Combining polarized Raman spectroscopy and micropillar compression to study microscale structure-property relationships in mineralized tissues

Kochetkova, Tatiana; Peruzzi, Cinzia; Braun, Oliver; Overbeck, Jan; Maurya, Anjani K.; Neels, Antonia; Calame, Michel; Michler, Johann; Zysset, Philippe; Schwiedrzik, Jakob

doi:10.1016/j.actbio.2020.10.034

Cited by 29 publications

(17 citation statements)

References 102 publications

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“…(24) Kochetkova and colleagues reported that the change of strain hardening to softening occurs around a 50 angle of the collagen with respect to the loading axis and the failure mechanism is highly dependent on this angle. (37) The heterogeneity of the micropillar compression curves in the present study may be explained first by the less regular osteonal organization of iliac crest bone compared with ovine tibia (long bone) and second by the twisted plywood organization of the lamellar architecture within a human osteon.…”

Section: Mechanical Properties In Oimentioning

confidence: 61%

“…It has been shown in several studies that the mechanical properties are dependent on the orientation of the collagen fibers. ( 18 , 24 , 25 , 37 , 39 ) Unfortunately, due to pillar fabrication using FIB (Gallium) and the destructive nature of the microcompression tests, it was not anymore possible to measure the collagen orientation in the specimen at the position of the micropillar using a technique such as polarized light. However, the careful orientation of our samples along the osteonal canals and the similar appearance/variability of the post‐yield behavior among the groups exclude a systematic bias of the mechanical properties due to collagen orientation.…”

Section: Discussionmentioning

confidence: 99%

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Compressive Strength of Iliac Bone ECM Is Not Reduced in Osteogenesis Imperfecta and Increases With Mineralization

Indermaur

Casari

Kochetkova

et al. 2020

Journal of Bone and Mineral Research

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Osteogenesis imperfecta (OI) is an inheritable, genetic, and collagen-related disorder leading to an increase in bone fragility, but the origin of its "brittle behavior" is unclear. Because of its complex hierarchical structure, bone behaves differently at various length scales. This study aims to compare mechanical properties of human OI bone with healthy control bone at the extracellular matrix (ECM) level and to quantify the influence of the degree of mineralization. Degree of mineralization and mechanical properties were analyzed under dry conditions in 12 fixed and embedded transiliac crest biopsies (control n = 6, OI type I n = 3, OI type IV n = 2, and OI type III n = 1). Mean degree of mineralization was measured by microcomputed tomography at the biopsy level and the mineral-tomatrix ratio was assessed by Raman spectroscopy at the ECM level. Both methods revealed that the degree of mineralization is higher for OI bone compared with healthy control. Micropillar compression is a novel technique for quantifying post-yield properties of bone at the ECM level. Micropillars (d = 5 μm, h = 10 μm) were fabricated using focused ion beam milling and quasi-statically compressed to capture key post-yield properties such as ultimate strength. The qualitative inspection of the stress-strain curves showed that both OI and healthy control bone have a ductile response at the ECM level. The quantitative results showed that compressive strength is not reduced in OI bone and is increasing with OI severity. Nanoindentation measurements revealed that OI bone tends to have a higher Young's modulus, hardness, and dissipated energy compared with healthy bone. Micropillar strength and indentation modulus increased linearly and significantly (p < .0001) with mineral-to-matrix ratio. In conclusion, this study indicates that compressive mechanical properties of dry OI bone at the iliac crest are not inferior to healthy control at the ECM level and increase with mineralization.

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Section: Mechanical Properties In Oimentioning

confidence: 61%

Section: Discussionmentioning

confidence: 99%

Compressive Strength of Iliac Bone ECM Is Not Reduced in Osteogenesis Imperfecta and Increases With Mineralization

Indermaur

Casari

Kochetkova

et al. 2020

Journal of Bone and Mineral Research

Self Cite

View full text Add to dashboard Cite

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“…The excitation wavelength was 532 nm, the objective lens was 50 × , and the scanning spectra ranged from 600 to 2000 cm −1 . The wave peaks of five spectral bands were extracted, and the range of each peak was determined by referring to other investigations, which was consistent in all spectra [ 33 , 34 ]. Finally, the ratio of each integral area was calculated.…”

Section: Methodsmentioning

confidence: 99%

Effects of continuous subcutaneous insulin infusion on the microstructures, mechanical properties and bone mineral compositions of lumbar spines in type 2 diabetic rats

Gong

Hou

et al. 2022

BMC Musculoskelet Disord

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Background Continuous subcutaneous insulin infusion (CSII) for the treatment of type 2 diabetes (T2D) can improve the structure and strength of femur of rats, but the effect of CSII treatment on the lumbar spine of T2D rats is unknown. The purpose of this study is to investigate the effects of CSII on the microstructure, multi-scale mechanical properties and bone mineral composition of the lumbar spine in T2D rats. Methods Seventy 6-week-old male Sprague–Dawley (SD) rats were divided into two batches, each including Control, T2D, CSII and Placebo groups, and the duration of insulin treatment was 4-week and 8-week, respectively. At the end of the experiment, the rats were sacrificed to take their lumbar spine. Microstructure, bone mineral composition and nanoscopic-mesoscopic-apparentand-macroscopic mechanical properties were evaluated through micro-computed tomography (micro-CT), Raman spectroscopy, nanoindentation test, nonlinear finite element analysis and compression test. Results It was found that 4 weeks later, T2D significantly decreased trabecular thickness (Tb.Th), nanoscopic-apparent and partial mesoscopic mechanical parameters of lumbar spine (P < 0.05), and significantly increased bone mineral composition parameters of cortical bone (P < 0.05). It was shown that CSII significantly improved nanoscopic-apparent mechanical parameters (P < 0.05). In addition, 8 weeks later, T2D significantly decreased bone mineral density (BMD), bone volume fraction (BV/TV) and macroscopic mechanical parameters (P < 0.05), and significantly increased bone mineral composition parameters of cancellous bone (P < 0.05). CSII treatment significantly improved partial mesoscopic-macroscopic mechanical parameters and some cortical bone mineral composition parameters (P < 0.05). Conclusions CSII treatment can significantly improve the nanoscopic-mesoscopic-apparent-macroscopic mechanical properties of the lumbar spine in T2D rats, as well as the bone structure and bone mineral composition of the lumbar vertebrae, but it will take longer treatment time to restore the normal level. In addition, T2D and CSII treatment affected bone mineral composition of cortical bone earlier than cancellous bone of lumbar spine in rat. Our study can provide evidence for clinical prevention and treatment of T2D-related bone diseases.

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“…In contrast, soft tissue components were more present in transition zone and trabecular bone, and were correlated with the distribution of lipids and blood, which arise from the bone marrow. In another very recent study, Kochetkova et al [155] combined polarized Raman spectroscopy with compression testing to assess the role of collagen alignment on tissue mechanical properties at the microscale. For this study, the authors used a confocal Raman microscope varying the polarization angles of the exciting laser to collect a series of polarized spectra from the same region of interest.…”

Section: Raman Spectral Analysis Of Bonementioning

confidence: 99%

Applications of Vibrational Spectroscopy for Analysis of Connective Tissues

2021

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Advances in vibrational spectroscopy have propelled new insights into the molecular composition and structure of biological tissues. In this review, we discuss common modalities and techniques of vibrational spectroscopy, and present key examples to illustrate how they have been applied to enrich the assessment of connective tissues. In particular, we focus on applications of Fourier transform infrared (FTIR), near infrared (NIR) and Raman spectroscopy to assess cartilage and bone properties. We present strengths and limitations of each approach and discuss how the combination of spectrometers with microscopes (hyperspectral imaging) and fiber optic probes have greatly advanced their biomedical applications. We show how these modalities may be used to evaluate virtually any type of sample (ex vivo, in situ or in vivo) and how “spectral fingerprints” can be interpreted to quantify outcomes related to tissue composition and quality. We highlight the unparalleled advantage of vibrational spectroscopy as a label-free and often nondestructive approach to assess properties of the extracellular matrix (ECM) associated with normal, developing, aging, pathological and treated tissues. We believe this review will assist readers not only in better understanding applications of FTIR, NIR and Raman spectroscopy, but also in implementing these approaches for their own research projects.

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Combining polarized Raman spectroscopy and micropillar compression to study microscale structure-property relationships in mineralized tissues

Cited by 29 publications

References 102 publications

Compressive Strength of Iliac Bone ECM Is Not Reduced in Osteogenesis Imperfecta and Increases With Mineralization

Compressive Strength of Iliac Bone ECM Is Not Reduced in Osteogenesis Imperfecta and Increases With Mineralization

Effects of continuous subcutaneous insulin infusion on the microstructures, mechanical properties and bone mineral compositions of lumbar spines in type 2 diabetic rats

Applications of Vibrational Spectroscopy for Analysis of Connective Tissues

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