Regeneration of Articular Cartilage in Lizard Knee from Resident Stem/Progenitor Cells

The epiphysis of femur and tibia in the lizard Podarcis muralis can extensively regenerate after injury. The process involves the articular cartilage and metaphyseal (growth) plate after damage. The secondary ossification center present between the articular cartilage and the growth plate is replaced by cartilaginous epiphyses after about one month of regeneration at high temperature. The present study analyzes the origin of the chondrogenic cells from putative stem cells located in the growing centers of the epiphyses. The study is carried out using immunocytochemistry for the detection of 5BrdU-labeled long retaining cells and for the localization of telomerase, an enzyme that indicates stemness. The observations show that putative stem cells retaining 5BrdU and positive for telomerase are present in the superficial articular cartilage and metaphyseal growth plate located in the epiphyses. This observation suggests that these areas represent stem cell niches lasting for most of the lifetime of lizards. In healthy long bones of adult lizards, the addition of new chondrocytes from the stem cells population in the articular cartilage and the metaphyseal growth plate likely allows for slow, continuous longitudinal growth. When the knee is injured in the adult lizard, new populations of chondrocytes actively producing chondroitin sulfate proteoglycan are derived from these stem cells to allow for the formation of completely new cartilaginous epiphyses, possibly anticipating the re-formation of secondary centers in later stages. The study suggests that in this lizard species, the regenerative ability of the epiphyses is a pre-adaptation to the regeneration of the articular cartilage.     

Role of iRhoms 1 and 2 in Endochondral Ossification

Growth of the axial and appendicular skeleton depends on endochondral ossification, which is controlled by tightly regulated cell–cell interactions in the developing growth plates. Previous studies have uncovered an important role of a disintegrin and metalloprotease 17 (ADAM17) in the normal development of the mineralized zone of hypertrophic chondrocytes during endochondral ossification. ADAM17 regulates EGF-receptor signaling by cleaving EGFR-ligands such as TGFα from their membrane-anchored precursor. The activity of ADAM17 is controlled by two regulatory binding partners, the inactive Rhomboids 1 and 2 (iRhom1, 2), raising questions about their role in endochondral ossification. To address this question, we generated mice lacking iRhom2 (iR2−/−) with floxed alleles of iRhom1 that were specifically deleted in chondrocytes by Col2a1-Cre (iR1∆Ch). The resulting iR2−/−iR1∆Ch mice had retarded bone growth compared to iR2−/− mice, caused by a significantly expanded zone of hypertrophic mineralizing chondrocytes in the growth plate. Primary iR2−/−iR1∆Ch chondrocytes had strongly reduced shedding of TGFα and other ADAM17-dependent EGFR-ligands. The enlarged zone of mineralized hypertrophic chondrocytes in iR2−/−iR1∆Ch mice closely resembled the abnormal growth plate in A17∆Ch mice and was similar to growth plates in Tgfα−/− mice or mice with EGFR mutations. These data support a model in which iRhom1 and 2 regulate bone growth by controlling the ADAM17/TGFα/EGFR signaling axis during endochondral ossification.     

Molecular Mechanism of Induction of Bone Growth by the C-Type Natriuretic Peptide

The skeletal development process in the body occurs through sequential cellular and molecular processes called endochondral ossification. Endochondral ossification occurs in the growth plate where chondrocytes differentiate from resting, proliferative, hypertrophic to calcified zones. Natriuretic peptides (NPTs) are peptide hormones with multiple functions, including regulation of blood pressure, water-mineral balance, and many metabolic processes. NPTs secreted from the heart activate different tissues and organs, working in a paracrine or autocrine manner. One of the natriuretic peptides, C-type natriuretic peptide-, induces bone growth through several mechanisms. This review will summarize the knowledge, including the newest discoveries, of the mechanism of CNP activation in bone growth.     

New columnar texture of carbonado: Cathodoluminescence study

We investigated a very unusual sample of carbonado by optical and electron microscopy, and cathodoluminescence imaging and spectroscopy. We observed two textures: a microporphyritic texture, and columnar transparent diamonds growing perpendicularly.We propose that this sample grew in three stages: the slow growth of large porphyrocrysts (step 1), followed by a fast growth period that produced the fine-grained cement (step 2), and ultimate growth of columnar diamond perpendicular to the initial material (step 3). The first two steps growth have been extensively documented in the literature, and the third one is described for the first time in carbonado. The classical texture suggests a growth from flowing fluids while the columnar texture evidences diamond development in an open fracture.The orange (N–V⁰ center) and green (N₂V center) cathodoluminescence observed around pores, grain boundaries and fractures throughout the sample is due to irradiation of uranium-rich fluids. As uranium was not mobile at the time of carbonado formation (between 3.8 and 2.6 Ga), we propose that irradiation occurred most likely long after diamond growth. The blue CL of the porphyrocrysts and of the columnar diamond, preserved from late transient irradiation, is due to N₃V centers (and, to a lesser extent, N₂V centers). It is a primary feature which reveals a certain degree of nitrogen aggregation. This is more likely related to some high-temperature events that remain to be determined.     

Impaired Differentiation of Highly Proliferative ICOS+-Tregs Is Involved in the Transition from Low to High Disease Activity in Systemic Lupus Erythematosus (SLE) Patients

Dysregulations in the differentiation of CD4⁺-regulatory-T-cells (Tregs) and CD4⁺-responder-T-cells (Tresps) are involved in the development of active systemic lupus erythematosus (SLE). Three differentiation pathways of highly proliferative inducible costimulatory molecule (ICOS)⁺- and less proliferative ICOS⁻-CD45RA⁺CD31⁺-recent-thymic-emigrant (RTE)-Tregs/Tresps via CD45RA⁻CD31⁺-memory-Tregs/Tresps (CD31⁺-memory-Tregs/Tresps), their direct proliferation via CD45RA⁺CD31⁻-mature naïve (MN)-Tregs/Tresps, and the production and differentiation of resting MN-Tregs/Tresp into CD45RA⁻CD31⁻-memory-Tregs/Tresps (CD31⁻-memory-Tregs/Tresps) were examined in 115 healthy controls, 96 SLE remission patients, and 20 active disease patients using six color flow cytometric analysis. In healthy controls an appropriate sequence of these pathways ensured regular age-dependent differentiation. In SLE patients, an age-independently exaggerated differentiation was observed for all Treg/Tresp subsets, where the increased conversion of resting MN-Tregs/Tresps particularly guaranteed the significantly increased ratios of ICOS⁺-Tregs/ICOS⁺-Tresps and ICOS⁻-Tregs/ICOS⁻-Tresps during remission. Changes in the differentiation of resting ICOS⁺-MN-Tresps and ICOS⁻-MN-Tregs from conversion to proliferation caused a significant shift in the ratio of ICOS⁺-Tregs/ICOS⁺-Tresps in favor of ICOS⁺-Tresps and a further increase in the ratio of ICOS⁻-Tregs/ICOS⁻-Tresps with active disease. The differentiation of ICOS⁺-RTE-Tregs/Tresps seems to be crucial for keeping patients in remission, where their limited production of proliferating resting MN-Tregs may be responsible for the occurrence of active disease flares.     

Post-Traumatic Caspase-3 Expression in the Adjacent Areas of Growth Plate Injury Site: A Morphological Study

The epiphyseal plate is a hyaline cartilage plate that sits between the diaphysis and the epiphysis. The objective of this study was to determine the impact of an injury in the growth plate chondrocytes through the study of histological morphology, immunohistochemistry, histomorphometry and Western Blot analyses of the caspase-3 and cleaved PARP-1, and levels of the inflammatory cytokines, Interleukin-6 (IL-6) and Tumor Necrosis Factor alpha (TNF-α), in order to acquire more information about post-injury reactions of physeal cell turnover. In our results, morphological analysis showed that in experimental bones, neo-formed bone trabeculae—resulting from bone formation repair—invaded the growth plate and reached the metaphyseal bone tissue (bone bridge), and this could result in some growth arrest. We demonstrated, by ELISA, increased expression levels of the inflammatory cytokines IL-6 and TNF-α. Immunohistochemistry, histomorphometry and Western Blot analyses of the caspase-3 and cleaved PARP-1 showed that the physeal apoptosis rate of the experimental bones was significantly higher than that of the control ones. In conclusion, we could assume that the inflammation process causes stress to chondrocytes that will die as a biological defense mechanism, and will also increase the survival of new chondrocytes for maintaining cell homeostasis. Nevertheless, the exact stimulus leading to the increased apoptosis rate, observed after injury, needs additional research to understand the possible contribution of chondrocyte apoptosis to growth disturbance.     

Positron annihilation spectroscopy of sputtered boron carbide films

Positron annihilation spectroscopy (PAS) was carried out on boron carbide films deposited by sputtering and the results correlated to the bombardment conditions during film growth. Films were deposited with substrate bias voltages in the range of 0 to −200 V with a working pressure of 5 mTorr of Ar. Films deposited with bias voltages from −100 to −200 V present the same type of defect and the defect concentration increased linearly with the bias voltage. This defect was ascribed to vacancies in agreement with Monte Carlo simulations of Ar⁺ bombardment of boron carbide. On the other hand, films deposited with 0 V bias presented a higher S parameter values, whose origin was tentatively attributed to a relatively more open nanosized columnar structure, as suggested by the structure zone model. Annealing up to 800 °C for 30 min did not change the defect content.     

The Roles of Sodium-Independent Inorganic Phosphate Transporters in Inorganic Phosphate Homeostasis and in Cancer and Other Diseases

Inorganic phosphate (Pi) is an essential nutrient for the maintenance of cells. In healthy mammals, extracellular Pi is maintained within a narrow concentration range of 0.70 to 1.55 mM. Mammalian cells depend on Na⁺/Pi cotransporters for Pi absorption, which have been well studied. However, a new type of sodium-independent Pi transporter has been identified. This transporter assists in the absorption of Pi by intestinal cells and renal proximal tubule cells and in the reabsorption of Pi by osteoclasts and capillaries of the blood–brain barrier (BBB). Hyperphosphatemia is a risk factor for mineral deposition, the development of diseases such as osteoarthritis, and vascular calcifications (VCs). Na⁺-independent Pi transporters have been identified and biochemically characterized in vascular smooth muscle cells (VSMCs), chondrocytes, and matrix vesicles, and their involvement in mineral deposition in the extracellular microenvironment has been suggested. According to the growth rate hypothesis, cancer cells require more phosphate than healthy cells due to their rapid growth rates. Recently, it was demonstrated that breast cancer cells (MDA-MB-231) respond to high Pi concentration (2 mM) by decreasing Na⁺-dependent Pi transport activity concomitant with an increase in Na⁺-independent (H⁺-dependent) Pi transport. This Pi H⁺-dependent transport has a fundamental role in the proliferation and migratory capacity of MDA-MB-231 cells. The purpose of this review is to discuss experimental findings regarding Na⁺-independent inorganic phosphate transporters and summarize their roles in Pi homeostasis, cancers and other diseases, such as osteoarthritis, and in processes such as VC.     

Studies on the use of chromium ferrocyanide gels in the treatment of radioactive wastes

The sorption and distribution of some radioisotopes on chromium ferrocyanide gels under different physical conditions has been investigated. The exchanger shows high selectivity for Tl⁺, Cs⁺, Cd²⁺ and Pb²⁺. A complete elution of Cs⁺, Tl⁺ and Rb⁺ is possible with suitable eluants whereas Pb²⁺ and Cd²⁺ cannot be desorbed. The uptake of monovalent ions is by an ion exchange process unlike the sorption of bivalent ions.     

The synthesis and characterization of Cu(II) phthalocyanine bearing peripheral monoazacrown ethers and a spectral investigation of its film forming character

The synthesis, liquid crystalline behavior and self-organizing properties of Cu(II)Pc bearing monoazacrown ether moieties that contain long alkyloxyphenyl N-pivotal groups are described. The Cu(II) phthalocyanine derivative has a hexagonal, columnar structure; optical absorption and vibrational spectroscopy revealed a predominantly co-facial interaction between the chromophores and their orientation parallel to a NaCl surface. It is proposed that such homeotropic alignment is attributable to the ability of the pendant monoazacrown ether groups to coordinate the Na⁺ ions within the NaCl surface.     

Antimicrobial resistance of clay polymer nanocomposites

Antimicrobial-resistant polymeric Na⁺–bentonite nanocomposites were prepared by treating Na⁺–bentonite (Na⁺–Bent) with polymeric ultra-thin films of poly(diallyldimethyl ammonium chloride) (PDADMAC), poly(methylmethacrylate) (PMMA) and poly(vinylidene chloride) (PVDC) by admicellar polymerization technique. The clay polymer nanocomposites (CPNs) were characterized by several techniques including Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), BET surface analysis, thermogravimetric analysis (TGA) and scanning electron microscopy (SEM). In additional, the antimicrobial resistance was studied by measuring the diameter of inhibition zone of growths of Escherichia coli and Salmonella typhimurium. The results showed an inhibitory effect of these CPN against microbial growth in inoculated samples. The CPN exhibited efficacy in the inhibition of bacterial growth.     

Luminescence of modified nonbridging oxygen hole centers in silica and alkali silicate glasses

This paper reports on the results of the investigation of the cathodoluminescence spectra of silica and alkali silicate glasses upon excitation with a pulsed electron beam (energy, 180 keV; current density, 700 A/cm²; pulse duration, 2 ns). The luminescence band observed in the energy range 2.4–2.6 eV is assigned to modified structural defects of the ≡Si-O·/Me ⁺ type. These defects are revealed under high-density electronic excitation and, unlike the known L centers in alkali silicate glasses, are interpreted as a variety of nonbridging oxygen hole centers (defects of the dangling bond type) subjected to a disturbing action of the nearest neighbor alkali metal cations. The cathodoluminescence of similar centers is observed in neutron-irradiated silica glasses with lithium impurities; alkali silicate glasses with Li, Na, and K cations; and glasses in the two-alkali Na-K systems. It is established that the energy of the radiative transition of a modified nonbridging oxygen hole center, namely, ≡Si-O·/Me ⁺, depends on the alkali cation type.     

The crystallization behaviours of reactive-plasma-sprayed TiCN coatings with different Ti/graphite powder ratios

TiCN coatings were reactive plasma sprayed with different Ti/graphite powder ratios. Their crystallization behaviours were investigated. The results showed that the TiN coating consisted of TiN and TiN_0.3O phases. With the decrease in the Ti/graphite ratios, the phases in the coatings changed from TiC_0.7N_0.3+TiC_0.3N_0.7+amorphous (Ti:graphite = (6-10):1) to TiC_0.3N_0.7 + TiC + amorphous (Ti:graphite = 4:1). The surface of the TiN coating exhibited the columnar crystals. With the decrease in the Ti/graphite ratios, the columnar crystals gradually skewed towards the surface. The cross-section crystalline morphologies of all the coatings exhibited the layer-layer columnar crystals with better bonding interfaces between the layers. The micro-hardness of the sprayed TiN coating was 1000 HV_0.1. The micro-hardness of the TiCN coatings increased with the decrease in the Ti/graphite ratios. The maximum microhardness (1315 HV_0.1) was observed at a Ti/graphite ratio of 4:1, approximately 30% higher than that of the TiN coating. The increase in the micro-hardness was correlated with the phase composition and crystalline morphologies.     

A thermodynamic study of the effect of small-scale electrolytes on equilibrium redox potentials

Expressions are derived to calculate the equilibrium oxidation-reduction potentials for the Al⁺³/Al, Cu⁺²/Cu, and Zn⁺²/Zn systems in small-scale electrolytes. The geometrical system consists of a droplet of electrolyte resting on a flat metal plate, and the metal is considered to be immersed in a solution of its own ions. When the radius of the drop is allowed to vary, both the size of the electrolyte and the size of the active metal beneath the droplet change simultaneously. The total free energy change for the system consists of both electrochemical and surface chemical contributions. The interfacial free energy for the solid/liquid interface has been estimated from the Girifalco-Good expression or from spreading pressure considerations. When the droplet becomes sufficiently small in radius, the surface chemical contributions become significant, and the calculated redox potential changes from its normal value to more negative values as the size of the system decreases. The magnitude of this effect depends on the particular system. For 2 M Cu⁺², the calculated redox potential for a 0.8 nm radius droplet is 0.259 V more negative than for the bulk electrolyte. The effect is much smaller for aluminum and zinc. In all three systems, calculated redox potentials approach values for the bulk solution for droplet radii of about 10 nm.     

Cationic RuII Cyclopentadienyl Complexes with Antifungal Activity against Several Candida Species

Fungal infections, including those caused by antifungal-resistant Candida, are a very challenging health problem worldwide. Whereas different ruthenium complexes were previously studied for their anti-Candida potential, Ru-cyclopentadienyl complexes were overlooked. Here, we report an antifungal activity assessment of three Ru-cyclopentadienyl complexes with some insights into their potential mode of action. Among these complexes, only the cationic species [Ru-ACN]⁺ and [Ru-ATZ]⁺ displayed a significant antifungal activity against different Candida strains, notably against the ones that did not respond to one of the most currently used antifungal drugs fluconazole (FCZ). However, no apparent activity was observed for the neutral species, Ru−Cl, thus indicating the important role of the cationic backbone of these complexes in their biological activity. We suggest that reactive oxygen species (ROS) generation might be involved in the mechanism of action of these complexes as, unlike neutral Ru−Cl, [Ru-ACN]⁺ and [Ru-ATZ]⁺ could generate intracellular concentration-dependent ROS. We also observed a correlation between the ruthenium cellular uptake, ROS generation and fungal growth inhibitory activity of the compounds. Furthermore, docking simulations showed that the CYP51 enzyme can form more energetically favorable complexes with [Ru-ATZ]⁺ than fluconazole (FCZ); this suggests that CYP51 inhibition could also be considered as a potential mode of action.     

Could BMPs Therapy Be Improved if BMPs Were Used in Composition Acting during Bone Formation in Endochondral Ossification?

The discovery of bone morphogenetic proteins (BMPs) inspired hope for the successful treatment of bone disorders, but side effects worsening the clinical effects were eventually observed. BMPs exert a synergistic effect, stimulating osteogenesis; however, predicting the best composition of growth factors for use in humans is difficult. Chondrocytes present within the growth plate produce growth factors stored in calcified cartilage adhering to metaphysis. These factors stimulate initial bone formation in metaphysis. We have previously determined the growth factors present in bovine calcified cartilage and produced by rat epiphyseal chondrocytes. The results suggest that growth factors stimulating physiological ossification are species dependent. The collection of human calcified cartilage for growth factors determination does not appear feasible, but chondrocytes for mRNA determination could be obtained. Their collection from young recipients, in view of the Academy of Medical Royal Colleges Recommendation, would be ethical. The authors of this review do not have facilities to conduct such a study and can only appeal to competent institutions to undertake the task. The results could help to formulate a better recipe for the stimulation of bone formation and improve clinical results.     

Tumor Extracellular Matrix Stiffness Promptly Modulates the Phenotype and Gene Expression of Infiltrating T Lymphocytes

The immune system is a fine modulator of the tumor biology supporting or inhibiting its progression, growth, invasion and conveys the pharmacological treatment effect. Tumors, on their side, have developed escaping mechanisms from the immune system action ranging from the direct secretion of biochemical signals to an indirect reaction, in which the cellular actors of the tumor microenvironment (TME) collaborate to mechanically condition the extracellular matrix (ECM) making it inhospitable to immune cells. TME is composed of several cell lines besides cancer cells, including tumor-associated macrophages, cancer-associated fibroblasts, CD4⁺ and CD8⁺ lymphocytes, and innate immunity cells. These populations interface with each other to prepare a conservative response, capable of evading the defense mechanisms implemented by the host’s immune system. The presence or absence, in particular, of cytotoxic CD8⁺ cells in the vicinity of the main tumor mass, is able to predict, respectively, the success or failure of drug therapy. Among various mechanisms of immunescaping, in this study, we characterized the modulation of the phenotypic profile of CD4⁺ and CD8⁺ cells in resting and activated states, in response to the mechanical pressure exerted by a three-dimensional in vitro system, able to recapitulate the rheological and stiffness properties of the tumor ECM.     

The Effect of Orally Supplemented Melatonin on Larval Performance and Skeletal Deformities in Farmed Gilthead Seabream (Sparus aurata)

The gilthead seabream larval rearing in continuous light is common in most Mediterranean hatcheries to stimulate larval length growth and increase food consumption. Several studies have shown that continuous light affects larval development and increases the prevalence of skeletal deformities. Melatonin is a crucial pineal neurohormone that displays daily secretion patterns, stimulates cell proliferation and embryonic development in Atlantic salmon and zebrafish, and improves osseointegration in mice and humans. However, no studies have examined the effects of orally supplemented melatonin on skeletal deformities in Sparus aurata larvae. We administered exogenous melatonin to gilthead seabream larvae via enriched rotifers and nauplii of Artemia. Exogenous melatonin induced bone deformities and stimulated parathyroid hormone-related protein-coding gene (PTHrP) mRNA expression. In addition to the melatonin-induced PTHrP high expression level, the recorded non coordinated function of skeletal muscle and bone during growth can be the fountainhead of bone deformities. Both myosin light chain 2 (mlc2) and bone gamma-carboxyglutamate protein-coding gene (bglap) expression levels were significantly affected by melatonin administration in an inverse dose–response manner during the exogenous melatonin administration. This is the first study to report the effect of inducing melatonin bone deformities on Sparus aurata larvae reared under ordinary hatchery conditions.     

Nuclear Magnetic Resonance Therapy Modulates the miRNA Profile in Human Primary OA Chondrocytes and Antagonizes Inflammation in Tc28/2a Cells

Nuclear magnetic resonance therapy (NMRT) is discussed as a participant in repair processes regarding cartilage and as an influence in pain signaling. To substantiate the application of NMRT, the underlying mechanisms at the cellular level were studied. In this study microRNA (miR) was extracted from human primary healthy and osteoarthritis (OA) chondrocytes after NMR treatment and was sequenced by the Ion PI Hi-Q™ Sequencing 200 system. In addition, T/C-28a2 chondrocytes grown under hypoxic conditions were studied for IL-1β induced changes in expression on RNA and protein level. HDAC activity an NAD⁽⁺⁾/NADH was measured by luminescence detection. In OA chondrocytes miR-106a, miR-27a, miR-34b, miR-365a and miR-424 were downregulated. This downregulation was reversed by NMRT. miR-365a-5p is known to directly target HDAC and NF-ĸB, and a decrease in HDAC activity by NMRT was detected. NAD⁺/NADH was reduced by NMR treatment in OA chondrocytes. Under hypoxic conditions NMRT changed the expression profile of HIF1, HIF2, IGF2, MMP3, MMP13, and RUNX1. We conclude that NMRT changes the miR profile and modulates the HDAC and the NAD⁽⁺⁾/NADH signaling in human chondrocytes. These findings underline once more that NMRT counteracts IL-1β induced changes by reducing catabolic effects, thereby decreasing inflammatory mechanisms under OA by changing NF-ĸB signaling.