A comparison of patient-reported outcomes in patients undergoing abdominal wall repair with either synthetic or biosynthetic mesh: a pilot study.

PURPOSE: Repair of midline ventral incisional hernias (VIHR) requires mesh reinforcement. Mesh types can be categorised into synthetic, biosynthetic, or biological. There is a lack of evidence to support one type of mesh over another. The aim of this pilot study was to compare mesh sensation in patients having undergone elective open repair with synthetic or biosynthetic mesh. METHODS: Four years of prospectively collected data were retrospectively reviewed on 40 patients who had undergone VIHR, using either biosynthetic or synthetic mesh placed in the retromuscular plane. The decision on type of mesh used was governed by patient characteristics. Patients were invited to complete the Carolinas Comfort Scale (CCS) questionnaire, the higher the score indicating a poorer quality of life. The maximum length of follow-up was 36 months. RESULTS: Twenty patients received permanent synthetic and 20 biosynthetic mesh. There was no clinical evidence of hernia recurrence in either group in the short to medium term. Overall, 97% (39/40) patients reported an average of either no or mild symptoms (mean CCS score 17.9 of 115). Patients with a biosynthetic repair had a significant lower CCS at ≥ 18 months (p < 0.05). CONCLUSION: After VIHR, patients have low CCS scores, indicating good quality of life outcomes, in the short to medium term irrespective of the mesh used. However, biosynthetic mesh had lower CCS scores in the medium term. This may help surgeons and patients make better informed decisions about which mesh to use in their individual circumstances.

Delphi consensus statement for understanding and managing the subcostal hernia: subcostal hernias collaborative report (scholar study).

INTRODUCTION: Subcostal hernias are categorized as L1 based on the European Hernia Society (EHS) classification and frequently involve M1, M2, and L2 sites. These are common after hepatopancreatic and biliary surgeries. The literature on subcostal hernias mostly comprises of retrospective reviews of small heterogenous cohorts, unsurprisingly leading to no consensus or guidelines. Given the limited literature and lack of consensus or guidelines for dealing with these hernias, we planned for a Delphi consensus to aid in decision making to repair subcostal hernias. METHODS: We adopted a modified Delphi technique to establish consensus regarding the definition, characteristics, and surgical aspects of managing subcostal hernias (SCH). It was a four-phase Delphi study reflecting the widely accepted model, consisting of: 1. Creating a query. 2. Building an expert panel. 3. Executing the Delphi rounds. 4. Analysing, presenting, and reporting the Delphi results. More than 70% of agreement was defined as a consensus statement. RESULTS: The 22 experts who agreed to participate in this Delphi process for Subcostal Hernias (SCH) comprised 7 UK surgeons, 6 mainland European surgeons, 4 Indians, 3 from the USA, and 2 from Southeast Asia. This Delphi study on subcostal hernias achieved consensus on the following areas-use of mesh in elective cases; the retromuscular position with strong discouragement for onlay mesh; use of macroporous medium-weight polypropylene mesh; use of the subcostal incision over midline incision if there is no previous midline incision; TAR over ACST; defect closure where MAS is used; transverse suturing over vertical suturing for closure of circular defects; and use of peritoneal flap when necessary. CONCLUSION: This Delphi consensus defines subcostal hernias and gives insight into the consensus for incision, dissection plane, mesh placement, mesh type, and mesh fixation for these hernias.

Assessment of a novel alternative to conventional formocresol-zinc oxide eugenol pulpotomy for the treatment of pulpally involved human primary teeth: diode laser-mineral trioxide aggregate pulpotomy.

OBJECTIVE: The purpose of this study was to investigate whether a diode laser pulpotomy with mineral trioxide aggregate (MTA) sealing could be an acceptable alternative to the conventional formocresol pulpotomy and zinc oxide eugenol (ZOE) sealing in human primary teeth. METHODS: A randomized, single-blind, split-mouth study was used with a sample of 16 children aged from 3 to 8 years (mean age=5.10 years). A total of 26 pairs of teeth from these 16 patients were selected based on clinical and radiographic criteria. One tooth from each pair was randomly assigned to either the laser-MTA pulpotomy group or the formocresol-ZOE pulpotomy group. All teeth were followed up clinically and radiographically at 2.3, 5.2, 9.5 and 15.7 months. All extracted failures were sectioned and photographed to assess possible reasons for this. RESULTS: A total of seven laser-MTA-treated teeth were deemed to be radiographic failures (mean time until failure=9.1 months) compared to three formocresol-ZOE treated teeth (mean time until failure=12.5 months). These results were not significant using Fisher's exact test (P>0.05). Six of the laser-MTA failures and all three formocresol-ZOE failures exhibited furcal and/or periapical radiolucencies with or without pathologic root resorption. One of the laser-MTA failures displayed premature root resorption and is being observed for exfoliation. Analysis of photographs of teeth available for extraction revealed errors in clinical technique in addition to expected signs of a disease process such as the presence of granulation tissue and areas of pathologic root resorption. CONCLUSIONS: The laser-MTA pulpotomy showed reduced radiographic success rates compared to the formocresol-ZOE pulpotomy at 15.7 months; however, these results were not statistically significant. Improved success rates among a larger patient sample and a longer follow-up period would be required for the laser-MTA pulpotomy to be considered a routine alternative to the conventional formocresol-ZOE procedure. Meticulous restorative techniques must be followed to ensure the success of laser-MTA pulpotomies.

Promotion of selective cell attachment by the RGD sequence in dentine matrix protein 1.

Dentine matrix protein 1 (DMP1) is an important component of the non-collagenous extracellular matrix of developing teeth and bones. Functions of DMP1 other than a putative role in the initiation of mineralization are largely unknown. A first report on the DNA and deduced amino acid sequence showed that DMP1 has a single Arg-Gly-Asp (RGD) sequence. Here, whether the RGD sequence functions as a cell-attachment domain was tested. Using site-directed mutagenesis, two mutant recombinant DMP1 proteins with specific alterations at the RGD site were created. In the first mutant protein the RGD sequence was altered to a RGE (RGE) sequence; in the second the RGD domain was deleted (DEL). Mutated proteins were confirmed to be DMP1 by partial protein sequencing and dot-blot analysis with an anti-DMP1 antibody. Attachment of RPC-C2A (dental pulp cells), MC3T3-E1 (calvarial cells) or CHO (Chinese hamster ovary cells) to non-tissue-culture plastic coated with either DMP1, RGE or DEL proteins was compared. Bovine serum albumin and fibronectin served as negative and positive controls, respectively. The RGD-containing native DMP1 protein effectively allowed cell attachment and spreading. The RGE and DEL proteins with the altered and deleted RGD sites were significantly less effective in promoting cell attachment than the recombinant DMP1. Both RPC-C2A pulp cells and MC3T3-E1 cells showed similar reductions in attachment to mutated proteins. Treatment of RPC-C2A cells with a RGD-containing peptide prior to plating on DMP1-coated chambers abolished DMP1-mediated cell attachment. In contrast to RPC-C2A and MC3T3-E1cells, CHO cells, which normally do not express DMP1, failed to attach to DMP1. These data demonstrate that DMP1 promotes cell attachment through the RGD domain and that the attachment is cell- and tissue-specific. A basis for these observations is proposed using computer-generated models of the polypeptides within the DMP1 protein containing the RGD, RGE or DEL sequences.

Role of mitochondrial membrane potential in concanavalin A-induced apoptosis in human fibroblasts.

Lectins induce apoptosis in a wide variety of cell types but the mechanisms of apoptotic induction are unknown. We examined the role of mitochondrial membrane potential (Psi m) in concanavalin A-induced apoptosis in human diploid fibroblasts. Cells were treated with Con A for 0.5, 1, 3, 5, and 24 h. Con A induced a time-dependent increase of the proportion of TUNEL+ ve cells over 24 h. Psi m was examined by staining cells with the mitochondria-specific fluorescent cationic dye JC-1. Comparison of JC-1 fluorescence within mitochondria by flow cytometry showed that after 3 h, Con A reduced Psi m in a subpopulation of apoptotic cells with smaller cell volume and with apoptotic nuclear morphology. In contrast, Psi m was unchanged in a separate population of viable cells with normal volume and normal nuclear morphology. Cyclosporin A protected cells against reduction of Psi m and also against nuclear condensation and morphological apoptosis. Measurement of intracellular calcium ion concentration ([Ca2+]i) by ratio fluorimetry of fura 2-loaded cells showed that Con A did not affect [Ca2+]i in viable cells but induced a progressive depletion of [Ca2+]i with generation of calcium oscillations in apoptotic cells. Assessment of Bcl-2 in Con A-treated cells demonstrated an initially strong increase in Bcl-2 protein and mRNA but the appearance of degraded Bcl-2 protein at 3 and 5 h after treatment, indicating an inadequate protective response to the Con A stimulation. Collectively, these data indicate that lectin-induced apoptosis in fibroblasts is associated with breakdown of Psi m, loss of [Ca2+]i homeostasis, and induced Bcl-2 expression.

Paclitaxel selectively induces mitotic arrest and apoptosis in proliferating bovine synoviocytes.

OBJECTIVE: Rheumatoid arthritis (RA) is characterized by chronic progressive destruction of joints involving several disease processes, such as villous hypertrophy, proliferation of synovial lining cells, and infiltration of inflammatory cells. Synovial cell activation and proliferation is thought to be a key step in the destruction of cartilaginous and bony tissues in RA joints. In view of the invasive properties of synoviocytes in RA, we conducted in vitro studies to determine the mechanism of action of paclitaxel (Taxol) on synoviocytes, which may account for the inhibition of joint destruction found when this agent is administered. METHODS: Cultured synovial cells were treated with various concentrations of paclitaxel and were evaluated by cell viability, fluorescence microscopy, flow cytometry of DAPI-stained cells, and electron microscopy. RESULTS: The data indicated that paclitaxel inhibited synoviocyte proliferation by a G2/M phase block and was toxic to synoviocytes by inducing apoptosis. Confluent cells such as chondroyctes and synoviocytes were not affected by paclitaxel. Synchronization of synovioyctes at the G1/S boundary effectively abolished paclitaxel-induced apoptosis. CONCLUSION: The data indicate that induction of apoptosis in synoviocytes might be dependent on transit through the cell cycle, specifically through G2 and mitosis. Further, paclitaxel was selectively toxic to proliferating synoviocytes but spared nonproliferating synoviocytes and chondrocytes. These results demonstrate that paclitaxel can inhibit synovial cell proliferation and pannus formation in RA joints in vivo. We suggest that paclitaxel be considered as a prototypical compound for a new class of potential chondroprotective agents.

Serum deprivation induces apoptotic cell death in a subset of Balb/c 3T3 fibroblasts.

Little is known about the regulation of apoptosis in fibroblasts although several model systems including serum deprivation and treatment with staurosporine or topoisomerase inhibitors have been used to induce apoptosis in vitro. To validate a reproducible in vitro model for the study of apoptosis in fibroblasts, we cultured density-inhibited monolayer cultures of Balb/c 3T3 fibroblasts in Dulbecco's modified essential medium plus 15% fetal calf serum and then withdrew serum. Time-lapse video microscopy demonstrated that within minutes of serum withdrawal, cells lost substrate attachment and floated to the top of the liquid growth medium. There was a time-dependent increase in the number of non-adherent cells. Some of these cells regained attachment and spread momentarily, but they eventually rounded up and lost attachment permanently. In contrast to serum-containing cultures in which similar morphological changes were followed by mitosis, in serum-free cultures repeated attempts at mitosis were followed by permanent attachment loss and presumably cell death. To assess whether all the non-adherent cells were in fact dead, the percentages of cells that continued to proliferate upon return to serum-supplemented conditions was computed. After various periods of serum starvation a decreasing proportion (approx. 75% at 30 minutes; < 2% at 24 hours) of the non-adherent cells could be rescued by addition of serum. Transmission electron microscopy of cells 3 hours after serum withdrawal showed that the majority (approximately 60%) of non-adherent cells exhibited marked intranuclear chromatin condensation but maintained integrity of cell and nuclear membranes and cell organelles, morphological changes consistent with those of apoptotic cell death. Scanning electron microscopy of cultures 3 hours following serum withdrawal showed rounded cells with marked surface blebbing. Fluorescence and confocal microscopy revealed increased intensity of nuclear staining with DAPI while actin filaments became indistinct or collapsed around the nucleus. After cycloheximide treatment to inhibit protein synthesis, there was no reduction of apoptosis. Gel electrophoresis of DNA from both control and 3 hour-serum-deprived cells showed intact DNA with no oligonucleosomal length fragmentation. After serum withdrawal, intracellular calcium was reduced by about 32% over 5 minutes as measured by fura2 ratio fluorimetry in single cells. Serum-starved cells showed a time-dependent shrinkage in mean cell diameter compared to trypsinized, adherent control cells (at 0 hours, mean diameter = 18.0 microns--viable; at 4 hours, mean diameter = 15.5 microns--apoptotic). Flow cytometric analysis showed increased propidium iodide staining and reduced fluorescein diacetate uptake over 3 hours, changes that were contemporaneous with the reduction of cell diameter.(ABSTRACT TRUNCATED AT 400 WORDS)