A rapid and low-cost platform for detection of bacterial based on microchamber PCR microfluidic chip.

Polymerase chain reaction (PCR) has been considered as the gold standard for detecting nucleic acids. The simple PCR system is of great significance for medical applications in remote areas, especially for the developing countries. Herein, we proposed a low-cost self-assembled platform for microchamber PCR. The working principle is rotating the chamber PCR microfluidic chip between two heaters with fixed temperature to solve the problem of low temperature variation rate. The system consists of two temperature controllers, a screw slide rail, a chamber array microfluidic chip and a self-built software. Such a system can be constructed at a cost of about US$60. The micro chamber PCR can be finished by rotating the microfluidic chip between two heaters with fixed temperature. Results demonstrated that the sensitivity of the temperature controller is 0.1℃. The relative error of the duration for the microfluidic chip was 0.02 s. Finally, we successfully finished amplification of the target gene of Porphyromonas gingivalis in the chamber PCR microfluidic chip within 35 min and on-site detection of its PCR products by fluorescence. The chip consisted of 3200 cylindrical chambers. The volume of reagent in each volume is as low as 0.628 nL. This work provides an effective method to reduce the amplification time required for micro chamber PCR.

De Novo Design of Entropy-Driven Polymers Resistant to Bacterial Attachment via Multicomponent Reactions.

Nonbactericidal polymers that prevent bacterial attachment are important for public health, environmental protection, and avoiding the generation of superbugs. Here, inspired by the physical bactericidal process of carbon nanotubes and graphene derivatives, we develop nonbactericidal polymers resistant to bacterial attachment by using multicomponent reactions (MCRs) to introduce molecular "needles" (rigid aliphatic chains) and molecular "razors" (multicomponent structures) into polymer side chains. Computer simulation reveals the occurrence of spontaneous entropy-driven interactions between the bacterial bilayers and the "needles" and "razors" in polymer structures and provides guidance for the optimization of this type of polymers for enhanced resistibility to bacterial attachment. The blending of the optimized polymer with commercially available polyurethane produces a film with remarkably superior stability of the resistance to bacterial adhesion after wear compared with that of commercial mobile phone shells made by the Sharklet technology. This proof-of-concept study explores entropy-driven polymers resistant to bacterial attachment via a combination of MCRs, computer simulation, and polymer chemistry, paving the way for the de novo design of nonbactericidal polymers to prevent bacterial contamination.

Stimuli-Responsive Multifunctional Phenylboronic Acid Polymers Via Multicomponent Reactions: From Synthesis to Application.

Stimuli-responsive polymers undergo changes under different environmental conditions. Among them, phenylboronic acid (PBA) containing polymers (PBA-polymers) are unique, because they can selectively react with diols to generate borates that are sensitive to pH, sugars, and H2 O2 , and can be effectively used to synthesize smart drug carriers and self-healing hydrogels. Recently, multifunctional PBA-polymers (MF-PBA-polymers) have been developed using multicomponent reactions (MCRs) to introduce PBA groups into polymer structures. These MF-PBA-polymers have features similar to those of traditional PBA-polymers; moreover, they exhibit additional properties, such as fluorescence, antimicrobial activity, and antioxidant capability, when different MCRs are used. In this mini review, the preparation of these MF-PBA-polymers are summarized and the new properties/functions that have been introduced into these polymers using different MCRs are discussed. The uses of these MF-PBA-polymers as fluorescent cell anticoagulants, drug carriers, and gelators of functional self-healing hydrogels have been discussed. Additionally, the challenges encountered during their preparation are discussed and also the future developments in this field are touched upon.

Double surfactants-assisted electromembrane extraction of cyromazine and melamine in surface water, soil and cucumber samples followed by capillary electrophoresis with contactless conductivity detection.

BACKGROUND: Cyromazine (CYR) and its main degradation product melamine (MEL) are attracting wide attention due to their potential hazards to the environment and humans. In this work, double surfactants-assisted electromembrane extraction (DS-EME) by Tween 20 and alkylated phosphate was firstly used for purification and extraction of CYR and MEL, and the extract was directly analyzed by capillary electrophoresis with capacitively coupled contactless conductivity detection. RESULTS: Under the optimum conditions, two targets could be well separated from the main interferences, including common biogenic amines and inorganic cations within 14 min. This developed method was successfully applied to the analyses of surface water, soil and cucumber samples, and the average recoveries were in the range 93.3-112%. DS-EME provided a synergistic purification and enrichment effect for CYR and MEL by adding Tween 20 and alkylated phosphate into donor phase and supporting liquid membrane, respectively. Satisfactory limits of detection [0.2-1.5 ng mL-1 , signal-to-noise ratio (S/N) = 3] could be obtained in the tested sample matrices, and the corresponding enrichment factors were up to 115∼123 times. CONCLUSION: This developed method provides an alternative for the simultaneous analysis of CYR and MEL in complex real-world samples. © 2019 Society of Chemical Industry.

Mebendazole elicits potent antimyeloma activity by inhibiting the USP5/c-Maf axis.

c-Maf is a critical oncogenic transcription factor that contributes to myelomagenesis. Our previous studies demonstrated that the deubiquitinase USP5 stabilizes c-Maf and promotes myeloma cell proliferation and survival; therefore, the USP5/c-Maf axis could be a potential target for myeloma therapy. As a concept of principle, the present study established a USP5/c-Maf-based luciferase system that was used to screen an FDA-approved drug library. It was found that mebendazole, a typical anthelmintic drug, preferentially induced apoptosis in c-Maf-expressing myeloma cells. Moreover, oral administration of mebendazole delayed the growth of human myeloma xenografts in nude mice but did not show overt toxicity. Further studies showed that the selective antimyeloma activity of mebendazole was associated with the inhibition of the USP5/c-Maf axis. Mebendazole downregulated USP5 expression and disrupted the interaction between USP5 and c-Maf, thus leading to increased levels of c-Maf ubiquitination and subsequent c-Maf degradation. Mebendazole inhibited c-Maf transcriptional activity, as confirmed by both luciferase assays and expression measurements of c-Maf downstream genes. In summary, this study identified mebendazole as a USP5/c-Maf inhibitor that could be developed as a novel antimyeloma agent.

Reactivation of Lysosomal Ca2+ Efflux Rescues Abnormal Lysosomal Storage in FIG4-Deficient Cells.

Loss of function of FIG4 leads to Charcot-Marie-Tooth disease Type 4J, Yunis-Varon syndrome, or an epilepsy syndrome. FIG4 is a phosphatase with its catalytic specificity toward 5'-phosphate of phosphatidylinositol-3,5-diphosphate (PI3,5P2). However, the loss of FIG4 decreases PI3,5P2 levels likely due to FIG4's dominant effect in scaffolding a PI3,5P2 synthetic protein complex. At the cellular level, all these diseases share similar pathology with abnormal lysosomal storage and neuronal degeneration. Mice with no FIG4 expression (Fig4(-/-)) recapitulate the pathology in humans with FIG4 deficiency. Using a flow cytometry technique that rapidly quantifies lysosome sizes, we detected an impaired lysosomal fission, but normal fusion, in Fig4(-/-) cells. The fission defect was associated with a robust increase of intralysosomal Ca(2+) in Fig4(-/-) cells, including FIG4-deficient neurons. This finding was consistent with a suppressed Ca(2+) efflux of lysosomes because the endogenous ligand of lysosomal Ca(2+) channel TRPML1 is PI3,5P2 that is deficient in Fig4(-/-) cells. We reactivated the TRPML1 channels by application of TRPML1 synthetic ligand, ML-SA1. This treatment reduced the intralysosomal Ca(2+) level and rescued abnormal lysosomal storage in Fig4(-/-) culture cells and ex vivo DRGs. Furthermore, we found that the suppressed Ca(2+) efflux in Fig4(-/-) culture cells and Fig4(-/-) mouse brains profoundly downregulated the expression/activity of dynamin-1, a GTPase known to scissor organelle membranes during fission. This downregulation made dynamin-1 unavailable for lysosomal fission. Together, our study revealed a novel mechanism explaining abnormal lysosomal storage in FIG4 deficiency. Synthetic ligands of the TRPML1 may become a potential therapy against diseases with FIG4 deficiency.

Nuclear-targeting TAT-PEG-Asp8-doxorubicin polymeric nanoassembly to overcome drug-resistant colon cancer.

AIM: Drug efflux-associated multidrug resistance (MDR) is a main obstacle to effective cancer chemotherapy. Large molecule drugs are not the substrates of P-glycoprotein, and can circumvent drug efflux and be retained inside cells. In this article we report a polymer-drug conjugate nanoparticulate system that can overcome MDR based on size-related exclusion effect. METHODS: Doxorubicin was coupled with the triblock polymeric material cell-penetrating TAT-PEG-poly(aspartic acid). The amphiphilic macromolecules (termed TAT-PEG-Asp8-Dox) could self-assemble into nanoparticles (NPs) in water. The antitumor activity was evaluated in drug-resistant human colon cancer HCT8/ADR cells in vitro and in nude mice bearing HCT8/ADR tumor. RESULTS: The self-assembling TAT-PEG-Asp8-Dox NPs were approximately 150 nm with a narrow particle size distribution, which not only increased the cellular uptake efficiency, but also bypassed P-glycoprotein-mediated drug efflux and improved the intracellular drug retention, thus yielding an enhanced efficacy for killing drug-resistant HCT8/ADR colon cancer cells in vitro. Importantly, the TAT-PEG-Asp8-Dox NPs enhanced the intranuclear disposition of drugs for grater inhibition of DNA/RNA biosynthesis. In nude mice bearing xenografted HCT8/ADR colon cancers, intravenous or peritumoral injection of TAT-PEG-Asp8-Dox NPs for 22 d effectively inhibited tumor growth. CONCLUSION: TAT-PEG-Asp8-Dox NPs can increase cellular drug uptake and intranuclear drug delivery and retain effective drug accumulation inside the cells, thus exhibiting enhanced anticancer activity toward the drug-resistant human colon cancer HCT8/ADR cells.

Walnut shell oil-bath torrefaction coupled with fast pyrolysis: Effect of torrefaction heating modes.

Lignocellulosic biomass, renewable with short growth cycle and diverse sources, can be substituted fossil fuel. However, low effective hydrogen-to-carbon ratio (H/Ceff) limits its applications. Torrefaction and co-pyrolysis with high H/Ceff feedstocks are promising technology. This paper investigated the effect of heating modes on oil-bath torrefaction of walnut shells, followed by fast co-pyrolysis. Six heating modes during oil-bath torrefaction were evaluated. Com1 (Microwave 67 %, Lightwave 33 %) yielded the lowest residual yield 84 wt%, while the highest gas production 495.47 mL/g which mainly composed of CO and CO2. Torrefied feedstock under Com1 had the highest H/Ceff. Decarboxylation and decarbonylation reactions dominated among oil-bath torrefaction. Com1 produced the most hydrocarbons and least oxygen-containing compounds. As microwave ratio decreased, the content of olefins, acids and phenols decreased, monocyclic aromatic hydrocarbons and alcohols was showed opposite tend. This study offers new ideas for microwave and lightwave torrefaction and promoting hydrocarbon production from lignocellulosic biomass.

Influences of input concentration, media particle size, metal cation valence, and ionic concentration on the transport, long-term release, and particle breakage of polyvinyl chloride nanoplastics in saturated porous media.

The environmental impact of nanoplastics has gradually attracted widespread attention; however, nanoplastics of polyvinyl chloride, one of the most commonly used plastics, have not yet been studied. In this study, we investigated the transport, long-term release behavior, and particle fracture of polyvinyl chloride nanoplastics (PVC NPs) in saturated quartz sand with different metal cations, ionic concentrations, input concentrations, and sand grain sizes by determining breakthrough, long-term release, and particle size distribution curves. The breakthrough curves and retention profiles were simulated by a mathematical model. The transport of PVC NPs increased with increased input concentration and sand grain size, which could be predicted by the Derjaguin-Landau-Verwey-Overbeek (DLVO) and colloid filtration theories. Increased ionic concentration and metal cation valence could restrain the transport of PVC NPs in saturated quartz sand owing to the decreased energy barrier between PVC NPs and sand grains. The total released amount of PVC NPs in the long-term release tests with different experimental conditions ranged from 3.91 to 21.95%. Increased sand grain size and decreased metal cation valence and ionic concentration resulted in an increased released amount of retained PVC NPs in saturated quartz sand, indicating increased release ability and mobility. The particle fracture results indicated that the PVC NPs were not broken down during long-term release under the experimental conditions of this research. This opens up a completely new and meaningful study of whether nanoplastics are broken down into smaller nanoplastics during their long-term release under various conditions.

Release characteristics of hydrochar-derived dissolved organic matter: Effects of hydrothermal temperature and environmental conditions.

Much research has been done on the preparation and application of hydrochars, but research on the release characteristics of hydrochar-derived dissolved organic matter (HDOM) is very limited; clarifying the release characteristics of HDOM is important for understanding and adjusting the environmental behaviour of hydrochar. Herein, the potential release of HDOM from rice straw-derived hydrochars prepared at different hydrothermal temperatures was investigated under various potential environmental conditions for the first time. The total release quantity and humification degree of HDOM decreased with increasing hydrothermal temperature. The critical dividing line for various hydrothermal reactions, decomposition and polymerization, was in the range of 240 °C-260 °C. Alkaline condition increased the HDOM release amount (up to 299 mg g-1), molecular weight (as high as 423 Da) and molecular diversity (8857 compounds) from rice straw-derived hydrochars. The unique substances of HDOM released under alkaline condition were mainly distributed in lipids-like substances, CRAM/lignins-like substances, aromatic structures, and tannins-like substances, while few unique substances were found under acidic condition. Additionally, CRAM/lignins-like substances were the most abundant in all HDOM samples, reaching 82%, which were relatively stable and could achieve carbon sequestration in different environments. The findings provided a new insight on understanding the potential environment behaviors of hydrochar.

Neurotrophin-3 gene-modified Schwann cells promote TrkC gene-modified mesenchymal stem cells to differentiate into neuron-like cells in poly(lactic-acid-co-glycolic acid) multiple-channel conduit.

Rapid progress in the field of nerve tissue engineering has opened up the way for new therapeutic strategies for spinal cord injury (SCI). Bone marrow-derived mesenchymal stem cells (MSCs) could be differentiated into neural lineages, which can be used as a potential cell source for nerve repair. Schwann cells (SCs) have been reported to support structural and functional recovery of SCI. In this study, we co-cultured neurotrophin-3 (NT-3) gene-modified SCs and NT-3 receptor tyrosine protein kinase C (TrkC) gene-modified MSCs in a three-dimensional porous poly(lactic-acid-co-glycolic acid) (PLGA) conduit with multiple channels in vitro for 14 days. Our results showed that more than 50% of the grafted MSCs were MAP2- and ß-III-tubulin-positive cells, and the MSCs expressed a high level of ß-III-tubulin detected by Western blotting, indicating a high rate of neuronal differentiation. Furthermore, immunostaining of PSD95 revealed the formation of a synapse-like structure, which was confirmed under electron microscopy. In conclusion, co-culture of NT-3 gene-modified SCs and TrkC gene-modified MSCs in the PLGA multiple-channeled conduit can promote MSCs' differentiation into neuron-like cells with synaptogenesis potential. Our study provides a biological basis for future application of this artificial MSCs/SCs/PLGA complex in the SCI treatment.

Magnetic Self-Healing Hydrogel from Difunctional Polymers Prepared via the Kabachnik-Fields Reaction.

The development of high quality magnetic self-healing hydrogels containing well-dispersed magnetic nanoparticle has been a challenging procedure due to unavailable methods of facilely introducing groups that can efficiently stabilize these magnetic nanoparticles in the self-healing hydrogels. In this research, a polymer containing both phenylboronic acid (PBA) and phosphonic acid (PA) groups has been developed by the Kabachnik-Fields (KF) reaction. This polymer well disperses iron oxide nanoparticles (IONPs) through the strong interactions between the PA groups and the surface of the IONPs; thus, this polymer effectively mixed IONPs and poly(vinyl alcohol) (PVA) to form a hydrogel containing well-dispersed IONPs. The resulting hydrogel is self-healing, owing to the dynamic borate ester linkages. Moreover, the presence of the IONPs endowed the hydrogel with magnetic properties, also making it heat-responsive in an alternating magnetic field and expanding its application as a contrast agent for magnetic resonance imaging. The magnetic self-healing hydrogel showed excellent biosafety properties in animal experiments, suggesting its potential as an injectable implant material for biological and medical applications. This research exploits a biocompatible magnetic self-healing hydrogel with well-dispersed IONPs, demonstrating the value of the KF reaction in the development of functional polymers and smart materials, which might prompt a broad study of multicomponent reactions in interdisciplinary fields.

Microwave catalytic co-pyrolysis of waste cooking oil and low-density polyethylene to produce monocyclic aromatic hydrocarbons: Effect of different catalysts and pyrolysis parameters.

It is promising to convert waste oil and plastics to renewable fuels and chemicals by microwave catalytic co-pyrolysis, enabling pollution reduction and resource recovery. The purpose of this study was to evaluate the effect of catalysts on the product selectivity of microwave-assisted co-pyrolysis of waste cooking oil and low-density polyethylene and optimize the pyrolysis process, including pyrolysis temperature, catalytic temperature, waste cooking oil to low-density polyethylene ratio, and catalyst to feedstocks ratio. The results indicated that catalysts had a great influence on the product distribution, and the yield of BTX (benzene, toluene, and xylenes), which increased in the following order: SAPO-34 < Hß < HY < HZSM-5. HZSM-5 was more active for the formation of light aromatic hydrocarbons as compared to others, where the concentrations of toluene, benzene and xylenes reached 252.59 mg/mL, 114.7 mg/mL and 132.91 mg/mL, respectively. The optimum pyrolysis temperature, catalytic temperature, waste cooking oil to low-density polyethylene ratio and catalyst to feedstocks ratio could be 550 °C, 450 °C, 1:1 and 1:2, respectively, to maximize the formation of BTX and inhibit the formation of polycyclic aromatic hydrocarbons.

Brominated and phosphate flame retardants from interior and surface dust of personal computers: insights into sources for human dermal exposure.

It remains unclear whether internal or external sources play the more significant role in flame retardant (FR) contamination of surface dust from personal computers (PCs), which may lead to bias on dermal exposure assessment of FRs. In the present study, the occurrence and profiles of several brominated and phosphate FRs were measured in the interior dust, and the upper surface (keyboard) and bottom surface (bottom cover) wipes of PCs. BDE 209 (639 ng/g), decabromodiphenyl ethane (DBDPE, 885 ng/g), and triphenyl phosphate (TPHP, 1880 ng/g) were the most abundant chemicals in interior PC dust, while tris(2-chloroisopropyl) phosphate (TCIPP), TPHP, and DBDPE were dominant on both surfaces of PCs. No significant correlation between interior dust and both PC surfaces was observed for concentrations of most FRs except BDE 183. Different sources of FRs for interior and surface dust of PCs were further revealed by principal component analysis (PCA). FRs from external sources, rather than emission from inner PC components, are likely the main contributor for FR profiles on PC surfaces. Exposure assessment results demonstrated a minor contribution from PC dermal contact, compared with hand-to-mouth uptake, to total exposure. The applicability of surface wipes to assess dermal exposure to FR-treated products needs to be further investigated.

Fabrication of claviform fluorescent polymeric nanomaterials containing disulfide bond through an efficient and facile four-component Ugi reaction.

Multicomponent reactions (MCRs) have attracted broad interest for preparation of functional nanomaterials especially for the synthesis of functional polymers. Herein, we utilized an "old" MCR, the four-component Ugi reaction, to synthesize disulfide bond containing poly(PEG-TPE-DTDPA) amphiphilic copolymers with aggregation-induced emission (AIE) feature. This four-component Ugi reaction was carried out under rather mild reaction conditions, such as room temperature, no gas protection and absent of catalysts. The amphiphilic poly(PEG-TPE-DTDPA) copolymers with high number-average molecular weight (up to 86,440 Da) can self-assemble into claviform fluorescent polymeric nanoparticles (FPNs) in aqueous solution, and these water-dispersed nanoparticles exhibited strong emission, large Stokes shift (142 nm), low toxicity and remarkable ability in cellular imaging. Moreover, owing to the introduction of 3,3'-dithiodipropionic acid with disulfide bond, the resultant AIE-active poly(PEG-TPE-DTDPA) could display reduction-responsiveness and be utilized for synthesis of photothermal agents in-situ. Therefore, the AIE-active poly(PEG-TPE-DTDPA) could be promising for controlled intracellular delivery of biological activity molecules and fabrication of multifunctional AIE-active materials. Therefore, these novel AIE-active polymeric nanoparticles could be of great potential for various biomedical applications, such as biological imaging, stimuli-responsive drug delivery and theranostic applications.

An antioxidant self-healing hydrogel for 3D cell cultures.

In this paper, an antioxidant self-healing hydrogel has been prepared. The Biginelli reaction was used to prepare a monomer containing phenylboronic acid (PBA) and 3,4-dihydropyrimidin-2(1H)-one (DHPM) groups. This PBA-DHPM monomer was copolymerized with poly(ethylene glycol methyl ether) methacrylate (PEGMA) to produce a water-soluble copolymer via radical polymerization. The resulting copolymer quickly crosslinked poly(vinyl alcohol) (PVA) through borate ester bonds to generate a self-healing hydrogel under mild conditions (pH ∼ 7.4, 25 °C). The prepared hydrogel showed an inherent antioxidant ability because of the DHPM moieties in the hydrogel structure. It also showed no cytotoxicity, and in an in vivo mouse model the hydrogel injected under the skin of a mouse hardly caused any adverse reactions, suggesting that this hydrogel could be used as an implantable biomaterial. This first report of an antioxidant self-healing hydrogel demonstrates a new application of the Biginelli reaction in materials science, which might prompt a broad study of multicomponent reactions in interdisciplinary fields.

High-throughput preparation of radioprotective polymers via Hantzsch's reaction for in vivo X-ray damage determination.

Radioprotectors for acute injuries caused by large doses of ionizing radiation are vital to national security, public health and future development of humankind. Here, we develop a strategy to explore safe and efficient radioprotectors by combining Hantzsch's reaction, high-throughput methods and polymer chemistry. A water-soluble polymer with low-cytotoxicity and an excellent anti-radiation capability has been achieved. In in vivo experiments, this polymer is even better than amifostine, which is the only approved radioprotector for clinical applications, in effectively protecting zebrafish embryos from fatally large doses of ionizing radiation (80 Gy X-ray). A mechanistic study also reveals that the radioprotective ability of this polymer originates from its ability to efficiently prevent DNA damage due to high doses of radiation. This is an initial attempt to explore polymer radioprotectors via a multi-component reaction. It allows exploiting functional polymers and provides the underlying insights to guide the design of radioprotective polymers.

Neurotrophin-3 released from implant of tissue-engineered fibroin scaffolds inhibits inflammation, enhances nerve fiber regeneration, and improves motor function in canine spinal cord injury.

Spinal cord injury (SCI) normally results in cell death, scarring, cavitation, inhibitory molecules release, etc., which are regarded as a huge obstacle to reconnect the injured neuronal circuits because of the lack of effective stimulus. In this study, a functional gelatin sponge scaffold was used to inhibit local inflammation, enhance nerve fiber regeneration, and improve neural conduction in the canine. This scaffold had good porosity and modified with neurotrophin-3 (NT-3)/fibroin complex, which showed sustained release in vitro. After the scaffold was transplanted into canine spinal cord hemisection model, hindlimb movement, and neural conduction were improved evidently. Migrating host cells, newly formed neurons with associated synaptic structures together with functional blood vessels with intact endothelium in the regenerating tissue were identified. Taken together, the results demonstrated that using bioactive scaffold could establish effective microenvironment stimuli for endogenous regeneration, providing a potential and practical strategy for treatment of spinal cord injury. © 2018 The Authors Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2158-2170, 2018.

A comparative study of gelatin sponge scaffolds and PLGA scaffolds transplanted to completely transected spinal cord of rat.

This study sought to investigate whether gelatin sponge (GS) scaffold would produce less acidic medium in injured spinal cord, as compared with poly(lactic-co-glycolic acid) (PLGA) scaffold, to determine which of the two scaffolds as the biomaterial is more suitable for transplantation into spinal cord. GS scaffold or PLGA scaffold was transplanted into a transected spinal cord in this study. Two months after transplantation of scaffolds, acid sensing ion channel 1a (ASIC1a) positive cells expressing microtubule associated protein 2 (Map2) were observed as well as expressing adenomatous polyposis coli (APC) in spinal cord. GFAP positive cells were distributed at the rostral and caudal of the injury/graft area in the GS and PLGA groups. Western blot showed ASIC1a and GFAP expression of injured spinal cord was downregulated in the GS group. The number of CD68 positive cells was fewer and NF nerve fibers were more in the GS group. Nissl staining and cell counting showed that the number of survival neurons was comparable between the GS and PLGA groups in the pyramidal layer of sensorimotor cortex and the red nucleus of midbrain. However, in the Clarke's nucleus at L1 spinal segment, the surviving neurons in the GS group were more numerous than that in the PLGA group. H&E staining showed that the tissue cavities in the GS group were smaller in size than that in the PLGA group. The results suggest that GS scaffold is more suitable for transplantation to promote the recovery of spinal cord injury compared with PLGA scaffold.

Bone marrow mesenchymal stem cells in a three-dimensional gelatin sponge scaffold attenuate inflammation, promote angiogenesis, and reduce cavity formation in experimental spinal cord injury.

Three-dimensional (3D) gelatin sponge (GS) scaffolds were constructed by ensheathing GS with a thin film of poly-(lactide-co-glycolide) (PLGA). Rat bone marrow-derived mesenchymal stem cells (MSCs) were isolated, cultured, and then seeded to the scaffolds. Distribution of cells and cell growth, survival, and proliferation within the scaffolds were then determined. Immunofluorescence and Western blot analysis were employed to detect the deposition of fibronectin to the scaffolds on day 3 and day 7 of culture. Scaffolds with or without MSCs were then transplanted into the transected rat spinal cord. One or 8 weeks following transplantation, cavity areas, activated macrophages/microglia, expression of TNF-α and IL-1ß, and neovascularization within the grafts were examined and quantified. Deposition of fibronectin (FN) and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) as potential inducing factors for angiogenesis were also examined. Results showed that 3D GS scaffolds allowed MSCs to adhere, survive, and proliferate and also FN to deposit. In vivo transplantation experiments demonstrated that these scaffolds were biocompatible, and MSCs seeded to the scaffolds played an important role in attenuating inflammation, promoting angiogenesis, and reducing cavity formation. Therefore, the GS scaffolds with MSCs may serve as promising supporting transplants for repairing spinal cord injury.