A Novel Photopharmacological Tool: Dual-Step Luminescence for Biological Tissue Penetration of Light and the Selective Activation of Photodrugs.

Conventional pharmacology lacks spatial and temporal selectivity in terms of drug action. This leads to unwanted side effects, such as damage to healthy cells, as well as other less obvious effects, such as environmental toxicity and the acquisition of resistance to drugs, especially antibiotics, by pathogenic microorganisms. Photopharmacology, based on the selective activation of drugs by light, can contribute to alleviating this serious problem. However, many of these photodrugs are activated by light in the UV-visible spectral range, which does not propagate through biological tissues. In this article, to overcome this problem, we propose a dual-spectral conversion technique, which simultaneously makes use of up-conversion (using rare earth elements) and down-shifting (using organic materials) techniques in order to modify the spectrum of light. Near-infrared light (980 nm), which penetrates tissue fairly well, can provide a "remote control" for drug activation. Once near-IR light is inside the body, it is up-converted to the UV-visible spectral range. Subsequently, this radiation is down-shifted in order to accurately adjust to the excitation wavelengths of light which can selectively activate hypothetical and specific photodrugs. In summary, this article presents, for the first time, a "dual tunable light source" which can penetrate into the human body and deliver light of specific wavelengths; thus, it can overcome one of the main limitations of photopharmacology. It opens up promising possibilities for the moving of photodrugs from the laboratory to the clinic.

Human-Centric Lighting: Rare-Earth-Free Photoluminescent Materials for Correlated Color Temperature Tunable White LEDs.

Artificial lighting is ubiquitous in modern society, with detrimental effects on sleep and health. The reason for this is that light is responsible not only for vision but also for non-visual functions, such as the regulation of the circadian system. To avoid circadian disruption, artificial lighting should be dynamic, changing throughout the day in a manner comparable to natural light in terms of both light intensity and associated color temperature. This is one of the main goals of human-centric lighting. Regarding the type of materials, the majority of white light-emitting diodes (WLEDs) make use of rare-earth photoluminescent materials; therefore, WLED development is at serious risk due to the explosive growth in demand for these materials and a monopoly on sources of supply. Photoluminescent organic compounds are a considerable and promising alternative. In this article, we present several WLEDs that were manufactured using a blue LED chip as the excitation source and two photoluminescent organic dyes (Coumarin 6 and Nile Red) embedded in flexible layers, which function as spectral converters in a multilayer remote phosphor arrangement. The correlated color temperature (CCT) values range from 2975 K to 6261 K, while light quality is preserved with chromatic reproduction index (CRI) values superior to 80. Our findings illustrate for the first time the enormous potential of organic materials for supporting human-centric lighting.

Retinal pigment epithelium degeneration caused by aggregation of PRPF31 and the role of HSP70 family of proteins.

BACKGROUND: Mutations in pre-mRNA splicing factor PRPF31 can lead to retinitis pigmentosa (RP). Although the exact disease mechanism remains unknown, it has been hypothesized that haploinsufficiency might be involved in the pathophysiology of the disease. METHODS: In this study, we have analyzed a mouse model containing the p.A216P mutation in Prpf31 gene. RESULTS: We found that mutant Prpf31 protein produces cytoplasmic aggregates in the retinal pigment epithelium and decreasing the protein levels of this splicing factor in the nucleus. Additionally, normal protein was recruited in insoluble aggregates when the mutant protein was overexpressed in vitro. In response to protein aggregation, Hspa4l is overexpressed. This member of the HSP70 family of chaperones might contribute to the correct folding and solubilization of the mutant protein, allowing its translocation to the nucleus. CONCLUSIONS: Our data suggests that a mechanism haploinsufficiency and dominant-negative is involved in retinal degeneration due to mutations in PRPF31. HSP70 over-expression might be a new therapeutic target for the treatment of retinal degeneration due to PRPF31 mutations.

Bioengineered tissue and cell therapy products are efficiently cryopreserved with pathogen-inactivated human platelet lysate-based solutions.

BACKGROUND: There remains much interest in improving cryopreservation techniques for advanced therapy medicinal products (ATMPs). Recently, human platelet lysate (hPL) has emerged as a promising candidate to replace fetal bovine serum (FBS) as a xeno-free culture supplement for the expansion of human cell therapy products. Whether hPL can also substitute for FBS in cryopreservation procedures remains poorly studied. Here, we evaluated several cryoprotective formulations based on a proprietary hPL for the cryopreservation of bioengineered tissues and cell therapy products. METHODS: We tested different xenogeneic-free, pathogen-inactivated hPL (ihPL)- and non-inactivated-based formulations for cryopreserving bioengineered tissue (cellularized nanostructured fibrin agarose hydrogels (NFAHs)) and common cell therapy products including bone marrow-derived mesenchymal stromal cells (BM-MSCs), human dermal fibroblasts (FBs) and neural stem cells (NSCs). To assess the tissue and cellular properties post-thaw of NFAHs, we analyzed their cell viability, identity and structural and biomechanical properties. Also, we evaluated cell viability, recovery and identity post-thaw in cryopreserved cells. Further properties like immunomodulation, apoptosis and cell proliferation were assessed in certain cell types. Additionally, we examined the stability of the formulated solutions. The formulations are under a bidding process with MD Bioproducts (Zurich, Switzerland) and are proprietary. RESULTS: Amongst the tissue-specific solutions, Ti5 (low-DMSO and ihPL-based) preserved the viability and the phenotype of embedded cells in NFAHs and preserved the matrix integrity and biomechanical properties similar to those of the standard cryopreservation solution (70% DMEM + 20% FBS + 10% DMSO). All solutions were stable at - 20 °C for at least 3 months. Regarding cell-specific solutions, CeA maintained the viability of all cell types > 80%, preserved the immunomodulatory properties of BM-MSCs and promoted good recovery post-thaw. Besides, both tested solutions were stable at - 20 °C for 18 months. Finally, we established that there is a 3-h window in which thawed NFAHs and FBs maintain optimum viability immersed in the formulated solutions and at least 2 h for BM-MSCs. CONCLUSIONS: Our results show that pathogen-inactivated solutions Ti5 allocated for bioengineered tissues and CeA allocated for cells are efficient and safe candidates to cryopreserve ATMPs and offer a xenogeneic-free and low-DMSO alternative to commercially available cryoprotective solutions.

Light Pollution and Circadian Misalignment: A Healthy, Blue-Free, White Light-Emitting Diode to Avoid Chronodisruption.

Sunlight has participated in the development of all life forms on Earth. The micro-world and the daily rhythms of plants and animals are strongly regulated by the light-dark rhythm. Human beings have followed this pattern for thousands of years. The discovery and development of artificial light sources eliminated the workings of this physiological clock. The world's current external environment is full of light pollution. In many electrical light bulbs used today and considered "environmentally friendly," such as LED devices, electrical energy is converted into short-wavelength illumination that we have not experienced in the past. Such illumination effectively becomes "biological light pollution" and disrupts our pineal melatonin production. The suppression of melatonin at night alters our circadian rhythms (biological rhythms with a periodicity of 24 h). This alteration is known as chronodisruption and is associated with numerous diseases. In this article, we present a blue-free WLED (white light-emitting diode) that can avoid chronodisruption and preserve circadian rhythms. This WLED also maintains the spectral quality of light measured through parameters such as CRI (color reproduction index).

Sunlike White Light-Emitting Diodes Based on Rare-Earth-Free Luminescent Materials.

Solid-state lighting (SSL) sources based on light-emitting diodes represent the new generation of highly efficient illumination systems that significantly impact energy-saving. The development of white light-emitting diodes (WLEDs) with a combination of high color rendering index (CRI) and high deep-red color rendering R9 is an important challenge in the field of solid-state lighting. On the other hand, most WLEDs use rare-earth inorganic luminescent materials. The annual demand for rare-earth metals has doubled to 125,000 tons in 15 years, and the demand is projected to reach 315,000 tons in 2030. The explosion in demand for these materials, combined with a monopolistic supply source, represents a real risk for the development of WLEDs in the next few years. Luminescent organic materials are a relevant and promising alternative. Here, we report a WLED with a very high CRI of 95.7 and R9 of 78.7, obtained using a combination of a blue LED chip (excitation source) and two organic luminescent dyes (Coumarin 6 and Lumogen Red) acting as spectral converters in a multilayer remote phosphor configuration. To the best of our knowledge, this is the first rare-earth-free WLED with such high values of CRI and R9.

Brain Organoids to Evaluate Cellular Therapies.

Animal models currently used to test the efficacy and safety of cell therapies, mainly murine models, have limitations as molecular, cellular, and physiological mechanisms are often inherently different between species, especially in the brain. Therefore, for clinical translation of cell-based medicinal products, the development of alternative models based on human neural cells may be crucial. We have developed an in vitro model of transplantation into human brain organoids to study the potential of neural stem cells as cell therapeutics and compared these data with standard xenograft studies in the brain of immunodeficient NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice. Neural stem cells showed similar differentiation and proliferation potentials in both human brain organoids and mouse brains. Our results suggest that brain organoids can be informative in the evaluation of cell therapies, helping to reduce the number of animals used for regulatory studies.

Human Neural Stem Cells for Cell-Based Medicinal Products.

Neural stem cells represent an attractive tool for the development of regenerative therapies and are being tested in clinical trials for several neurological disorders. Human neural stem cells can be isolated from the central nervous system or can be derived in vitro from pluripotent stem cells. Embryonic sources are ethically controversial and other sources are less well characterized and/or inefficient. Recently, isolation of NSC from the cerebrospinal fluid of patients with spina bifida and with intracerebroventricular hemorrhage has been reported. Direct reprogramming may become another alternative if genetic and phenotypic stability of the reprogrammed cells is ensured. Here, we discuss the advantages and disadvantages of available sources of neural stem cells for the production of cell-based therapies for clinical applications. We review available safety and efficacy clinical data and discuss scalability and quality control considerations for manufacturing clinical grade cell products for successful clinical application.