Evaluation of Surgical Treatment of Oroantral Fistulae in Smokers Versus Non-Smokers.

Background and Objectives: Smoking has been found to interfere with wound healing processes. Therefore, the purpose of this study was to compare surgical treatment of oroantral fistulae (OAFs) in smokers and non-smokers. Materials and Methods: Medical records of all consecutive patients who underwent surgical closure of OAFs between 2003 and 2016 at the oral and maxillofacial surgery department, Rabin Medical Center, Israel were reviewed. Patients' demographic data, preoperative signs and symptoms, surgical method of repair, and postoperative complications were recorded. Results: The cohort consisted of 38 smokers and 59 non-smokers. Age and gender distributions were similar in both groups. The main etiology in both groups was tooth extraction, followed by pre-prosthetic surgery in smokers and odontogenic infection in non-smokers (p = 0.02). Preoperative conditions were not significantly different between smokers and non-smokers in terms of size of soft tissue fistula and bony defect, chronic sinusitis and foreign bodies inside the sinus. OAFs were repaired by local soft tissue flaps without consideration of smoking status. Smokers experienced more moderate-severe postoperative pain (p = 0.05) and requested more weak opioids (p = 0.06). Postoperative complications included infection, delayed wound healing, residual OAF, pain, sensory disturbances and sino nasal symptoms. These were mostly minor and tended to be more frequent in smokers (p = 0.35). Successful closure of OAFs was obtained in all patients except one smoker who required revision surgery. Conclusions: Smokers may be more susceptible to OAFs secondary to preprosthetic surgery. In this cohort, there was no statistically significant difference in outcome between smokers and non-smokers in terms of failure. However, smokers tended to have more severe postoperative pain and discomfort and to experience more postoperative complications. Further studies with larger sample sizes should be conducted to validate these results.

Identification of OAF and PVRL1 as candidate genes for an ocular anomaly characterized by Peters anomaly type 2 and ectopia lentis.

Keratolenticular dysgenesis (KLD) and ectopia lentis are congenital eye defects. The aim of this study is the identification of molecular genetic alterations responsible for those ocular anomalies with neurologic impairment in an individual with a de novo balanced chromosome translocation t(11;18)(q23.3;q11.2)dn. Disruption of OAF, the human orthologue of the Drosophila oaf, by the 11q23.3 breakpoint results in reduced expression of this transcriptional regulator. Furthermore, four most likely nonfunctional chimeric transcripts comprising up to OAF exon 3, derived from the der(11) allele, have also been identified. This locus has been implicated by publicly available genome-wide association data in corneal disease and corneal topography. The expression of the poliovirus receptor-related 1(PVRL1) or nectin cell adhesion molecule 1 (NECTIN1), a paralogue of nectin cell adhesion molecule 3 (PVRL3) associated with congenital ocular defects, situated 500 kb upstream from 11q23.3 breakpoint, is increased. The 18q11.2 breakpoint is localized between cutaneous T-cell lymphoma-associated antigen 1(CTAGE1) and retinoblastoma binding protein 8 (RBBP8) genes. Genomic imbalance that could contribute to the observed phenotype was excluded. Analysis of gene expression datasets throughout normal murine ocular lens embryogenesis suggests that OAF expression is significantly enriched in the lens from early stages of development through adulthood, whereas PVRL1 is lens-enriched until E12.5 and then down-regulated. This contrasts with the observation that the proposita's lymphoblastoid cell lines exhibit low OAF and high PVRL1 expression as compared to control, which offers further support that the alterations described above are most likely responsible for the clinical phenotype. Finally, gene interaction topology data for PVRL1 also agree with our proposal that disruption of OAF by the translocation breakpoint and misregulation of PVRL1 due to a position effect contribute to the observed ocular and neurological phenotype.

Yeast 14-3-3 protein functions as a comodulator of transcription by inhibiting coactivator functions.

In eukaryotes combinatorial activation of transcription is an important component of gene regulation. In the budding yeast Saccharomyces cerevisiae, Adr1-Cat8 and Adr1-Oaf1/Pip2 are pairs of activators that act together to regulate two diverse sets of genes. Transcription activation of both sets is regulated positively by the yeast AMP-activated protein kinase homolog, Snf1, in response to low glucose or the presence of a non-fermentable carbon source and negatively by two redundant 14-3-3 isoforms, Bmh1 and Bmh2. Bmh regulates the function of these pairs at a post-promoter binding step by direct binding to Adr1. However, how Bmh regulates transcription after activator binding remains unknown. In the present study we analyzed Bmh-mediated regulation of two sets of genes activated independently by these pairs of activators. We report that Bmh inhibits mRNA synthesis when the second activator is absent. Using gene fusions we show that Bmh binding to the Adr1 regulatory domain inhibits an Adr1 activation domain but not a heterologous activation domain or artificially recruited Mediator, consistent with Bmh acting at a step in transcription downstream of activator binding. Bmh inhibits the assembly and the function of a preinitiation complex (PIC). Gene expression studies suggest that Bmh regulates Adr1 activity through the coactivators Mediator and Swi/Snf. Mediator recruitment appeared to occur normally, but PIC formation and function were defective, suggesting that Bmh inhibits a step between Mediator recruitment and PIC activation.

Development of Recombinant PLC-Zeta Protein as a Therapeutic Intervention for the Clinical Treatment of Oocyte Activation Failure.

The sperm-specific phospholipase C zeta (PLCζ) protein is widely considered as the predominant physiological stimulus for initiating the Ca2+ release responsible for oocyte activation during mammalian fertilization. The increasing number of genetic and clinical reports that directly link PLCζ defects and/or deficiencies with oocyte activation failure (OAF) necessitates the use of a powerful therapeutic intervention to overcome such cases of male factor infertility. Currently, in vitro fertilization (IVF) clinics treat OAF cases after intracytoplasmic sperm injection (ICSI) with Ca2+ ionophores. Despite their successful use, such chemical agents are unable to trigger the physiological pattern of Ca2+ oscillations. Moreover, the safety of these ionophores is not yet fully established. We have previously demonstrated that recombinant PLCζ protein can be successfully used to rescue failed oocyte activation, resulting in efficient blastocyst formation. Herein, we produced a maltose binding protein (MBP)-tagged recombinant human PLCζ protein capable of inducing Ca2+ oscillations in mouse oocytes similar to those observed at fertilization. Circular dichroism (CD) experiments revealed a stable, well-folded protein with a high helical content. Moreover, the recombinant protein could retain its enzymatic properties for at least up to 90 days after storage at -80 °C. Finally, a chick embryo model was employed and revealed that exposure of fertilized chicken eggs to MBP-PLCζ did not alter the embryonic viability when compared to the control, giving a first indication of its safety. Our data support the potential use of the MBP-PLCζ recombinant protein as an effective therapeutic tool but further studies are required prior to its use in a clinical setting.

Mineralization and collagen orientation throughout aging at the vertebral endplate in the human lumbar spine.

The human vertebral body and intervertebral disc interface forms the region where the cartilaginous endplate, annulus fibrosis and bone of the vertebral body are connected through an intermediate calcified cartilage layer. While properties of both the vertebral body and components of the disc have been extensively studied, limited quantitative data exists describing the microstructure of the vertebral body-intervertebral disc interface in the spine throughout development and degeneration. Quantitative backscattered scanning electron and second harmonic generation confocal imaging were used to collect quantitative data describing the mineral content and collagen fiber orientation across the interface, respectively. Specimens spanned ages 56 days to 84 years and measurements were taken across the vertebral endplate at the outer annulus, inner annulus and nucleus pulposis. In mature and healthy endplates, collagen fibers span the calcified cartilage layer in all regions, including the endplate adjacent to the central nucleus pulposis. We also observed an abrupt transition from high mineral volume fractions (35-50%) to 0% over short distances measuring 3-15 microns in width across the transition from calcified cartilage to unmineralized cartilage. The alignment of collagen fibers at the outer annulus and thickness of the CC layer indicated that collagen fiber mineralization adjacent to the bone may serve to anchor the soft tissue without a gradual change in material properties. Combining backscattered scanning electron microscopy and second harmonic generation imaging on the same sections thus enable a novel assessment of morphology and properties in both mineralized and soft tissues at the vertebral body-intervertebral disc throughout development and aging.

Decellularized matrix for repairing intervertebral disc degeneration: Fabrication methods, applications and animal models.

Intervertebral disc degeneration (IDD)-induced low back pain significantly influences the quality of life, placing a burden on public health systems worldwide. Currently available therapeutic strategies, such as conservative or operative treatment, cannot effectively restore intervertebral disc (IVD) function. Decellularized matrix (DCM) is a tissue-engineered biomaterial fabricated using physical, chemical, and enzymatic technologies to eliminate cells and antigens. By contrast, the extracellular matrix (ECM), including collagen and glycosaminoglycans, which are well retained, have been extensively studied in IVD regeneration. DCM inherits the native architecture and specific-differentiation induction ability of IVD and has demonstrated effectiveness in IVD regeneration in vitro and in vivo. Moreover, significant improvements have been achieved in the preparation process, mechanistic insights, and application of DCM for IDD repair. Herein, we comprehensively summarize and provide an overview of the roles and applications of DCM for IDD repair based on the existing evidence to shed a novel light on the clinical treatment of IDD.

PRIMUS: Comprehensive proteomics of mouse intervertebral discs that inform novel biology and relevance to human disease modelling.

Mice are commonly used to study intervertebral disc (IVD) biology and related diseases such as IVD degeneration. Discs from both the lumbar and tail regions are used. However, little is known about compartmental characteristics in the different regions, nor their relevance to the human setting, where a functional IVD unit depends on a homeostatic proteome. Here, we address these major gaps through comprehensive proteomic profiling and in-depth analyses of 8-week-old healthy murine discs, followed by comparisons with human. Leveraging on a dataset of over 2,700 proteins from 31 proteomic profiles, we identified key molecular and cellular differences between disc compartments and spine levels, but not gender. The nucleus pulposus (NP) and annulus fibrosus (AF) compartments differ the most, both in matrisome and cellularity contents. Differences in the matrisome are consistent with the fibrous nature required for tensile strength in the AF and hydration property in the NP. Novel findings for the NP cells included an enrichment in cell junction proteins for cell-cell communication (Cdh2, Dsp and Gja1) and osmoregulation (Slc12a2 and Wnk1). In NP cells, we detected heterogeneity of vacuolar organelles; where about half have potential lysosomal function (Vamp3, Copb2, Lamp1/2, Lamtor1), some contain lipid droplets and others with undefined contents. The AF is enriched in proteins for the oxidative stress responses (Sod3 and Clu). Interestingly, mitochondrial proteins are elevated in the lumbar than tail IVDs that may reflect differences in metabolic requirement. Relative to the human, cellular and structural information are conserved for the AF. Even though the NP is more divergent between mouse and human, there are similarities at the level of cell biology. Further, common cross-species markers were identified for both NP (KRT8/19, CD109) and AF (COL12A1). Overall, mouse is a relevant model to study IVD biology, and an understanding of the limitation will facilitate research planning and data interpretation, maximizing the translation of research findings to human IVDs.

A pedicled palatal periosteal flap for the closure of oro-antral fistula.

Various surgical techniques have been developed for oro-antral fistula (OAF) closure, all of which have some drawback. Twenty consecutive patients with an OAF were enrolled in this prospective study. A trapezoid full-thickness flap extending from the palatal area to the buccal gingiva was raised, including the fistula at its centre. The palatal free end aspect was split into two layers and the deep periosteal layer was folded deep to the flap over the bony defect, thereby sealing the fistula. The superficial layer was returned to its primary position and sutured. The patients were followed for 3 months. Nineteen patients showed immediate OAF closure. One patient showed a residual oro-antral communication of 0.5mm in diameter that resolved spontaneously within 4 weeks. The pain level (on a visual analogue scale) was highest at the first follow-up week, with a mean score of 5.5, which decreased to a mean level of 2.5 in the second week and 0 in the fourth week. The mean satisfaction level was 9.85 on a scale of 0-10 (10 representing total satisfaction). The pedicled palatal periosteal flap is a simple and effective surgical technique with high predictability and patient satisfaction levels, offering one more alternative for the treatment of OAF.

Evaluation of a New Surgical Technique for Closing Oroantral Fistula Using Auto-transplanted Upper Third Molar: A 1-Year Follow-Up Study.

BACKGROUND: Oroantral fistula (OAF) is considered a frequent complication in dental practice. Many surgical techniques/methods have been proposed to close it. The aim of this study was to evaluate the auto-transplantation of upper third molar for closing OAF. MATERIALS AND METHODS: Twenty patients participated in this study aged between 20 and 40 years old. The OAF was closed by auto-transplantation of upper third molar placed directly in the socket of the extracted tooth. Results were evaluated clinically and radiographically through the period of observation which lasted for 1 year. RESULTS: Final results showed that the success rate of closing OAF was 95% while the success rate of upper third molar auto-transplantation was 90%. CONCLUSION: This technique is simple, applicable, provides immediate replacement of the missing tooth, and does not require complicated instruments or procedures.

Evaluation of different treatment modalities for closure of oro-antral communications and formulation of a rational approach.

Oro-antral communication is a common occurrence following removal of maxillary premolars and molars because of anatomic proximity of root apices of these teeth and maxillary antrum. Various methods have been described in literature for closure of these communications which vary from simple local methods like buccal advancement flap to complex distal flaps and grafts. Out of these plethora of the treatment modalities available for the treatment of oro antral fistula, the most simple and commonly used ones are either the buccal flap or the buccal pad of fat. In our study we compared the results, advantages and disadvantages of using buccal advancement technique and buccal fat pad individually and also in combination. With this paper, we aim to shed light on the efficacy of buccal pad of fat and the buccal flap, either alone or together, for the closure of OAF of various regions. We also aim to provide a systematic and rational approach for repair of oro-antral communications.