Platelet-Rich Fibrin Facilitates One-Stage Cartilage Repair by Promoting Chondrocytes Viability, Migration, and Matrix Synthesis.

The main aim of this study is to develop a one-stage method to combine platelet-rich fibrin (PRF) and autologous cartilage autografts for porcine articular cartilage repair. The porcine chondrocytes were treated with different concentrations of PRF-conditioned media and were evaluated for their cell viability and extracellular glycosaminoglycan (GAG) synthesis during six day cultivation. The chemotactic effects of PRF on chondrocytes on undigested cartilage autografts were revealed in explant cultures. For the in vivo part, porcine chondral defects were created at the medial femoral condyles of which were (1) left untreated, (2) implanted with PRF combined with hand-diced cartilage grafts, or (3) implanted with PRF combined with device-diced cartilage grafts. After six months, gross grades, histological, and immunohistochemical analyses were compared. The results showed that PRF promotes the viability and GAG expression of the cultured chondrocytes. Additionally, the PRF-conditioned media induce significant cellular migration and outgrowth of chondrocytes from undigested cartilage grafts. In the in vivo study, gross grading and histological scores showed significantly better outcomes in the treatment groups as compared with controls. Moreover, both treatment groups showed significantly more type II collagen staining and minimal type I collagen staining as compared with controls, indicating more hyaline-like cartilage and less fibrous tissue. In conclusion, PRF enhances the viability, differentiation, and migration of chondrocytes, thus, showing an appealing capacity for cartilage repair. The data altogether provide evidences to confirm the feasibility of a one-stage, culture-free method of combining PRF and cartilage autografts for repairing articular cartilage defects. From translational standpoints, these advantages benefit clinical applications by simplifying and potentiating the efficacy of cartilage autograft transplants.

Overexpression of AKR1B10 predicts tumor recurrence and short survival in oral squamous cell carcinoma patients.

BACKGROUND: Aldo-keto reductase family 1 member B10 (AKR1B10) is an enzyme implicated in physiological xenobiotic detoxification and also in pathological carcinogenesis. Overexpression of AKR1B10 has been reported in oral squamous cell carcinoma (OSCC), but its correlation with clinical prognosis is controversial. The aim of this study was to investigate and clarify the role of AKR1B10 in OSCC carcinogenesis. METHODS: Tumor tissue specimens were surgically obtained from 107 patients with OSCC. The expression of AKR1B10 was analyzed by immunohistochemistry to explore the relationship between the level of AKR1B10 and clinicopathological features of OSCC patients. Kaplan-Meier survival and Cox proportional hazard analysis were used to determine the prognostic value of AKR1B10 in OSCC. RESULTS: High expression of AKR1B10 was found to be associated with tumor size (P = 0.043), perineural invasion (P = 0.012), and recurrence (P = 0.001) in OSCC. Cox model analysis revealed that high expression of AKR1B10 is significantly associated with poor overall and disease-free survival in OSCC patients. With the combination of clinicopathological factors in analysis, we found that the expression level of AKR1B10 was a practical indicator that could categorize OSCC patients into different risk groups. High expression of AKR1B10 was associated with a reduced survival in patients with well and moderately differentiated OSCC and even a high incidence of tumor recurrence in the patients with late-stage (III and IV) disease. CONCLUSION: We validated and expanded data on the expression of AKR1B10 in OSCC, suggesting that it is a valuable biomarker for prognostic prediction of recurrence and survival in OSCC.

Outcomes of necrotic immature open-apex central incisors treated by MTA apexification using poly(ε-caprolactone) fiber mesh as an apical barrier.

BACKGROUND/PURPOSE: Although unset mineral trioxide aggregate (MTA) has some cytotoxicity, MTA is still a biocompatible material suitable for doing apexification. This study assessed the outcomes for 8 necrotic immature open-apex permanent maxillary central incisors treated by MTA apexification using poly(ε-caprolactone) fiber mesh (PCL-FM) as an apical barrier (so-called PCL-FM/MTA apexification) to prevent extrusion of MTA materials into the periapical tissues of open-apex teeth. METHODS: Eight necrotic immature open-apex permanent maxillary central incisors with the open apices measuring 2.5 mm-3.5 mm in diameter in 8 patients (6 boys and 2 girls; age range, 8-10 years) were first cleaned using ultrasonic activated irrigation with 2.5% sodium hypochlorite solution and then treated by PCL-FM/MTA apexification procedure. RESULTS: All the 8 permanent maxillary central incisors showed successful outcomes after PCL-FM/MTA apexification procedure. The mean duration for apical hard tissue barrier formation of the 8 incisors was 6.8 ± 0.5 weeks (range 6-7 weeks). The mean increased root length was 1.8 ± 0.7 mm (range 1-3 mm) at 7 weeks and 3.1 ± 0.6 mm (range 2-4 mm) at 3 months. The mean increased dentinal wall thickness at the most apical portion of the root was 1.3 ± 0.5 mm (range 1-2 mm) at 7 weeks and 2.4 ± 0.6 mm (range 1.5-3 mm) at 3 months. None of the teeth treated by PCL-FM/MTA apexification showed tooth discoloration after a follow-up period of 3 months. CONCLUSION: PCL-FM/MTA apexification is an excellent technique for treatment of necrotic immature open-apex permanent maxillary central incisors.

Serous degeneration of bone marrow mimics spinal tumor.

OBJECTIVE: To present a rare case of serous degeneration of bone marrow which resembles primary spinal tumor or bony metastasis to spine. Serous degeneration of bone marrow or gelatinous marrow transformation is a rare disease characterized by focal marrow hypoplasia, fat atrophy, and accumulation of extracellular mucopolysaccharides abundant in hyaluronic acid. Few literature was reviewed and few clinical case was presented. METHODS: Two cases of serous marrow transformation were reported. RESULTS: In the first case, a 29-year-old man suffered from severe left buttock pain. Bone metastasis was impressed in radiology examinations. Percutaneous endoscopic lumbar discectomy was performed along with bone biopsy. In the second case, a 49-year-old man presented lower back pain with radiation to bilateral lower legs. Magnetic resonance imaging revealed a water-like signal lesion in sacrum. Serous marrow transformation was confirmed pathologically in both cases. CONCLUSION: To the best of our knowledge, a case of serous degeneration of bone marrow resembling malignancy has not been reported in the literature. In this report, two cases demonstrate serous transformation of bone marrow mimics spinal tumor.

Platelet-Rich Fibrin Facilitates Rabbit Meniscal Repair by Promoting Meniscocytes Proliferation, Migration, and Extracellular Matrix Synthesis.

Although platelet-rich fibrin (PRF) has been used in clinical practice for some time, to date, few studies reveal its role as a bioactive scaffold in facilitating meniscal repair. Here, the positive anabolic effects of PRF on meniscocytes harvested from the primary culture of a rabbit meniscus were revealed. The rabbit meniscocytes were cultured with different concentrations of PRF-conditioned medium, and were evaluated for their ability to stimulate cell migration, proliferation, and extracellular matrix formation. In vivo, meniscal defects were created via an established rabbit animal model and were evaluated by a histology-based four-stage scoring system to validate the treatment outcome three months postoperatively. The in vitro results showed that PRF could induce cellular migration and promote proliferation and meniscocyte extracellular matrix (ECM) synthesis of cultured meniscocytes. In addition, PRF increased the formation and deposition of cartilaginous matrix produced by cultured meniscocytes. Morphological and histological evaluations demonstrated that PRF could facilitate rabbit meniscal repair. The data highlight the potential utility of using PRF in augmenting the healing of meniscal injuries. These advantages would benefit clinical translation, and are a potential new treatment strategy for meniscal repair.

Effect of nanostructured thin film on minimally invasive surgery devices applications: characterization, cell cytotoxicity evaluation and an animal study in rat.

BACKGROUND: Minimally invasive surgery is performed using an endoscope and other instruments including the electrosurgical units. However, concerns including surgical smoke, tissue sticking and thermal injury are remaining in electrosurgery. AIMS: Accordingly, a newly developed electrosurgical electrode coating with hydrogenated Cu-incorporated diamond-like carbon (DLC-Cu) film is purposed to improve the instrument performance. METHODS: The morphologies of DLC-Cu surfaces were characterized using transmission electron microscopy, scanning electron microscopy and atomic force microscopy. In this study, lesions were made on the liver lobes of adult rats, using a monopolar electrosurgical unit equipped with untreated stainless steel electrodes or treated-electrodes. Animals were killed for evaluations at 0, 3, 7 and 28 days postoperatively. RESULTS: Treated-electrodes generate less sticking tissues and adhesive blood cells. Thermography revealed that the surgical temperature in liver tissue from the treated-electrode was significantly lower than the untreated-electrode. Total injury area of livers treated with treated-electrodes was significantly smaller than the untreated-electrodes treatment. Moreover, treated-electrodes caused a relatively smaller area of lateral thermal injury, a smaller area of fibrotic tissue and a faster process of remodeling than the untreated-electrodes. Western blot analysis showed that rats treated with treated-electrode expressed lower levels of NF-κB, caspase-3 and MMP-9 than untreated-electrode. Immunofluorescence staining for caspase-3 revealed that the untreated-electrode caused more serious injury. CONCLUSIONS: This study reveals that the plating of electrodes with hydrogenated Cu-incorporated diamond-like carbon film is an efficient method for improving the performance of electrosurgical units, and should benefit wound remodeling. However, more tests must be carried out to confirm these promising findings in human patients.

Osseointegration of titanium implants with SLAffinity treatment: a histological and biomechanical study in miniature pigs.

PURPOSE: Electrochemical oxidation following sandblasting and acid-etching (SLA) treatment has received interest as a surface modification procedure for titanium (Ti) implants (denoted as an SLAffinity surface); however, little information is available on its impacts on the in vivo performance of SLAffinity-Ti implants. The present study evaluated the osseointegration and biomechanical bone-tissue response to SLAffinity-Ti implants with micro- and nanoporous oxide layers. MATERIALS AND METHODS: The interaction between blood and the tested implants was examined. In total, 144 implants with the following surfaces were used: a standard machined (M-Ti), an SLA-Ti, and an SLAffinity-Ti surface. For each animal, four implants (one M-Ti, one SLA-Ti, and two SLAffinity-Ti) were inserted into the mandibular canine-premolar area for histomorphometric observations and another four implants were inserted into the flat surface on the anteromedial aspect of the rear tibia for removal torque (RT) tests. After 2, 4, and 8 weeks of implantation, histomorphometric and RT tests were conducted. RESULTS: Interactions between blood and implants were better for implants with the SLAffinity-Ti surface. RT tests showed a significant improvement in fixation strength for SLAffinity-Ti implants (84.5 ± 8.7 N-cm) after 8 weeks compared to M-Ti (62.95 ± 11.5 N-cm) and SLAffinity-Ti (76.1 ± 6.6 N-cm) implants. A histological evaluation showed that osseous integration had occurred with all implants after 8 weeks. SLAffinity-Ti implants exhibited 28.5 ± 6.2 % bone-to-implant contact (BIC) at 2 weeks and 84.3 ± 8.1 % at 8 weeks. M-Ti implants exhibited BIC levels of 17.0 ± 5.4 and 76.5 ± 6.3 %, whereas SLA-Ti implants exhibited BIC levels of 28.5 ± 6.2 and 81.1 ± 8.4 % at corresponding time intervals. In terms of the peri-implant bone area (BA), values for SLAffinity-Ti implants ranged from 29.5 ± 4.1 to 88.3 ± 3.0 %. For M-Ti implants, values ranged from 20.3 ± 5.5 to 81.7 ± 4.2 %. For SLA-Ti implants, values ranged from 23.0 ± 3.5 to 84.0 ± 3.6 %. CONCLUSIONS: Electrochemical oxidation increased the oxide layers and improved the blood interaction with SLAffinity-Ti implants, resulting in significantly higher bone apposition with the SLAffinity-Ti implants after 2 and 8 weeks of healing. An increase in resistance for the RT of SLAffinity-Ti implants over the 8-week healing period was also observed. CLINICAL RELEVANCE: The use of SLAffinity-Ti implants has potential for improvement of early osseointegration.

Research of StemBios Cell Therapy on Dental Implants Containing Nanostructured Surfaces: Biomechanical Behaviors, Microstructural Characteristics, and Clinical Trial.

PURPOSE: The aim of the present study was to examine the osseointegration in low-density bone tissue for SLAffinity-treated implants with StemBios (SB) cell therapy. MATERIALS AND METHODS: The morphologies of SLAffinity-treated surfaces were characterized using scanning electron microscopy. In the animal model, implants were installed in the mandibular canine-premolar area of 12 miniature pigs. Each pig received 3 implants of machine, sand blasted, large grit, and acid etched, and SLAffinity-treated implants. In the clinical trial, 10 patients received 1 SLAffinity-treated implant in the maxilla in the posterior area and 1 patient with low bone tissue density received 2 SLAffinity-treated implants with SB cell therapy. Resonance frequency analysis and computed tomography were assessed monthly over the first 3 months after implant placement. RESULTS: The results demonstrated that surface treatment significantly affected early osseointegration in patients who received SB cell therapy. SB cell therapy transferred the stress caused by the implant more uniformly, and the stress decreased with healing time. SLAffinity-treated implants also proved clinically successful after the 3 months. CONCLUSION: The SLAffinity treatments enhanced osseointegration significantly, especially at early stages of bone tissue healing with SB cell therapy.

Comparison of clinical outcomes for 40 necrotic immature permanent incisors treated with calcium hydroxide or mineral trioxide aggregate apexification/apexogenesis.

BACKGROUND/PURPOSE: Traumatic injury often results in pulp necrosis of immature permanent incisors in children. This study compared clinical outcomes for 40 necrotic immature permanent incisors treated with calcium hydroxide [Ca(OH)2] or mineral trioxide aggregate (MTA) apexification/apexogenesis. METHODS: Forty necrotic open-apex incisors from 40 children aged 6.5-10 years were divided evenly into four groups with each group containing teeth of similar type and similar root apex width in patients of similar age. Group 1 incisors were treated with ultrasonic filing and MTA placement; Group 2 were treated with ultrasonic filing and Ca(OH)2 medication; Group 3 were treated with hand filing and MTA placement; and Group 4 were treated with hand filing and Ca(OH)2 medication. RESULTS: Group 1 incisors needed the shortest mean duration (5.4 ± 1.1 weeks) for apical hard tissue barrier formation, followed by Group 3 incisors (7.8 ± 1.8 weeks), Group 2 incisors (11.3 ± 1.3 weeks), and Group 4 incisors (13.1 ± 1.5 weeks). Group 1 incisors had a significantly shorter mean elongated root length (2.1 ± 0.2 mm) after treatment than Group 2 incisors (3.5 ± 0.3 mm, p < 0.001), and Group 3 incisors had a significantly shorter mean elongated root length (2.1 ± 0.1 mm) after treatment than Group 4 incisors (3.7 ± 0.3 mm, p < 0.001). CONCLUSION: Necrotic open-apex incisors treated with ultrasonic filing plus MTA placement need the shortest mean duration for apical hard tissue barrier formation. For elongation of apical root length, Ca(OH)2 apexification/apexogenesis is better than MTA apexification/apexogenesis, regardless if either ultrasonic or hand filing are used.

Ginseng extract and ginsenoside Rb1 attenuate carbon tetrachloride-induced liver fibrosis in rats.

BACKGROUND: Ginsenosides, the major bioactive compounds in ginseng root, have been found to have antioxidant, immunomodulatory and anti-inflammatory activities. This study investigated the effects of ginsenosides on carbon tetrachloride (CCl4)-induced hepatitis and liver fibrosis in rats. METHODS: Male Sprague-Dawley rats were randomly divided into four groups: control, CCl4, CCl4 + 0.5 g/kg Panax ginseng extract and CCl4 + 0.05 g/kg ginsenoside Rb1 groups. The treated groups were orally given Panax ginseng extract or ginsenoside Rb1 two weeks before the induction of liver injury for successive 9 weeks. Liver injury was induced by intraperitoneally injected with 400 ml/l CCl4 at a dose of 0.75 ml/kg body weight weekly for 7 weeks. The control group was intraperitoneally injected with olive oil. RESULTS: The pathological results showed that ginsenoside Rb1 decreased hepatic fat deposition (2.65 ± 0.82 vs 3.50 ± 0.75, p <0.05) and Panax ginseng extract lowered hepatic reticular fiber accumulation (1.05 ± 0.44 vs 1.60 ± 0.39, p <0.01) increased by CCl4. Plasma alanine aminotransferase and aspartate aminotransferase activities were increased by CCl4 (p <0.01), and aspartate aminotransferase activity was decreased by Panax ginseng extract at week 9 (p <0.05). Exposure to CCl4 for 7 weeks, the levels of plasma and hepatic triglycerides (p <0.01), hepatic cholesterol (p <0.01), interleukin-1ß (p <0.01), prostaglandin E2 (p <0.05), soluble intercellular adhesion molecule-1 (p <0.05), hydroxyproline (p <0.05), matrix metalloproteinase-2 (p <0.05) and tissue inhibitor of metalloproteinase-1 (TIMP-1) (p <0.01) were elevated, however, hepatic interleukin-10 level was lowered (p <0.05). Both Panax ginseng extract and ginsenoside Rb1 decreased plasma and hepatic triglyceride, hepatic prostaglandin E2, hydroxyproline and TIMP-1 levels, and Panax ginseng extract further inhibited interleukin-1ß concentrations (p <0.05). CONCLUSIONS: Panax ginseng extract and ginsenoside Rb1 attenuate plasma aminotransferase activities and liver inflammation to inhibit CCl4-induced liver fibrosis through down-regulation of hepatic prostaglandin E2 and TIMP-1.

A Rare Gastric Subepithelial Lesion Removed through Submucosal Tunneling Endoscopic Resection: Case Report and Literature Review.

Gastric subepithelial lesions are common. However, their diagnosis and management can pose a challenge. Herein, we present the case of a 49-year-old man who was incidentally discovered to have a gastric subepithelial lesion that increased in size during follow-up. Submucosal tunneling endoscopic resection was performed, and the tumor was successfully removed en bloc. The pathological and immunohistochemical findings were consistent with a gastric globus tumor. Although rare, glomus tumors should be considered when gastric subepithelial lesions are discovered. Resection with an endoscopic technique can be used to preserve the stomach and can be considered an alternative to surgical removal. However, such procedures should only be performed by experienced therapeutic endoscopists.

The effect of titanium with electrochemical anodization on the response of the adherent osteoblast-like cell.

PURPOSE: The osseointegration of titanium dental implants is essential for successful therapy, and this is strongly affected by the surface chemistry and roughness. In this study, using electrochemical anodization after sand blasting, and acid etching of Ti surface (SLA), SLA specimens with the high wettability of the thick TiO2 layer (SLAffinity) surface was developed to superimpose the nanoscale topographies on the microscale roughness of SLA surface without greatly altering the surface features. MATERIALS: The surface characteristics of SLAffinity specimens were investigated using scanning electron microscopy, atomic force microscopy, and x-ray photoelectron spectroscopy. The viability and proliferation of osteoblast-like cells (MG63 cells), which were seeded on SLAffinity specimens, were analyzed. RESULTS: Such electrochemically anodized specimens were predominantly composed of bioactive TiO2. The cell culturing tests revealed that the microscale roughness in combination with the nanoscale structures and bioactive properties improved osteoblast viability and differentiation on the SLAffinity surface. CONCLUSION: The favorable biological response of SLAffinity surfaces to MG63 osteoblast-like cells suggested that electrochemical anodization after SLA treatments is a potential procedure for better osseointegration in vivo.

Cellularity and its relation with capsular characteristics as an influencing factor for operative strategies of pleomorphic adenoma.

INTRODUCTION: This study investigates the relationship between cellularity and capsular characteristics of pleomorphic adenoma and its influence on operative strategies. MATERIAL AND METHODS: The capsular characteristics and clinical data of patients with pleomorphic adenomas were reviewed according to Seifert's definition: (1) classic type with balanced amount of cells and stroma, (2) myxoid type with abundant ground substance, interspersed spindle cells, and (3) cellular type with predominance of ductal trabecular structures and little stroma. The immunoreactivity of cellular proliferation (Ki-67) was semi-quantitatively measured using immunohistochemistry. Variables were analyzed using Fisher's test and one-way ANOVA, with (p < 0.05) considered statistically significant. RESULTS: The duration of presence was associated with cellularity (p = 0.01). In terms of capsular characteristics, satellite nodules and positive resection margins were not related to cellularity, except for incomplete capsules (p = 0.03). There was no difference in the staining scores of Ki-67 (p = 0.12). CONCLUSION: Lower cellularity reflects higher probability of an incomplete capsule, requiring more consideration for operative strategies to prevent recurrence.

INTRODUCCIÓN: Este estudio investiga la relación entre la celularidad y las características capsulares del adenoma pleomórfico y su influencia en las estrategias operativas. MATERIAL Y MÉTODOS: Se revisaron las características capsulares y los datos clínicos de los pacientes con adenomas pleomórficos según la definición de Seifert: 1) tipo clásico con cantidad equilibrada de células y estroma, 2) tipo mixoide con abundante sustancia fundamental, células fusiformes intercaladas y 3) tipo celular con predominio de estructuras trabeculares ductales y poco estroma. La inmunorreactividad de la proliferación celular (Ki-67) se midió semicuantitativamente usando inmunohistoquímica. Las variables se analizaron mediante la prueba de Fisher y ANOVA de una vía, considerándose significativo un valor de p inferior a 0.05. RESULTADOS: La duración de la presencia se asoció con la celularidad (p = 0.01). En cuanto a las características capsulares, los nódulos satélites y los márgenes de resección positivos no se relacionaron con la celularidad, a excepción de las cápsulas incompletas (p = 0.03). No hubo diferencia en las puntuaciones de tinción de Ki-67 (p = 0.12). CONCLUSIONES: La celularidad más baja refleja una mayor probabilidad de una cápsula incompleta, lo que requiere una mayor consideración de las estrategias quirúrgicas para prevenir la recurrencia.

2,3,5,4'-Tetrahydroxystilbene-2-O-ß-glucoside Attenuates Reactive Oxygen Species-Dependent Inflammation and Apoptosis in Porphyromonas gingivalis-Infected Brain Endothelial Cells.

We recently reported that the periodontopathic bacteria Porphyromonas gingivalis (P. gingivalis) initiates an inflammatory cascade that disrupts the balance of reactive oxygen species (ROS), resulting in apoptotic cell death in brain endothelial cells. An extract from Polygonum multiflorum Thunb., 2,3,5,4'-Tetrahydroxystilbene-2-O-ß-glucoside (THSG) has been well-reported to diminish the inflammation in many disease models. However, the effects of THSG in the area of the brain-oral axis is unknown. In this study, we examined the effects of THSG in P. gingivalis-stimulated inflammatory response and apoptotic cell death in brain endothelial cells. THSG treatment remarkably lessened the upregulation of IL-1ß and TNF-α proteins in bEnd.3 cells infected with P. gingivalis. Treatment of THSG further ameliorated brain endothelial cell death, including apoptosis caused by P. gingivalis. Moreover, the present study showed that the inhibitory effects on NF-κB p65 and antiapoptotic properties of THSG is through inhibiting the ROS pathway. Importantly, the ROS inhibitory potency of THSG is similar to a ROS scavenger N-Acetyl-L-Cysteine (NAC) and NADPH oxidase inhibitor apocynin. Furthermore, the protective effect of THSG from P. gingivalis infection was further confirmed in primary mouse brain endothelial cells. Taken together, this study indicates that THSG attenuates an ROS-dependent inflammatory response and cell apoptosis in P. gingivalis-infected brain endothelial cells. Our results also suggest that THSG could be a potential herbal medicine to prevent the risk of developing cerebrovascular diseases from infection of periodontal bacteria.

Porphyromonas gingivalis Induces Proinflammatory Cytokine Expression Leading to Apoptotic Death through the Oxidative Stress/NF-κB Pathway in Brain Endothelial Cells.

Porphyromonas gingivalis, a periodontal pathogen, has been proposed to cause blood vessel injury leading to cerebrovascular diseases such as stroke. Brain endothelial cells compose the blood-brain barrier that protects homeostasis of the central nervous system. However, whether P. gingivalis causes the death of endothelial cells and the underlying mechanisms remain unclear. This study aimed to investigate the impact and regulatory mechanisms of P. gingivalis infection in brain endothelial cells. We used bEnd.3 cells and primary mouse endothelial cells to assess the effects of P. gingivalis on endothelial cells. Our results showed that infection with live P. gingivalis, unlike heat-killed P. gingivalis, triggers brain endothelial cell death by inducing cell apoptosis. Moreover, P. gingivalis infection increased intracellular reactive oxygen species (ROS) production, activated NF-κB, and up-regulated the expression of IL-1ß and TNF-α. Furthermore, N-acetyl-L-cysteine (NAC), a most frequently used antioxidant, treatment significantly reduced P. gingivalis-induced cell apoptosis and brain endothelial cell death. The enhancement of ROS production, NF-κB p65 activation, and proinflammatory cytokine expression was also attenuated by NAC treatment. The impact of P. gingivalis on brain endothelial cells was also confirmed using adult primary mouse brain endothelial cells (MBECs). In summary, our results showed that P. gingivalis up-regulates IL-1ß and TNF-α protein expression, which consequently causes cell death of brain endothelial cells through the ROS/NF-κB pathway. Our results, together with the results of previous case-control studies and epidemiologic reports, strongly support the hypothesis that periodontal infection increases the risk of developing cerebrovascular disease.

Chicken model of steroid-induced bone marrow adipogenesis using proteome analysis: a preliminary study.

BACKGROUND: Steroid-induced adipogenesis increases fat-cell volume and pressure in bone marrow. This may be a contributing factor in some forms of osteonecrosis. In this observational study, we aimed to determine the protein expression relating to steroid-induced adipogenesis of femoral bone marrow with use of a chicken model. We compared the histologic features of the femoral marrow of eight methylprednisolone (MP)-treated chickens with those of three control chickens and assessed differential proteins with 2-dimensional gel electrophoresis and differential proteins were identified by MALDI-TOF MS. RESULTS: One MP-induced chicken died of overdose anesthesia. Methylprednisolone-induced proliferation of adipose tissue and new bone formation were found on histologic examination. In our study, 13 proteins in the control and MP-induced groups were differently expressed and nine protein spots showed marked threefold downregulation after 19 weeks of MP treatment. These were serum amyloid P-component precursor, zinc finger protein 28, endothelial zinc finger protein 71, T-box transcription factor 3, cyclin-dependent kinase inhibitor 1, myosin 1D, dimethylaniline monooxygenase, and two uncharacterized proteins. CONCLUSIONS: Proteomic profiling can be a useful dynamic approach for detecting protein expression in MP-induced adipogenesis of the femur in chickens.

High-Performance Sample Substrate of Gold Nanoparticle Multilayers for Surface-Assisted Laser Desorption/Ionization Mass Spectrometry.

The development of a sample substrate with superior performance for desorption and ionization of analyte is the key issue to ameliorate the quality of mass spectra for measurements of small molecules in surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). Herein, the homogeneous sample substrate of gold nanoparticle multilayers (AuNPs-ML) with hexagonal lattice was successfully prepared by self-assembly technique. With strong surface plasmon resonance absorption and superior photothermal effect, the sample substrate of AuNPs-ML exhibited high signal sensitivity and low background noise for the detection of model analyte of glucose without additional matrixes in SALDI-MS. Furthermore, compared to merchant matrixes of α-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB), the sample substrate of AuNPs-ML was demonstrated to ameliorate the quality of mass spectra, including signal strength, background interference and signal/noise (S/N) ratio. The sucrose and tryptophan were also measured to show the extensive applications of AuNPs-ML sample substrate for the detections of small molecules in SALDI-MS. Most importantly, the remarkable reproducibility of glucose mass spectra with relative signal of 7.3% was obtained by the use of AuNPs-ML sample substrate for SALDI-MS. The homogeneous sample substrate of AuNPs-ML greatly improved the quality of mass spectra because of its strong absorption of laser energy, low specific heat, high heat conductivity and extraordinary homogeneity. We believe that AuNPs-ML could be a practical sample substrate for small molecule detection in SALDI-MS.

Enhanced expression of transforming growth factor-beta 1 in inflammatory cells, alpha-smooth muscle actin in stellate cells, and collagen accumulation in experimental granulomatous hepatitis caused by Toxocara canis in mice.

Although toxocaral granulomatous hepatitis (TGH) characterized with a dominant-Th2 type immune response is a self-limiting disease, little is known concerning the role of fibrosis-related cytokine transforming growth factor-beta 1 (TGF-beta 1) in pathogenesis of TGH. A detailed histological and quantitatively immunohistochemical analysis of TGF-beta 1, alpha-smooth muscle actins (alpha-SMA), and collagen was performed on the liver tissues from mice infected with Toxocara canis as assessed between day 1 and 42 weeks post-infection (DPI or WPI). TGF-beta1 was detected mainly in infiltrating leukocytes in lesions with strong expressions from 4 to 16 WPI. Larvae per se also exhibited strong TGF-beta 1-like molecule expressions in the trial. Alpha-SMA was detected predominantly in hepatic stellate cells (HSC) which surrounded the lesions with moderate expressions largely throughout the period of the entire experiment. Collagen was observed to accumulate in inflammatory lesions and biliary basement with moderate to strong expressions from 1 WPI onwards in the trial. Since many evidences have indicated that leukocytes have the potential to influence HSC by producing TGF-beta 1 which can affect HSC to increase collagen synthesis in various liver diseases, we may propose that persistently elevated TGF-beta 1 expression in infiltrating leukocytes and active HSC with marked alpha-SMA expressions may contribute to healing of injured sites through up-stimulation of collagen deposition; in contrast, abnormally persistent collagen accumulation may cause irreversible fibrotic injury in the TGH.

Brain injury-associated biomarkers of TGF-beta1, S100B, GFAP, NF-L, tTG, AbetaPP, and tau were concomitantly enhanced and the UPS was impaired during acute brain injury caused by Toxocara canis in mice.

BACKGROUND: Because the outcomes and sequelae after different types of brain injury (BI) are variable and difficult to predict, investigations on whether enhanced expressions of BI-associated biomarkers (BIABs), including transforming growth factor beta1 (TGF-beta1), S100B, glial fibrillary acidic protein (GFAP), neurofilament light chain (NF-L), tissue transglutaminases (tTGs), beta-amyloid precursor proteins (AbetaPP), and tau are present as well as whether impairment of the ubiquitin-proteasome system (UPS) is present have been widely used to help delineate pathophysiological mechanisms in various BIs. Larvae of Toxocara canis can invade the brain and cause BI in humans and mice, leading to cerebral toxocariasis (CT). Because the parasitic burden is light in CT, it may be too cryptic to be detected in humans, making it difficult to clearly understand the pathogenesis of subtle BI in CT. Since the pathogenesis of murine toxocariasis is very similar to that in humans, it appears appropriate to use a murine model to investigate the pathogenesis of CT. METHODS: BIAB expressions and UPS function in the brains of mice inoculated with a single dose of 250 T. canis embryonated eggs was investigated from 3 days (dpi) to 8 weeks post-infection (wpi) by Western blotting and RT-PCR. RESULTS: Results revealed that at 4 and 8 wpi, T. canis larvae were found to have invaded areas around the choroid plexus but without eliciting leukocyte infiltration in brains of infected mice; nevertheless, astrogliosis, an indicator of BI, with 78.9~142.0-fold increases in GFAP expression was present. Meanwhile, markedly increased levels of other BIAB proteins including TGF-beta1, S100B, NF-L, tTG, AbetaPP, and tau, with increases ranging 2.0~12.0-fold were found, although their corresponding mRNA expressions were not found to be present at 8 wpi. Concomitantly, UPS impairment was evidenced by the overexpression of conjugated ubiquitin and ubiquitin in the brain. CONCLUSION: Further studies are needed to determine whether there is an increased risk of CT progression into neurodegenerative disease because neurodegeneration-associated AbetaPP and phosphorylated tau emerged in the brain.

Low dietary n-6/n-3 polyunsaturated fatty acid ratio prevents induced oral carcinoma in a hamster pouch model.

Dietary fatty acid patterns have been linked to the prevalence of certain cancers, however in oral carcinoma is limited. Thus, we investigated the chemopreventive effects of various dietary n-6 and n-3 fatty acids in a 9,10-dimethyl-1,2-benz[a]-anthracene (DMBA)- and betel quid extract (BQE) -induced hamster oral cancer model. Thirty 6-week-old adult male hamsters were housed and divided into normal, low, and high dietary n-6 and n-3 fatty acid groups under DMBA + BQE treatment for 16 weeks. The right buccal pouch of all hamsters were evaluated by tumor number, volume, burden and selected inflammatory parameters. The results indicate that the low dietary n-6/n-3 fatty acid group exhibited a significantly lower tumor number, volume, and burden than those of the other groups. Furthermore, this group had significantly lower nuclear factor-κB, proliferating cell nuclear antigen, and cyclin D1 expression in the right buccal pouch tissue. In conclusion, the lower dietary n-6/n-3 fatty acid ratio exerted chemopreventive effects in the DMBA- and BQE-induced hamster oral cancer model.