Spatially Offset Raman Spectroscopy toward In Vivo Assessment of the Adipose Tissue in Cardiometabolic Pathologies.

Spatially offset Raman spectroscopy (SORS) enhanced the capabilities of Raman spectroscopy for the depth-resolved analysis of biological and diffusely scattering samples. This technique offers selective probing of subsurface layers, providing molecular insights without invasive procedures. While SORS has found application in biomedical research, up to now, studies have focused mainly on the detection of mineralization of bones and tissues. Herein, for the first time, SORS is used to assess the soft, organic tissue beneath the skin's surface. In this study, we demonstrate the diagnostic utility of a hand-held SORS device for evaluating the chemical composition of the adipose tissue. We compared perigonadal white adipose tissue (gWAT) in a murine model of atherosclerosis, heart failure, and high-fat diet (HFD) induced obesity. Our results reveal distinct chemical differences in gWAT between HFD-fed and control mice, showcasing the potential of SORS for intravital adipose tissue phenotype characterization. Furthermore, our findings underscore the effectiveness of SORS as a valuable tool for noninvasive assessment of the adipose tissue composition, holding potential diagnostic significance for metabolic disorders.

Increased obesogenic action of palmitic acid during early stage of adipogenesis.

The functional differences between preadipocytes and fully differentiated mature adipocytes derived from stromal vascular fraction stem cells, as well as primary adipocytes have been analysed by evaluating their response to the obesogenic factor (a saturated fatty acid) and TNF-triggered inflammation. The analysis of single adipocytes shows that the saturated fatty acid (palmitic acid) accumulation is accompanied by inflammation and considerably dependent on the stage of the adipogenesis. In particular, preadipocytes show the exceptional potential for palmitic acid uptake resulting in their hypertrophy and the elevated cellular expression of the inflammation marker (ICAM-1). Our research provides new information on the impact of obesogenic factors on preadipocytes that is important in the light of childhood obesity prevention.

Imaging of perivascular adipose tissue in cardiometabolic diseases by Raman spectroscopy: Towards single-cell analysis.

Perivascular adipose tissue (PVAT) has emerged as a dynamic organ influencing vascular function and cardiovascular health. In this brief review, an overview of the recent research in the investigation of PVAT is presented, ranging from in vivo studies to single-cell methodologies, in particular those based on Raman spectroscopy. The strengths and limitations of each, emphasizing their contributions to the current understanding of PVAT biology were discussed. Ultimately, the integration of these diverse methodologies promises to uncover new therapeutic targets and diagnostic biomarkers, including those emerging from simple Raman spectroscopy-based measurements of alterations in lipid unsaturation degree, invariably associated with PVAT dysfunction.

Direct oral and fiber-derived butyrate supplementation as an anti-obesity treatment via different targets.

BACKGROUND & AIMS: Butyric (one of the short-chain fatty acids), a major byproduct of the fermentation of non-digestible carbohydrates (e.g. fiber), is supposed to have anti-obesity and anti-inflammatory properties. However, butyrate's potential and mechanism in preventing obesity and the efficient form of administration remain to be clarified. METHODS: Hence, we studied the effect of oral supplementation with 5% (w/w) sodium butyrate and 4% (w/w) ß-glucan (fiber) on young male mice (C57BL/6J) with high-fat diet-induced obesity (HFD: 60 kcal% of fat + 1% of cholesterol). Six weeks old mice were fed diets based on HFD or control (AIN-93G) diet with/without supplements for 4 weeks. The unique, interdisciplinary approach combining several Raman-based techniques (including Raman microscopy and fiber optic Raman spectroscopy) and next-generation sequencing was used to ex vivo analyze various depots of the adipose tissue (white, brown, perivascular) and gut microbiome, respectively. RESULTS: The findings demonstrate that sodium butyrate more effectively prevent the pathological increase in body weight caused by elevated saturated fatty acids influx linked to a HFD in comparison to ß-glucan, thereby entirely inhibiting diet-induced obesity. Moreover, butyrate significantly affects the white adipose tissue (WAT) reducing the epididymal WAT mass in comparison to HFD without supplements, and decreasing lipid saturation in the epididymal WAT and perivascular adipose tissue of the thoracic aorta. Contrarily, ß-glucan significantly changes the composition and diversity of the gut microbiome, reversing the HFD effect, but shows no effect on the epididymal WAT mass and therefore the weight gain inhibition is not as effective as with sodium butyrate. CONCLUSIONS: Here, oral supplementation with sodium butyrate and ß-glucan (fiber) has been proven to have an anti-obesity effect through two different targets. Administration-dependent effects that butyrate imposes on the adipose tissue (oral administration) and microbiome (fiber-derived) make it a promising candidate for the personalized treatment of obesity.

Efficient delivery of carotenoids to adipocytes with albumin.

Carotenoids are very effectively delivered by albumin to adipocytes. The uptake of carotenoids to the cells occurs in the form of self-aggregates that localize in the vicinity of the adipocyte membrane, as shown by high spatial resolution Raman spectroscopy. The binding of carotenoids to albumin and the mechanism of their transport were elucidated with the help of chiroptical spectroscopies, in tandem with molecular docking and molecular dynamics simulations. In particular, apart from the recognized high affinity pocket of albumin that binds a carotenoid monomer in domain I, we have identified a hydrophobic periphery area in domain IIIB that loosely bounds the self-aggregated carotenoid in aqueous media and enables its easy detachment in hydrophobic environments. This explains the effectiveness of albumins as nanocarriers of carotenoids to adipocytes in vitro.

Raman studies of the adipose tissue: Current state-of-art and future perspectives in diagnostics.

The last decades revealed that the adipose tissue shows an unexplored therapeutic potential. In particular, targeting the perivascular adipose tissue (PVAT), that surrounds blood vessels, can prevent cardiovascular pathologies and browning of the adipose tissue can become an effective strategy against obesity. Therefore, new analytical tools are necessary to analyze this tissue. This review reports on the recent developments of various Raman-based techniques for the identification and quantification of the adipose tissue compared to conventional analytical methods. In particular, the emphasis is on analysis of PVAT, investigation of pathological changes of the adipose tissue in model systems and possibilities for its characterization in the clinical context. Overall, the review critically discusses the potential and limitations of Raman techniques in adipose tissue-targeted diagnostics and possible future anti-obesity therapies.

Formation of carotenoid supramolecular aggregates in nanocarriers monitored via aggregation-sensitive chiroptical output of enantiopure (3S,3'S)-astaxanthin.

The aggregation-sensitive chiroptical (ECD and RROA) output, provided by enantiopure (3S,3'S)-astaxanthin, was used to investigate and control the assembling processes of the carotenoid in Pluronic F-127 nanoparticles. The process of carotenoid J-aggregation inside nanocarriers is interfered with by the formation of kinetically stabilized H1 self-assemblies outside the micelles. Nanocarriers with encapsulated stable J-aggregates provide controlled release of carotenoid molecules to primary murine adipocytes.

The distinct phenotype of primary adipocytes and adipocytes derived from stem cells of white adipose tissue as assessed by Raman and fluorescence imaging.

Spectroscopy-based analysis of chemical composition of cells is a tool still scarcely used in biological sciences, although it provides unique information about the cell identity accessible in vivo and in situ. Through time-lapse spectroscopic monitoring of adipogenesis in brown and white adipose tissue-derived stem cells we have demonstrated that considerable chemical and functional changes occur along with cells differentiation and maturation, yet yielding mature adipocytes with a similar chemical composition, independent of the cellular origin (white or brown adipose tissue). However, in essence, these stem cell-derived adipocytes have a markedly different chemical composition compared to mature primary adipocytes. The consequences of this different chemical (and, hence, functional) identity have great importance in the context of selecting a suitable methodology for adipogenesis studies, particularly in obesity-related research.