peak laser energy to the plasma electrons, enabling long (~1 mm), collimated (divergence angle ~2°) transport of fast electrons inside the dielectric medium at relativistic speeds (~0.66c). In comparison, the laser system with a contrast of ~106 has a large pre-plasma, limiting the coupling of laser energy to the solid and yielding limited fast electron injection into the dielectric. In the lower contrast case, bulk of the electrons expand as a cloud inside the medium with an order of magnitude lower speed than that of the fast electrons obtained with the high contrast laser. The expansion speed of the plasma towards vacuum is similar for the two contrasts.

Contrasting levels of absorption of intense femtosecond laser pulses by solids.

The absorption of ultraintense, femtosecond laser pulses by a solid unleashes relativistic electrons, thereby creating a regime of relativistic optics. This has enabled exciting applications of relativistic particle beams and coherent X-ray radiation, and fundamental leaps in high energy density science and laboratory astrophysics. Obviously, central to these possibilities lies the basic problem of understanding and if possible, manipulating laser absorption. Surprisingly, the absorption of intense light largely remains an open question, despite the extensive variations in target and laser pulse structures. Moreover, there are only few experimental measurements of laser absorption carried out under very limited parameter ranges. Here we present an extensive investigation of absorption of intense 30 femtosecond laser pulses by solid metal targets. The study, performed under varying laser intensity and contrast ratio over four orders of magnitude, reveals a significant and non-intuitive dependence on these parameters. For contrast ratio of 10(-9) and intensity of 2 × 10(19)W cm(-2), three observations are revealed: preferential acceleration of electrons along the laser axis, a ponderomotive scaling of electron temperature, and red shifting of emitted second-harmonic. These point towards the role of J × B absorption mechanism at relativistic intensity. The experimental results are supported by particle-in-cell simulations.

Enhanced x-ray emission from nano-particle doped bacteria.

Recently, it has been greatly appreciated that intense light matter interaction is modified due to the nano- and microstructures in the target by--surface plasmons, laser energy localization scattering etc. Extreme laser intensities produce dense plasmas and collective mechanisms generate energetic electrons, ions and hard x-rays. Recently, it is postulated that the anharmonic electron motion, driven by ultrashort, high-intensity laser pulses, provides a universal mechanism for the laser absorption. Here, we provide the first demonstration of anharmonic-resonance-aided high laser-absorption in a biological system. At intensities of ∼ 10¹6⁻¹8 W/cm², 40 fs pulses excite a plasma formed with E. coli bacteria. The density-inhomogeneities due to the micro- and nanostructures in the bacterial target increase anharmonic resonance (AHR) heating and result in a 104-fold enhancement in the hard x-ray yield compared to plain solid targets. These observations lead to novel high-energy x-ray sources that have implications to lithography, imaging and medical applications.

Terahertz acoustics in hot dense laser plasmas.

We present a hitherto unobserved facet of hydrodynamics, namely the generation of an ultrahigh frequency acoustic disturbance in the terahertz frequency range, whose origins are purely hydrodynamic in nature. The disturbance is caused by differential flow velocities down a density gradient in a plasma created by a 30 fs, 800 nm high-intensity laser (∼5×10(16) W/cm(2)). The picosecond scale observations enable us to capture these high frequency oscillations (1.9±0.6 THz) which are generated as a consequence of the rapid heating of the medium by the laser. Adoption of two complementary techniques, namely pump-probe reflectometry and pump-probe Doppler spectrometry provides unambiguous identification of this terahertz acoustic disturbance. Hydrodynamic simulations well reproduce the observations, offering insight into this process.

Ultrafast optics of solid density plasma using multicolor probes.

We present time-resolved reflectivity and transmissivity of hot, overdense plasma by employing a multicolor probe beam, consisting of harmonics at wavelengths of 800 nm, 400 nm and 266 nm. The hot-dense plasma, formed by exciting a fused silica target with a 30 fs, 2 × 10(17) W cm(-2) intensity pulse, shows a sub-picosecond transition in reflectivity (transmissivity), and a wavelength-dependent fall (rise) in the reflected (transmitted) signal. A simple model of probe absorption in the plasma via inverse bremsstrahlung is used to determine electron-ion collision frequency at different plasma densities.