A Novel Double‐Network Hydrogel Induces Spontaneous Articular Cartilage Regeneration in vivo in a Large Osteochondral Defect

We have developed a novel method to induce spontaneous hyaline cartilage regeneration in vivo for a large osteochondral defect by implanting a plug made from a double‐network hydrogel composed of poly(2‐acrylamido‐2‐methylpropanesulfonic acid) and poly(N,N′‐dimethylacrylamide) at the bottom of the defect, leaving the cavity vacant. In cells regenerated in the treated defect, type‐2 collagen, Aggrican, and SOX9 mRNAs were highly expressed and the regenerated matrix was rich in proteoglycan and type‐2 collagen at 4 weeks. This fact gave a significant modification to the commonly established concept that hyaline cartilage tissue cannot regenerate in vivo. This study prompted an innovative strategy in the field of joint surgery to repair an osteochondral defect using an advanced, high‐function hydrogel.

     

Ion chemistry of 1H-1,2,3-triazole

A combination of experimental methods, photoelectron-imaging spectroscopy, flowing afterglow-photoelectron spectroscopy and the flowing afterglow-selected ion flow tube technique, and electronic structure calculations at the B3LYP/6-311++G(d,p) level of density functional theory (DFT) have been employed to study the mechanism of the reaction of the hydroxide ion (HO-) with 1H-1,2,3-triazole. Four different product ion species have been identified experimentally, and the DFT calculations suggest that deprotonation by HO- at all sites of the triazole takes place to yield these products. Deprotonation of 1H-1,2,3-triazole at the N1-H site gives the major product ion, the 1,2,3-triazolide ion. The 335 nm photoelectron-imaging spectrum of the ion has been measured. The electron affinity (EA) of the 1,2,3-triazolyl radical has been determined to be 3.447 +/- 0.004 eV. This EA and the gas-phase acidity of 2H-1,2,3-triazole are combined in a negative ion thermochemical cycle to determine the N-H bond dissociation energy of 2H-1,2,3-triazole to be 112.2 +/- 0.6 kcal mol-1. The 363.8 nm photoelectron spectroscopic measurements have identified the other three product ions. Deprotonation of 1H-1,2,3-triazole at the C5 position initiates fragmentation of the ring structure to yield a minor product, the ketenimine anion. Another minor product, the iminodiazomethyl anion, is generated by deprotonation of 1H-1,2,3-triazole at the C4 position, followed by N1-N2 bond fission. Formation of the other minor product, the 2H-1,2,3-triazol-4-ide ion, can be rationalized by initial deprotonation of 1H-1,2,3-triazole at the N1-H site and subsequent proton exchanges within the ion-molecule complex. The EA of the 2H-1,2,3-triazol-4-yl radical is 1.865 +/- 0.004 eV.     

Incorporation of water-soluble drugs in PLGA microspheres

Poly(lactide-co-glycolide) (PLGA) microspheres containing blue dextran, as a model of water-soluble drugs, were prepared from w₁/o/w₂ emulsions by using a microhomogenizer and a solvent evaporation method. Effects of preparation conditions, such as, concentration of poly(vinyl alcohol) (PVA) in w₂ phase, viscosity of inner soluble water phase, volume ratio of oil phase to w₁ phase in primary emulsion, PLGA concentration in oil phase, and molecular weight or composition of PLGA, upon the properties of PLGA microspheres containing water-soluble drugs were examined. Concentration of poly(vinyl alcohol) (PVA), the dispersant dissolved in w₂ phase of secondary emulsion did not show any effects on the final particle size. On the other hand, volume ratio of oil phase to water one in primary emulsion affected the final particle size, which seemed to be related to the local PLGA concentration in w₁/o emulsions. That is, the particle size increased as the volume ratio of w₁ phase against oil phase, w₁/o (v/v), increased. The loading efficiency, however, was not affected by the volume ratio of w₁/o (v/v), but affected by blue dextran concentration in w₁ phase. Higher loading efficiency was observed in PLGA microspheres prepared from w₁ phase containing lower concentration of blue dextran. Blue dextran solution (inner water phase) with the lower viscosity may result in the lower leakage ratio of blue dextran during the preparation procedure. Increases in concentration and molecular weight of PLGA made particle size larger.     

Automatic F positron source supply system for a monoenergetic positron beam

A system which supplies an intense ¹⁸F (half life 110 min) positron source produced by an AVF cyclotron through ¹⁸O(p,n)¹⁸F reaction has been constructed. Produced ¹⁸F is transferred to a low background experiment hall through a capillary. It is electro-deposited on a graphite rod and used for a source of a slow positron beam. In the meantime the next batch of target ¹⁸O water is loaded and proton irradiation proceeds. This system makes it possible to perform continuous positron beam experiments using the ¹⁸F positron source.     

Matrix-assisted laser desorption/ionization imaging mass spectrometry revealed traces of dental problem associated with dental structure

Periodontal disease is a serious dental problem because it does not heal naturally and leads to tooth loss. In periodontal disease, inflammation at periodontal tissue is thought as predominant, and its effect against tooth itself remains unclear. In this study, we applied matrix-assisted laser desorption/ionization imaging mass spectrometry (IMS) to teeth for the first time. By comparing anatomical structure of tooth affected with periodontal disease with normal ones, we analyzed traces of the disease on tooth. We found signals characteristic of enamel, dentin, and dental pulp, respectively, in mass spectra obtained from normal teeth. Ion images reconstructed using these signals showed anatomical structures of the tooth clearly. Next, we performed IMS upon teeth of periodontal disease. Overall characteristic of the mass spectrum appeared similar to normal ones. However, ion images reconstructed using signals from the tooth of periodontal disease revealed loss of periodontal ligament visualized together with dental pulp in normal teeth. Moreover, ion image clearly depicted an accumulation of signal at m/z 496.3 at root surface. Such an accumulation that cannot be examined only from mass spectrum was revealed by utilization of IMS. Recent studies about inflammation revealed that the signal at m/z 496.3 reflects lyso-phosphatidylcholine (LPC). Infiltration of the signal is statistically significant, and its intensity profile exhibited the influence has reached deeply into the tooth. This suggests that influence of periodontal disease is not only inflammation of periodontal tissue but also infiltration of LPC to root surface, and therefore, anti-inflammatory treatment is required besides conventional treatments.     

Structures of Stoichiometric Sodium Oxide Cluster Cations Studied by Ion Mobility Mass Spectrometry

Structures of stable compositions of sodium oxide cluster cations (Na_nO_m⁺,n≤11) have been investigated by ion mobility mass spectrometry. Stoichiometric compositions series, Na(Na₂O)_(n-1)/2⁺ (n=3, 5, 7, 9, and 11), were observed as stable composition series, and NaO(Na₂O)_(n-1)/2⁺ series (n=5, 7, 9, and 11) were observed as secondary stable series in the mass spectra. To assign the structures of these cluster ion series, collision cross sections between the ions and helium buffer gas were determined experimentally from the ion mobility measurements. Theoretical collision cross sections were also calculated for optimized structures of these compositions. Finally, the structures of Na(Na₂O)_(n-1)/2⁺ and NaO(Na₂O)_(n-1)/2⁺ were assigned to those having similar structural frames for each n except for n=9. All bonds in the assigned structures of Na(Na₂O)_(n-1)/2⁺ were between sodium and oxygen. On the other hand, there was one O-O bond in addition to Na-O bonds in NaO(Na₂O)_(n-1)/2⁺. This result indicates that NaO(Na₂O)_(n-1)/2⁺ have a peroxide ion (O₂^2-) as a substitute for an oxide ion (O^2-) of Na(Na₂O)_(n-1)/2⁺. As a result, both stable series, Na(Na₂O)_(n-1)/2⁺ and NaO(Na₂O)_(n-1)/2⁺, are closed-shell compositions. These closed-shell characteristics have a strong influence on the stability of sodium oxide cluster cations.     

Novel separation medium spongy monolith for high throughput analyses

Sponge-like material was utilized as novel chromatographic media for high throughput analyses. The pore size of the sponge-like material was several dozen micrometer, and was named spongy monolith because it consists of continuous structured copolymers, which was made of poly(ethylene-co-vinyl acetate), such as monolithic materials including silica monoliths and organic polymer monoliths. The spongy monolith was packed into a stainless steel column (100 mm × 4.6 mm I.D.) and evaluated in liquid chromatography (LC) with an on-line column-switching LC concentration system. The results indicate that the packed column could be used with high flow rates and low back pressure (9.0 mL/min at 0.5 MPa). Furthermore, bisphenol A was quantitatively recovered by on-line column-switching LC concentration with the spongy monolithic column. Additionally, the adsorption capacity and physical strength of the media was enhanced via chemical modification of spongy monoliths using glycerol dimethacrylate. The results compared with original spongy monolith demonstrated that a higher adsorption capacity was achieved on a shorter column, and a stable low back pressure was obtained at high throughput elution even with a longer column.     

Mesoporous silica nanoparticles for 19F magnetic resonance imaging,fluorescence imaging,and drug delivery

Multifunctional mesoporous silica nanoparticles (MSNs) are good candidates for multimodal applications in drug delivery, bioimaging, and cell targeting. In particular, controlled release of drugs from MSN pores constitutes one of the superior features of MSNs. In this study, a novel drug delivery carrier based on MSNs, which encapsulated highly sensitive ¹⁹F magnetic resonance imaging (MRI) contrast agents inside MSNs, was developed. The nanoparticles were labeled with fluorescent dyes and functionalized with small molecule-based ligands for active targeting. This drug delivery system facilitated the monitoring of the biodistribution of the drug carrier by dual modal imaging (NIR/¹⁹F MRI). Furthermore, we demonstrated targeted drug delivery and cellular imaging by the conjugation of nanoparticles with folic acid. An anticancer drug (doxorubicin, DOX) was loaded in the pores of folate-functionalized MSNs for intracellular drug delivery. The release rates of DOX from the nanoparticles increased under acidic conditions, and were favorable for controlled drug release to cancer cells. Our results suggested that MSNs may serve as promising ¹⁹F MRI-traceable drug carriers for application in cancer therapy and bio-imaging.     

Cadinane‐Type Sesquiterpenoids,Strobilols E – K,from the Liquid Culture of Strobilurus ohshimae

New cadinane sequiterpenoids, strobilols E–K ( 5 – 11 , resp.) have been isolated from a liquid culture of Strobilurus ohshimae. Their structures have been established on the basis of spectral analyses.