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Objectives To evaluate dimensional ridge alterations and sequential healing processes following ridge augmentation after tooth extraction in damaged extraction sockets with buccal-bone-deficiency. Material and methods Bilateral dental roots of three mandibular premolars were extracted with entire removal of the buccal-bone plate in eight beagle dogs. Unilateral sites were grafted with biomaterials (test group) and contralateral sites were healed without grafting (control group). Observations were made after 1, 2, 4, and 8 weeks, and all sites were distributed evenly (n = 6 for each group and period). Radiographic/histomorphometric analyses were performed. Results In spontaneous healing of damaged extraction sockets, the dimension of regenerated alveolar ridge gradually increased until 4 weeks and then remained stable, but radiographic/histomorphometric analyses revealed evident dimensional shrinkage compared to the pristine tissue at 8 weeks in the coronal and middle areas. Bone grafting retained the pristine dimension of alveolar ridge, and newly formed bone area within the augmented space continuously expanded during the observational period to the outermost border of the space. Conclusions Spontaneous healing of damaged extraction sockets caused substantial dimensional shrinkage. However, ridge augmentation can provide space into which new bone may grow continuously, resulting in the final dimensions comparable to those of the pristine alveolar ridge.

The aims of the present study were (i) to determine the short-term effects of putty-type porcine bone substitute material (PB) with recombinant bone morphogenetic protein-2 (rhBMP-2) for alveolar ridge preservation (ARP) and (ii) evaluate the early healing of labial overaugmentation. Two groups were randomly assigned to the extraction sockets in mandibular incisors of eight dogs: labial overaugmentation with (i) PB (control) or (ii) rhBMP-2-loaded PB (BMP). Microcomputed tomography (micro-CT), and histologic and histomorphometric analyses were performed after 4 weeks. Micro-CT revealed that some of the overaugmented PB was dispersed in both groups. The new bone volume was significantly larger in the BMP group than in the control group (18.4 ± 3.3 vs. 15.5 ± 3.0 mm3, mean ± SD, P

Defect-specific bone regeneration using 3-dimensional (3D) printing of block bone has been developed. Polycaprolactone (PCL) is biocompatible polymer that can be used as 3D scaffold. The aim of this study is to assess the biocompatibility and osteogenic efficacy of 3D printed PCL scaffold and to evaluate the effectiveness of β-tricalcium phosphate (β-TCP) addition in PCL scaffold. In this work, four circular defects (diameter: 8 mm) in rabbit calvarium were randomly assigned to (1) negative control (control), (2) PCL block (PCL), (3) PCL mixed with 10 wt% β-TCP (PCL/β-TCP), and (4) PCL/β-TCP plus collagen membrane (PCL/β-TCP + M). Animals were euthanized at 2 (n = 5) and 8 weeks (n = 5). Results indicated that in micro-CT, PCL/β-TCP + M showed the highest total augmented volume and new bone volume at 8 weeks, but there was no significant difference among four groups. Histomorphometrically, PCL, PCL/β-TCP, and PCL/β-TCP + M showed the significantly higher total augmented area compared to the control. PCL/β-TCP + M showed the highest new bone area but not statistically higher than the control. New bone formation deep inside the scaffold was observed only in β-TCP added scaffold. PCL showed high biocompatibility with great volume maintenance. Addition of β-TCP to PCL seemed to increase hydrophilicity and osteoconductivity. Developments in 3D-printed PCL material are expected. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1254-1263, 2019.

OBJECTIVES To histologically determine the alteration in horizontal mucosal thickness at sites that received guided bone regeneration (GBR) with additional use of collagen matrix and to assess whether bone formation is affected by adding collagen matrix at GBR sites at 8 weeks of healing. MATERIALS AND METHODS Eight weeks after bilateral extraction of maxillary premolars, standardized defects were created on buccal side of edentulous ridges in four beagle dogs. One side was randomly allocated as control (biphasic calcium phosphate plus collagen membrane; GBR only), while contralateral side was allocated as test (biphasic calcium phosphate plus collagen membrane plus an additional layer of collagen matrix). Histologic observations, histomorphometric and micro-computed tomography analyses were performed after 8 weeks. RESULTS Membrane complex comprising residual collagen membrane and adjacent dense connective tissue was observed at both control and test sites. The thickness in the histologic analysis were 1.69 ± 0.23 mm (control) and 1.76 ± 0.07 mm (test) in histologic analysis and were 2.03 ± 0.26 mm (control) and 2.14 ± 0.24 mm (test) in radiographic analysis. The thickness of the membrane complex in soft-tissue layer were 723.0 ± 241.6 μm (control) and 984.6 ± 334.4 μm (test). The percentage of new bone formation were 22.30 ± 5.92% (control) and 25.50 ± 8.08% (test). All measured outcome did not show significant differences between control and test groups. CONCLUSION The addition of collagen matrix on top of standard GBR procedure did not increase the soft tissue thickness and dense connective tissue formation at 8 weeks of healing. Bone regeneration was not affected by the addition of collagen matrix. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 00B: 000-000, 2018.

Backgrounds Clinical benefits in bone grafting of intact extraction socket have been widely known, but limited evidence is available for the procedure in damaged extraction sockets due to periodontal disease. Purpose This study aimed to determine the dimensional alteration of alveolar ridge following bone grafting of damaged extraction sockets, and compare the outcomes of using deproteinized bovine (DBBM) versus porcine bone mineral (DPBM) in the damaged sockets. Materials and Methods One hundred patients (n = 50 for each group) with periodontitis‐induced damaged extraction socket were included in this randomized, single‐blind trial. After removal of tooth and granulation tissue, sites were grafted with either DBBM (DBBM group) or DPBM (DPBM group), and covered with collagen membrane. Linear/volumetric analyses of hard and soft‐tissue dimensions were performed on reconstructed/superimposed computed tomography and scanned cast images, taken immediately and 4 months after surgery. Results The two groups showed comparable hard tissue augmentation with minimal reductions in the grafted volume, as well as in vertical (1.22 ± 2.16 and 1.45 ± 1.92 mm for DPBM and DBBM group, respectively) and horizontal (1.43 ± 3.40 and 1.83 ± 2.85 mm on the central section, respectively) dimensions at 4 months after surgery. However, several cases showed large variations in maintenance of the grafted volume. None of the measured parameters in hard and soft tissue dimensions differed significantly between DBBM and DPBM sites. Conclusions DBBM and DPBM can comparably augment damaged extraction sockets with minimal postoperative reduction of the grafted volume. However, the large variations in the results should be further evaluated for application in routine dental clinics.

Variable lymphocyte receptors B (VLRBs) are non-immunoglobulin components of the humoral immune system in jawless vertebrates including hagfish (Eptatretus burgeri) and lamprey (Petromyzon marinus). Hagfish VLRBs consist of leucine rich repeat (LRR) modules with a superhydrophobic C-terminal tail, the latter of which leads to extremely low expression levels in recombinant protein technology. Here, we present an artificially oligomerized VLRB (arVLRB) that conjugates via the C4bp oligomerization domain derived from human C4b-binding protein (hC4bp) rather than the superhydrophobic tail. The resulting arVLRB had a tightly multimerized form with seven monomeric VLRB arms and showed high expression and secretion levels in a mammalian expression system. To isolate antigen-specific arVLRB, we constructed large VLRB libraries from hagfish immunized with the fish pathogen, viral hemorrhagic septicemia virus (VHSV). The selected arVLRBs were found to recognize various types of antigens, including the recombinant target protein, purified viruses, and progeny viruses, with high antigen binding abilities and specificities. We also performed in vitro affinity maturation of the arVLRBs through LRRCT mutagenesis, and found that this enhanced their antigen-binding properties by at least 125-fold. Our epitope mapping analysis revealed that 37DWDTPL42, which is located in a region conserved among the glycoproteins of all VHSV isolates, is the recognition epitope of the arVLRBs. Thus, our newly developed arVLRB could prove useful in the development of universal diagnostic tools and/or therapeutic agents for the virus. Together, our novel findings provide valuable insights into hagfish VLRB and its potential use as a novel alternative to conventional antibodies for biotechnological applications.

This study is designed to determine the effect of collagen membrane (CM) soaked with bone morphogenetic protein-2 (rhBMP-2) for the treatment of peri-implant dehiscence defects. Material and Methods. Three treatment groups were allocated at each defect in 5 dogs: (i) collagenated synthetic bone (OC) and CM soaked with rhBMP-2 (BMP group), (ii) OC and CM soaked with saline (nonBMP group), and (iii) no further treatment (control group). Titanium pins were used to stabilize the membranes in two dogs. Radiographic and histomorphometric analyses were performed 4 weeks later. Results. The median augmented volumes were 4.27 mm3, 6.24 mm3, and 2.75 mm3 in the BMP, nonBMP, and control groups, respectively; the corresponding median first bone-to-implant contact (fBIC) distances were 3.25 mm, 3.08 mm, and 2.56 mm (P>0.05). The placement of pins (with the BMP and nonBMP groups pooled) significantly improved bone regeneration: the augmented volumes were 17.60 mm3 with pins and 3.68 mm3 without pins (P=0.024), with corresponding fBIC distances of 2.25 mm and 3.31 mm, respectively (P<0.001). Conclusions. The addition of rhBMP-2 to CM failed to improve bone regeneration of peri-implant dehiscence defects compared to using an unsoaked CM after 4 weeks. However, the stabilization of CMs using pins positively influenced the outcomes.

This study aimed to determine the cellular characteristics and behaviors of human bone marrow stromal cells (hBMSCs) expanded in media in a hypoxic or normoxic condition and with or without fibroblast growth factor-2 (FGF-2) treatment. hBMSCs isolated from the vertebral body and expanded in these four groups were evaluated for cellular proliferation/migration, colony-forming units, cell-surface characterization, in vitro differentiation, in vivo transplantation, and gene expression. Culturing hBMSCs using a particular environmental factor (hypoxia) and with the addition of FGF-2 increased the cellular proliferation rate while enhancing the regenerative potential, modulated the multipotency-related processes (enhanced chondrogenesis-related processes/osteogenesis, but reduced adipogenesis), and increased cellular migration and collagen formation. The gene expression levels in the experimental samples showed activation of the hypoxia-inducible factor-1 pathway and glycolysis in the hypoxic condition, with this not being affected by the addition of FGF-2. The concurrent application of hypoxia and FGF-2 could provide a favorable condition for culturing hBMSCs to be used in clinical applications associated with bone tissue engineering, due to the enhancement of cellular proliferation and regenerative potential. Keywords: Bone marrow stromal cells, Hypoxia, Fibroblast growth factor, Tissue regeneration, Microenvironment interactions

Outer membrane vesicles (OMVs) containing various bacterial compounds are released from mainly gram-negative bacteria. Secreted OMVs play important roles in the ability of a bacterium to defend itself, and thus contribute to the survival of bacteria in a community. In this study, we collected OMVs from β-lactam antibiotic-resistant Escherichia coli established by conjugation assay and the parental β-lactam antibiotic-susceptible strain, and performed comparative proteomic analysis to examine whether these OMVs carried β-lactam-resistant compounds. We also investigated whether both types of OMVs could protect susceptible cells from β-lactam-induced death and/or directly degrade β-lactam antibiotics. Several proteins that can be involved in degrading β-lactam antibiotics were more abundant in OMVs from β-lactam-resistant E. coli, and thus OMVs from β-lactam resistant E. coli could directly and dose-dependently degrade β-lactam antibiotics and fully rescue β-lactam-susceptible E. coli and other bacterial species from β-lactam antibiotic-induced growth inhibition. Taken together, present study demonstrate that OMVs from β-lactam-resistant E. coli play important roles in survival of antibiotic susceptible bacteria against β-lactam antibiotics. This finding may pave the way for new efforts to combat the current global spread of antibiotic resistances, which is considered to be a significant public health threat.

This study aimed to quantify the healing following vertical augmentation of allogenic bone blocks with/without recombinant human bone morphogenetic protein-2 (rhBMP-2) on rabbit calvaria. Experiments were performed using allogenic bone blocks which were grafted bilaterally with or without rhBMP-2 on 20 rabbit calvaria, and these animals were divided to four groups according to the use of rhBMP-2 and healing periods (2 and 8 weeks; n = 10 in each group). Onlay-type bone blocks (8 mm in diameter and 5 mm high) were fixed with a self-tapping screw after removing the cortex in the control group, and the same protocol was applied with the addition of soaking the bone blocks with rhBMP-2 for 15 min in the test group. Radiographic and histologic analyses were performed after 2 or 8 weeks to evaluate the volumetric stability and bone regeneration within the grafted area. The radiographic analysis revealed that the height of the allogenic bone block decreased but its volume was maintained from 2 to 8 weeks in both the control and test groups. The histologic results demonstrated a statistically significant increase in new bone area in the test group, especially in the lower region adjacent to the preexisting calvarial floor. The amount of newly formed bone in all regions of the augmented bone blocks in both the control and test groups was greater at 8 weeks than at 2 weeks. In conclusion, the vertically grafted allogenic bone block maintained its volume with new bone formation, and this was accelerated by the addition of rhBMP-2. These findings indicate that allogenic bone block soaked with rhBMP-2 could be a useful candidate biomaterial for vertical augmentation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2700-2707, 2018.

The aim of the present study was to evaluate the physiochemical properties and resorption progress of two cross-linked, porcine skin-derived collagen membranes and compare their features with those of a membrane without cross-linking (Bio-Gide® [BG], Geistlich Biomaterials, Wolhusen, Switzerland). Three porcine skin-derived collagen membranes, dehydrothermally (DHT) cross-linked (experimental), DHT and 1-ethyl-3(3-dimethylaminopropyl)-carbodiimide (DHT/EDC) cross-linked (experimental) and BG were investigated for their morphology, enzyme resistance, and tensile strength in vitro and biodegradation in vivo. DHT and DHT/EDC membranes exhibited irregular, interconnected macro- and micropores that formed a 3D mesh, whereas BG exhibited individual collagen fibrils interlaced to form coarse collagen strands. In enzyme resistance and tensile strength tests, DHT and DHT/EDC membranes demonstrated good resistance and mechanical properties compared with BG. In vivo, all three membranes were well integrated into the surrounding connective tissue. Thus, the DHT membrane exhibited its potential as a barrier membrane for guided bone and tissue regeneration.

To retrospectively determine the profile of the sinus membrane (SM), potential factors affecting SM thickening (SMT), and the correlation between SMT and sinus augmentation (SA) complications.In the patients who received lateral SA, SMT was classified in sagittal sections of cone-beam computed tomography according to its thickness and morphology. The correlation between SMT and the following factors was analyzed: age, sex, endodontic and periodontic statuses of neighboring teeth, and shape of the sinus inferior border. The association between SMT and SA complications was investigated.SMT of ≤2 mm was prevalent (60%). Irregular SM was mostly observed for SMT of2 mm. There was no statistically significant association between SMT and the included factors. SMT did not significantly correlate with either perforation or postoperative complications. There was a statistically significant increase in implant failure when SMT was2 mm, but it was hard to determine that the failure was solely affected by SMT.SMT was not influenced by the factors included in this study, and it might not be a risk factor for SA and implant failure.

The objective of this study was to determine the validity of a graft-free sinus floor elevation (SFE) procedure with simultaneous placement of recombinant morphogenetic protein-2 (rhBMP-2)-coated implants compared to uncoated control implants. In 10 rabbits, SFE was performed on both sides. Dental implants were randomly placed in the sinus filled with a blood clot. Test implants were coated with rhBMP-2, whereas in the control group, implants were uncoated. Micro-computed tomographic and histomophometric analyses were performed at 4 and 8 weeks, including measurement for newly formed bone height (NBHm). Bone formation was evident along the implant surfaces up to the apex in test, but limited in control implants at 4 weeks. NBHm amounted to 5.1 mm (Q1 = 4.1; Q3 = 5.3) for test implants and to 3.4 mm (2.6; 3.7) for control implants at 4 weeks. NBHm then decreased to 8 weeks (3.4 mm (3.3; 3.7)) for test implants, whereas in control sites, NBHm increased slightly to 4.4 mm (4.1; 4.5) (p = 0.1250; p = 0.6250). Implants coated with rhBMP-2 presented a strong osteogenic reaction at 4 weeks with more favorable outcomes in terms of bone formation along the implant surface up to the apex compared to uncoated control implants. Remodeling and resorption process between 4 and 8 weeks did not further improve the outcomes in the test, but in the control group. The use of rhBMP-2-coated implants in a graft-free SFE might show an advantage in early implant stability to prevent collapse of membrane. However, a potential clinical benefit still needs to be proven.

The biophysical parameters governing nanoparticle (NP)-cell interactions significantly affect biological responses, particularly in the application of NP-based immunotherapeutics. Modulation of the surface biophysical character of NPs can be achieved via introduction of amino acids, which offer the ability to fine tune a range of biophysical parameters of interest. We employed this approach using monodisperse silica NPs coated with numerous poly(amino acid)s (PAAs). The NPs were incubated with dendritic cells (DCs) in conjunction with TLR ligands and production of IL-1β from DCs and IFNγ from T cells primed by these DCs were measured. These key cytokines can prognosticate the efficacy of the NP platform as a potential vaccine or active cellular immunotherapy carrier. IL-1β production showed a correlation with both NP size and degree of hydrophobicity. High IFNγ secretion from T cells was shown to be correlated with both the hydrophobicity and charge of the NPs used to activate the DCs. Other cytokines were also screened in order to compare the immune responses. The results of this study highlight the importance of nanoparticle biophysical parameters and the selection of TLR ligands to the rational design of nanoparticle-based vaccines and immunotherapies. Statement of Significance The manuscript describes a systematic investigation into the effects of biophysical parameters of nanoparticles (NPs) on immune cells. Modulation of the biophysical character of the NP surface can be achieved by introduction of amino acids on monodisperse silica NPs, introducing a range of tunable biophysical parameters of interest, i.e. distinct sizes, different surface charges and varying degrees of surface hydrophobicity. We examine internalization of the NP in dendritic cells (DCs) and measure a myriad of cytokines, including IL-1β and IFNγ, which prognosticate the efficacy of the NPs as a potential vaccine (IL-1β metric) or active cellular immunotherapy carrier (IFNγ metric). Two different TLR ligands (a viral TLR3 ligand and a bacterial TLR4 ligand) were used along with the PAA NPs to compare their costimulatory immunogenicity. We strongly believe that this study will provide crucial information to many readers of Acta Biomaterialia and further drive the use of nanoparticle platforms in modulating immune responses.

The T cell receptor (TCR) is the binding site of antigen and is responsible for specifically activating the adaptive immune response. CD3, an essential component of the CD3-TCR complex, is known to be composed of γδ and ε chains in teleost. However, there are few monoclonal antibodies (mAb) available to identify these molecules on T cells, so we aimed to produce a mAb against CD3ε to improve our understanding of T cell immune response in olive flounder (Paralichthys olivaceus). CD3ε recombinant protein was expressed in yeast, the expression of which was confirmed by SDS-PAGE, MALDI-TOF/TOF MS and Western blot analysis. A CD3ε-specific mAb 4B2 was selected, the specificity of which was examined by confocal microscopy, flow cytometry and RT-PCR, and the mAb was subsequently used to examine the CD3ε lymphocyte population in several different immune organs, with relatively high percentages of these cells seen in trunk-kidney and spleen, while lower percentages were seen in the liver and peripheral blood of olive flounder. During a viral hemorrhagic septicemia virus (VHSV) infection in olive flounder, the number of CD3ε lymphocytes was seen to gradually increase in the liver, spleen and trunk-kidney of infected fish until 7 days post infection (dpi). In peripheral blood, on the other hand, the increase in CD3ε lymphocyte numbers peaked by 3 dpi. These results suggest that CD3ε lymphocytes might be involved in the immune response against VHSV.

A Typhoid Conjugate Vaccine (TCV) is expected to acquire WHO prequalification soon, which will pave the way for its use in many low- and middle-income countries where typhoid fever is endemic. Thus it is critical to forecast future vaccine demand to ensure supply meets demand, and to facilitate vaccine policy and introduction planning. We forecasted introduction dates for countries based on specific criteria and estimated vaccine demand by year for defined vaccination strategies in 2 scenarios: rapid vaccine introduction and slow vaccine introduction. In the rapid introduction scenario, we forecasted 17 countries and India introducing TCV in the first 5 y of the vaccine's availability while in the slow introduction scenario we forecasted 4 countries and India introducing TCV in the same time period. If the vaccine is targeting infants in high-risk populations as a routine single dose, the vaccine demand peaks around 40 million doses per year under the rapid introduction scenario. Similarly, if the vaccine is targeting infants in the general population as a routine single dose, the vaccine demand increases to 160 million doses per year under the rapid introduction scenario. The demand forecast projected here is an upper bound estimate of vaccine demand, where actual demand depends on various factors such as country priorities, actual vaccine introduction, vaccination strategies, Gavi financing, costs, and overall product profile. Considering the potential role of TCV in typhoid control globally; manufacturers, policymakers, donors and financing bodies should work together to ensure vaccine access through sufficient production capacity, early WHO prequalification of the vaccine, continued Gavi financing and supportive policy.

Background Various biomaterials have been introduced for vertical ridge augmentation to replace autogenous block bone grafting. Purpose This retrospective study radiographically evaluated dimensional alterations of the vertically augmented alveolar ridge using collagen membrane and 3 types of materials: autogenous bone block, allogenous bone block, and particulated bone substitute. Materials and methods The electronic medical records of 32 patients who received vertical ridge augmentation using 3 types of materials were searched: 9 for autogenous bone block, 12 for allogenous bone block, and 11 for particulated bone substitutes. The vertical bone gain, progression of bone resorption, and peri-implant marginal bone loss after prosthetic loading were measured on follow-up radiographs. Results The alveolar ridge was vertically augmented by 5.13 ± 1.61, 4.54 ± 2.48, and 3.90 ± 0.85 mm (mean ± standard deviation) after grafting with autogenous bone block, allogenous bone block, and particulated bone substitute, respectively. The radiographic vertical height of the augmented ridge that received autogenous bone block reduced continuously during the first year but was stable thereafter. Sites that received allogenous bone block or particulated bone substitute exhibited dimensional shrinkage for up to 1.5 years postsurgery. However, the peri-implant marginal bone loss did not exceed 1 mm throughout the observational periods in all groups. Conclusions The clinical findings of this study suggest that the alveolar ridge can be vertically augmented using either allogenous bone block or particulated bone substitute. However, they require a longer healing period to ensure dimensional stability compared to the autogenous bone block.

The cellular DEAD-box helicase DDX41, which functions as an initial sensor for cytoplasmic DNA, is involved in the activation of type I interferon (IFN-1) immune response in olive flounder. A plasmid encoding DDX41 (pEF-D) was introduced into flounder cells in vitro and in vivo. Immune responses induced by DDX41 were evaluated by relative quantification value (ΔΔCt) method of RT-qPCR using specific IFN-related gene primers. Results in in vitro, transcript levels of IFN-1, IRF-3, ISG-15 and IL-1β were significantly higher in pEF-D- than pEF-A-transfected cells, with 15-, 4-, 10- and 32-fold changes, respectively. In vivo, elevated expression of these genes was observed in the kidney of pEF-D group on days 1 and 3 post-treatment. The viral challenge test revealed higher survival rate in fish treated with pEF-D (67.5%) than controls, PBS- and pEF-A-treated fish (45%). Conclusively DDX41 elicits a robust IFN-mediated immune response, validating its adjuvant property.

Purpose The aim of this study was to radiographically compare horizontal and vertical alterations of the alveolar ridge after ridge preservation using different combinations of biomaterials. Materials and methods The patients were randomly assigned into two groups: (a) the test group, which received collagenated porcine bone plus cross-linked collagen membrane (n = 15), and (b) the control group, which received collagenated bovine bone plus non-cross-linked collagen membrane (n = 15). Cone beam computed tomography (CBCT) was performed immediately after ridge preservation and 4 months thereafter to compare the horizontal and vertical alterations of the ridge. Intention-to-treat (ITT) and per protocol (PP) analyses were performed. Results In both analyses, there was no statistically significant difference in the horizontal alteration of the alveolar ridge between the test and control groups: −1.3 ± 0.6 (ITT)/1.2 ± 0.5 (PP) vs. −1.5 ± 0.9 mm at the 1-mm level; −1.2 ± 0.7 (ITT and PP) vs. −1.2 ± 0.7 mm at the 3-mm level; and −0.9 ± 0.7 (ITT)/ −0.9 ± 0.7 (PP) vs. −0.9 ± 0.9 mm at the 5-mm level, whereas the vertical height in the midfacial area was significantly reduced in the test group compared with the control group: −-1.1 ± 2.8 (ITT)/1.5 ± 3.0 (PP) vs. −0.7 ± 1.8 mm. Conclusions The horizontal ridge alteration in ridge preservation did not differ significantly between using the test the control groups, but the vertical ridge alteration was more pronounced when using the test group (KCT0001709).

This study aimed to compare the accuracy and reliability of digital versus conventional clinical measurements of the width of keratinized tissue. To this end, the keratinized tissue width was measured at 110 tooth sites in 5 pig jaws. The measurements were made at each site using three-dimensional (3D) scanned images and a periodontal probe. The actual keratinized tissue width was subsequently measured on histologic slides prepared from the same sites, and differences between the histologic slides and the digital and clinical measurements were analyzed to determine their accuracy in two measurement rounds. Furthermore, intrarater and interrater reliabilities were evaluated using the intraclass correlation coefficient (ICC). Here we show that the mean differences (and lower/upper limits of agreement) between the histologic and the digital/clinical measurements were 0.10 mm (−1.34/1.54 mm) and 1.11 mm (−0.69/2.92 mm), respectively, in the first round of measurements (p p p

Poly(ethylene glycol) (PEG) is widely used as a gold standard in bioconjugation and nanomedicine to prolong blood circulation time and improve drug efficacy. The conjugation of PEG to proteins, peptides, oligonucleotides (DNA, small interfering RNA (siRNA), microRNA (miRNA)) and nanoparticles is a well-established technique known as PEGylation, with PEGylated products have been using in clinics for the last few decades. However, it is increasingly recognized that treating patients with PEGylated drugs can lead to the formation of antibodies that specifically recognize and bind to PEG (i.e., anti-PEG antibodies). Anti-PEG antibodies are also found in patients who have never been treated with PEGylated drugs but have consumed products containing PEG. Consequently, treating patients who have acquired anti-PEG antibodies with PEGylated drugs results in accelerated blood clearance, low drug efficacy, hypersensitivity, and, in some cases, life-threatening side effects. In this succinct review, we collate recent literature to draw the attention of polymer chemists to the issue of PEG immunogenicity in drug delivery and bioconjugation, thereby highlighting the importance of developing alternative polymers to replace PEG. Several promising yet imperfect alternatives to PEG are also discussed. To achieve asatisfactory alternative, further joint efforts of polymer chemists and scientists in related fields are urgently needed to design, synthesize and evaluate new alternatives to PEG.

The variable lymphocyte receptor (VLR) mediates the humoral immune response in jawless vertebrates, including lamprey (Petromyzon marinus) and hagfish (Eptatretus burgeri). Hagfish VLRBs are composed of leucine-rich repeat (LRR) modules, conjugated with a superhydrophobic C-terminal tail, which contributes to low levels of expression in recombinant protein technology. In this study, we screened Ag-specific VLRBs from hagfish immunized with nervous necrosis virus (NNV). The artificially multimerized form of VLRB was constructed using a mammalian expression system. To enhance the level of expression of the Ag-specific VLRB, mutagenesis of the VLRB was achieved in vitro through domain swapping of the LRR C-terminal cap and variable LRR module. The mutant VLRB obtained, with high expression and secretion levels, was able to specifically recognize purified and progeny NNV, and the Ag binding ability of this mutant was increased by at least 250-fold to that of the nonmutant VLRB. Furthermore, preincubation of the Ag-specific VLRB with NNV reduced the infectivity of NNV in E11 cells in vitro, and in vivo experiment. Our results suggest that the newly developed Ag-specific VLRB has the potential to be used as diagnostic and therapeutic reagents for NNV infections in fish.

Oral formulations of insulin are typically designed to improve its intestinal absorption and increase its blood bioavailability. Here we show that polymerized ursodeoxycholic acid, selected from a panel of bile-acid polymers and formulated into nanoparticles for the oral delivery of insulin, restored blood-glucose levels in mice and pigs with established type 1 diabetes. The nanoparticles functioned as a protective insulin carrier and as a high-avidity bile-acid-receptor agonist, increased the intestinal absorption of insulin, polarized intestinal macrophages towards the M2 phenotype, and preferentially accumulated in the pancreas of the mice, binding to the islet-cell bile-acid membrane receptor TGR5 with high avidity and activating the secretion of glucagon-like peptide and of endogenous insulin. In the mice, the nanoparticles also reversed inflammation, restored metabolic functions and extended animal survival. When encapsulating rapamycin, they delayed the onset of diabetes in mice with chemically induced pancreatic inflammation. The metabolic and immunomodulatory functions of ingestible bile-acid-polymer nanocarriers may offer translational opportunities for the prevention and treatment of type 1 diabetes.

The occurrence of CD4 helper T cells has already been established for a number of teleost species, though, it has not been possible to analyze these responses at a cellular level due to a large lack of appropriate monoclonal antibodies (mAbs). In the present study, we produced a mAb against olive flounder (Paralichthys olivaceus) CD4-1 lymphocyte to investigate the functional activity of the cells to improve our understanding of the T cell response in this species. This mAb is specifically able to detect CD4-1 lymphocytes in olive flounder proved by immunofluorescence staining and RT-PCR analysis. In flow cytometry analysis, the number of CD4-1-positive lymphocytes was observed to gradually increase from 3 days post infection (dpi) and then reach peak at 7 dpi against two viruses challenge. As a conclusion, both the basic properties of CD4-1 T cells and its response to viral infections in olive flounder are very similar to the helper T cells in terrestrial animals.

Objectives This study histologically analyzed biopsy samples obtained from sites of damaged extraction socket grafting using deproteinized bovine bone mineral (DBBM) or deproteinized porcine bone mineral (DPBM) with coverage by a collagen membrane. Material and methods One hundred patients participated in this randomized controlled clinical trial of extraction socket grafts performed in cases of periodontally compromised teeth. All participants were blinded to their group allocations, and each material was grafted with coverage by collagen membranes after extraction of the tooth and removal of granulation tissue. At implant placement at 4 months, a biopsy was harvested at the implant site using a trephine was analyzed histologically. Results Eighty-five biopsy samples were acquired, of which 81 were finally included in the histologic analysis (42 in DBBM and 39 in DPBM group). Both DBBM and DPBM groups showed comparable proportions of residual biomaterial (12.37 ± 5.67% and 12.21 ± 5.75%, respectively), newly formed bone (15.07 ± 10.52% and 18.47 ± 11.47%, respectively), and nonmineralized tissue (72.56 ± 10.07% and 71.55 ± 15.47%, respectively). There were no significant differences in these histologic parameters between the two groups with different biomaterials. Conclusion Comparable histologic bone formation was found in both socket grafted groups with DBBM or DPBM covered by collagen membranes in periodontally damaged extraction sockets. However, a wide variation in new bone formation was found after 4 months of postsurgical healing and a tendency of higher new bone formation was shown at damaged sockets that had an intact unilateral residual wall regardless of buccal or lingual side.

Background Calcium phosphate (CaP)-coated surface showed enhanced contact osteogenesis around dental implant and finally accelerate osseointegration in early healing phase. Purpose The aim of this randomized controlled trial was to compare the peri-implant marginal bone level around uncoated and CaP-coated sandblasted, large-grit, acid-etched (SLA) surface implants during the first year after placement. Materials and methods This study was performed upon 34 patients with randomized and double-blinded design. Clinical and radiographic examinations were performed immediately after implant placement, at re-entry after 3 months, and after 12 months to evaluate the initial stability and change in the marginal bone level. The distance from the implant shoulder to the top of the bone-to-implant contact was defined as the marginal bone level, and its alteration was measured at 1 year after implant installation. Results None of the implants failed, and most of them showed a marginal bone loss of less than 1 mm. Small changes in the bone level were noted at all sites in the control and test groups, and there were no clinically significant differences in the changes in the marginal bone. Conclusion Both CaP-coated and uncoated SLA surface implants showed comparably successful marginal bone stability without any complications during the first year after placement.

Purpose To histologically characterize periodontal healing at 8 weeks in surgically created dehiscence defects in beagle dogs that received a collagen matrix with periodontal ligament (PDL) progenitor cells. Methods The bilateral maxillary premolars and first molars in 6 animals were used. Standardized experimental dehiscence defects were made on the buccal side of 3 premolars, and primary culturing of PDL progenitor cells was performed on the molars. Collagen matrix was used as a scaffold and a delivery system for PDL progenitor cells. The experimental sites were grafted with collagen matrix (COL), PDL progenitor cells with collagen matrix (COL/CELL), or left without any material (CTL). Histologic and histomorphometric analyses were performed after 8 weeks. Results The defect height from the cementoenamel junction to the most apical point of cementum removal did not significantly differ across the CTL, COL, and COL/CELL groups, at 4.57±0.28, 4.56±0.41, and 4.64±0.27 mm (mean ± standard deviation), respectively; the corresponding values for epithelial adhesion were 1.41±0.51, 0.85±0.29, and 0.30±0.41 mm (P, Graphical Abstract

Background: Group A Streptococcus (Strep A) causes a broad spectrum of disease manifestations, ranging from benign symptoms including throat or skin infections, to fatal illness such as rheumatic heart disease, or chronic renal failure. Currently, there is no vaccine available against Strep A infections. Despite the high burden of Strep A-associated infections worldwide, little attention has been paid to the research of these diseases, including standardized surveillance programs, resulting in a lack of economic evaluations for prevention efforts. This study aims at identifying existing cost-effectiveness analyses (CEA) on any Strep A infections. Methods: A systematic literature review was conducted by searching the PubMed electronic database. Results: Of a total of 321, 44 articles met the criteria for inclusion. Overall, CEA studies on Strep A remain limited in number. In particular, a number of available CEA studies on Strep A are disproportionately lower in low-income countries than in high-income countries. Decision-analytic models were the most popular choice for CEA on Strep A. A majority of the models considered pharyngitis and acute rheumatic fever, but it was rare to observe a model which covered a wide range of disease manifestations. Conclusions: Future research is needed to address missing clinical outcomes, imbalance on study locations by income group, and the transmission dynamic of selected diseases.

Purpose Optical coherence tomography (OCT) is a noninvasive diagnostic technique that may be useful for both qualitative and quantitative analyses of the periodontium. Micro-computed tomography (micro-CT) is another noninvasive imaging technique capable of providing submicron spatial resolution. The purpose of this study was to present periodontal images obtained using ex vivo dental OCT and to compare OCT images with micro-CT images and histologic sections. Methods Images of ex vivo canine periodontal structures were obtained using OCT. Biologic depth measurements made using OCT were compared to measurements made on histologic sections prepared from the same sites. Visual comparisons were made among OCT, micro-CT, and histologic sections to evaluate whether anatomical details were accurately revealed by OCT. Results The periodontal tissue contour, gingival sulcus, and the presence of supragingival and subgingival calculus could be visualized using OCT. OCT was able to depict the surface topography of the dentogingival complex with higher resolution than micro-CT, but the imaging depth was typically limited to 1.2–1.5 mm. Biologic depth measurements made using OCT were a mean of 0.51 mm shallower than the histologic measurements. Conclusions Dental OCT as used in this study was able to generate high-resolution, cross-sectional images of the superficial portions of periodontal structures. Improvements in imaging depth and the development of an intraoral sensor are likely to make OCT a useful technique for periodontal applications., Graphical Abstract

Streptococcus iniae causes severe mortalities among cultured marine species, especially in the olive flounder (Paralichthys olivaceus), which is economically important in Korea and Japan. Recently, there has been growing concern regarding the emergence of S. iniae as a zoonotic pathogen. Here, 89 S. iniae isolates obtained from diseased olive flounders collected from 2003 to 2008 in Jeju Island, South Korea, were characterized using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The results were aligned both with the available Bruker Daltonics data-base and with a new set of S. iniae data entries developed in our laboratory, and the results were compared. When we used the Bruker Daltonics database, the 89 isolates yielded either "no reliable identification" or were incorrectly identified as Streptococcus pyogenes at the genus level. When we used the new data entries from our laboratory, in contrast, all of the isolates were correctly identified as S. iniae at the genus (100%) and species (96.6%) levels. We performed proteomic analysis, divided the 89 isolates into cluster I (51.7%), cluster II (20.2%), and cluster III (28.1%), and then used the MALDI Biotyper software to identify specific mass peaks that enabled discrimination between clusters and between Streptococcus species. Our results suggest that the use of MALDI TOF MS could outperform the conventional methods, proving easier, faster, cheaper and more efficient in properly identifying S. iniae. This strategy could facilitate the epidemiological and taxonomical study of this important fish pathogen.

A practical strategy to strengthen the inherent tissue adhesiveness of bioglues was investigated without compromising their chemical structure. A simple blending of α-cyclodextrins (α-CDs) in gelatin glues substantially improved the adhesiveness of the glues upon contact with porcine skin. Interestingly, the adhesiveness was even further enhanced when the glues were supplemented with γ-CDs through the formation of multivalent supramolecular networks inside of the glue. In contrast, the effect of β-CDs was rather limited because their water solubility is relatively low. A model study using modified gold substrates demonstrated that CDs also interacted with biomolecules naturally present in skin, thereby improving the adhesiveness. Altogether, we suggest that the supramolecular networking of CDs, both inside the glue and at the glue-skin interface, could reinforce tissue adhesiveness of bioglues, which allows them to be of great relevance to a primary treatment modality for larger skin incisions.

Subluxation of the extensor tendon results from a disruption to the sagittal band at the metacarpophalangeal joint. When conservative treatment fails to correct the subluxation, surgical treatment may be necessary. Surgical techniques for chronic cases vary in graft source and graft pathway. We present a surgical technique to recentralize and stabilize the extensor tendon using a residual ruptured sagittal band. This technique is simple and effective without donor site morbidity and seems to provide potential biomechanical advantages by restoring nearly normal anatomy.

Immunoglobulins (Ig) are heterodimeric proteins that play critical roles in the adaptive immune system of vertebrates. Because of their plasticity, teleostean Igs are more diverse, and thus do not conform to mammalian classifications. Because of this, mammalian-based Ig cell markers cannot be used successfully to study immune responses in fish. There is therefore a need to produce Ig-specific cell markers for fish. Here, we attempted to identify the specific isotype detected by an Ig light chain-specific monoclonal antibody (anti-olive flounder IgL-mAb: M7C3-4) that we had previously produced [11]. Three newly identified sequences of the Ig light chain from olive flounder were classified according to their isotypes. Subsequent analyses revealed that M7C3-4 was able to specifically detect lymphocytes expressing one of the κ chains (Igκ-a) in olive flounder. Interestingly, Igκ-a+ B cells were more abundant in spleen and trunk-kidney than in peripheral blood, indicating a distribution different from that of IgM+ B cells. Our work reveals interesting aspects of B cell distribution and differentiation, and may aid in the production of suitable and effective cell markers for olive flounder.

Aim The aim of this study was to evaluate the effect of fibroblast growth factor-2 (FGF-2) in combination with porcine collagen matrix (CM) for coverage of gingival recession defects in dogs. Materials and Methods In five male mongrel dogs, labial gingival recession defects were surgically created in the lower-third incisors bilaterally. The defects were randomly assigned to the CM/FGF-2 group (experimental) or the CM-only group (control). Standardised clinical photographs and silicone impressions were taken at 4 and 16 weeks of healing to calculate the remaining recession area. The dogs were euthanised after 16 weeks for histometric analysis. Results At 4 weeks, the recession area of the CM/FGF-2 group was significantly smaller than that of the CM-only group (2.55 ± 0.66 and 4.92 ± 1.05 mm2, respectively, p=0.023), However, no significant difference was observed at 16 weeks. Histometrically, the amount of newly formed cementum was larger in the CM/FGF-2 group than in the CM-only group at 16 weeks (1.55 ± 0.44 and 0.88 ± 0.51 mm, respectively, p=0.024). Conclusions The combination of CM/FGF-2 may enhance the early wound healing and may promote cementum formation to a higher extent compared to the use of CM alone. This article is protected by copyright. All rights reserved.

Centrifugation based on density gradients is a general methodology for isolating human bone marrow (hBM)-derived mesenchymal stem cells (hBMSCs). The mononuclear cell (MNC) layer can be obtained using a density gradient solution in the conventional protocol, but it is not suitable for direct transplantation due to the possible toxicity of this solution. The results obtained are also influenced by the skill level when applying the technique, which involves time-consuming processes. We have developed a novel protocol for isolating hBMSCs using hyaluronic acid (HA), which is the most widely used injectable biomaterial in clinical settings and a major component of the extracellular matrix. Laying hBM over the HA and then applying centrifugation yielded three separate layers, with the HA layer, including MNCs being the most superficial one. Increasing the volume of HA and/or its crosslinking rate enhanced the yield of MNCs from hBM, and the cell yield was also significantly higher for a lower centrifugal acceleration (530 g) than for a higher one (1500 g). Isolated hBMSCs by HA exhibited similar biological characteristics such as in terms of their proliferation rate, fibroblast-like morphology, cell-cycle status, immunophenotype, and multipotency. The use of either type of hBMSC confirmed the regenerative potential of bone and bone marrow-like tissue in ectopic transplantation models. This is the first report of a novel protocol for isolating hBMSCs that utilize HA. We suggest that this novel isolation technique can be used for the direct application of autogenous MSCs with advantages of being less time-consuming and involving steps that are easier to perform.

The beneficial effects of a strength exercise program and a combined exercise program of strength training plus walking were examined in overweight with chronic back pain patients. The participants were randomly placed in the strength exercise group (SEG, n=15), combined exercise group (CEG, n=15), and control group (CG, n=6). All subjects performed exercise twice per week, 50 min per session with a professional instructors for 12 weeks. In order to evaluate exercise intervention effects, lumbar function was measured by back strength and flexibility. Roland-Morris disability questionnaire (RMDQ) and visual analogue scale (VAS) were used to evaluate pain level. Fat and muscle mass were measured to compare body composition changes. All measurements were performed before and after 12 weeks of exercise program. Lumbar function: Back strength was significantly different over time, and significant time×group differences were found between SEG and CG and, CEG and CG. Pain disorder degree: VAS showed a significant group difference, and significant time×group differences were shown between SEG and CG, and CEG and CG. Also, RMDG showed a significant difference between CEG and CG. Body composition: Fat mass was significantly different over time×group between SEG and CG. In conclusion, participating in strength and walking exercises were beneficial to improve lumbar function. Also, the combined exercise program was more effective for reducing pain levels than the strength exercise. Finally, fat mass was reduced in this study and this may play a possible role in the improvement of lumbar function and reduction in low back pain.

Submerged plasma irradiation (SPI)-based advanced oxidation processes have been studied for the oxidation of recalcitrant organic compounds because of their various physical and chemical properties. However, SPI technologies still have a few drawbacks such as relatively low efficiency for wastewater treatment and high energy consumption. In order to overcome these drawbacks, in this study, we proposed the combination of SPI and the Cu(II)-catalyzed Fenton-like system. The removal of methylene blue (MB) by the SPI system was significantly enhanced upon the addition of H2O2. The pseudo-first-order rate constants of MB removal increased with the increase of applied voltage. In addition, the optimum H2O2 dose and initial solution pH were 100 mM and 9, respectively. The reactive oxidants responsible for MB removal in copper electrode-based SPI/H2O2 systems are likely to be hydroxyl radicals (OH) or cupryl ion (Cu(III)), wherein Cu(III) is especially important. Furthermore, the copper electrode-based SPI/H2O2 system is a novel advanced oxidation process capable of oxidizing water recalcitrant and toxic organic pollutants at neutral pH.

Aims The objectives of this study were to evaluate bone regeneration beneath a newly devised bone substitute combined with collagen membrane (called a bone patch) lying over a concomitantly placed mini-implant following sinus floor elevation and verify its usefulness as a carrier system for recombinant human bone morphogenetic protein-2 (rhBMP-2) in rabbits. Materials and methods The sinus floor elevation procedure was performed bilaterally in five rabbits. Either a plain bone patch (control group) or an rhBMP-2-loaded patch (experimental group) was randomly placed beneath the elevated sinus membrane (SM) of both sinuses, where the mini-implants were concomitantly placed. Micro-computed tomographic and histologic analyses were performed at 4 weeks post-surgery. Results In micro-computed tomography, the median values of the total augmented volume and the mineralized bone volume were significantly higher in the experimental group than in the control group (161 vs. 122 mm3 [P

Background Deproteinized porcine bone mineral (DPBM) was recently developed and commercially available in maxillary sinus grafting, in which demineralized bovine bone mineral (DBBM) was widely used. Objectives The present randomized controlled clinical trial aimed to compare histological bone quality and radiographic volume stability in maxillary sinuses grafted with DPBM and DBBM. Materials and Methods Twenty sinuses in 16 participants were enrolled and randomly allocated to control and test groups using sequentially numbered, sealed envelopes; laterally approached sinus grafting with DBBM and DPBM, respectively. All participants were blinded to the group assignment during the entire experiment. After standardized osteotomy at the lateral wall of the maxillary sinus, the sinus membrane was elevated, and the control or test biomaterial was grafted. Computed tomography (CT) images were taken immediately after surgery, and another CT and trephine biopsy was taken for radiographic and histological analyses after 6 months. The histological bone quality was measured as a primary outcome, and changes in the height and volume of the graft were evaluated in the reconstructed CT images as secondary outcomes. Results Fifteen sites (7 and 8 sites for control and test group) in 11 participants were finally included in the per protocol (PP) analysis, and 16 sites (7 and 9 sites, respectively) in 12 participants were included in the intention-to-treat (ITT) analysis; there were four drop-outs and one minor protocol violation. In both statistical analyses, the test groups showed comparable new bone formation and residual biomaterials in histology, and both groups exhibited minimal volume/height changes in radiographies. However, smaller sizes of residual biomaterials were observed in the histological samples from the test compared to control sites, despite the use of the same sizes of both biomaterials. Conclusions The results suggested that DPBM might produce comparable bone formation and volumetric stability with DBBM in maxillary sinus grafting, however, further clinical study with longer-term periods and larger sample sizes should be needed for confirming this suggestion.

Histone deacetylase inhibitors such as sodium butyrate (SB) have been suggested to be promising candidate small molecules for bone regeneration. In this study, the capacity of SB loaded onto calcium sulfate (CaS) to enhance bone formation was investigated in a rabbit sinus model. Following preparation of the sinus access window on a randomly selected side, SB loaded onto CaS (CaS/SB) was grafted in five rabbits, and CaS alone (control) was grafted in another five rabbits. The animals were euthanized after 4 weeks for radiographic, histometric, and immunohistochemical analyses. There was a statistically significant difference in the total augmented volume between the groups in the radiographic analysis (158.22 ± 39.31 mm3 and 107.09 ± 39.69 mm3 , respectively, p = 0.040). The CaS/SB group showed a larger portion of mature lamellar bone and a higher level of mineralization of bone trabeculae, characterized by more intense labeling with osteocalcin compared with the control group in the immunohistochemical analysis. The number of osteocalcin-positive cells within the central area of the augmented sinus was significantly higher in the CaS/SB group than in the control group (179 ± 26.0 mm2 and 123 ± 33.2 mm2 , respectively, p = 0.027). In conclusion, CaS/SB exhibited superior osteogenic potential, especially in the central portion of the augmented sinus as well as improvement of the volume maintenance for sinus augmentation. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1916-1923, 2017.

Objectives The aim of this study was to evaluate the hybrid technique, which was modified from the lateral approach in sinus elevation by combining transalveolar access. Materials and methods Sixty patients (mean age of 53.3 ± 11.6 years) with 105 implants placed in the posterior maxilla with the average remaining bone height (RBH) of 4.3 ± 1.8 mm (range of 1.2–9.2 mm) were enrolled in the hybrid group and followed up for 96 months. The control group included 36 patients with 44 implants placed in the corresponding site with sufficient RBH. Cumulative survival rate (CSR), radiographic measurements in RBH, augmented bone height (ABH), and marginal bone loss (MBL) were analyzed during the follow-up period. Results Cumulative survival rate in the hybrid group was 98.1%, which was comparable to the control group (97.7%). MBL of the mesial and distal aspects in the hybrid group (0.28 ± 0.66 mm and 0.41 ± 0.93 mm, respectively) was comparable to the control group (0.84 ± 1.11 mm and 0.76 ± 0.88 mm, respectively). In the hybrid group, the membrane perforation rate during the surgery was 4.5%. ABH was 8.3 ± 1.8 mm after the surgery and maintained consistently during the follow-up period (7.4 ± 1.9 mm after 4 years). Conclusion The hybrid technique is a simple and predictable surgical method for simultaneous placement of a dental implant with sinus floor elevation regardless of the RBH.