Induced regeneration of articular cartilage - identification of a dormant regeneration program for a non-regenerative tissue.

A mouse organoid culture model was developed to regenerate articular cartilage by sequential treatment with BMP2 and BMP9 (or GDF2) that parallels induced joint regeneration at digit amputation wounds in vivo. BMP9-induced chondrogenesis was used to identify clonal cell lines for articular chondrocyte and hypertrophic chondrocyte progenitor cells from digit fibroblasts. A protocol that includes cell aggregation enhanced by BMP2 followed by BMP9-induced chondrogenesis resulted in the differentiation of organized layers of articular chondrocytes, similar to the organization of middle and deep zones of articular cartilage in situ, and retained a differentiated phenotype following transplantation. In addition, the differentiation of a non-chondrogenic connective tissue layer containing articular chondrocyte progenitor cells demonstrated that progenitor cell sequestration is coupled with articular cartilage differentiation at a clonal level. The studies identify a dormant endogenous regenerative program for a non-regenerative tissue in which fibroblast-derived progenitor cells can be induced to initiate morphogenetic and differentiative programs that include progenitor cell sequestration. The identification of dormant regenerative programs in non-regenerative tissues such as articular cartilage represents a novel strategy that integrates regeneration biology with regenerative medicine.

Deficiency and excess of groundwater iodine and their health associations.

More than two billion people worldwide have suffered thyroid disorders from either iodine deficiency or excess. By creating the national map of groundwater iodine throughout China, we reveal the spatial responses of diverse health risks to iodine in continental groundwater. Greater non-carcinogenic risks relevant to lower iodine more likely occur in the areas of higher altitude, while those associated with high groundwater iodine are concentrated in the areas suffered from transgressions enhanced by land over-use and intensive anthropogenic overexploitation. The potential roles of groundwater iodine species are also explored: iodide might be associated with subclinical hypothyroidism particularly in higher iodine regions, whereas iodate impacts on thyroid risks in presence of universal salt iodization exhibit high uncertainties in lower iodine regions. This implies that accurate iodine supply depending on spatial heterogeneity and dietary iodine structure optimization are highly needed to mitigate thyroid risks in iodine-deficient and -excess areas globally.

Understanding the Importance of Labile Fe(III) during Fe(II)-Catalyzed Transformation of Metastable Iron Oxyhydroxides.

Transformation of metastable Fe(III) oxyhydroxides is a prominent process in natural environments and can be significantly accelerated by the coexisting aqueous Fe(II) (Fe(II)aq). Recent evidence points to the solution mass transfer of labile Fe(III) (Fe(III)labile) as the primary intermediate species of general importance. However, a mechanistic aspect that remains unclear is the dependence of phase outcomes on the identity of the metastable Fe(III) oxyhydroxide precursor. Here, we compared the coupled evolution of Fe(II) species, solid phases, and Fe(III)labile throughout the Fe(II)-catalyzed transformation of lepidocrocite (Lp) versus ferrihydrite (Fh) at equal Fe(III) mass loadings with 0.2-1.0 mM Fe(II)aq at pH = 7.0. Similar to Fh, the conversion of Lp to product phases occurs by a dissolution-reprecipitation mechanism mediated by Fe(III)labile that seeds the nucleation of products. Though for Fh we observed a transformation to goethite (Gt), accompanied by the transient emergence and decline of Lp, for initial Lp we observed magnetite (Mt) as the main product. A linear correlation between the formation rate of Mt and the effective supersaturation in terms of Fe(III)labile concentration shows that Fe(II)-induced transformation of Lp into Mt is governed by the classical nucleation theory. When Lp is replaced by equimolar Gt, Mt formation is suppressed by opening a lower barrier pathway to Gt by heterogeneous nucleation and growth on the added Gt seeds. The collective findings add to the mechanistic understanding of factors governing phase selections that impact iron bioavailability, system redox potential, and the fate and transport of coupled elements.

Effects of pre-oxidation on residual dissolved aluminum in coagulated water: A pilot-scale study.

Residual dissolved aluminum (Al) in drinking water is becoming a serious concern due to its high potential risks to human health. However, the mechanism by which residual dissolved Al forms is yet to be elucidated in detail. In this study, the effects of pre-oxidation by ozonation and chlorination on the properties of dissolved organic matter (DOM) and residual Al concentrations remaining in solution after coagulation were explored in a pilot-scale test. Changes in the DOM properties caused by the water treatment process were characterized by ultraviolet-visible absorbance spectroscopy. Theprotonation-active sites, carboxylic- and phenolic-type groupsof DOM were quantified by spectral parameter DlnA400 (differential log-transformed spectra at wavelength 400 nm) in combination with the revised non-ideal competitive adsorption model. The results show that ozonation and chlorination significantly affect the properties of DOM and the amount of residual dissolved Al in coagulated drinking water. This effect was associated with the changes in the carboxylic- and phenolic-type groups in DOM. Results of the study show that residual dissolved Al in coagulated water can be controlled by affecting theAl binding sites in DOM by pre-oxidation before coagulation. The nature of pre-oxidation agent and its dosage should be selected depending on the quality of the raw water to be treated. Ozonation was concluded to be preferable pre-oxidation agent for the water in examined this study.

Interpreting main features of the differential absorbance spectra of chlorinated natural organic matter: Comparison of the experimental and theoretical spectra of model compounds.

This study compared chlorination-induced changes of the properties of natural organic matter (NOM) represented by standard humic substances and NOM present in pristine and anthropogenically-affect reservoirs, rivers, groundwater and seawater. The chlorination-induced changes of NOM properties were quantified using the differential absorbance spectra (DAS) which were processed via numeric deconvolution. Six Gaussian bands were found to comprise the DAS of all examined waters. These bands (denoted as A0, A1, A2, A3, A4 and A5, respectively) have maxima located at ca. 200, 240, 276, 316, 385 and 547 nm. The bands A1-A4 were observed in the DAS of representative model chlorinated compounds. Quantum chemical (QC) calculations were carried out to examine the intrinsic nature of these bands and electronic transitions associated with them. QC data demonstrate that bands A1 and A2 are present in almost all aromatic organic species, A3 is likely to be associated with acetophenone- and/or styrene-like groups. A4 can be attributed to the engagement of m-hydroxyaromatic and flavone-type groups typical for the polyphenolic moiety in NOM and known to be the key precursors of disinfection by-product (DBP) formation. Thus, the intensity of band A4 is predicted to be an especially strong predictor of DBP formation.

Recovery of phosphorus and metallic nickel along with HCl production from electroless nickel plating effluents: The key role of three-compartment photoelectrocatalytic cell system.

A three-compartment photoelectrocatalytic (PEC) cell system combined with ion exchange and chemical precipitation was proposed to recover phosphorus and nickel from electroless nickel plating effluents containing hypophosphite (H2PO2-) and nickel ions (Ni2+). Ion exchange was used to concentrate and separate Ni2+ and H2PO2-. As a key unit, the established PEC system consisted of TiO2/Ni-Sb-SnO2 photoanode and Ti cathode. With 25.8 mM NaH2PO2 and 500 mM NiCl2, 100 % H2PO2- was oxidized to PO43- in the anode cell, 78 % Ni2+ was recovered as metallic Ni in the cathode cell, and 900 mM HCl was obtained in the middle cell within 24 h at 3.0 V. Based on quenching experiments and ESR technique, OH radicals were mainly responsible for H2PO2- oxidation. In situ Raman spectroscopy indicated that Ni2+ initially reacted with OH- to form α-Ni(OH)2, which was gradually reduced to metallic Ni. Fortunately, a slight pH decrease in the cathode cell in the three-compartment cell system was beneficial for Ni2+ reduction to Ni°. The obtained PO43- was recovered by chemical precipitation. Finally, recovery of phosphorus and metallic nickel along with HCl production from an actual electroless nickel plating effluents in terms of efficiency, cost-benefit, and stability assessment were demonstrated.

Comparing electron donating/accepting capacities (EDC/EAC) between crop residue-derived dissolved black carbon and standard humic substances.

Although dissolved black carbon (DBC) is an important component of dissolved organic matter (DOM), little is known about the electron transfer properties of DBC and their correlation with the structural properties. This study determined the electron donating/accepting capacities (EDC/EAC) of six DBC collected from water extracts of black carbon derived by pyrolyzing crop residues (soybean, wheat, rice, sorghum, peanut, and corn) using an electrochemical approach, along with their chemical and spectroscopic properties. The EDC of the tested DBC ranged from 2.42 to 7.10 mmole-(gC)-1and was generally much higher than the EDC (1.31-3.78 mmole-(gC)-1) of the four standard dissolved humic substances (DHS). In contrast, with the exception of the sorghum DBC, the EAC of all DBC (0.40-0.81 mmole-(gC)-1) was apparently lower than the EAC of DHS (0.87-1.68 mmole-(gC)-1). For the whole pool of DBC and DHS, a strong positive correlation (r = 0.94, P < 0.05) existed between the EDC and the phenolic group content quantified by base titration, suggesting that phenolic groups were the major electron-donating groups. The EAC and EDC of the tested DOM pool were further correlated with the components identified from the fluorescence excitation emission matrices and parallel factor (EEM-PARAFAC) analysis. This is the first study to show that DBC has significantly different electron transfer properties from humic substances and thus likely behaviors differently in many geochemical and environmental aspects. Capsule: Dissolved black carbon (DBC) has significantly different electron-donating/accepting capacities (EDC/EAC) from humic substances.

Macrophages are required to coordinate mouse digit tip regeneration.

In mammals, macrophages are known to play a major role in tissue regeneration. They contribute to inflammation, histolysis, re-epithelialization, revascularization and cell proliferation. Macrophages have been shown to be essential for regeneration in salamanders and fish, but their role has not been elucidated in mammalian epimorphic regeneration. Here, using the regenerating mouse digit tip as a mammalian model, we demonstrate that macrophages are essential for the regeneration process. Using cell-depletion strategies, we show that regeneration is completely inhibited; bone histolysis does not occur, wound re-epithelialization is inhibited and the blastema does not form. Although rescue of epidermal wound closure in the absence of macrophages promotes blastema accumulation, it does not rescue cell differentiation, indicating that macrophages play a key role in the redifferentiation of the blastema. We provide additional evidence that although bone degradation is a component, it is not essential to the overall regenerative process. These findings show that macrophages play an essential role in coordinating the epimorphic regenerative response in mammals.

Deposition behavior of residual aluminum in drinking water distribution system: Effect of aluminum speciation.

Finished drinking water usually contains some residual aluminum. The deposition of residual aluminum in distribution systems and potential release back to the drinking water could significantly influence the water quality at consumer taps. A preliminary analysis of aluminum content in cast iron pipe corrosion scales and loose deposits demonstrated that aluminum deposition on distribution pipe surfaces could be excessive for water treated by aluminum coagulants including polyaluminum chloride (PACl). In this work, the deposition features of different aluminum species in PACl were investigated by simulated coil-pipe test, batch reactor test and quartz crystal microbalance with dissipation monitoring. The deposition amount of non-polymeric aluminum species was the least, and its deposition layer was soft and hydrated, which indicated the possible formation of amorphous Al(OH)3. Al13 had the highest deposition tendency, and the deposition layer was rigid and much less hydrated, which indicated that the deposited aluminum might possess regular structure and self-aggregation of Al13 could be the main deposition mechanism. While for Al30, its deposition was relatively slower and deposited aluminum amount was relatively less compared with Al13. However, the total deposited mass of Al30 was much higher than that of Al13, which was attributed to the deposition of particulate aluminum matters with much higher hydration state. Compared with stationary condition, stirring could significantly enhance the deposition process, while the effect of pH on deposition was relatively weak in the near neutral range of 6.7 to 8.7.

Study of iron and aluminum binding to Suwannee River fulvic acid using absorbance and fluorescence spectroscopy: comparison of data interpretation based on NICA-Donnan and Stockholm humic models.

This study examined the evolution of absorbance and fluorescence spectra of standard Suwannee River fulvic acid (SRFA) induced by its interactions with iron and aluminum. The results show that changes of SRFA absorbance are associated with a consistent response of the carboxylic and phenolic functional groups to iron and aluminum forming bonds with these groups, and their deprotonation induced by such binding. The observed changes of SRFA absorbance were quantified via the use of DSlope325-375 parameter that determines the behavior of the slope of logarithms of SRFA absorbance in the range of wavelengths 325-375 nm in the presence of varying concentrations of iron or aluminum. DSlope325-375 values were correlated linearly with the concentration of SRFA-bound iron and aluminum determined using either NICA-Donnan or Stockholm Humic Model (SHM) but the correlation was stronger for the former model (R(2) > 0.98). The slopes of these correlations were similar for both iron and aluminum concentrations <10.0 µM and at a wide pH range. Fluorescence of SRFA was responsive to metal binding but it changed less consistently in the presence of the examined metals, especially in the case of aluminum. The combination of these techniques can help explore in more detail manifestations of DOM site specificity at realistically low concentrations of DOM and metal ions.

Optical property of iron binding to Suwannee River fulvic acid.

In this work, absorbance and fluorescence spectra were used to study iron binding to standard Suwannee River fulvic acid (SRFA). The differential logarithm-transformed absorbance and fluorescence spectra of SRFA induced by iron binding were processed to examine the nature of the observed phenomena and to investigate the contributions of discrete binding sites present in SRFA. Both the Fe-differential log-transformed absorbance and fluorescence were well correlated to the bound iron concentrations predicted based on the Non-ideal Competitive Adsorption (NICA-Donnan) model at iron concentrations below 10.0µM (R(2)>0.99 for absorbance and R(2)>0.97 for fluorescence) and over a wide pH range of 3.5-8.0. At pH3.5, both the Fe-differential log-transformed absorbance and fluorescence vs. iron bound spectra exhibited significantly lower slopes than those at pH5.0, 7.0, and 8.0. These results suggest that a different set of complexation-active chromophores and fluorophores are responsible for iron binding at low pH values or that the NICA-Donnan model is limited at low pH. Because phenolic and carboxylic complex sites of different fluorophores respond to iron quenching, the fluorescence data indicate three stages of iron binding to phenolic, carboxylic, and Donnan gels (electrostatic interactions) in SRFA (with R(2)>0.99 at each stage). The agreement between observations from spectroscopic indices and established metal-binding models shows that the absorbance and fluorescence spectra provide important information about the involvement of metal complexation of specific functional groups typical for fulvic acids.

Mouse embryonic diastema region is an ideal site for the development of ectopically transplanted tooth germ.

The anterior eye chamber and the kidney capsule of the mouse have been traditionally used for long-term culture of tooth germ grafts. However, although these sites provide an excellent growth environment, they do not represent real in situ sites for the development of a grafted tooth germ. Here, we describe a protocol to transplant a tooth germ into the mandibular diastema region of mouse embryos using exo utero surgery. Our results demonstrate that the mouse embryonic diastema region represents a normal physiological environment for the development of transplanted tooth germs. Transplanted tooth germs developed synchronically with and became indistinguishable from the endogenous ones. These ectopic teeth were vascularized and surrounded with nerve fibers, and were able to erupt normally. Thus, the exo utero transplantation approach will provide a new avenue to study tooth development and regeneration.

Shox2 is required for chondrocyte proliferation and maturation in proximal limb skeleton.

Mutations in the short stature homeobox gene SHOX lead to growth retardation associated with Turner, Leri-Weill dyschondrosteosis, and Langer mesomelic dysplasia syndromes, which marked the shortening of the forearms and lower legs. We report here that in contrast to the SHOX mutations in humans, Shox2 deficiency in mice leads to a virtual elimination of the stylopod in the developing limbs, while the zeugopod and autopod appear relatively normal. This phenotype is consistent with the restriction of the Shox2 expression to the proximal mesenchyme in the limb bud and later to chondrocytes associated with the forming stylopod. In the Shox2(-/-) embryo, the mesenchymal condensation for the stylopod initiates normally but the cartilaginous element subsequently fails in growth, chondrogenesis and endochondral ossification. A dramatic down-regulation of Runx2 and Runx3 could account for the lack of chondrocyte hypertrophy, while a down-regulation of Ihh expression may be responsible for a significant reduction in chondrocyte proliferation in the mutant stylopod. We further demonstrate that an enhanced and ectopic Bmp4 expression in the proximal limb of the Shox2 embryo may underlie the down-regulation of Runx2, as ectopically applied exogenous BMP4 represses Runx2 expression in the early limb bud. Moreover, we show that mouse Shox2, similar to human SHOX, can perform opposite roles on gene expression: either as a transcription activator or a repressor in different cell types. Our results establish a key role for Shox2 in regulating the growth of stylopod by controlling chondrocyte maturation via Runx2 and Runx3.