Hydrophilic nanofibers with aligned topography modulate macrophage-mediated host responses via the NLRP3 inflammasome.

Successful biomaterial implantation requires appropriate immune responses. Macrophages are key mediators involved in this process. Currently, exploitation of the intrinsic properties of biomaterials to modulate macrophages and immune responses is appealing. In this study, we prepared hydrophilic nanofibers with an aligned topography by incorporating polyethylene glycol and polycaprolactone using axial electrospinning. We investigated the effect of the nanofibers on macrophage behavior and the underlying mechanisms. With the increase of hydrophilicity of aligned nanofibers, the inflammatory gene expression of macrophages adhering to them was downregulated, and M2 polarization was induced. We further presented clear evidence that the inflammasome NOD-like receptor thermal protein domain associated protein 3 (NLRP3) was the cellular sensor by which macrophages sense the biomaterials, and it acted as a regulator of the macrophage-mediated response to foreign bodies and implant integration. In vivo, we showed that the fibers shaped the implant-related immune microenvironment and ameliorated peritendinous adhesions. In conclusion, our study demonstrated that hydrophilic aligned nanofibers exhibited better biocompatibility and immunological properties.

Single cell protein production from methane in a gas-delivery membrane bioreactor.

Single cell protein (SCP, or microbial protein) is one of the emerging alternative protein sources to address the global challenge of food insecurity. Recently, the SCP produced from methane has attracted substantial attention since methane is a renewable resource attainable from anaerobic digestion. However, the supply of methane, an insoluble gas in water, is one of the major challenges in producing methane-based SCP. This work developed a novel bioreactor configuration, in which hollow fiber membrane was used for efficient methane supply while microorganisms were growing in the suspended form favourable for the biomass harvest. Over a 312-day operation, the impacts of three critical parameters on the SCP production were investigated, including the ratio of methane loading to ammonium loading, the ratio of methane loading to oxygen loading, and the sludge retention time (SRT). Under the condition of 4 g CH4/g NH4+, 4 g O2/g CH4, and SRT of 4 days, the highest SCP production yield was observed and determined to be 1.36 g SCP/g CH4 and 5.05 g SCP/g N, respectively. The protein content was up to 67 %, which is higher than the majority of reported values to date. Moreover, the methane and ammonium utilization efficiencies were both close to 100 %, suggesting the highly efficient utilization of substrates in this new bioreactor configuration. A high relative abundance of essential amino acids (EAA) above 42 % was achieved, representing the highest EAA content reported. These findings provide valuable insights into SCP production using methane as a feedstock.

Superior mainstream partial nitritation in an acidic membrane-aerated biofilm reactor.

Shortcut nitrogen removal holds significant economic appeal for mainstream wastewater treatment. Nevertheless, it is too difficult to achieve the stable suppression of nitrite-oxidizing bacteria (NOB), and simultaneously maintain the activity of ammonia-oxidizing bacteria (AOB). This study proposes to overcome this challenge by employing the novel acid-tolerant AOB, namely "Candidatus Nitrosoglobus", in a membrane-aerated biofilm reactor (MABR). Superior partial nitritation was demonstrated in low-strength wastewater from two aspects. First, the long-term operation (256 days) under the acidic pH range of 5.0 to 5.2 showed the successful NOB washout by the in situ free nitrous acid (FNA) of approximately 1 mg N/L. This was evidenced by the stable nitrite accumulation ratio (NAR) close to 100 % and the disappearance of NOB shown by 16S rRNA gene amplicon sequencing and fluorescence in situ hybridization. Second, oxygen was sufficiently supplied in the MABR, leading to an unprecedentedly high ammonia oxidation rate (AOR) at 2.4 ± 0.1 kg N/(m3 d) at a short hydraulic retention time (HRT) of a mere 30 min. Due to the counter diffusion of substrates, the present acidic MABR displayed a significantly higher apparent oxygen affinity (0.36 ± 0.03 mg O2/L), a marginally lower apparent ammonia affinity (14.9 ± 1.9 mg N/L), and a heightened sensitivity to FNA and pH variations, compared with counterparts determined by flocculant acid-tolerant AOB. Beyond supporting the potential application of shortcut nitrogen removal in mainstream wastewater, this study also offers the attractive prospect of intensifying wastewater treatment by markedly reducing the HRT of the aerobic unit.

Nitrogen removal from wastewater by coupling anammox and methane-dependent denitrification in a membrane biofilm reactor.

This work demonstrates, for the first time, the feasibility of nitrogen removal by using the synergy of anammox and denitrifying anaerobic methane oxidation (DAMO) microorganisms in a membrane biofilm reactor (MBfR). The reactor was fed with synthetic wastewater containing nitrate and ammonium. Methane was delivered from the interior of hollow fibres in the MBfR to the biofilm that grew on the fiber's outer wall. After 24 months of operation, the system achieved a nitrate and an ammonium removal rate of about 190 mgN L(-1) d(-1) (or 86 mgN m(-2) d(-1), with m(2) referring to biofilm surface area) and 60 mgN L(-1) d(-1) (27 mgN m(-2) d(-1)), respectively. No nitrite accumulation was observed. Fluorescence in situ hybridization (FISH) analysis indicated that DAMO bacteria (20-30%), DAMO archaea (20-30%) and anammox bacteria (20-30%) jointly dominated the microbial community. Based on the known metabolism of these microorganisms, mass balance, and isotope studies, we hypothesize that DAMO archaea converted nitrate, both externally fed and produced by anammox, to nitrite, with methane as the electron donor. Anammox and DAMO bacteria jointly removed the nitrite produced, with ammonium and methane as the electron donor, respectively. The process could potentially be used for anaerobic nitrogen removal from wastewater streams containing ammonium and nitrate/nitrite.

Advanced Polymer-Based Drug Delivery Strategies for Meniscal Regeneration.

The meniscus plays a critical role in maintaining knee joint homeostasis. Injuries to the meniscus, especially considering the limited self-healing capacity of the avascular region, continue to be a challenge and are often treated by (partial) meniscectomy, which has been identified to cause osteoarthritis. Currently, meniscus tissue engineering focuses on providing extracellular matrix (ECM)-mimicking scaffolds to direct the inherent meniscal regeneration process, and it has been found that various stimuli are essential. Numerous bioactive factors present benefits in regulating cell fate, tissue development, and healing, but lack an optimal delivery system. More recently, bioengineers have developed various polymer-based drug delivery systems (PDDSs), which are beneficial in terms of the favorable properties of polymers as well as novel delivery strategies. Engineered PDDSs aim to provide not only an ECM-mimicking microenvironment but also the controlled release of bioactive factors with release profiles tailored according to the biological concerns and properties of the factors. In this review, both different polymers and bioactive factors involved in meniscal regeneration are discussed, as well as potential candidate systems, with examples of recent progress. This article aims to summarize drug delivery strategies in meniscal regeneration, with a focus on novel delivery strategies rather than on specific delivery carriers. The current challenges and future prospects for the structural and functional regeneration of the meniscus are also discussed. Impact statement Meniscal injury remains a clinical Gordian knot owing to the limited healing potential of the region, restricted surgical approaches, and risk of inducing osteoarthritis. Existing tissue engineering scaffolds that provide mechanical support and a favorable microenvironment also lack biological cues. Advanced polymer-based delivery strategies consisting of polymers incorporating bioactive factors have emerged as a promising direction. This article primarily reviews the types and applications of biopolymers and bioactive factors in meniscal regeneration. Importantly, various carrier systems and drug delivery strategies are discussed with the hope of inspiring further advancements in this field.

Decreasing microbially influenced metal corrosion using free nitrous acid in a simulated water injection system.

Microbially influenced corrosion (MIC) is the main cause of metal corrosion in anoxic environments. Biocides are often dosed to the corrosive media to inhibit and kill the microbes which cause MIC. In this study, intermittent dosages of free nitrous acid (FNA), which was previously found to be a biocide, were applied to a simulated water injection system containing carbon steel coupons with mature biofilm, to study the effect of FNA on mitigation of metal corrosion. In each treatment, 0.49 mg-N/L FNA was dosed using 200 mg-N/L nitrite at pH 6 for 24 h. The corrosion properties were monitored by open circuit potential (OCP), electrochemical impedance spectroscopy (EIS), linear polarization resistance (LPR), 3D optical profiling, and direct weight measurement. The biofilm viability was monitored by measuring cellular ATP level. The general corrosion rate (calculated by weight-loss measurement) was decreased by up to 31%, which was supported by LPR tests and reduced ATP levels of the corrosion-inducing biofilm. The 3D optical profiling results showed that FNA decreased the average pitting corrosion rate by 59%, with 2 intermittent treatments and 82-day interval over 304 days. Intermittent dosing of FNA has strong potential to be an effective and efficient strategy for controlling MIC in oil recovery infrastructure.

Rebar corrosion and its interaction with concrete degradation in reinforced concrete sewers.

Concrete corrosion, as a major issue in sewer management, has attracted considerable research. In comparison, the corrosion of reinforcing steel bar (rebar) is not well understood. Particularly, fundamental knowledge of rebar corrosion and its interactions with concrete corrosion/cracking is largely lacking. This study investigated rebar corrosion and concrete degradation using reinforced concrete coupons exposed in a pilot sewer system. The physical-chemical corrosion characteristics were investigated in local regions; the nature of rebar rusts was analyzed using the advanced mineral analytical techniques, including Scanning Electron Microscope (SEM), Energy Dispersive X-ray Spectroscopy (EDS) and X-ray Diffraction (XRD); further, the interactions between rebar corrosion and concrete corrosion/cracking were elucidated by characterizing the microstructure and element distribution in interfacial areas using Mineral Liberation Analysis (MLA). The rebar corrosion products were found to be iron oxides, oxyhydroxides, chlorides, sulfides and sulfates. The predominant rebar corrosion reactions varied with exposure time and the development of concrete corrosion. When concrete corrosion reached rebar surface, the cracking of the concrete cover was influenced by multiple effects, including the macro-cracking induced by the corrosion products expansion, and the micro-cracking accelerated by the dissolution, diffusion and deposition of Fe derived from rebar rusts at the concrete corrosion front. A conceptual model elucidating rebar corrosion and the complex interactions between rebar corrosion and concrete degradation is proposed to support the development of corrosion prevention and refurbishment strategies for reinforced concrete sewers.

Preclinical evaluation of a mini-arthroplasty implant based on polyetheretherketone and Ti6AI4V for treatment of a focal osteochondral defect in the femoral head of the hip.

A mini or partial arthroplasty may offer the advantages of reduced pain, shorter hospital stay, and increased range of motion, which are beneficial for the treatment of large-sized focal osteochondral defects. We aimed to evaluate the in vivo histologic response and function of our nonresorbable, composite structure implant, developed using polyetheretherketone (PEEK) and Ti6AI4V alloy, as a treatment for full-thickness osteochondral defects in the femoral head of the hip using a goat model. The gross and imaging appearance and histologic results were compared to those of a similar size cobalt-chromium-molybdenum (CoCrMo) alloy implant placed in a defect in the femoral head and evaluated up to 12 weeks. The X radiographs showed that there was no evidence of loosening of the implants for both the PEEK-Ti6AI4V and CoCrMo groups. Magnetic resonance imaging results showed no inflammatory signal findings in both PEEK-Ti6AI4V and CoCrMo implants. Macroscopically and histologically, there was lesser cartilage degeneration in the PEEK-Ti6AI4V implant than in the CoCrMo implant. The modified macroscopic articular evaluation score was lower in the PEEK-Ti6AI4V group than in the CoCrMo group (p < 0.05), and the histological score of the periprosthetic and acetabular cartilage was lower in the PEEK-Ti6AI4V group than in the CoCrMo group (P < 0.05). The micro-computed tomography results showed that the uncemented PEEK-Ti6AI4V implant has better osseointegration and higher bone-implant contact than the cemented CoCrMo implant. The peri-implant bone mass was higher in the PEEK-Ti6AI4V implant(p < 0.05). Meanwhile, the optical profile analytical results showed that the surface roughness of the cartilage in the acetabulum was higher in the CoCrMo group. In conclusion, the mini-arthroplasty implant based on PEEK-Ti6AI4V was superior to an identical CoCrMo alloy implant as a treatment for local osteochondral defect in the femoral head, owing to its in vivo cartilage protection and better osseointegration.

Understanding the properties of aerobic sludge granules as hydrogels.

Aerobic sludge granules are larger, denser microbial aggregates than activated sludge flocs with a smoother and more regular surface, which facilitates greater wastewater treatment intensity. Factors important in their growth are still poorly understood, which is an impediment to the construction and operation of full-scale aerobic sludge granule processes. Data in this article obtained with granules treating an abattoir wastewater provide evidence that aerobic sludge granules are hydrogels. The results also demonstrate a method for characterizing macromolecular associations. The rheological profile of these granules was found to be analogous with that of typical polymer gels. Water uptake or swelling reflects an equilibrium between granule elastic modulus and osmotic pressure, whereby uptake is increased by reducing solute concentration or the elastic modulus. A weakening of the extracellular polymeric substance (EPS) matrix as demonstrated with mechanical spectroscopy was induced by several environmental factors including temperature, pH and ionic strength. Uniform and elastic deformation was observed at low strain. Enzymatic degradation studies indicate that proteins and alpha-polysaccharides were the major granule structural materials. The aerobic sludge granules in the current study were therefore protein-polysaccharide composite physical hydrogels. While aerobic sludge granules treating an abattoir wastewater are used as a case study, many of the fundamental principles detailed here are relevant to other granulation processes. The paradigm established in this study can potentially be applied to better understand the formation of aerobic sludge granules and thus overcome a hurdle in the acceptance of aerobic sludge granulation as an alternative to more traditional wastewater treatment processes.

PCL-MECM-Based Hydrogel Hybrid Scaffolds and Meniscal Fibrochondrocytes Promote Whole Meniscus Regeneration in a Rabbit Meniscectomy Model.

Regeneration of an injured meniscus continues to be a scientific challenge due to its poor self-healing potential. Tissue engineering provides an avenue for regenerating a severely damaged meniscus. In this study, we first investigated the superiority of five concentrations (0%, 0.5%, 1%, 2%, and 4%) of meniscus extracellular matrix (MECM)-based hydrogel in promoting cell proliferation and the matrix-forming phenotype of meniscal fibrochondrocytes (MFCs). We found that the 2% group strongly enhanced chondrogenic marker mRNA expression and cell proliferation compared to the other groups. Moreover, the 2% group showed the highest glycosaminoglycan (GAG) and collagen production by day 14. We then constructed a hybrid scaffold by 3D printing a wedge-shaped poly(ε-caprolactone) (PCL) scaffold as a backbone, followed by injection with the optimized MECM-based hydrogel (2%), which served as a cell delivery system. The hybrid scaffold (PCL-hydrogel) clearly yielded favorable biomechanical properties close to those of the native meniscus. Finally, PCL scaffold, PCL-hydrogel, and MFCs-loaded hybrid scaffold (PCL-hydrogel-MFCs) were implanted into the knee joints of New Zealand rabbits that underwent total medial meniscectomy. Six months postimplantation we found that the PCL-hydrogel-MFCs group exhibited markedly better gross appearance and cartilage protection than the PCL scaffold and PCL-hydrogel groups. Moreover, the regenerated menisci in the PCL-hydrogel-MFCs group had similar histological structures, biochemical contents, and biomechanical properties as the native menisci in the sham operation group. In conclusion, PCL-MECM-based hydrogel hybrid scaffold seeded with MFCs can successfully promote whole meniscus regeneration, and cell-loaded PCL-MECM-based hydrogel hybrid scaffold may be a promising strategy for meniscus regeneration in the future.

Characterisation of polyhydroxyalkanoate copolymers with controllable four-monomer composition.

Polyhydroxyalkanoate (PHA) copolymers comprising the four monomers 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 3-hydroxy-2-methylvalerate (3HMV) and 3-hydroxy-2-methylbutyrate (3HMB) were generated using the recently discovered Defluviicoccus vanus-related glycogen accumulating organisms (DvGAOs) under anaerobic conditions without applying any nutrient limitations. The composition could be manipulated in a defined range by modifying the ratio of propionate and acetate provided in the feed stream. The PHAs produced were characterised as random copolymers (from propionate alone) or a mixture of random copolymers (from mixture of propionate and acetate) through microstructure analysis using 13C NMR spectroscopy. The sequence distribution of all eight comonomer pairs in the carbonyl region of 3HB and 3HV was identified and assigned with confidence utilising two-dimensional heteronuclear multiple bond coherence (HMBC) spectroscopy. Weight average molecular weights were in the range 390-560 kg/mol. Differential scanning calorimetry (DSC) traces showed that the melting temperature (Tm) varied between 70 and 161 degrees C and glass transition temperature (Tg) ranged from -8 to 0 degrees C. The incorporation of considerable amounts of 3HMV and 3HMB monomer units introduced additional "defects" into the PHBV copolymer structure and hence greatly lowered the crystallinity. The data indicate the potential of these four-monomer PHAs to be employed for practical applications, considering their favourable properties and the cost-effective production process using a mixed culture and simple carbon sources.

Achieving high-level nitrogen removal in mainstream by coupling anammox with denitrifying anaerobic methane oxidation in a membrane biofilm reactor.

To achieve energy neutral wastewater treatment, mainstream anaerobic ammonium oxidation (anammox) has attracted extensive attention in the past decade. However, the relatively high effluent nitrogen concentration (>10 mg N L-1) remains a significant barrier hindering its practical implementation. A novel technology integrating the anammox and denitrifying anaerobic methane oxidation (DAMO) reactions in a membrane biofilm reactor (MBfR) was developed in this study to enhance the mainstream anammox process. With the hydraulic retention time (HRT) progressively decreased from 12 to 4 h, the total nitrogen (TN) removal rate increased stepwise from 0.09 to 0.28 kg N m-3 d-1, with an effluent TN concentration below 3.0 mg N L-1 achieved. Mass balance analysis showed that 30-60% of the nitrate produced by the anammox reaction was reduced back to nitrite by DAMO archaea, and the anammox and DAMO bacteria were jointly responsible for nitrite removal with contributions of >90% and <10%, respectively. Additionally, the established MBfR was robust and achieved consistently high effluent quality with >90% TN removal when the influent nitrite to ammonium molar ratio varied in the range of 1.17-1.55. Fluorescence in situ hybridization (FISH) and 16S rRNA gene sequencing indicated that anammox bacteria, DAMO bacteria and DAMO archaea jointly dominated the biofilm, and were likely the key contributors to nitrogen removal. This is the first study that a high nitrogen removal rate (>0.2 kg N m-3 d-1) and satisfactory effluent quality (∼3 mg TN L-1) were achieved simultaneously by integrating anammox and DAMO reactions in mainstream wastewater treatment.

An electrospun fiber reinforced scaffold promotes total meniscus regeneration in rabbit meniscectomy model.

Low vascularization in meniscus limits its regeneration ability after injury, and tissue engineering is the most promising method to achieve meniscus regeneration. In this study, we fabricated a kind of composite scaffold by decellularized meniscus extracellular matrix/polycaprolactone (DMECM/PCL) electrospinning fibers and porous DMECM, in which DMECM/PCL fibers were used as reinforcing component. The tensile modulus of the composite scaffold in longitudinal and crosswise directions were 8.5 ±â€¯1.9 and 2.3 ±â€¯0.3 MPa, respectively. Besides that, the DMECM/PCL electrospinning fibers enhanced suture resistance of the composite scaffold more than 5 times than DMECM scaffold effectively. In vitro cytocompatibility showed that the porous structure provided by DMECM component facilitated meniscus cells' proliferation. DMECM was also the main component to regulate cell behaviors, which promoted meniscus cells expressing extracellular matrix related genes such as COL I, COL II, SOX9 and AGG. Rabbits with total meniscectomy were used as animal model to evaluated the composited scaffolds performance in vivo at 3 and 6 months. Results showed that rabbits with scaffold implanting could regenerate neo-menisci in both time points. The neo-menisci had similar histology structure and biochemical content with native menisci. Although neo-menisci had inferior tensile modulus than native ones, its modulus was improved with implanting time prolonging. MRI imaging showed the signal of neo-meniscus in the body is clear, and X-ray imaging of knee joints demonstrated the implantation of scaffolds could relief joint space narrowing. Moreover, rabbits with neo-menisci had better cartilage condition in femoral condyle and tibial plateau compared than meniscectomy group. STATEMENT OF SIGNIFICANCE: We fabricated the meniscus scaffold by combining porous decellularized meniscus extracellular matrix (DMECM) and DMECM/PCL electrospinning fibers together, which used the porous structure of DMECM, and the good tensile property of electrospinning fibers. We believe single material cannot satisfy increasing needs of scaffold. Therefore, we combined not only materials but also fabrication methods together to develop scaffold to make good use of each part. DMECM in electrospinning fibers also made these two components possible to be integrated through crosslinking. Compared to existing meniscus scaffold, the composite scaffold had (1) soft structure and extrusion would not happen after implantation, (2) ability to be trimmed to suitable shape during surgery, and (3) good resistance to suture.

Production of targeted poly(3-hydroxyalkanoates) copolymers by glycogen accumulating organisms using acetate as sole carbon source.

One of the main limitations in bacterial polyhydroxyalkanoate (PHA) production with mixed cultures is the fact that primarily polyhydroxybutyrate (PHB) homopolymers are generated from acetate as the main carbon source, which is brittle and quite fragile. The incorporation of different 3-hydroxyalkanoate (HA) components into the polymers requires the addition of additional carbon sources, leading to extra costs and complexity. In this study, the production of poly(3-hydroxybutyrate (3HB)-co-3-hydroxyvalerate (3HV)-co-3-hydroxy-2-methylvalerate (3HMV)), with 7-35C-mol% of 3HV fractions from acetate as the only carbon source was achieved with the use of glycogen accumulating organisms (GAOs). An enriched GAO culture was obtained in a lab-scale reactor operated under alternating anaerobic and aerobic conditions with acetate fed at the beginning of the anaerobic period. The production of PHAs utilizing the enriched GAO culture was investigated under both aerobic and anaerobic conditions. A polymer content of 14-41% of dry cell weight was obtained. The PHA product accumulated by GAOs under anaerobic conditions contained a relatively constant proportion of non-3HB monomers (30+/-5C-mol%), irrespective of the amount of acetate assimilated. In contrast, under aerobic conditions, GAOs only produced 3HB monomers from acetate causing a gradually decreasing 3HV fraction during this aerobic feeding period. The PHAs were characterized by gel permeation chromatography (GPC) and differential scanning calorimetry (DSC). The data demonstrated that the copolymers possessed similar characteristics to those of commercially available poly(3HB-co-3HV) (PHBV) products. The PHAs produced under solely anaerobic conditions possessed lower melting points and crystallinity, higher molecular weights, and narrower molecular-weight distributions, compared to the aerobically produced polymers. This paper hence demonstrates the significant potential of GAOs to produce high quality polymers from a simple and cheap carbon source, contributing considerably to the growing research body on bacterial PHA production by mixed cultures.

Endogenous metabolism of Candidatus Accumulibacter phosphatis under various starvation conditions.

The endogenous processes of Candidatus Accumulibacter phosphatis (referred to as Accumulibacter), a known polyphosphate-accumulating organism (PAO) responsible for enhanced biological phosphorus removal systems (EBPR), were characterized during 8-day starvation under anaerobic, anoxic, aerobic and intermittent aerobic-anaerobic conditions. A lab-scale EBPR culture with Accumulibacter representing over 85% of the entire bacterial population as quantified with fluorescence in-situ hybridization was used in the study. Cell decay rates were found to be negligible under anaerobic and anoxic conditions and may be ignored in activated sludge models. The decay rate under aerobic conditions was determined to be 0.03/d at 22 degrees C, considerably lower than the values commonly used in activated sludge modeling. Polyphosphate and glycogen were utilized simultaneously under anaerobic and anoxic conditions for maintenance energy production, with glycogen being the primary energy source until the glycogen content reached very low levels. Glycogen was used by Accumulibacter as the primary source of energy for maintenance under aerobic conditions in the absence of polyhydroxyalkanoates. However, Accumulibacter did not seem to use polyphosphate for energy production during aerobic starvation, clearly contrasting the anaerobic and particularly the anoxic case. Intermittent aerobic-anaerobic storage resulted in not only negligible cell decay rate, but also slower rates of glycogen and polyphosphate utilization, and may therefore be an effective strategy for long-term storage of EBPR sludge.

AMECM/DCB scaffold prompts successful total meniscus reconstruction in a rabbit total meniscectomy model.

Tissue-engineered meniscus regeneration is a very promising treatment strategy for meniscus lesions. However, generating the scaffold presents a huge challenge for meniscus engineering as this has to meet particular biomechanical and biocompatibility requirements. In this study, we utilized acellular meniscus extracellular matrix (AMECM) and demineralized cancellous bone (DCB) to construct three different types of three-dimensional porous meniscus scaffold: AMECM, DCB, and AMECM/DCB, respectively. We tested the scaffolds' physicochemical characteristics and observed their interactions with meniscus fibrochondrocytes to evaluate their cytocompatibility. We implanted the three different types of scaffold into the medial knee menisci of New Zealand rabbits that had undergone total meniscectomy; negative control rabbits received no implants. The reconstructed menisci and corresponding femoral condyle and tibial plateau cartilage were all evaluated at 3 and 6 months (n = 8). The in vitro study demonstrated that the AMECM/DCB scaffold had the most suitable biomechanical properties, as this produced the greatest compressive and tensile strength scores. The AMECM/DCB and AMECM scaffolds facilitated fibrochondrocyte proliferation and the secretion of collagen and glycosaminoglycans (GAGs) more effectively than did the DCB scaffold. The in vivo experiments demonstrated that both the AMECM/DCB and DCB groups had generated neomeniscus at both 3 and 6 months post-implantation, but there was no obvious meniscus regeneration in the AMECM or control groups, so the neomeniscus analysis could not perform on AMECM and control group. At both 3 and 6 months, histological scores were better for regenerated menisci in the AMECM/DCB than in the DCB group, and significantly better for articular cartilage in the AMECM/DCB group compared with the other three groups. Knee MRI scores (Whole-Organ Magnetic Resonance Imaging Scores (WORMS)) were better in the AMECM/DCB group than in the other three groups at both 3 and 6 months. At both 3 and 6 months, RT-PCR demonstrated that aggrecan, Sox9, and collagen II content was significantly higher, and mechanical testing demonstrated greater tensile strength, in the AMECM/DCB group neomenisci compared with the DCB group.

Evaluating the Role of Microbial Internal Storage Turnover on Nitrous Oxide Accumulation During Denitrification.

Biological wastewater treatment processes under a dynamic regime with respect to carbon substrate can result in microbial storage of internal polymers (e.g., polyhydroxybutyrate (PHB)) and their subsequent utilizations. These storage turnovers play important roles in nitrous oxide (N2O) accumulation during heterotrophic denitrification in biological wastewater treatment. In this work, a mathematical model is developed to evaluate the key role of PHB storage turnovers on N2O accumulation during denitrification for the first time, aiming to establish the key relationship between N2O accumulation and PHB storage production. The model is successfully calibrated and validated using N2O data from two independent experimental systems with PHB storage turnovers. The model satisfactorily describes nitrogen reductions, PHB storage/utilization, and N2O accumulation from both systems. The results reveal a linear relationship between N2O accumulation and PHB production, suggesting a substantial effect of PHB storage on N2O accumulation during denitrification. Application of the model to simulate long-term operations of a denitrifying sequencing batch reactor and a denitrifying continuous system indicates the feeding pattern and sludge retention time would alter PHB turnovers and thus affect N2O accumulation. Increasing PHB utilization could substantially raise N2O accumulation due to the relatively low N2O reduction rate when using PHB as carbon source.

The effect of free nitrous acid on key anaerobic processes in enhanced biological phosphorus removal systems.

In this study, the effect of nitrite/FNA on the anaerobic metabolism of polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs) is investigated. The results clearly show that FNA has a detrimental effect on the acetate uptake rate by both PAOs and GAOs, but this adverse effect is much stronger on PAOs than on GAOs. Also, when FNA was increased, phosphate release to acetate uptake ratio by PAOs increased substantially (250-300% compared to control), which was accompanied by decreases (40-60%) in glycogen degradation and PHA production to VFA uptake. In contrast, these ratios for GAOs remained constant or increased slightly towards the highest FNA concentration applied. These results indicate that the anaerobic metabolism of PAOs is more adversely affected than that of GAOs when FNA is present. This might provide a competitive advantage to GAOs over PAOs in enhanced biological phosphorus removal systems when nitrite is present.

Gel-forming exopolysaccharides explain basic differences between structures of aerobic sludge granules and floccular sludges.

The sol-gel transition of extracellular polymeric substances (EPS) derived from sludge flocs and granules is investigated in order to explain basic differences between the two aggregates. A reversible, pH dependent sol-gel transition was observed at pH 9.0-12.0 in EPS extracted from granules. At pH <9 granule EPS existed as a strong gel, indicating that their EPS exist in a gel state at normal operating pH of a wastewater treatment system (i.e. 6.0-8.5). This characteristic transition from solution to strong gel was not observed in any of the EPS samples derived from floccular sludges. A transition to a weak gel was however, observed at pH 4.0-5.0. Enriched exopolysaccharides from the granular EPS exhibited rheological behaviour analogous to the granules and the granule EPS. The critical overlap concentration (c*) of the exopolysaccharide concentrate was 0.33% w/w, similar to the c* of other known bacterial exopolysaccharides. Additionally, the protein content was found to be not contributing to the storage modulus of granule EPS gels. These factors suggest that exopolysaccharides or glycosides were the gelling agent in aerobic sludge granules. Given that EPS derived from aerobic sludge granules and flocs are distinguished by such a sol-strong gel transition, these exopolysaccharides therefore likely play an important role in granulation.

Proposed modifications to metabolic model for glycogen-accumulating organisms under anaerobic conditions.

Filipe et al. (2001) proposed an anaerobic metabolic model for glycogen-accumulating organisms (GAO) in which the succinate-propionate pathway was used to describe the production of propionyl-CoA. However, propionyl-CoA is only an intermediate product in the above pathway. Stopping at propionyl-CoA instead of propionate (the end product of the pathway) results in the consumption of one ATP from succinate to succinyl-CoA, which was not accounted for in the model of Filipe et al. (2001). This resulted in significant errors in the stoichiometric coefficients in the final metabolic model. A modified model is presented in this communication and is shown to fit the experimental data significantly better than the original model.