This study investigates the influence of osteoarthritis (OA) disease severity on the bio-composition of the osteochondral junction at the human tibial plateau using Raman microspectroscopy. We specifically aim to analyze the spatial composition of mineralized osteochondral tissues, i.e., calcified cartilage (CC) and subchondral bone plate (SBP) from unfixed, hydrated specimens. We hypothesize that the mineralization of CC and SBP decreases in advanced OA. Twenty-eight cylindrical osteochondral samples (d = 4 mm) from tibial plateaus of seven cadaveric donors were harvested and sorted into three groups following histopathological grading: healthy (n = 5), early OA (n = 8), and advanced OA (n = 15). Raman spectra were subjected to multivariate cluster analyses to identify different tissues. Finally, the tissue-specific composition was analyzed, and the impact of OA was statistically evaluated with linear mixed models. Cluster analyses of Raman spectra successfully distinguished CC and SBP as well as a tidemark region and uncalcified cartilage. CC was found to be more mineralized and the mineral was more crystalline compared with SBP. Both tissues exhibited similar compositional changes as a function of histopathological OA severity. In early OA, the mineralization tends to increase, and the mineral contains fewer carbonate substitutions. Compared with early OA, mineral crystals are rich in carbonate while the overall mineralization decreases in advanced OA. This Raman spectroscopic study advances the methodology for investigating the complex osteochondral junction from native tissue. The developed methodology can be used to elucidate detailed tissue-specific changes in the chemical composition with advancing OA. STATEMENT OF SIGNIFICANCE: In this study, Raman microspectroscopy was utilized to investigate the influence of osteoarthritic degeneration on the tissue-specific biochemical composition of the human osteochondral junction. Multivariate cluster analyses allowed us to characterize subtle compositional changes in the calcified cartilage and subchondral bone plate as well as in the tidemark region. The compositional differences found between the calcified cartilage and subchondral bone plate in both organic and mineral phases will serve as critical benchmark parameters when designing biomaterials for osteochondral repair. We found tissue-specific changes in the mineralization and carbonate substitution as a function of histopathological OA severity. Our developed methodology can be used to investigate the metabolic changes in the osteochondral junction associated with osteoarthritis.

Osteoarthritis (OA) is the most common joint disease, where articular cartilage degradation is often accompanied with sclerosis of the subchondral bone. However, the association between OA and tissue mineralization at the nanostructural level is currently not understood. In particular, it is technically challenging to study calcified cartilage, where relevant but poorly understood pathological processes such as tidemark multiplication and advancement occur. Here, we used state-of-the-art microfocus small-angle X-ray scattering with a 5-μm spatial resolution to determine the size and organization of the mineral crystals at the nanostructural level in human subchondral bone and calcified cartilage. Specimens with a wide spectrum of OA severities were acquired from both medial and lateral compartments of medial compartment knee OA patients (n = 15) and cadaver knees (n = 10). Opposing the common notion, we found that calcified cartilage has thicker and more mutually aligned mineral crystals than adjoining bone. In addition, we, for the first time, identified a well-defined layer of calcified cartilage associated with pathological tidemark multiplication, containing 0.32 nm thicker crystals compared to the rest of calcified cartilage. Finally, we found 0.2 nm thicker mineral crystals in both tissues of the lateral compartment in OA compared with healthy knees, indicating a loading-related disease process because the lateral compartment is typically less loaded in medial compartment knee OA. In summary, we report novel changes in mineral crystal thickness during OA. Our data suggest that unloading in the knee might be involved with the growth of mineral crystals, which is especially evident in the calcified cartilage. © 2022 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Dental caries is the most common oral disease that causes demineralization of enamel and later in dentin. The depth-wise assessment of the demineralization process could be used to help treatment planning. In this study, we aimed to provide baseline information for Raman probe development by characterizing the mineral composition of the dental tissues from large composition maps (6x3 mm² with 15 µm step size) using Raman microspectroscopy. Ten human wisdom teeth with different stages of dental caries lesions were examined. All of the teeth were cut in half at representative locations of the caries lesions and then imaged with a Raman imaging microscope. The pre-processed spectral maps were combined into a single data matrix, and the spectra of enamel, dentin, and caries were identified by using K-means cluster analysis. Our results showed that unsupervised identification of dental caries is possible with the K-means clustering. The compositional analysis revealed that the carious lesions are less mineralized than healthy enamel, and when the lesions extend to dentin, they are even less mineralized. Furthermore, there were more carbonate imperfections in the mineral crystal lattice of the caries tissues than in healthy tissues. Interestingly, we observed gradients in the sound enamel showing higher mineralization and greater mineral crystal perfection towards the tooth surface. To conclude, our results provide a baseline for methodological development aimed for clinical diagnostic of early detection of active caries lesions.

The influence of joint degeneration on the biomechanical properties of calcified cartilage and subchondral bone plate at the osteochondral junction is relatively unknown. Common experimental difficulties include accessibility to and visualization of the osteochondral junction, application of mechanical testing at the appropriate length scale, and availability of tissue that provides a consistent range of degenerative changes. This study addresses these challenges. A well-established bovine patella model of early joint degeneration was employed, in which micromechanical testing of fully hydrated osteochondral sections was carried out in conjunction with high-resolution imaging using differential interference contrast (DIC) optical light microscopy. A total of forty-two bovine patellae with different grades of tissue health ranging from healthy to mild, moderate, and severe cartilage degeneration, were selected. From the distal-lateral region of each patella, two adjacent osteochondral sections were obtained for the mechanical testing and the DIC imaging, respectively. Mechanical testing was carried out using a robotic micro-force acquisition system, applying compression tests over an array (area: 200 μm × 1000 μm, step size: 50 μm) across the osteochondral junction to obtain a stiffness map. Morphometric analysis was performed for the DIC images of fully hydrated cryo-sections. The levels of cartilage degeneration, DIC images, and the stiffness maps were used to associate the mechanical properties onto the specific tissue regions of cartilage, calcified cartilage, and subchondral bone plate. The results showed that there were up to 20% and 24% decreases (p < 0.05) in the stiffness of calcified cartilage and subchondral bone plate, respectively, in the severely degenerated group compared to the healthy group. Furthermore, there were increases (p < 0.05) in the number of tidemarks, bone spicules at the cement line, and the mean thickness of the subchondral bone plate with increasing levels of degeneration. The decreasing stiffness in the subchondral bone plate coupled with the presence of bone spicules may be indicative of a subchondral remodeling process involving new bone formation. Moreover, the mean thickness of the subchondral bone plate was found to be the strongest indicator of mechanical and associated structural changes in the osteochondral joint tissues.

Selenium (Se), an important micronutrient with several biological effects, exhibits improved stability, biocompatibility, and safety through nanoparticle (NP) formulation. We present here a novel approach for producing smaller and structurally stable biogenic selenium nanoparticles (SeNPs) with bioactive coating. Poly(allylamine hydrochloride)-coated SeNPs were synthesized biogenically using Trachyspermum ammi (TA) seed extract, in parallel to chemically synthesized SeNPs. NPs were characterized and assessed for their in vitro antioxidant/radical scavenging potential through multiple complementary chemical assays along with the intracellular reactive oxygen species (ROS) scavenging and biocompatibility assay using human dermal fibroblasts. Both NPs were spherical and positively charged, with TA-SeNPs having a smaller size and a more amorphous nature. X-ray photoelectron spectroscopy confirmed the presence of elemental Se. Fourier transform infrared spectroscopy (FTIR) indicated the functional groups common to both NPs, with unique aliphatic, aromatic and glycosidic linkages, and phenolic compounds in TA-SeNPs, attributed to the TA extract. Both NPs showed dose-dependent antioxidative properties, with TA-SeNPs exhibiting significantly increased antioxidant activity among all the tested samples and showing similar to relatively higher activity relative to L-ascorbic acid (AA) and TA extract in several assays. Viability assays revealed that both NPs were nontoxic at low concentrations; however, cell viability is decreased at high concentrations, indicating dose-dependent cytotoxicity. TA-SeNPs suppressed reactive oxygen species more effectively at low concentrations, whereas SeNPs showed stronger activity at high concentrations. Our findings showed that the TA extract-mediated biosynthesis offers a cost-effective and biologically favorable route to produce SeNPs, with potential for safer antioxidant nanomedicine applications.

In craniofacial research and routine dental clinical procedures, multifunctional materials with antimicrobial properties are in constant demand. Ionic liquids (ILs) are one such multifunctional intelligent material. Over the last three decades, ILs have been explored for different biomedical applications due to their unique physical and chemical properties, high task specificity, and sustainability. Their stable physical and chemical characteristics and extremely low vapor pressure make them suitable for various applications. Their unique properties, such as density, viscosity, and hydrophilicity/hydrophobicity, may provide higher performance as a potential dental material. ILs have functionalities for optimizing dental implants, infiltrate materials, oral hygiene maintenance products, and restorative materials. They also serve as sensors for dental chairside usage to detect oral cancer, periodontal lesions, breath-based sobriety, and dental hard tissue defects. With further optimization, ILs might also make vital contributions to craniofacial regeneration, oral hygiene maintenance, oral disease prevention, and antimicrobial materials. This review explores the different advantages and properties of ILs as possible dental material.