Osseointegration Potential Assessment of Bone Graft Materials Loaded with Mesenchymal Stem Cells in Peri-Implant Bone Defects.

Many studies have been exploring the use of bone graft materials (BGMs) and mesenchymal stem cells in bone defect reconstruction. However, the regeneration potential of Algipore (highly purified hydroxyapatite) and Biphasic (hydroxyapatite/beta-tricalcium phosphate) BGMs combined with bone marrow-derived mesenchymal stem cells (BMSCs) remains unclear. Therefore, we evaluated their osseointegration capacities in reconstructing peri-implant bone defects. The cellular characteristics of BMSCs and the material properties of Algipore and Biphasic were assessed in vitro. Four experimental groups-Algipore, Biphasic, Algipore+BMSCs, and Biphasic+BMSCs-were designed in a rabbit tibia peri-implant defect model. Implant stability parameters were measured. After 4 and 8 weeks of healing, all samples were evaluated using micro-CT, histological, and histomorphometric analysis. In the energy-dispersive X-ray spectroscopy experiment, the Ca/P ratio was higher for Algipore (1.67) than for Biphasic (1.44). The ISQ values continuously increased, and the PTV values gradually decreased for all groups during the healing period. Both Algipore and Biphasic BGM promoted new bone regeneration. Higher implant stability and bone volume density were observed when Algipore and Biphasic BGMs were combined with BMSCs. Biphasic BGM exhibited a faster degradation rate than Algipore BGM. Notably, after eight weeks of healing, Algipore with BSMCs showed more bone-implant contact than Biphasic alone (p < 0.05). Both Algipore and Biphasic are efficient in reconstructing peri-implant bone defects. In addition, Algipore BGM incorporation with BSMCs displayed the best performance in enhancing implant stability and osseointegration potential.

Assessment of the immunogenicity and protection of a Nipah virus soluble G vaccine candidate in mice and pigs.

Nipah virus (NiV) is a newly emerged extremely dangerous zoonotic pathogen highly fatal to humans. Currently, no approved vaccine is available against NiV. This study employed a mammalian eukaryotic system to express NiV soluble G glycoprotein (NiV-sG), using CpG oligodeoxynucleotides (CpG)/Aluminum salt (Alum) as adjuvants to obtain a recombinant subunit vaccine candidate. We also evaluated the immunogenicity and efficacy of the protein in mice and pigs. The results showed that humoral and cellular immune responses were induced in all the vaccination groups in two animal models. The levels of specific and neutralizing antibodies and the proliferation levels of T helper(Th) cells were significantly higher than those in the control group. The protective efficacy of the subunit vaccines evaluated in the pseudovirus in vivo infection mouse model strongly suggested that this vaccine could provide protective immunity against NiV. A neoadjuvant (HTa) based on liposomes and cholera toxin combined with CpG/Alum was exploited and evaluated in mice. The neoadjuvant group showed a more protective efficacy than the CpG/Alum group. The aforementioned results indicated that the subunit vaccine could be used as a promising candidate vaccine for preventing Nipah virus infection.

Multiplexed Molecular Imaging Strategy Integrated with RNA Sequencing in the Assessment of the Therapeutic Effect of Wharton's Jelly Mesenchymal Stem Cell-Derived Extracellular Vesicles for Osteoporosis.

INTRODUCTION: Osteoporosis is a result of an imbalance in bone remodeling. Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have been considered as a potentially promising treatment for osteoporosis. However, the therapeutic effect, genetic alterations, and in vivo behavior of exogenous EVs for osteoporosis in mice models remain poorly understood. METHODS: A multiplexed molecular imaging strategy was constructed by micro-positron emission tomography (µPET)/computed tomography (CT), µCT, and optical imaging modality which reflected the osteoblastic activity, microstructure, and in vivo behavior of EVs, respectively. RNA sequencing was used to analyze the cargo of EVs, and the bone tissues of ovariectomized (OVX) mice post EV treatment. RESULTS: The result of [18F]NaF µPET showed an increase in osteoblastic activity in the distal femur of EV-treated mice, and the bone structural parameters derived from µCT were also improved. In terms of in vivo behavior of exogenous EVs, fluorescent dye-labeled EVs could target the distal femur of mice, whereas the uptakes of bone tissues were not significantly different between OVX mice and healthy mice. RNA sequencing demonstrated upregulation of ECM-related genes, which might associate with the PI3K/AKT signaling pathway, in line with the results of microRNA analysis showing that mir-21, mir-29, mir-221, and let-7a were enriched in Wharton's jelly-MSC-EVs and correlated to the BMP and PI3K/AKT signaling pathways. CONCLUSION: The therapeutic effect of exogenous WJ-MSC-EVs in the treatment of osteoporosis was successfully assessed by a multiplexed molecular imaging strategy. The RNA sequencing demonstrated the possible molecular targets in the regulation of bone remodeling. The results highlight the novelty of diagnostic and therapeutic strategies of EV-based treatment for osteoporosis.

Damping Factor as a Diagnostic Parameter for Assessment of Osseointegration during the Dental Implant Healing Process: An Experimental Study in Rabbits.

The purpose of this study was to evaluate the possibility of using damping factor (DF) analysis to provide additional information on osseointegration of dental implants during the healing period. A total of 30 dental implants were installed in the bilateral femoral condyles of 15 rabbits. A DF analyzer detected with an impulse-forced vibration method and a commercialized dental implant stability analyzer based on resonance frequency (RF) analysis were used to measure the implant stability immediately after implant placement and 1, 2, 4, and 8 weeks post-surgically. Results of DF and RF analyses at different time points were compared with the corresponding osseointegration performance of dental implants via micro-computed tomography (micro-CT), histological and histomorphometrical analysis. The DF values revealed a decrease with time and reached 0.062 ± 0.007 at 8 weeks after implantation, which is almost 50% lower than the initial value. Moreover, highly significant correlations between DF values and bone volume densities (R 2 = 0.9797) and percentages of bone-to-implant contact measured at trabecular bone area (R 2 = 0.9773) were also observed. These results suggested that DF analysis combined with RF analysis results in a more sensitive assessment of changes in the dental implant/bone complex during the healing period than RF analysis alone.

Summarizing and quantifying multilocus linkage disequilibrium patterns with multi-order Markov chain models.

Studies on linkage disequilibrium (LD) are important in mapping disease genes. A novel statistical method, the multi-order Markov chain model has been recently developed to quantify the complexity level of multilocus LD patterns among single nucleotide polymorphism markers (Kim et al., 2008). In this study, mathematical relationships between two types of LD measures are derived to understand the Markov chain model parameters in terms of conventional LD measures. Statistical sample properties of the Markov chain order estimates are investigated by simulations. Two published data sets are reanalyzed to illustrate the proposed approach.

Measuring and partitioning the high-order linkage disequilibrium by multiple order Markov chains.

A map of the background levels of disequilibrium between nearby markers can be useful for association mapping studies. In order to assess the background levels of linkage disequilibrium (LD), multilocus LD measures are more advantageous than pairwise LD measures because the combined analysis of pairwise LD measures is not adequate to detect simultaneous allele associations among multiple markers. Various multilocus LD measures based on haplotypes have been proposed. However, most of these measures provide a single index of association among multiple markers and does not reveal the complex patterns and different levels of LD structure. In this paper, we employ non-homogeneous, multiple order Markov Chain models as a statistical framework to measure and partition the LD among multiple markers into components due to different orders of marker associations. Using a sliding window of multiple markers on phased haplotype data, we compute corresponding likelihoods for different Markov Chain (MC) orders in each window. The log-likelihood difference between the lowest MC order model (MC0) and the highest MC order model in each window is used as a measure of the total LD or the overall deviation from the gametic equilibrium for the window. Then, we partition the total LD into lower order disequilibria and estimate the effects from two-, three-, and higher order disequilibria. The relationship between different orders of LD and the log-likelihood difference involving two different orders of MC models are explored. By applying our method to the phased haplotype data in the ENCODE regions of the HapMap project, we are able to identify high/low multilocus LD regions. Our results reveal that the most LD in the HapMap data is attributed to the LD between adjacent pairs of markers across the whole region. LD between adjacent pairs of markers appears to be more significant in high multilocus LD regions than in low multilocus LD regions. We also find that as the multilocus total LD increases, the effects of high-order LD tends to get weaker due to the lack of observed multilocus haplotypes. The overall estimates of first, second, third, and fourth order LD across the ENCODE regions are 64, 23, 9, and 3%.

Determination of duloxetine in human plasma via LC/MS and subsequent application to a pharmacokinetic study in healthy Chinese volunteers.

BACKGROUND: The pharmacokinetics of duloxetine hydrochloride have been well studied after its approval for clinical use. However, few such data have been reported in the English language literature. We developed a method to determine the pharmacokinetics of duloxetine enteric-coated capsules in healthy Chinese volunteers. METHODS: A rapid and sensitive liquid chromatography-mass spectrometric (LC/MS) method for the determination of duloxetine in human plasma using flupentixol as the internal standard (I.S.) was developed and validated. Sample preparation of the plasma involved deproteination with acetonitrile twice, repeatedly. Samples were then analyzed by HPLC on a Thermo Hypersil-Hypurity C18 column (150 x 2.1 mm, 5 microm). A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M+H](+) ions at 298 m/z for duloxetine and at 435 m/z for the internal standard. RESULTS: Pharmacokinetics were measured in 12 healthy Chinese male volunteers (6 males and 6 females) who received a single regimen with 3 different dosages at 22.4, 44.8 and 67.2 mg of duloxetine enteric-coated capsules. CONCLUSION: A sensitive and specific method for quantifying duloxetine levels in human plasma has been devised and successfully applied to a clinic pharmacokinetic study of an enteric-coated capsule of duloxetine hydrochloride administered as a single oral dose.