The inability to produce perfusable microvasculature networks capable of supporting tissue survival and of withstanding physiological pressures without leakage is a fundamental problem facing the field of tissue engineering. Microvasculature is critically important for production of bioengineered lung (BEL), which requires systemic circulation to support tissue survival and coordination of circulatory and respiratory systems to ensure proper gas exchange. To advance our understanding of vascularization after bioengineered organ transplantation, we produced and transplanted BEL without creation of a pulmonary artery anastomosis in a porcine model. A single pneumonectomy, performed 1 month before BEL implantation, provided the source of autologous cells used to bioengineer the organ on an acellular lung scaffold. During 30 days of bioreactor culture, we facilitated systemic vessel development using growth factor-loaded microparticles. We evaluated recipient survival, autograft (BEL) vascular and parenchymal tissue development, graft rejection, and microbiome reestablishment in autografted animals 10 hours, 2 weeks, 1 month, and 2 months after transplant. BEL became well vascularized as early as 2 weeks after transplant, and formation of alveolar tissue was observed in all animals ( = 4). There was no indication of transplant rejection. BEL continued to develop after transplant and did not require addition of exogenous growth factors to drive cell proliferation or lung and vascular tissue development. The sterile BEL was seeded and colonized by the bacterial community of the native lung.

The eukaryotic cell compartmentalizes into spatially confined, membrane-enclosed, intracellular structures (e.g., organelles, endosomes, and vesicles). Here, peculiar physicochemical properties of the local environment occur and participate in the regulation of ongoing molecular processes. In spite of the huge amount of available environmental probes, experiments on subcellular structures are severely challenged by their three-dimensional (3D) movement. This bottleneck is tackled here by focusing an excitation light beam in a periodic orbit around the structure of interest. The recorded signal is used as feedback to localize the structure position at high temporal resolution: microseconds along the orbit, milliseconds between orbits. The lysosome is selected as the intracellular target, together with 6-acetyl-2dimethylaminonaphthalene (ACDAN) as probe of the physicochemical properties of the intralysosomal environment. Generalized polarization (GP) analysis of ACDAN emission is used to get a quantitative view on intralysosomal solvent dipolar relaxation. Thus, raster image correlation spectroscopy (RICS) analysis reveals that the ACDAN GP signal is fluctuating in the micro-to-millisecond time range during natural organelle 3D trafficking. We show that ACDAN GP fluctuations are characteristic of lysosomes in living cells, are selectively abolished by lysosomal basification, and depend on metabolic energy in the form of ATP. We argue that intralysosomal ACDAN GP fluctuates according to the ongoing organelle metabolism. Indeed, we report alterations in amplitude and timing of GP fluctuations in a cellular model of lysosomal storage disorder (LSD). The strategy proposed provides insight into the elusive local environment of a trafficking lysosome and supports similar molecular investigations at the subcellular level.

Diabetes has no well-established cure; thus, its management is critical for avoiding severe health complications involving multiple organs. This requires frequent glycaemia monitoring, and the gold standards for this are fingerstick tests. During the last decades, several blood-withdrawal-free platforms have been being studied to replace this test and to improve significantly the quality of life of people with diabetes (PWD). Devices estimating glycaemia level targeting blood or biofluids such as tears, saliva, breath and sweat, are gaining attention; however, most are not reliable, user-friendly and/or cheap. Given the complexity of the topic and the rise of diabetes, a careful analysis is essential to track scientific and industrial progresses in developing diabetes management systems. Here, we summarize the emerging blood glucose level (BGL) measurement methods and report some examples of devices which have been under development in the last decades, discussing the reasons for them not reaching the market or not being really non-invasive and continuous. After discussing more in depth the history of Raman spectroscopy-based researches and devices for BGL measurements, we will examine if this technique could have the potential for the development of a user-friendly, miniaturized, non-invasive and continuous blood glucose-monitoring device, which can operate reliably, without inter-patient variability, over sustained periods.