Regeneration of Intrabony Defects with Nano Hydroxyapatite Graft, Derived from Eggshell along with Periosteum as Barrier Membrane under Magnification—An Interventional Study

Intrabony defects can be treated by various approaches. Use of GTR along with bone grafts is said to enhance the outcome. The periosteum has been claimed to increase the regeneration. The egg-shell-derived nano hydroxyapatite (EnHA) has shown a scope as alloplastic graft. Thus, the following study was undertaken to combine the periosteal pedicle along with EnHA for the treatment of intrabony defects under magnification to achieve optimal bone regeneration. A total of 21 patients, having intrabony defects with ≥6 mm probing depth (PD) and two or three wall defects as detected on CBCT, satisfying inclusion criteria were enrolled. The sites were randomly allocated as Group A, B and C (n = 7). The following parameters, defect density and defect fill in CBCT (at baseline and 6 months), PPD, RAL, Plaque index (PI), Gingival index (GI) and Gingival Bleeding Index (GBI) were recorded at baseline, 1, 3 and 6 months. p < 0.05 is considered as statistically significant. Bone density and bone fill values were found to be much higher in pedicle with EnHA and EnHA alone group and the values showed statistically significant results. The current clinical research showed that periosteal pedicle along with EnHA and EnHA as stand-alone therapy gave superior results compared to OFD alone, which is an innovative and feasible treatment option

Tuning the critical gelation temperature of thermo-responsive diblock copolymer worm gels

Amphiphilic diblock copolymer nano-objects can be readily prepared using reversible addition–fragmentation chain transfer (RAFT) polymerization. For example, poly(glycerol monomethacrylate) (PGMA) chain transfer agents (CTA) can be chain-extended using 2-hydroxypropyl methacrylate (HPMA) via RAFT aqueous dispersion polymerization to form well-defined spheres, worms or vesicles at up to 25% solids. The worm morphology is of particular interest, since multiple inter-worm contacts lead to the formation of soft free-standing gels, which undergo reversible degelation on cooling to sub-ambient temperatures. However, the critical gelation temperature (CGT) for such thermo-responsive gels is ≤20 °C, which is relatively low for certain biomedical applications. In this work, a series of new amphiphilic diblock copolymers are prepared in which the core-forming block comprises a statistical mixture of HPMA and di(ethylene glycol) methyl ether methacrylate (DEGMA), which is a more hydrophilic monomer than HPMA. Statistical copolymerizations proceeded to high conversion and low polydispersities were achieved in all cases (Mw/Mn < 1.20). The resulting PGMA-P(HPMA-stat-DEGMA) diblock copolymers undergo polymerization-induced self-assembly at 10% w/w solids to form free-standing worm gels. SAXS studies indicate that reversible (de)gelation occurs below the CGT as a result of a worm-to-sphere transition, with further cooling to 5 °C affording weakly interacting copolymer chains with a mean aggregation number of approximately four. This corresponds to almost molecular dissolution of the copolymer spheres. The CGT can be readily tuned by varying the mean degree of polymerization and the DEGMA content of the core-forming statistical block. For example, a CGT of 31 °C was obtained for PGMA59-P(HPMA91-stat-DEGMA39). This is sufficiently close to physiological temperature (37 °C) to suggest that these new copolymer gels may offer biomedical applications as readily-sterilizable scaffolds for mammalian cells, since facile cell harvesting can be achieved after a single thermal cycle

Monophasic synovial sarcoma presenting as a primary ileal mass: a case report and review of the literature

<p>Abstract</p> <p>Introduction</p> <p>Synovial sarcoma is a rare malignant mesenchymal tumor mainly arising in the peri-articular tissue in young adults. There are few cases reported in other areas.</p> <p>Case presentation</p> <p>We report the case of a 29-year-old Saudi woman of Arabian ethnicity with synovial sarcoma arising primarily from the ileum who presented with abdominal pain, a palpable mass and incomplete intestinal obstruction. A literature review was performed to gather information on this rare gastrointestinal tract sarcoma.</p> <p>Conclusions</p> <p>Although it is a rare tumor of the pre-articular tissues, synovial sarcoma can present, in exceedingly rare cases, in unusual anatomical sites such as the gastrointestinal tract. We believe the reporting of all rare or unexpected presentations of sarcoma will eventually improve our understanding of this relatively unusual malignancy.</p

Serine phosphorylation regulates paxillin turnover during cell migration

BACKGROUND: Paxillin acts as an adaptor protein that localizes to focal adhesion. This protein is regulated during cell migration by phosphorylation on tyrosine, serine and threonine residues. Most of these phosphorylations have been implicated in the regulation of different steps of cell migration. The two major phosphorylation sites of paxillin in response to adhesion to an extracellular matrix are serines 188 and 190. However, the function of this phosphorylation event remains unknown. The purpose of this work was to determine the role of paxillin phosphorylation on residues S188 and S190 in the regulation of cell migration. RESULTS: We used NBT-II epithelial cells that can be induced to migrate when plated on collagen. To examine the role of paxillin serines 188/190 in cell migration, we constructed an EGFP-tagged paxillin mutant in which S188/S190 were mutated into unphosphorylatable alanine residues. We provide evidence that paxillin is regulated by proteasomal degradation following polyubiquitylation of the protein. During active cell migration on collagen, paxillin is protected from proteasome-dependent degradation. We demonstrate that phosphorylation of serines 188/190 is necessary for the protective effect of collagen. In an effort to understand the physiological relevance of paxillin protection from degradation, we show that cells expressing the paxillin S188/190A interfering mutant spread less, have reduced protrusive activity but migrate more actively. CONCLUSION: Our data demonstrate for the first time that serine-regulated degradation of paxillin plays a key role in the modulation of membrane dynamics and consequently, in the control of cell motility

Chlamydial Pre-Infection Protects From Subsequent Herpes Simplex Virus-2 Challenge in a Murine Vaginal Super-Infection Model

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Chlamydia trachomatis and Herpes Simplex Virus-2 (HSV-2) genital tract co-infections have been reported in humans and studied in vitro but the clinical consequences are unknown. Limited epidemiologic evidence suggests that these co-infections could be more severe than single infections of either pathogen, but the host-pathogen interactions during co-infection remain uncharacterized. To determine whether disease progression and/or pathogen shedding differs between singly-infected and super-infected animals, we developed an in vivo super-infection model in which female BALB/c mice were vaginally infected with Chlamydia muridarum (Cm) followed later by HSV-2. Pre-infection with Chlamydia 3 or 9 days prior to HSV-2 super-infection conferred significant protection from HSV-2-induced neurologic disease and significantly reduced viral recovery compared to HSV-2 singlyinfected controls. Neither protection from mortality nor reduced viral recovery were observed when mice were i) super-infected with HSV-2 on day 27 post Cm; ii) infected with UV-irradiated Cm and super-infected with HSV-2; or iii) azithromycin-treated prior to HSV-2 super-infection. Therefore, protection from HSV-2-induced disease requires active infection with viable chlamydiae and is not observed after chlamydial shedding ceases, either naturally or due to antibiotic treatment. Thus, Chlamydia-induced protection is transient and requires the continued presence of chlamydiae or their components. These data demonstrate that chlamydial pre-infection can alter progression of subsequent HSV-2 infection, with implications for HSV-2 transmission from co-infected humans

Monocyte Differentiation into Destructive Macrophages on In Vitro Administration of Gingival Crevicular Fluid from Periodontitis Patients

Background: Periodontitis is an inflammatory condition of the tooth-supporting structures initiated and perpetuated by pathogenic bacteria present in the dental plaque biofilm. In periodontitis, immune cells infiltrate the periodontium to prevent bacterial insult. Macrophages derived from monocytes play an important role in antigen presentation to lymphocytes. However, they are also implicated in causing periodontal destruction and bystander damage to the host tissues. Objectives: The objective of the present study was to quantify the cytokine profile of gingival crevicular fluid (GCF) samples obtained from patients with periodontitis. The study further aimed to assess if GCF of periodontitis patients could convert CD14+ monocytes into macrophages of destructive phenotype in an in vitro setting. The secondary objectives of the study were to assess if macrophages that resulted from GCF treatment of monocytes could affect the synthetic properties, stemness, expression of extracellular matrix proteins, adhesion molecules expressed by gingival stem cells, gingival mesenchymal stromal cells, and osteoblasts. Methods: GCF, blood, and gingival tissue samples were obtained from periodontitis subjects and healthy individuals based on specific protocols. Cytokine profiles of the GCF samples were analyzed. CD14+ monocytes were isolated from whole blood, cultured, and treated with the GCF of periodontitis patients to observe if they differentiated into macrophages. Further, the macrophages were assessed for a phenotype by surface marker analysis and cytokine assays. These macrophages were co-cultured with gingival stem cells, epithelial, stromal cells, and osteoblasts to assess the effects of the macrophages on the synthetic activity of the cells. Results: The GCF samples of periodontitis patients had significantly higher levels of IFN gamma, M-CSF, and GM-CSF. Administration of the GCF samples to CD14+ monocytes resulted in their conversion to macrophages that tested positive for CD80, CD86, and CD206. These macrophages produced increased levels of IL-1β, TNF-α, and IL-6. Co-culture of the macrophages with gingival stem cells, epithelial cells, and stromal cells resulted in increased cytotoxicity and apoptotic rates to the gingival cells. A reduced expression of markers related to stemness, extracellular matrix, and adhesion namely OCT4, NANOG, KRT5, POSTN, COL3A1, CDH1, and CDH3 were seen. The macrophages profoundly affected the production of mineralized nodules by osteoblasts and significantly reduced the expression of COL1A1, OSX, and OCN genes. Conclusion: In periodontitis patients, blood-derived monocytes transform into macrophages of a destructive phenotype due to the characteristic cytokine environment of their GCF. Further, the macrophages affect the genotype and phenotype of the resident cells of the periodontium, aggravate periodontal destruction, as well as jeopardize periodontal healing and resolution of inflammation

Preliminary Evaluation of Proliferation, Wound Healing Properties, Osteogenic and Chondrogenic Potential of Dental Pulp Stem Cells Obtained from Healthy and Periodontitis Affected Teeth

Background: Dental pulp tissue within the central cavity of the tooth is composed of dental pulp stem cells (DPSC). These mesenchymal stem cells have good proliferative as well as differentiation potential. DPSC has been isolated even from teeth with inflamed pulps and is found to retain their proliferative and differentiation potential. Little research is available about the viability and differentiation potential of DPSC obtained from teeth with periodontitis. In the present study, the aim was to compare the morphological features, stem cell marker (MSC) expression, proliferation rate, migratory and wound healing properties, osteogenic and chondrogenic differentiation potential of DPSCs obtained from periodontally healthy teeth (hDPSCs) and periodontitis affected teeth (pDPSCs). Methods: Dental pulp tissue was obtained from periodontally healthy volunteers (n = 3) and patients with periodontitis undergoing extraction of mobile teeth (n = 3). DPSC were isolated using the explant technique and cultured. All the experiments were performed at early passage (Passage 2), late passage (Passage 6) and after cryopreservation. Morphological features of the hDPSCs and pDPSCs were ascertained using microscopy. The expression of cell surface stem cell markers was assessed by the flow cytometry method. The proliferation and growth rate of the cells were assayed by plotting a growth curve from 0–13 days of culture. The migratory characteristics were assessed by wound scratch assay. Osteogenic and chondrogenic differentiation of the cells was assessed using standard protocols with and without induction. Results: DPSCs were successfully obtained from periodontally healthy teeth (hDPSC) and periodontitis-affected teeth (pDPSCs). The data suggests that there were no morphological differences observed in early passage cells between the two cohorts. Cryopreservation did change the morphology of pDSPCs. There was no significant difference in the positive expression of mesenchymal markers CD73, CD90 and CD105 in early passage cells. However, serial passaging and cryopreservation affected the marker expression in pDPSCs. A faint expression of hematopoietic stem cell markers CD34, CD45 and MHC class II antigen HLA-DR was observed in both the cell types. The expression of HLA-DR is upregulated in pDPSCs compared to hDPSC. A significantly slower growth rate and slower wound healing properties was observed in pDPSCs compared to hDPSC. In late passage and after cryopreservation, the migratory ability of pDPSCs was found to be increased drastically. There was no significant difference in osteogenic potential between the two cell types. However, the chondrogenic potential of pDPSCs was significantly lower compared to hDPSc. Yet, pDPSCs showed enhanced osteogenesis and chondrogenesis at late passage as well as after cryopreservation. Conclusion: The results of this novel study shed light on the isolation of viable DPSC from periodontitis-affected teeth. These cells exhibit a slower growth rate and migratory characteristics compared to their healthy counterparts. There was no difference in osteogenic potential but a reduction in chondrogenic potential was seen in pDPSCs compared to hDPSC. The findings reveal that DPSC from periodontitis-affected teeth presents an easy and viable option for regenerative medicine application. Some additional nutritive factors and protocols may be required to attain better regenerative benefits while using pDPSCs