Time of flight secondary ion mass spectrometry of bone—Impact of sample preparation and measurement conditions

Bone histology of decalcified or undecalcified samples depends on the investigation. However, in research each method provides different information to answer the scientific question. Decalcification is the first step after sample fixation and governs what analysis is later feasible on the sections. Besides, decalcification is favored for immunostaining and in situ hybridization. Otherwise, sample decalcification can be damaging to bone biomaterials implants that contains calcium or strontium. On the other hand, after decalcification mineralization cannot be assessed using histology or imaging mass spectrometry. The current study provides a solution to the hardship caused by material presence within the bone tissue. The protocol presents a possibility of gaining sequential and alternating decalcified and undecalcified sections from the same bone sample. In this manner, investigations using histology, protein signaling, in situ hybridization, and mass spectrometry on the same sample can better answer the intended research question. Indeed, decalcification of sections and grindings resulted in well-preserved sample and biomaterials integrity. Immunostaining was comparable to that of classically decalcified samples. The study offers a novel approach that incites correlative analysis on the same sample and reduces the number of processed samples whether clinical biopsies or experimental animals.

show abstract

“…Bi 3 + was used as primary ion species. A detailed description of the method can be found elsewhere [14–16]. …”

Section: Methodsmentioning

confidence: 99%

Postembedding Decalcification of Mineralized Tissue Sections Preserves the Integrity of Implanted Biomaterials and Minimizes Number of Experimental Animals

Khassawna

Daghma

Stoetzel

et al. 2017

BioMed Research International

Self Cite

View full text Add to dashboard Cite

show abstract

“…The obtained mass resolution m/Dm was for all measurements better than 3000 for the Ca þ mass signal. More detailed images of smaller areas were taken in the delayed extraction mode: 512 Â 512 pixel, 10 shots/pixel, 50 scans (1024 Â 1024 pixel, 1 shots/pixel, 100 scans) with a delay time of 0.135 ms and a current of 0.08-0.1 pA. More detailed information on SIMS measurements of bone is described in [26,37].…”

Section: Time-of-flight Secondary Ion Mass Spectrometrymentioning

confidence: 99%

Investigation of strontium transport and strontium quantification in cortical rat bone by time-of-flight secondary ion mass spectrometry

Kern

Quade

Ray

et al. 2019

J. R. Soc. Interface.

Self Cite

View full text Add to dashboard Cite

Next-generation bone implants will be functionalized with drugs for stimulating bone growth. Modelling of drug release by such functionalized biomaterials and drug dispersion into bone can be used as predicting tool for biomaterials testing in future. Therefore, the determination of experimental parameters to describe and simulate drug release in bone is essential. Here, we focus on Sr 2+ transport and quantification in cortical rat bone. Sr 2+ dose-dependently stimulates bone-building osteoblasts and inhibits bone-resorbing osteoclasts. It should be preferentially applied in the case of bone fracture in the context of osteoporotic bone status. Transport properties of cortical rat bone were investigated by dipping experiments of bone sections in aqueous Sr 2+ solution followed by time-of-flight secondary ion mass spectrometry (ToF-SIMS) depth profiling. Data evaluation was carried out by fitting a suitable mathematical diffusion equation to the experimental data. An average diffusion coefficient of D = (1.68 ± 0.57) · 10 −13 cm 2 s −1 for healthy cortical bone was obtained. This value differed only slightly from the value of D = (4.30 ± 1.43) · 10 −13 cm 2 s −1 for osteoporotic cortical bone. Transmission electron microscopy investigations revealed a comparable nano- and ultrastructure for both types of bone status. Additionally, Sr 2+ -enriched mineralized collagen standards were prepared for ToF-SIMS quantification of Sr 2+ content. The obtained calibration curve was used for Sr 2+ quantification in cortical and trabecular bone in real bone sections. The results allow important insights regarding the Sr 2+ transport properties in healthy and osteoporotic bone and can ultimately be used to perform a simulation of drug release and mobility in bone.

show abstract

“…Nuclear magnetic resonance (NMR) has been utilized for structural investigations on bones and teeth, including studies of proteins (29)(30)(31)(32)(33)(34)(35)(36). Time-of-flight secondary ion mass spectroscopy (ToF-SIMS) has proven to be a useful method for the chemical analyses of HAP in enamel and bone tissue, as well as for analyses of organic molecules, such as protein fragments (33,(37)(38)(39)(40)(41)(42)(43)(44)(45).…”

mentioning

confidence: 99%

Molecular insights into hypomineralized enamel

Malmberg

Norén

Bernin

2019

European J Oral Sciences

View full text Add to dashboard Cite

Hypomineralized enamel may be found in connection with the condition molar incisor hypomineralization (MIH), which has a prevalence of around 15% in most parts of the world. Molar incisor hypomineralization is associated with extensive objective and subjective problems, such as hypersensitivity of the affected teeth, enamel breakdown, and problems with retention of restorations. The etiology behind MIH has not yet been elucidated, but a number of possible factors, which affect the same or different functions of ameloblasts during their different stages of maturation, have been suggested. The aim of this study was to utilize multi‐nuclear, solid‐state nuclear magnetic resonance (ss‐NMR) and time‐of‐flight secondary ion mass spectroscopy (ToF‐SIMS) to elucidate any differences, at a molecular level, between enamel powder prepared from normal, healthy teeth and enamel powder prepared from teeth diagnosed with MIH. 31P and 23Na ss‐NMR confirmed the presence of HPO42− and two different Na+ sites in hypomineralized enamel, suggesting a heterogeneous chemical composition. The content of organic components was higher in hypomineralized enamel, as shown by both 13C ss‐NMR and ToF‐SIMS, indicating the presence of higher numbers of proteins and phospholipids. The interplay between both is necessary for the formation and mineralization of enamel, which might be disturbed or halted in hypomineralized enamel.

show abstract

Time of flight secondary ion mass spectrometry of bone—Impact of sample preparation and measurement conditions

Cited by 19 publications

References 24 publications

Postembedding Decalcification of Mineralized Tissue Sections Preserves the Integrity of Implanted Biomaterials and Minimizes Number of Experimental Animals

Postembedding Decalcification of Mineralized Tissue Sections Preserves the Integrity of Implanted Biomaterials and Minimizes Number of Experimental Animals

Investigation of strontium transport and strontium quantification in cortical rat bone by time-of-flight secondary ion mass spectrometry

Molecular insights into hypomineralized enamel

Contact Info

Product

Resources

About