Combination of Metal-Phenolic Network-Based Immunoactive Nanoparticles and Bipolar Irreversible Electroporation for Effective Cancer Immunotherapy.

To circumvent the limitations of conventional cancer immunotherapy, it is critical to prime antigen-presenting cells (APCs) to initiate the cancer-immune cycle. Here, the authors develop a metal-phenolic network (MPN)-based immunoactive nanoparticle in combination with irreversible electroporation (IRE) for an effective cancer immunotherapy. The MPN nanoparticles are synthesized by coordinating tannic acid with manganese (Mn) ions, and subsequent coating with CpG-oligodeoxynucleotides (CpG-ODNs) via hydrogen bonding. The CpG-ODN-coated Mn-phenolic network (CMP) nanoparticles are effectively internalized into macrophages, a type of APCs, and successfully trigger M1 polarization to promote release of proinflammatory cytokines. Notably, the CMP nanoparticles demonstrate an extended retention time period than the free CpG-ODN in the tumor. The tumor microenvironment tailored bipolar IRE, enhances the therapeutic efficacy by significantly broadening the ablation zone, which further increases immunogenic cell death (ICD). Ultimately, the simultaneous CMP nanoparticles and IRE treatment successfully inhibit tumor growth and prolong survival in a mouse tumor model. Thus, CMP nanoparticles are empowered with Mn and CpG-ODN immunomodulators and the tumor microenvironment tailored bipolar IRE will be a new tool for effective cancer immunotherapy to treat intractable malignancies.

A Small Molecule That Promotes Cellular Senescence Prevents Fibrogenesis and Tumorigenesis.

Uncontrolled proliferative diseases, such as fibrosis or cancer, can be fatal. We previously found that a compound containing the chromone scaffold (CS), ONG41008, had potent antifibrogenic effects associated with EMT or cell-cycle control resembling tumorigenesis. We investigated the effects of ONG41008 on tumor cells and compared these effects with those in pathogenic myofibroblasts. Stimulation of A549 (lung carcinoma epithelial cells) or PANC1 (pancreatic ductal carcinoma cells) with ONG41008 resulted in robust cellular senescence, indicating that dysregulated cell proliferation is common to fibrotic cells and tumor cells. The senescence was followed by multinucleation, a manifestation of mitotic slippage. There was significant upregulation of expression and rapid nuclear translocation of p-TP53 and p16 in the treated cancer cells, which thereafter died after 72 h confirmed by 6 day live imaging. ONG41008 exhibited a comparable senogenic potential to that of dasatinib. Interestingly, ONG41008 was only able to activate caspase-3, 7 in comparison with quercetin and fisetin, also containing CS in PANC1. ONG41008 did not seem to be essentially toxic to normal human lung fibroblasts or primary prostate epithelial cells, suggesting ONG41008 can distinguish the intracellular microenvironment between normal cells and aged or diseased cells. This effect might occur as a result of the increased NAD/NADH ratio, because ONG41008 restored this important metabolic ratio in cancer cells. Taken together, this is the first study to demonstrate that a small molecule can arrest uncontrolled proliferation during fibrogenesis or tumorigenesis via both senogenic and senolytic potential. ONG41008 could be a potential drug for a broad range of fibrotic or tumorigenic diseases.

Prognostic factors for the survival of primary molars following pulpotomy with mineral trioxide aggregate: a retrospective cohort study.

OBJECTIVES: The aim of the present study was to evaluate potential factors influencing the success rates of mineral trioxide aggregate (MTA) pulpotomy performed in primary molars. MATERIALS AND METHODS: A total of 347 teeth treated between March 2012 and December 2016 in 258 patients, with a mean age of 5.3 ± 1.7 years, were included in the analysis. Kaplan-Meier analyses were used to analyze were used time to failure. Multivariate Cox regression analysis with shared frailty was used to evaluate the clinical factors associated with failures. RESULTS: The mean (standard deviation) follow-up period was 35.8 (19.6) months. Within 84 months, the survival rate was 87.1%. In multivariate Cox regression, treatment performed in lower primary molars had a lower survival rate than upper primary molars (hazard ratio [HR] = 3.38, P = 0.012). Caries extension below the cemento-enamel junction had more risk of failure (HR = 10.9, P < 0.001). Final restoration using resin-modified glass ionomer or amalgam (direct filling) had a lower survival rate than stainless steel crown (HR = 5.62, P = 0.002). CONCLUSIONS: Clinical variables such as arch type, degree of caries extension, and type of final restoration may affect the survival of primary molars following MTA pulpotomy. CLINICAL RELEVANCE: The results of this study indicate that specific clinical variables can be used to predict the prognosis of MTA pulpotomy in primary teeth, and estimate the risk of treatment failure. Assessments of these variables should be performed in the context of evidence-based clinical decision making.

Human umbilical cord blood mesenchymal stem cells expansion via human fibroblast-derived matrix and their potentials toward regenerative application.

Large expansion of human mesenchymal stem cells (MSCs) is of great interest for clinical applications. In this study, we examine the feasibility of human fibroblast-derived extracellular matrix (hFDM) as an alternative cell expansion setting. hFDM is obtained from decellularized extracellular matrix (ECM) derived from in vitro cultured human lung fibroblasts. Our study directly compares conventional platforms (tissue culture plastic (TCP), fibronectin (FN)-coated TCP) with hFDM using umbilical cord blood-derived MSCs (UCB-MSCs). Early cell morphology shows a rather rounded shape on TCP but highly elongated morphology on hFDM. Cell proliferation demonstrates that MSCs on hFDM were significantly better compared to the others in both 10 and 2% serum condition. Cell migration assay suggests that cell motility was improved and a cell migration marker CXCR4 was notably up-regulated on hFDM. MSCs differentiation into osteogenic lineage on hFDM was also very effective as examined via gene expression, von Kossa staining and alkaline phosphatase activity. In addition, as the MSCs were expanded on each substrate, transferred to 3D polymer mesh scaffolds and then cultivated for a while, the data found better cell proliferation and more CXCR4 expression with MSCs pre-conditioned on hFDM. Moreover, higher gene expression of stemness and engraftment-related markers was noticed with the hFDM group. Furthermore when UCB-MSCs expanded on TCP or hFDM were injected into emphysema (a lung disease) animal model, the results indicate that MSCs pre-conditioned on hFDM (with 2% serum) retain more advanced therapeutic efficacy on the improvement of emphysema than those on TCP. Current works demonstrate that compared to the conventional platforms, hFDM can be a promising source of cell expansion with a naturally derived biomimetic ECM microenvironment and may find some practical applications in regenerative medicine.

Correlation between Dental Maturity and Body Mass Index in Korean Children.

Objective: The aim of this study was to determine relationships between dental maturity and body mass index (BMI) in Korean children. Study Design: 600 Korean children aged between 5 and 10 years for whom panoramic radiographs have been obtained between 2010 and 2017 were selected. Subjects were divided into four weight-status groups: underweight, normal weight, overweight, and obese. Five lower-left permanent teeth were observed and rated. The stage of each tooth was converted into a score using the table suggested by Demirjian, and the sum of these scores was designated as the 'maturity score'. Results: This study found statistically significant differences in dental maturity between the weight groups (analysis of variance, P=0.003), with the maturity score being higher in the obese group than in normal-weight subjects (Tukey's post-hoc test, P=0.004). The linear regression showed a positive association between BMI and the maturity score after adjusting for sex and age (ß=0.34, P<0.001). The linear regression coefficient was higher in girls (ß=0.61, P<0.001) than in boys (ß=0.31, P=0.02). Conclusions: These data suggest that dental maturation is positively associated with BMI in Korean children. Since many treatment decisions are made in relation to dental maturity, these findings may have implications for pediatric dental care.

Multi-lineage differentiation of human mesenchymal stromal cells on the biophysical microenvironment of cell-derived matrix.

We obtained fibroblast- (FDM) and preosteoblast- (PDM) derived matrices in vitro from their respective cells. Our hypothesis was that these naturally occurring cell-derived matrices (CDMs) would provide a better microenvironment for the multi-lineage differentiation of human mesenchymal stromal cells (hMSCs) than those based on traditional single-protein-based platforms. Cells cultured for 5-6 days were decellularized with detergents and enzymes. The resulting matrices showed a fibrillar surface texture. Under osteogenic conditions, human bone-marrow-derived stromal cells (HS-5) exhibited higher amounts of both mineralized nodule formation and alkaline phosphatase (ALP) expression than those cultured on plastic or gelatin. Osteogenic markers (Col I, osteopontin, and cbfa1) and ALP activity from cells cultured on PDM were notably upregulated at 4 weeks. The use of FDM significantly improved the cellular expression of chondrogenic markers (Sox 9 and Col II), while downregulating that of Col I at 4 weeks. Both CDMs were more effective in inducing cellular synthesis of glycosaminoglycan content than control substrates. We also investigated the effect of matrix surface texture on hMSC (PT-2501) differentiation; soluble matrix (S-matrix)-coated substrates exhibited a localized fibronectin (FN) alignment, whereas natural matrix (N-matrix)-coated substrates preserved the naturally formed FN fibrillar alignment. hMSCs cultured for 4 weeks on N-matrices under osteogenic or chondrogenic conditions deposited a greater amount of calcium and proteoglycan than those cultured on S-matrices as assessed by von Kossa and Safranin O staining. In contrast to the expression levels of lineage-specific markers for cells cultured on gelatin, FN, or S-matrices, those cultured on N-matrices yielded highly upregulated levels. This study demonstrates not only the capacity of CDM for being an effective inductive template for the multi-lineage differentiation of hMSCs, but also the critical biophysical role that the matrix fibrillar texture itself plays on the induction of stem cell differentiation.

Synthesis and electroluminescent properties of a novel electroluminescence material of bis-2-(4-(diphenylphosphino)phenyl)benzo[d]oxazole (DPB).

A new light-emissive material, bis-2-(4-(diphenylphosphino)phenyl)benzo[d]oxazole (DPB), has been synthesized and characterized by FT-NMR, FT-IR, UV-Vis and elemental analysis. DPB has the band gap of 4.3 eV between HOMO and LUMO levels. The photoluminescence (PL) of DPB was measured at 410 nm from the chloroform solution. The electroluminescent (EL) devices with structures of ITO/NPB/DPB/LiF/Al and ITO/NPB/DPB/Alq3/LiF/Al were constructed and showed maximum emission at 540 nm. The device using DPB as emitting material showed the luminance of 1000 cd/m2 at 11 V. The CIE chromaticity of the device showed near the region of white color emission.

Dual growth factor delivery using biocompatible core-shell microcapsules for angiogenesis.

An optimized electrodropping system produces homogeneous core-shell microcapsules (C-S MCs) by using poly(L-lactic-co-glycolic acid) (PLGA) and alginate. Fluorescence imaging clearly shows the C-S domain in the MC. For release control, the use of high-molecular-weight PLGA (HMW 270 000) restrains the initial burst release of protein compared to that of low-MW PLGA (LMW 40 000). Layer-by-layer (LBL) assembly of chitosan and alginate on MCs is also useful in controlling the release profile of biomolecules. LBL (7-layer) treatment is effective in suppressing the initial burst release of protein compared to no LBL (0-layer). The difference of cumulative albumin release between HMW (7-layer LBL) and LMW (0-layer LBL) PLGA is determined to be more than 40% on day 5. When dual angiogenic growth factors (GFs), such as platelet-derived GF (PDGF) and vascular endothelial GF (VEGF), are encapsulated separately in the core and shell domains, respectively, the VEGF release rate is much greater than that of PDGF, and the difference of the cumulative release percentage between the two GFs is about 30% on day 7 with LMW core PLGA and more than 45% with HMW core PLGA. As for the angiogenic potential of MC GFs with human umbilical vein endothelial cells (HUVECs), the fluorescence signal of CD31+ suggests that the angiogenic sprout of ECs is more active in MC-mediated GF delivery than conventional GF delivery, and this difference is significant, based on the number of capillary branches in the unit area. This study demonstrates that the fabrication of biocompatible C-S MCs is possible, and that the release control of biomolecules is adjustable. Furthermore, MC-mediated GFs remain in an active form and can upregulate the angiogenic activity of ECs.

Biocompatibility and mineralization potential of new calcium silicate cements.

Background/purpose: As calcium silicate cements (CSCs) have been successfully used in various types of vital pulp therapy, many new CSC products have been developed. The aim of this study was to evaluate the biocompatibilities and mineralization potential of new CSC. The experimental materials were NeoMTA Plus and EndoSequence Root Repair Material-Fast Set Putty (ERRM-FS) which were compared to ProRoot MTA. Materials and methods: In vitro, the effects of the new CSC on stem cells were evaluated. Each CSC was prepared for cell viability testing, alkaline phosphatase (ALP) assay, and calcium ion release assay. In vivo, the exposed pulp model was used for the partial pulpotomy procedure. Thirty-six teeth were treated with three materials: ProRoot MTA, NeoMTA Plus, or ERRM-FS. After four weeks, the teeth were extracted and processed for histologic analysis. Dentin bridge formation, pulp inflammation, and odontoblastic cell layer were evaluated and the area of newly formed calcific barrier of each group was measured. Results: Three CSCs demonstrated similar cell viability on stem cells and the levels of ALP and calcium release were not significantly different between tested materials. ProRoot MTA and ERRM-FS showed better tissue healing process than NeoMTA Plus after partial pulpotomy, in terms of quality of calcific barrier and pulp inflammation. The outcomes from measuring newly formed calcific area demonstrated no significant differences between the materials. Conclusion: NeoMTA Plus and ERRM-FS displayed similar biocompatibilities and mineralization potential compared to ProRoot MTA. Therefore, these new CSCs can be used as desirable alternatives to ProRoot MTA.

Assessment of Real-Time Active Noise Control Devices in Dental Treatment Conditions.

Dental clinics are exposed to various uncomfortable noises. The aim of this study was to quantify the effectiveness of active noise control devices in dental treatment conditions. Two types of commercial headsets (Airpods Pro, QC30) and two types of dental headsets (Alltalk, Quieton Dental) were used for the experiment. Three sounds (high-speed handpiece, low-speed handpiece, and suction system) were measured at three different distances from the dental teeth model, typodont. The distances of 10, 40, and 70 cm reflected the positions of the patient, assistant, and practitioner's ears, respectively. Sound analysis was performed, and the significance of differences in the maximum noise level using each device was determined with the Kruskal−Wallis test. Dental noise was characterized by the peak in sound pressure level (SPL) at 4−5 kHz and >15 kHz frequencies. The commercial headsets efficiently blocked 1 kHz and 10 kHz of noise. The dental headsets efficiently reduced 4−6 and >15 kHz noise. Quieton had the highest maximum SPL in all situations and positions among the four devices. For a better dental clinic, however, active noise control devices more suitable for the characteristics of dental noise should be developed.

Novel 3D Printed Resin Crowns for Primary Molars: In Vitro Study of Fracture Resistance, Biaxial Flexural Strength, and Dynamic Mechanical Analysis.

This study evaluated the fracture resistance, biaxial flexural strength (BFS), and dynamic mechanical analysis (DMA) of three-dimensional (3D) printing resins for the esthetic restoration of primary molars. Two 3D printing resins, Graphy (GP) and NextDent (NXT), and a prefabricated zirconia crown, NuSmile (NS), were tested. GP and NXT samples were 3D printed using the workflow recommended by each manufacturer. Data were collected and statistically analyzed. As a result of the fracture resistance test of 0.7-mm-thick 3D printed resin crowns with a thickness similar to that of the NS crown, there was no statistically significant difference among GP (1491.6 ± 394.6 N), NXT (1634.4 ± 289.3 N), and NS (1622.8 ± 323.9 N). The BFS of GP was higher for all thicknesses than that of NXT. Both resins showed high survival probabilities (more than 90%) when subjected to 50 and 150 MPa. Through DMA, the glass transition temperatures of GP and NXT were above 120 °C and the rheological behavior of GP and NXT according to temperature and frequency were analyzed. In conclusion, GP and NXT showed optimum strength to withstand bite forces in children, and 3D printed resin crowns could be an acceptable option for fixed prostheses of primary teeth.

Image-guided in situ cancer vaccination with combination of multi-functional nano-adjuvant and an irreversible electroporation technique.

Cancer immunotherapy is a next-generation treatment strategy; however, its side effects limit its clinical translation. Here, a novel combination of a multi-functional nano-adjuvant (M-NA) prepared with an iron oxide/gold core and a cationic polymer shell via multilayer synthesis with CpG oligodeoxynucleotide (CpG-ODN) electrostatically complexed on its surface, and irreversible electroporation (IRE) technique was developed for effective image-guided in situ cancer vaccination. The M-NA can be retained long-term in the dense tumoral extracellular matrix after intratumoral injection and internalized by antigen-presenting cells (APCs). The IRE can induce immunogenic cell death. Indeed, in a mouse tumor model, the M-NA showed longer tumor retention time than free CpG-ODN. Compared with other treatments, the combined treatment significantly inhibited tumor growth with 100% survival rate for ∼60 days. The therapy induced the activation of cytotoxic lymphocytes and the maturation of APCs in vivo. This treatment could be effective in image-guided local cancer immunotherapy.

Production and purification of human papillomavirus type 33 L1 virus-like particles from Spodoptera frugiperda 9 cells using two-step column chromatography.

The major capsid protein L1 of human papillomavirus (HPV) is essential in construction of recombinant antigen vaccines against cervical cancer. HPV type 33 accounts for about 10% of all HPV infections in Asia. The gene encoding the major capsid protein L1 of the high-risk HPV type 33 was isolated from a Korean patient and expressed in Sf-9 insect cells using a baculovirus expression system. HPV33 L1 protein was isolated by two-step chromatographic purification using strong-cation exchange and ceramic hydroxyapatite chromatography. Strong-cation-exchange chromatography was performed to achieve initial purification of HPV33 L1 and to remove most contaminating proteins, and secondary ceramic hydroxyapatite chromatography yielded pure HPV33 L1 virus-like particles (VLPs). Ceramic hydroxyapatite columns are particularly useful in the purification of antibodies, antigens, human viruses, and VLPs, and we thus used this system. The expression of HPV L1 protein in Sf-9 cells was examined by SDS-PAGE, Western-blotting, and ELISA analyses, and the data showed that HPV33 L1 VLPs were determined to > 98% purity and 58.7% recovery by a quantitative immuno-ELISA assay. Transmission electron microscopy analysis revealed that the HPV VLPs were approximately 50-60 nm in diameter and created by self-assembly of HPV L1 protein. The efficient and simple purification process described here should be useful in production of a cervical cancer vaccine.

Expression and characterization of a second L-amino acid deaminase isolated from Proteus mirabilis in Escherichia coli.

L-amino acid deaminases catalyze the deamination of natural L-amino acids. Two types of L-amino acid deaminase have been identified in Proteus species. One exhibits high levels of activity toward a wide range of aliphatic and aromatic L-amino acids, typically L-phenylalanine, whereas the other acts on a relatively narrow range of basic L-amino acids, typically L-histidine. In this study, we cloned, expressed, and characterized a second amino acid deaminase, termed Pm1, from P. mirabilis KCTC 2566. Homology alignment of the deduced amino acid sequence of Pm1 demonstrated that the greatest similarity (96%) was with the L-amino acid deaminase (LAD) of P. vulgaris, and that homology with Pma was relatively low (72%). Also, similar to LAD, Pm1 was most active on L-histidine, indicating that Pm1 belongs to the second type of amino acid deaminase. In agreement with this conclusion, the V(max) and K(m) values of Pm1 were 119.7 (µg phenylpyruvic acid/mg/min) and 31.55 mM phenylalanine, respectively, values lower than those of Pma. The Pml deaminase will be very useful industrially in the preparation of commercially valuable materials including urocanic acid and α-oxoglutarate.

Human nasal septal chondrocytes (NSCs) preconditioned on NSC-derived matrix improve their chondrogenic potential.

BACKGROUND: Extracellular matrix (ECM) has a profound effect on cell behaviors. In this study, we prepare a decellularized human nasal septal chondrocyte (NSC)-derived ECM (CHDM), as a natural (N-CHDM) or soluble form (S-CHDM), and investigate their impact on NSCs differentiation. METHODS: N-CHDM, S-CHDM were obtained from NSC. To evaluate function of NSC cultured on each substrate, gene expression using chondrogenic marker, and chondrogenic protein expression were tested. Preconditioned NSCs-loaded scaffolds were transplanted in nude mice for 3 weeks and analyzed. RESULTS: When cultivated on each substrate, NSCs exhibited similar cell spread area but showed distinct morphology on N-CHDM with significantly lower cell circularity. They were highly proliferative on N-CHDM than S-CHDM and tissue culture plastic (TCP), and showed more improved cell differentiation, as assessed via chondrogenic marker (Col2, Sox9, and Aggrecan) expression and immunofluorescence of COL II. We also investigated the effect of NSCs preconditioning on three different 2D substrates while NSCs were isolated from those substrates, subsequently transferred to 3D mesh scaffold, then cultivated them in vitro or transplanted in vivo. The number of cells in the scaffolds was similar to each other at 5 days but cell differentiation was notably better with NSCs preconditioned on N-CHDM, as assessed via real-time q-PCR, Western blot, and immunofluorescence. Moreover, when those NSCs-loaded polymer scaffolds were transplanted subcutaneously in nude mice for 3 weeks and analyzed, the NSCs preconditioned on the N-CHDM showed significantly advanced cell retention in the scaffold, more cells with a chondrocyte lacunae structure, and larger production of cartilage ECM (COL II, glycosaminoglycan). CONCLUSIONS: Taken together, a natural form of decellularized ECM, N-CHDM would present an advanced chondrogenic potential over a reformulated ECM (S-CHDM) or TCP substrate, suggesting that N-CHDM may hold more diverse signaling cues, not just limited to ECM component.

Comparative study of pulpal responses to ProRoot MTA, Vitapex, and Metapex in canine teeth.

BACKGROUND/PURPOSE: ProRoot MTA, Vitapex, and Metapex are widely used for pulp treatment of primary tooth. The aim of this study was to compare the pulpal responses to ProRoot MTA, Vitapex, and Metapex in a canine model of pulpotomy. MATERIALS AND METHODS: Pulpotomy procedure was performed to 34 teeth (21 incisors and 13 premolars) and ProRoot MTA, Vitapex or Metapex was applicated to artificially exposed pulp tissues. After 13 weeks, the teeth were extracted and processed with hematoxylin-eosin staining for histologic evaluation. All specimens were evaluated in several categorys related to calcific barrier, inflammatory responses and the area of calcific barrier formation was measured. RESULTS: Most of the specimens in the ProRoot MTA group developed a calcific barrier at the pulp amputation site and showed a low level of inflammatory response. However, in comparison to ProRoot MTA group, a small amount of calcific barrier formed in Vitapex and Metapex groups. CONCLUSION: This in vivo study found that Vitapex and Metapex induced similar pulpal responses but showed poor outcomes compared with using ProRoot MTA. Vitapex and Metapex are therefore not good substitutes for ProRoot MTA in direct pulp capping and pulpotomy.

The Piston Elastic: A Novel Device for Treating Entrapped Ectopic Permanent Molars.

Ectopic eruption of the permanent molar may absorb the distal root of the primary second molar and may result in a decreased arch length or delayed eruption of the permanent tooth, requiring timely treatment. Therefore, we devised an effective and convenient method to unlock the entrapped tooth using a novel device called a "piston-elastic". This case report aims to explain the design and clinical application of this piston-elastic and to describe successful cases. Three patients (aged 6, 13, and 16 years) with ectopically erupted maxillary and mandibular molars, respectively, were treated with a piston-elastic. It was bound to the locked molar to improve the eruption path. After a certain time period, the repulsive force pushed the surface of the adjacent tooth, improving the eruption path of the entrapped tooth. The piston-elastic is a novel device that simply and effectively changes the direction of eruption of ectopically entrapped molars. As it can be manufactured and attached to the chair side, impression acquisition on a model cast and laboratory procedures are unnecessary. Compared to existing methods, the piston-elastic can be easily produced and delivered, causes little irritation, and is inexpensive.

In Vivo Evaluation of Decellularized Human Tooth Scaffold for Dental Tissue Regeneration.

Conventional root canal treatment may result in loss of tooth vitality, which can lead to unfavorable treatment outcomes. Notably, a ceased tooth development of immature permanent teeth with open apices, regeneration of periodontal ligaments (PDL), and pulp is highly expected healing process. For regeneration, the scaffold is one of the critical components that carry biological benefits. Therefore, this study evaluated a decellularized human tooth as a scaffold for the PDL and pulp tissue regeneration. A tooth scaffold was fabricated using an effective decellularization method as reported in previous studies. PDL stem cells (PDLSCs) and dental pulp stem cells (DPSCs) obtained from human permanent teeth were inoculated onto decellularized scaffolds, then cultured to transplant into immunosuppressed mouse. After 9 weeks, PDLSCs and DPSCs that were inoculated onto decellularized tooth scaffolds and cultured in an in vivo demonstrated successful differentiation. In PDLSCs, a regeneration of the cementum/PDL complex could be expected. In DPSCs, the expression of genes related to revascularization and the hard tissue regeneration showed the possibility of pulp regeneration. This study suggested that the potential possible application of decellularized human tooth could be a scaffold in regeneration PDL and pulp tissue along with PDLSCs and DPSCs, respectively, as a novel treatment method.

Overexpression of Newcastle disease virus (NDV) V protein enhances NDV production kinetics in chicken embryo fibroblasts.

Newcastle disease virus (NDV) is not only one of the most economically important pathogen of poultry but also has a potential as anticancer virotherapy. The role of NDV V protein in virus-production kinetics was investigated using DF-1 cell-based production system. The presence of an anti-interferon (IFN)-alpha antibody resulted in enhanced NDV production kinetics in a dose-dependent manner by blocking binding of NDV-induced IFN to its receptor. To prepare DF-1 cell whose cellular IFN signaling is blocked efficiently, stable cell lines expressing either lentogenic or velogenic NDV V protein known as an IFN antagonist were established. The overexpression of NDV V protein enhanced NDV production kinetics and expedited the rate of NDV production, while it had no effect on Japanese encephalitis virus production. NDV V protein functions as an IFN antagonist by inhibiting the increase in type I IFNs by NDV infection. The IFN signals in cells expressing NDV V protein were weakened by decreased activation or expression of the dsRNA-activated enzymes. These IFN antagonist activities enhance rapid virus replication and spread in the early phase of viral infection and will be useful in improving the production of viral vaccine strains.

Colorimetric pH measurement of animal cell culture media.

Most animal cell culture media can be buffered using bicarbonate and high pressure CO(2) in a closed system. However, in an open system, the pH of the culture media increases continuously due to the marked difference in CO(2) pressure between the culture media and the atmosphere. Therefore, it is important to measure the exact pH of the culture media in an intact closed system. In this study, a pH measurement method was developed using visible light. The pH was calculated from light absorbance by the cells and by the culture media. This method was successfully applied to both suspension and anchorage-dependent cell cultures.