Nitrite-driven anaerobic ethane oxidation.

Ethane, the second most abundant gaseous hydrocarbon in vast anoxic environments, is an overlooked greenhouse gas. Microbial anaerobic oxidation of ethane can be driven by available electron acceptors such as sulfate and nitrate. However, despite nitrite being a more thermodynamically feasible electron acceptor than sulfate or nitrate, little is known about nitrite-driven anaerobic ethane oxidation. In this study, a microbial culture capable of nitrite-driven anaerobic ethane oxidation was enriched through the long-term operation of a nitrite-and-ethane-fed bioreactor. During continuous operation, the nitrite removal rate and the theoretical ethane oxidation rate remained stable at approximately 25.0 mg NO2 -N L-1 d-1 and 11.48 mg C2H6 L-1 d-1, respectively. Batch tests demonstrated that ethane is essential for nitrite removal in this microbial culture. Metabolic function analysis revealed that a species affiliated with a novel genus within the family Rhodocyclaceae, designated as 'Candidatus Alkanivoras nitrosoreducens', may perform the nitrite-driven anaerobic ethane oxidation. In the proposed metabolic model, despite the absence of known genes for ethane conversion to ethyl-succinate and succinate-CoA ligase, 'Ca. A. nitrosoreducens' encodes a prospective fumarate addition pathway for anaerobic ethane oxidation and a complete denitrification pathway for nitrite reduction to nitrogen. These findings advance our understanding of nitrite-driven anaerobic ethane oxidation, highlighting the previously overlooked impact of anaerobic ethane oxidation in natural ecosystems.

Impact of hydrogen sulfide on anammox and nitrate/nitrite-dependent anaerobic methane oxidation coupled technologies.

The coupling between anammox and nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) has been considered a sustainable technology for nitrogen removal from sidestream wastewater and can be implemented in both membrane biofilm reactor (MBfR) and granular bioreactor. However, the potential influence of the accompanying hydrogen sulfide (H2S) in the anaerobic digestion (AD)-related methane-containing mixture on anammox/n-DAMO remains unknown. To fill this gap, this work first constructed a model incorporating the C/N/S-related bioprocesses and evaluated/calibrated/validated the model using experimental data. The model was then used to explore the impact of H2S on the MBfR and granular bioreactor designed to perform anammox/n-DAMO at practical levels (i.e., 0∼5% (v/v) and 0∼40 g/S m3, respectively). The simulation results indicated that H2S in inflow gas did not significantly affect the total nitrogen (TN) removal of the MBfR under all operational conditions studied in this work, thus lifting the concern about applying AD-produced biogas to power up anammox/n-DAMO in the MBfR. However, the presence of H2S in the influent would either compromise the treatment performance of the granular bioreactor at a relatively high influent NH4+-N/NO2--N ratio (e.g., >1.0) or lead to increased energy demand associated with TN removal at a relatively low influent NH4+-N/NO2--N ratio (e.g., <0.7). Such a negative effect of the influent H2S could not be attenuated by regulating the hydraulic residence time and should therefore be avoided when applying the granular bioreactor to perform anammox/n-DAMO in practice.

Nitrate-dependent anaerobic methane oxidation coupled to Fe(III) reduction as a source of ammonium and nitrous oxide.

'Candidatus Methanoperedens nitroreducens' is an archaeal methanotroph with global importance that links carbon and nitrogen cycles and great potential for sustainable operation of wastewater treatment. It has been reported to mediate the anaerobic oxidation of methane through a reverse methanogenesis pathway while reducing nitrate to nitrite. Here, we demonstrate that 'Ca. M. nitroreducens' reduces ferric iron forming ammonium (23.1 %) and nitrous oxide (N2O, 46.5 %) from nitrate. These results are supported with the upregulation of genes coding for proteins responsible for dissimilatory nitrate reduction to ammonium (nrfA), N2O formation (norV, cyt P460), and multiple multiheme c-type cytochromes for ferric iron reduction. Concomitantly, an increase in the N2O-reducing SJA-28 lineage and a decrease in the nitrite-reducing 'Candidatus Methylomirabilis oxyfera' are consistent with the changes in 'Ca. M. nitroreducens' end products. These findings demonstrate the highly flexible physiology of 'Ca. M. nitroreducens' in anaerobic ecosystems with diverse electron acceptor conditions, and further reveals its roles in linking methane oxidation to global biogeochemical cycles. 'Ca. M. nitroreducens' could significantly affect the bioavailability of nitrogen sources as well as the emission of greenhouse gas in natural ecosystems and wastewater treatment plants.

Different oxygen affinities of methanotrophs and Comammox Nitrospira inform an electrically induced symbiosis for nitrogen loss.

Aerobic methanotrophs establish a symbiotic association with denitrifiers to facilitate the process of aerobic methane oxidation coupled with denitrification (AME-D). However, the symbiosis has been frequently observed in hypoxic conditions continuing to pose an enigma. The present study has firstly characterized an electrically induced symbiosis primarily governed by Methylosarcina and Hyphomicrobium for the AME-D process in a hypoxic niche caused by Comammox Nitrospira. The kinetic analysis revealed that Comammox Nitrospira exhibited a higher apparent oxygen affinity compared to Methylosarcina. While the coexistence of comammox and AME-D resulted in an increase in methane oxidation and nitrogen loss rates, from 0.82 ± 0.10 to 1.72 ± 0.09 mmol CH4 d-1 and from 0.59 ± 0.04 to 1.30 ± 0.15 mmol N2 d-1, respectively. Furthermore, the constructed microbial fuel cells demonstrated a pronounced dependence of the biocurrents on AME-D due to oxygen competition, suggesting the involvement of direct interspecies electron transfer in the AME-D process under hypoxic conditions. Metagenomic and metatranscriptomic analysis revealed that Methylosarcina efficiently oxidized methane to formaldehyde, subsequently generating abundant NAD(P)H for nitrate reduction by Hyphomicrobium through the dissimilatory RuMP pathway, leading to CO2 production. This study challenges the conventional understanding of survival mechanism employed by AME-D symbionts, thereby contributing to the characterization responsible for limiting methane emissions and promoting nitrogen removal in hypoxic regions.

Revisiting the Engineering Roadmap of Nitrate/Nitrite-Dependent Anaerobic Methane Oxidation.

Nitrate/nitrite-dependent anaerobic oxidation of methane (n-DAMO) is a recently discovered process, which provides a sustainable perspective for simultaneous nitrogen removal and greenhouse gas emission (GHG) mitigation by using methane as an electron donor for denitrification. However, the engineering roadmap of the n-DAMO process is still unclear. This work constitutes a state-of-the-art review on the classical and most recently discovered metabolic mechanisms of the n-DAMO process. The versatile combinations of the n-DAMO process with nitrification, nitritation, and partial nitritation for nitrogen removal are also clearly presented and discussed. Additionally, the recent advances in bioreactor development are systematically reviewed and evaluated comprehensively in terms of methane supply, biomass retention, membrane requirement, startup time, reactor performance, and limitations. The key issues including enrichment and operation strategy for the scaling up of n-DAMO-based processes are also critically addressed. Moreover, the challenges inherent to implementing the n-DAMO process in practical applications, including application scenario recognition, GHG emission mitigation, and operation under realistic conditions, are highlighted. Finally, prospects as well as opportunities for future research are proposed. Overall, this review provides a roadmap for potential applications and further development of the n-DAMO process in the field of wastewater treatment.

Evidence for Nitrous Oxide Emissions by Nitrite-Dependent Anaerobic Methane Oxidizing Bacteria.

Nitrite-dependent anaerobic methane oxidizing (n-DAMO) bacteria generally convert nitrite to dinitrogen and bypass the nitrous oxide (N2O) formation step. However, N2O is often detected in n-DAMO bacteria dominated cultures and it remains an open question as to the microbial origin of N2O in these enrichments. Using a stable nitrite consuming microbial community enriched for n-DAMO bacteria, we demonstrated that N2O production was coupled to methane oxidation and the higher initial nitrite concentrations led to increased quantities of N2O being formed. Moreover, continuous exposure of the enrichment culture to about 5 mg of N L-1 nitrite resulted in constant N2O being produced (12.5% of nitrite was reduced to N2O). Metatranscriptomic analyses revealed that nitrite reductase (nirS) and nitric oxide reductase (norZ) transcripts from n-DAMO bacteria increased in response to nitrite exposure. No other bacteria significantly expressed nor genes under these conditions, suggesting n-DAMO bacteria are responsible for N2O being produced. In a 35-day bioreactor experiment, N2O produced by the n-DAMO bacteria accumulated when nitrite was in excess; this was found to be up to 3.2% of the nitrogen that resulted from nitrite removal. Together, these results suggested that excess nitrite is an important driver of N2O production by n-DAMO bacteria. To this end, proper monitoring and control of nitrite levels in wastewater treatment plants would be effective strategies for mitigating N2O emissions to the atmosphere.

Microbial Niche Differentiation during Nitrite-Dependent Anaerobic Methane Oxidation.

Nitrite-dependent anaerobic methane oxidation (n-DAMO) has been demonstrated to play important roles in the global methane and nitrogen cycle. However, despite diverse n-DAMO bacteria widely detected in environments, little is known about their physiology for microbial niche differentiation. Here, we show the microbial niche differentiation of n-DAMO bacteria through long-term reactor operations combining genome-centered omics and kinetic analysis. With the same inoculum dominated by both species "Candidatus Methylomirabilis oxyfera" and "Candidatus Methylomirabilis sinica", n-DAMO bacterial population was shifted to "Ca. M. oxyfera" in a reactor fed with low-strength nitrite, but shifted to "Ca. M. sinica" with high-strength nitrite. Metatranscriptomic analysis showed that "Ca. M. oxyfera" harbored more complete function in cell chemotaxis, flagellar assembly, and two-component system for better uptake of nitrite, while "Ca. M. sinica" had a more active ion transport and stress response system, and more redundant function in nitrite reduction to mitigate nitrite inhibition. Importantly, the half-saturation constant of nitrite (0.057 mM vs 0.334 mM NO2-) and inhibition thresholds (0.932 mM vs 2.450 mM NO2-) for "Ca. M. oxyfera" vs "Ca. M. sinica", respectively, were highly consistent with genomic results. Integrating these findings demonstrated biochemical characteristics, especially the kinetics of nitrite affinity and inhibition determine niche differentiation of n-DAMO bacteria.

Instrumental role of bioreactors in nitrate/nitrite-dependent anaerobic methane oxidation-based biotechnologies for wastewater treatment: A review.

Recently, the nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) processes have become a research hotspot in the field of wastewater treatment. The n-DAMO processes could not only mitigate direct and indirect carbon emissions from wastewater treatment plants but also strengthen biological nitrogen removal. However, the applications of n-DAMO-based biotechnologies face practical difficulties mainly caused by the distinctive properties of n-DAMO microorganisms and the limited/availability of methane with poor solubility. In this sense, the choice of bioreactors will play important roles that influence the growth and functioning of n-DAMO microorganisms, thus enabling dedicated development of the n-DAMO processes and efficient applications of n-DAMO-based biotechnologies. Therefore, this paper aims to discuss the three commonly-applied types of bioreactors, covering the individual working principle and state-of-the-art removal performance of nitrogen as well as dissolved methane observed when adopted for n-DAMO-based biotechnologies. With noted limitations for each bioreactor type, several key perspectives were proposed which hopefully would inspire future investigation and practical applications of the n-DAMO processes.

Deciphering the Inhibition of Ethane on Anaerobic Ammonium Oxidation.

Anaerobic ammonium oxidation (anammox) and nitrification, two common biological ammonium oxidation pathways, are critical for the microbial nitrogen cycle. Short chain alkanes (C2-C8) have been well-known as inhibitors for nitrification through interaction with the ammonia monooxygenase, while whether these alkanes affect anammox is an open question. Here, this work demonstrated significant inhibition of ethane on anammox and revealed the inhibitory mechanism. The acute inhibition of ethane on anammox was concentration-dependent and reversible; 0.86 mM dissolved ethane caused 50% inhibition (IC50), and 1.72 mM ethane almost completely inhibited anammox. After long-term exposure to 0.09 mM ethane for 30 days, the ammonium (nitrite) removal rate dropped from 202 (267) mg N L-1 d-1 to 1 (1) mg N L-1 d-1, and the abundance of anammox bacteria decreased from 61.9% to 9.5%. The intercellular ammonium concentration of anammox bacteria decreased after ethane exposure, while metatranscriptome analysis showed significant upregulation of genes for ammonium transport of anammox bacteria. Thus, ethane could suppress ammonium uptake resulting in the inhibition of anammox activities. As ethane is the second most prevalent alkane after methane in various anoxic environments, ethane may have an important effect on the nitrogen cycle driven by anammox that should be investigated in future research.

A mechanistic model for denitrifying anaerobic methane oxidation coupled to dissimilatory nitrate reduction to ammonium.

Nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) is an important process linking nitrogen and carbon cycle. It is recently demonstrated that n-DAMO archaea are able to couple n-DAMO to dissimilatory nitrate reduction to ammonium (DNRA). In this work, a mathematical model is developed to describe DNRA by n-DAMO archaea for the first time. The anabolic and catabolic processes of n-DAMO archaea, n-DAMO bacteria and anaerobic ammonium oxidation (Anammox) bacteria are involved. The different impacts of exogenous and endogenous nitrite on DNRA and n-DAMO microbes are considered. The developed model is calibrated and validated using experimental data collected from a sequencing batch reactor (SBR) and a counter-diffusion membrane biofilm bioreactor (MBfR). The model outputs fit well with the profiles of nitrogen (N) dynamics and biomass changes in both reactors, demonstrating its good predictive ability. The developed model is further used to simulate the counter-diffusion MBfR incorporating n-DAMO and Anammox process to treat sidestream wastewater. The simulated distribution profiles of N removal/production rates by different microbes along biofilm depth reveal that DNRA by n-DAMO archaea plays an important role in N transformation of the integrated n-DAMO and Anammox process. It is further suggested that the counter-diffusion MBfR under the investigated conditions should be operated at proper hydraulic retention times (HRTs) (i.e. 6h and 8h) with exogenous NO2- in the range of 0-10 mg N/L or at HRTs >3h with the absence of exogenous NO2- in order to achieve dischargeable effluent.

Simultaneous nitrate and sulfate dependent anaerobic oxidation of methane linking carbon, nitrogen and sulfur cycles.

ANaerobic MEthanotrophic (ANME) archaea are critical microorganisms mitigating methane emission from anoxic zones. In previous studies, sulfate-dependent anaerobic oxidation of methane (AOM) and nitrate-dependent AOM, performed by different clades of ANME archaea, were detected in marine sediments and freshwater environments, respectively. This study shows that simultaneous sulfate- and nitrate-dependent AOM can be mediated by a clade of ANME archaea, which may occur in estuaries and coastal zones, at the interface of marine and freshwater environments enriched with sulfate and nitrate. Long-term (~1,200 days) performance data of a bioreactor, metagenomic analysis and batch experiments demonstrated that ANME-2d not only conducted AOM coupled to reduction of nitrate to nitrite, but also coupled to the conversion of sulfate to sulfide, in collaboration with sulfate-reducing bacteria (SRB). Sulfide was oxidized back to sulfate by sulfide-oxidizing autotrophic denitrifiers with nitrate or nitrite as electron acceptors, in turn alleviating sulfide accumulation. In addition, dissimilatory nitrate reduction to ammonium performed by ANME-2d was detected, providing substrates to Anammox. Metatranscriptomic analysis revealed significant upregulation of flaB in ANME-2d and pilA in Desulfococcus, which likely resulted in the formation of unique nanonets connecting cells and expanding within the biofilm, and putatively providing structural links between ANME-2d and SRB for electron transfer. Simultaneous nitrate- and sulfate-dependent AOM as observed in this study could be an important link between the carbon, nitrogen and sulfur cycles in natural environments, such as nearshore environments.

Anaerobic Oxidation of Methane Coupled with Dissimilatory Nitrate Reduction to Ammonium Fuels Anaerobic Ammonium Oxidation.

Nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) is critical for mitigating methane emission and returning reactive nitrogen to the atmosphere. The genomes of n-DAMO archaea show that they have the potential to couple anaerobic oxidation of methane to dissimilatory nitrate reduction to ammonium (DNRA). However, physiological details of DNRA for n-DAMO archaea were not reported yet. This work demonstrated n-DAMO archaea coupling the anaerobic oxidation of methane to DNRA, which fueled Anammox in a methane-fed membrane biofilm reactor with nitrate as only electron acceptor. Microelectrode analysis revealed that ammonium accumulated where nitrite built up in the biofilm. Ammonium production and significant upregulation of gene expression for DNRA were detected in suspended n-DAMO culture with nitrite exposure, indicating that nitrite triggered DNRA by n-DAMO archaea. 15N-labeling batch experiments revealed that n-DAMO archaea produced ammonium from nitrate rather than from external nitrite. Localized gradients of nitrite produced by n-DAMO archaea in biofilms induced ammonium production via the DNRA process, which promoted nitrite consumption by Anammox bacteria and in turn helped n-DAMO archaea resist stress from nitrite. As biofilms predominate in various ecosystems, anaerobic oxidation of methane coupled with DNRA could be an important link between the global carbon and nitrogen cycles that should be investigated in future research.

Denitrifying Anaerobic Methane Oxidation and Anammox Process in a Membrane Aerated Membrane Bioreactor: Kinetic Evaluation and Optimization.

Denitrifying anaerobic methane oxidation (DAMO) coupled to anaerobic ammonium oxidation (anammox) is a promising technology for complete nitrogen removal with economic and environmental benefit. In this work, a model framework integrating DAMO and anammox process was constructed based on suspended-growth systems. The proposed model was calibrated and validated using experimental data from a sequencing batch reactor and a membrane aerated membrane bioreactor (MAMBR). The model managed to describe removal rates of ammonium (NH4+), nitrite (NO2-), and total nitrogen, as well as biomass changes of DAMO archaea, DAMO bacteria, and anaerobic ammonium oxidizing bacteria (AnAOB) in both reactors. The estimated parameter values revealed that DAMO archaea possessed properties of faster growth and higher biomass yield in suspended-growth systems compared to those in attached-growth systems (e.g., biofilm). Model simulation demonstrated that solid retention time (SRT) was effective in washing out DAMO bacteria, but retaining DAMO archaea and AnAOB in the MAMBR. The optimal SRT and nitritation efficiency (the ratio of the NO2- to the sum of NH4+ and NO2- in the MAMBR influent) were simulated so that 99% of total nitrogen was removed to meet the discharge standard. MAMBR further suggested to be operated with SRT between 15 and 30 days so that the optimal nitritation efficiency could be minimized to 49% for cost savings.

Growth Factor Gene-Modified Mesenchymal Stem Cells in Tissue Regeneration.

There have been marked changes in the field of stem cell therapeutics in recent years, with many clinical trials having been conducted to date in an effort to treat myriad diseases. Mesenchymal stem cells (MSCs) are the cell type most frequently utilized in stem cell therapeutic and tissue regenerative strategies, and have been used with excellent safety to date. Unfortunately, these MSCs have limited ability to engraft and survive, reducing their clinical utility. MSCs are able to secrete growth factors that can support the regeneration of tissues, and engineering MSCs to express such growth factors can improve their survival, proliferation, differentiation, and tissue reconstructing abilities. As such, it is likely that such genetically modified MSCs may represent the next stage of regenerative therapy. Indeed, increasing volumes of preclinical research suggests that such modified MSCs expressing growth factors can effectively treat many forms of tissue damage. In the present review, we survey recent approaches to producing and utilizing growth factor gene-modified MSCs in the context of tissue repair and discuss its prospects for clinical application.

Operation strategies of n-DAMO and Anammox process based on microbial interactions for high rate nitrogen removal from landfill leachate.

Nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) coupling to Anaerobic ammonium oxidation (Anammox) provides an opportunity for simultaneous nitrogen removal and methane emissions mitigation from wastewater. However, to achieve high nitrogen removal rate in such a process remains a critical challenge in practical application. This work investigated the interactions between n-DAMO and Anammox in membrane biofilm reactor (MBfR) and then developed operational strategies of MBfR for high rate nitrogen removal from landfill leachate. Initially, influent containing nitrate and ammonium facilitated the development of n-DAMO and Anammox microorganisms in MBfR, but nitrogen removal performance is hard to be further improved even deteriorated. Detailed investigations of interactions among n-DAMO and Anammox microorganisms confirmed that extra addition of nitrite into MBfR fed with nitrate and ammonium not only stimulated the activities of Anammox bacteria, but also enhanced the activities of n-DAMO archaea from 172.3 to 356.9 mg NO3--N L-1 d-1. Functional gene analysis also indicated that mcrA and hzsA genes increased after nitrite addition. Based on this finding, influent containing NO3-, NO2- and NH4+ enabled nitrogen removal rates of MBfR increase from 224.9 to 888.2 mg N L-1 d-1. Finally, nitrate in the influent was gradually replaced with nitrite to mimic the effluent from partial nitriation of landfill leachate, but maintain the nitrate availability for n-DAMO archaea through increasing nitrate production from Anammox. These operation strategies enabled MBfR achieve the steady state with a nitrogen removal rate of 6.1 kg N m-3 d-1. Microbial community analysis revealed n-DAMO archaea, n-DAMO bacteria and Anammox bacteria jointly dominated the biofilm, and their relative abundance dynamically shifted with feeding regime. This work provides promising operational strategies for high rate of nitrogen removal from landfill leachate through integrating n-DAMO and Anammox process.

High performance nitrogen removal through integrating denitrifying anaerobic methane oxidation and Anammox: from enrichment to application.

Integrating denitrifying anaerobic methane oxidation (DAMO) with Anammox provides alternative solutions to simultaneously remove nitrogen and mitigate methane emission from wastewater treatment. However, the practical application of DAMO has been greatly limited by slow-growing DAMO microorganisms living on low-solubility gaseous methane. In this work, DAMO and Anammox co-cultures were fast enriched using high concentration of mixed sludges from various environments, and achieved nitrogen removal rate of 76.7 mg NH4+-N L-1 d-1 and 87.9 mg NO3--N L-1 d-1 on Day 178. Subsequently, nitrogen removal rate significantly decreased but recovered quickly through increasing methane flushing frequency, indicating methane availability could be the limiting factor of DAMO activity. Thus, this work developed a novel Membrane Aerated Membrane Bioreactor (MAMBR), which equipped with gas permeable membrane for efficient methane delivery and ultrafiltration membrane for complete biomass retention. After inoculated with enriched sludge, nitrogen removal rates of MAMBR were significantly enhanced to 126.9 mg NH4+-N L-1 d-1 and 158.8 mg NO3--N L-1 d-1 by membrane aeration in batch test. Finally, the MAMBR was continuously fed with synthetic wastewater containing ammonium and nitrite to mimic the effluent from partial nitritation. When steady state with nitrogen loading rate of 2500 mg N L-1 d-1 was reached, the MAMBR achieved total nitrogen removal of 2496.7 mg N L-1 d-1, with negligible nitrate in effluent (~6.5 mg NO3--N L-1). 16S rRNA amplicon sequencing and fluorescence in situ hybridization revealed the microbial community dynamics during enrichment and application. The high performance of nitrogen removal (2.5 kg N m-3 d-1) within 200 days operation and excellent biomass retention capacity (8.67 kg VSS m-3) makes the MAMBR promising for practical application of DAMO and Anammox in wastewater treatment.

Three-dimensional (3D) Printing Technology Assisted by Minimally Invasive Surgery for Pubic Rami Fractures.

The feasibility of three-dimensional (3D) printing technology combined with minimally invasive surgery in the treatment of pubic rami fractures was explored. From August 2015 to October 2017, a series of 30 patients who underwent surgical stabilization of their anterior pelvic ring (all utilizing the 3D printing technology) by one surgeon at a single hospital were studied. The minimally invasive incisions were made through anterior inferior cilia spine and pubic nodule. Data collected included the operative duration, the blood loss, the damage of the important tissue, the biographic union and the recovery of the function after the operation. Measurements on inlet and outlet pelvic cardiograph were made immediately post-operation and at all follow-up clinic visits. The scores of reduction and function were measured during follow-up. Results showed that the wounds of 30 patients were healed in the first stage, and there was no injury of important structures such as blood vessels and nerves. According to the Matta criteria, excellent effectiveness was obtained in 22 cases and good in 8 cases. According to the functional evaluation criteria of Majeed, excellent effectiveness was obtained in 21 cases and good in 9 cases. It was suggested that the 3D printing technology assisted by minimally invasive surgery can better evaluate the pelvic fracture before operation, which was helpful in plate modeling, and can shorten surgery duration and reduce intraoperative blood loss and complications. The positioning accuracy was improved, and better surgical result was finally achieved.

Novel three-dimensional nerve tissue engineering scaffolds and its biocompatibility with Schwann cells.

OBJECTIVE: To develop a novel scaffolding method for the copolymers poly lactide-co-glycolide acid (PLGA) to construct a three-dimensional (3-D) scaffold and explore its biocompatibility through culturing Schwann cells (SCs) on it. METHODS: The 3-D scaffolds were made by means of melt spinning, extension and weaving. The queueing discipline of the micro-channels were observed under a scanning electronic microscope (SEM).The sizes of the micropores and the factors of porosity were also measured. Sciatic nerves were harvested from 3-day-old Sprague Dawley (SD) rats for culture of SCs. SCs were separated, purified, and then implanted on PLGA scaffolds, gelatin sponge and poly-L-lysine (PLL)-coated tissue culture polystyrene (TCPS) were used as biomaterial and cell-supportive controls, respectively. The effect of PLGA on the adherence, proliferation and apoptosis of SCs were examined in vitro in comparison with gelatin sponge and TCPS. RESULTS: The micro-channels arrayed in parallel manners, and the pore sizes of the channels were uniform. No significant difference was found in the activity of Schwann cells cultured on PLGA and those on TCPS (P larger than 0.05), and the DNA of PLGA scaffolds was not damaged. CONCLUSION: The 3-D scaffolds developed in this study have excellent structure and biocompatibility, which may be taken as a novel scaffold candidate for nerve-tissue engineering.