Raman microspectroscopy identifies fibrotic tissues in collagen-related disorders via deconvoluted collagen type I spectra

Becker, Lucas; Lu, Chuan-En; Montes-Mojarro, Ivonne A.; Layland, Shannon L.; Khalil, Suzan; Nsair, Ali; Duffy, Garry P.; Fend, Falko; Marzi, Julia; Schenke‐Layland, Katja

doi:10.1016/j.actbio.2023.03.016

Cited by 9 publications

(7 citation statements)

References 82 publications

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“…In contrast, myosin (pink) was assigned by its morphology and increased Raman intensities at 827 and 853 cm −1 representatives for C-C in proline and ring-breathing mode in tyrosine ( 33 , 36 ). Based on in-house Raman measurements validated by colocalization of IF signals, αSMA was assigned to the fifth spectral component identified by TCA (red) ( 22 ). The potential of RMS being utilized for noninvasive, and marker-independent imaging of tissue structures was identified as Raman images and IF images showed similar morphological features and comparable results in the quantification of the amounts of COL I and αSMA.…”

Section: Resultsmentioning

confidence: 99%

“…COL I average Raman spectra were cropped to the amide I region (1,508–1,780 cm −1 ) and normalized to 1 for better comparability. By spectral deconvolution, five substructural peaks located at 1,562, 1,588, 1,608, 1,637, and 1,667 cm −1 could be identified based on the shape of the amide I region and literature ( 22 , 27 ). The subpeaks were assigned to tryptophane, phenylalanine, tyrosine, β-sheets as well β-turns ( 37 , 38 ).…”

Section: Resultsmentioning

confidence: 99%

“…Spectral deconvolution was performed as previously described ( 22 ). In short, with the software WITec project 5, spectral deconvolution was used to calculate the peak width and peak area of the substructural bands of the amide I region (1,550–1,720 cm −1 ) of COL I. Collagen maps were extracted from the Raman data and then cropped to the amide I region before normalization to the peak at 1,667 cm −1 to 1.…”

Section: Methodsmentioning

confidence: 99%

“…To provide information on peak ratios of all individual collagen spectra within one Raman map, sum filter images of collagen maps were created using WITec project 5 software as previously described ( 22 ). Sum filter images were collected at 1,667 ± 25 cm −1 and at 1,608 ± 10 cm −1 and then divided before exporting them to MATLAB 2019 b.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, our group used Raman microspectroscopy to identify epigenetic 5mC alterations in human colon carcinoma cells ( 25 ). Furthermore, it has recently been demonstrated that fibrotic collagen changes can be identified by spectral deconvolution in various human tissues ( 22 ).…”

Section: Introductionmentioning

confidence: 99%

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Exploring the relationship between epigenetic DNA methylation and cardiac fibrosis through Raman microspectroscopy

Becker

Montes-Mojarro

Layland

et al. 2023

American Journal of Physiology-Cell Physiology

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Cardiomyopathies are associated with fibrotic remodeling of the heart, which is characterized by the excessive accumulation of collagen type I (COL I) due to chronic inflammation and suspected epigenetic influences. Despite the severity and high mortality rate of cardiac fibrosis, current treatment options are often inadequate, underscoring the importance of gaining a deeper understanding of the disease's underlying molecular and cellular mechanisms. In this study, the extracellular matrix (ECM) and nuclei in fibrotic areas of different cardiomyopathies were molecularly characterized by Raman microspectroscopy and imaging and compared to the control myocardium. Patient samples were obtained from heart tissue affected by ischemia, hypertrophy, and dilated cardiomyopathy and analyzed for fibrosis through conventional histology and marker-independent Raman microspectroscopy (RMS). Prominent differences between control myocardium and cardiomyopathies were revealed by spectral deconvolution of COL I Raman spectra. Statistically significant differences were identified in the amide I region of spectral subpeak at 1608 cm-1, which is a representative endogenous marker for alterations in the structural conformation of COL I fibers. Moreover, epigenetic 5mC DNA modification was identified within cell nuclei by multivariate analysis. A statistically significant increase in signal intensities of spectral features indicative of DNA methylation was detected in cardiomyopathies in accordance with immunofluorescence 5mC staining. Overall, RMS is a versatile technology in the discrimination of cardiomyopathies based on molecular evaluation of COL I and nuclei, while providing insights into the pathogenesis of the diseases.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Exploring the relationship between epigenetic DNA methylation and cardiac fibrosis through Raman microspectroscopy

Becker

Montes-Mojarro

Layland

et al. 2023

American Journal of Physiology-Cell Physiology

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Deciphering UVA/Riboflavin Collagen Crosslinking: A Pathway to Improve Biomedical Materials

Fan,

Jung,

Herrmann

et al. 2024

Adv Funct Materials

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Collagen crosslinking employing ultraviolet A rays and riboflavin (UVA/R) has emerged as a pivotal technique in clinical therapies, especially in ophthalmology since the 1990s. Despite its clinical adoption, the lack of clarity of the detailed mechanism and the imperative for a refined manufacturing process necessitates further investigation. This study advances the understanding of UVA/R crosslinked collagen, concentrating on identifying the primary crosslinking sites using seven synthetic peptides and exploring the pathways of riboflavin‐mediated crosslinking. The results demonstrate that tyrosine residues are key crosslinking sites, and riboflavin plays a dual role as both a catalyst and a competitive inhibitor in the crosslinking process. Furthermore, the UVA/R crosslinked collagen matrix exhibits a more harmonious balance between stability and degradability compared with chemically crosslinked collagen matrices, coupled with superior mechanical properties and augmented biocompatibility. In vivo experiments further validate its excellent biocompatibility, reduced tissue inflammation, and promotion of tissue regeneration. The research provides crucial insights into collagen crosslinking mechanisms, paving the way for the development of sophisticated collagen‐based biomaterials tailored for biomedical applications.

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A comprehensive analysis of metabolomics and lipidomics in areca nut mediated oral submucous fibrosis progression through LCMS and Raman spectroscopy

Verma,

Adhikari,

Singh

et al. 2024

J Raman Spectroscopy

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Habitual consumption of areca/betel nut in Southeast Asia has been associated with oral submucous fibrosis (OSMF) and its malignant transformation to oral squamous cell carcinoma. The current study aimed to assess the molecular alterations in fibroblast cell lines after treatment with areca nut. Areca nut extract (ANE) was prepared and characterized for its active ingredient, arecoline. ANE‐treated cells were subjected to cell viability, proliferation, migration, morphologic changes, and transcript of OSMF genes (col1a1, col1a2, hsp47, timp1, timp2, timp3, and timp4). Further, liquid chromatography–mass spectrometry (LCMS) of cell lysate and Raman microspectroscopy (RMS) of fixed cells were performed for metabolomics/lipidomics and biomolecular alterations in the nucleus and periphery of ANE‐treated cells. We compared and integrated the data from both techniques and observed that the cells treated with ANE mimicked OSMF and could be used as an in vitro model. LCMS showed a significant alteration in 17 metabolites and 165 lipid molecules in the cells treated with ANE. Further, 40 molecules in the nucleus and 29 in the periphery were found to be altered in these cells when subjected to RMS. Molecules associated with pathways such as the transfer of acetyl groups into mitochondria, amino sugar metabolism, and the Warburg effect were modulated the most upon ANE treatment. Acetyl CoA was found to be common in most of the altered pathways. Besides, the pathways affecting carbohydrate metabolism were also altered significantly. Targeting these molecules and pathways can help to prevent the malignant transformation of OSMF.

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Raman microspectroscopy identifies fibrotic tissues in collagen-related disorders via deconvoluted collagen type I spectra

Cited by 9 publications

References 82 publications

Exploring the relationship between epigenetic DNA methylation and cardiac fibrosis through Raman microspectroscopy

Exploring the relationship between epigenetic DNA methylation and cardiac fibrosis through Raman microspectroscopy

Deciphering UVA/Riboflavin Collagen Crosslinking: A Pathway to Improve Biomedical Materials

A comprehensive analysis of metabolomics and lipidomics in areca nut mediated oral submucous fibrosis progression through LCMS and Raman spectroscopy

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