Myoconductive and osteoinductive free-standing polysaccharide membranes

Free-standing (FS) membranes have increasing applications in the biomedical field as drug delivery systems for wound healing and tissue engineering. Here, we studied the potential of free-standing membranes made by the layer-by-layer assembly of chitosan and alginate to be used as a simple biomimetic system of the periosteum. The design of a periosteum-like membrane implies the elaboration of a thick membrane suitable for both muscle and bone formation. Our aim was to produce well-defined ∼50 μm thick polysaccharide membranes that could be easily manipulated, were mechanically resistant, and would enable both myogenesis and osteogenesis in vitro and in vivo. The membranes were chemically crosslinked to improve their mechanical properties. Crosslinking chemistry was followed via Fourier transform infrared spectroscopy and the mechanical properties of the membranes were assessed using dynamic mechanical analysis. The loading and release of the potent osteoinductive growth factor bone morphogenetic protein 2 (BMP-2) inside and outside of the FS membrane was followed by fluorescence spectroscopy in a physiological buffer over 1 month. The myogenic and osteogenic potentials of the membranes in vitro were assessed using BMP-2-responsive skeletal myoblasts. Finally, their osteoinductive properties in vivo were studied in a preliminary experiment using a mouse ectopic model. Our results showed that the more crosslinked FS membranes enabled a more efficient myoblast differentiation in myotubes. In addition, we showed that a tunable amount of BMP-2 can be loaded into and subsequently released from the membranes, depending on the crosslinking degree and the initial BMP-2 concentration in solution. Only the more crosslinked membranes were found to be osteoinductive in vivo. These polysaccharide-based membranes have strong potential as a periosteum-mimetic scaffold for bone tissue regeneration.This work was financially supported by the Foundation for Science and Technology (FCT) through the scholarship SFRH/BPD/96797/2013, Fundo Social Europeu (FSE), and Programa Diferencial de Potencial Human (POPH) granted to Sofia G. Caridade. C.M. is indebted to the Association Francaise contre les Myopathies for financial support via a post-doctoral fellowship (AFM project 16673). J.A. acknowledges the Whitaker International Fellows and Scholars Program for support via a post-doctoral fellowship. This work was supported by the European Commission (FP7 program) via a European Research Council starting grant (BIOMIM, GA 259370 to C.P.) and by the AFM (grant Microtiss, 16530). We thank Isabelle Paintrand for her technical help with the confocal apparatus

Diagenesis of archaeological bone and tooth

An understanding of the structural complexity of mineralised tissues is fundamental for exploration into the field of diagenesis. Here we review aspects of current and past research on bone and tooth diagenesis using the most comprehensive collection of literature on diagenesis to date. Environmental factors such as soil pH, soil hydrology and ambient temperature, which influence the preservation of skeletal tissues are assessed, while the different diagenetic pathways such as microbial degradation, loss of organics, mineral changes, and DNA degradation are surveyed. Fluctuating water levels in and around the bone is the most harmful for preservation and lead to rapid skeletal destruction. Diagenetic mechanisms are found to work in conjunction with each other, altering the biogenic composition of skeletal material. This illustrates that researchers must examine multiple diagenetic pathways to fully understand the post-mortem interactions of archaeological skeletal material and the burial environment

The Mineral Phase: Calcified Microsphere Populations in Young and Ageing, Hard and Soft Tissues

There is the tendency with time for hard tissues to soften and soft tissues to harden 'With potential pathological consequences. Comparison of the mineral phase in bot.h may' provide novel insight into associated untreatable conditions of aging. The human proximal femur (the most vulnerable skeletal site) from nonfracture (osteoarthritis), fracture (osteoporosis) and normal subjects was examined, together 'With the human neonatal-TMJ and bone from functionally different skeletal locations in the rat, as wen as human dentine; soft tissues included human and sheep aorta, and sheep intervertebral disc and trachea. Histological methods encompassed plain and polarised light microscopy, epi-fluorescence microscopy, laser confocal microscopy with density gradient mapping, histochemistry and immunohistochemistry, together 'With SEM and TEM and EDXmicroanalysis of elements. These were applied to resin-embedded undecalcified sections, cryosectiGus' and mineral particle isolates separated from the tissue by hydrazine or sodium hydroxide digestion, or prepared synthetically. The results support the view that bone mineral does not consist of random uniform crystals. Population of calcified microspheres, about 1Ilm diameter, occurred in sections and isolates, singly, in bridged assemblies (10IlID long) and compressed into irregular domains or rounded macrosphere domains (30llm diameter). Apparently derived from dense nanospheres (50nm diameter) the microspheres are cell fabrications enclosing a filamentous substructure of 5nm beads. They varied with pathology, being larger (l-5Ilm) in osteoarthritis, and smaller (O.5-1Ilm) in osteoporosis, and the extent of their bridging increased in weight-bearing locations relative to non-weight-bearing sites. They generally lacked the uniform electron density and crystal-like nature of synthetic preparations, except in traces which increase with pathology or chemical exposure. There were some differences of degree with calcified microspheres populations in soft tissues. For example the CaIP ratio was more variable (1.17-2.59 cf. 1.52-1.54) while Mg, Si and Fe were regularly associated with both, Mg was particularly abundant in the calcified particles in the aorta and intervertebral disc. They did not permeate the collagen fibres but in their assemblies and convoluted domains looped around them to form a remarkably varied composite. Of special interest in this regard was the periosteum. Known to be as proactive in development of the skeleton, the results suggested it is a key determinant of its mass and maintenance as a periosteumSharpey's fibre-endosteum integrated system, rich in collagen type III that may function as a sensory avenue for musculoskeletal exchange. Contrasting features of this system are considered in osteoporosis, where it seems too tenuous and in osteoarthritis where it seems too robust. It was concluded that in the connective tissues neither the hard inorganic,phase nor the soft collagenous matrix are the homogeneous substances they are regularly portrayed to be. Their partnership in the young is optimal and beneficial in terms of structural strength and metabolic exchange but with age minor discord in this new histological facet to bone biology may lead to major disability and the challenge this presents for an aging populace.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

A new tool for early diagnosis of rheumatoid arthritis using combined biomarkers; synovial MAGE-1 mRNA and serum anti-CCP and RF

Introduction: rheumatoid arthritis (RA) is a common autoimmune disease with unknown etiology and pathogenesis. Biomarkers have the potential to aid in the clinical diagnosis of the disease, or to provide means of detecting early signs of the disease. Evaluating Melanoma associated antigen genes (MAGE-1) mRNA expression rate in synovial fluid cells and serum levels of anti-cyclic citrullinated peptides (anti-CCP) and rheumatoid factor (RF) for RA early diagnosis. Methods: a total of 213 subjects were enrolled in the study, 135 RA patients and 78 normal subjects with traumatic knee joints (control group). Serum RF and anti-CCP were estimated quantitatively using ELISA. MAGE-1 mRNA expression rate was analyzed by RT-PCR. Results: a significant increase in serum levels of RF IgM and anti-CCP in RA patients compared to the controls. A positively significant correlation was found between serum anti-CCP and RF IgM. The expression rate of MAGE-1 mRNA was 100% in RA patients versus the controls (0%). The specificity and the sensitivity of the three biomarkers was 100%. Conclusion: the high expression rate of MAGE-1 in synovial fluid cells of RA patients is encouraging its utilization as a diagnostic biomarker for RA. The combined use of MAGE-1 transcript in synovial fluid cells, serum RF and anti-CCP is recommended for improving early diagnostic ability of RA

Bone Age Determination of Epiphyseal Union Around Wrist Joint and its Correlation with Chronological Age: A Radiological Study in a Jordanian Population

Age determination is one of the most important parameters for human identification. Radiographic changes during epiphyseal union provide an important means to estimate the age of adolescent and young adult skeletons. This study aims to investigate the relationship between stage of epiphyseal union at the wrist joint and chronological age in a Jordanian population. The study was carried out in a total of 101 antero-posterior radiographs of healthy subjects (45 girls and 56 boys) aging from 12 to 22 years. The obtained results from the radiographs enrolled in the current study revealed that the complete union of lower end of radius is seen at 20-21 years. The complete union of lower end of ulna is seen at 20-21 years. Females were consistently developing epiphyseal union at a younger age than their male counterparts, with a two years difference. The results suggested that the ages of epiphyseal union are found to vary greatly all over the world indicating the need for separate standards of ages of epiphyseal union for separate regions

A new hypothesis may explain human parthenogenesis and ovarian teratoma: A review study

Abstract Parthenogenesis (PG) is a rare phenomenon occurring in humans, and understanding this may help us develop an explanation for such occurrences. Moreover, it may help reveal the cause of idiopathic ovarian teratoma (OT). We aim to explain the occurrence of PG and OT in humans based on a new hypothesis. Previous literature has been searched through relevant scientific websites and international journals on the causes and mechanisms of PG and OT in humans. The previous literature on human PG was sparse and mostly contained case reports. It appears that human PG is not as rare as previously reported but may occur spontaneously, resulting in OT formation. The difference between PG and sexual reproduction is that PG has no embryonic diversity. The biopsied embryonic samples in the PG correspond exclusively to those of the maternal side. Spontaneous PG in humans often degrades or leads to formation of OT. The cause and mechanism of spontaneous PG remain unclear in the available literature. Here, we hypothesized that in some cases the secondary oocyte and first polar body enclosed in the zona pellucida may fuse together to form a single cell that restores the diploid number of chromosomes and initiates cell division to form PG. It may go unnoticed or be represented by the OT. Future studies are recommended to investigate this hypothesis

VP37 Protein Inhibitors for Mpox Treatment: Highlights on Recent Advances, Patent Literature, and Future Directions

Monkeypox disease (Mpox) has threatened humankind worldwide since mid-2022. The Mpox virus (MpoxV) is an example of Orthopoxviruses (OPVs), which share similar genomic structures. A few treatments and vaccines are available for Mpox. OPV-specific VP37 protein (VP37P) is a target for developing drugs against Mpox and other OPV-induced infections such as smallpox. This review spotlights the existing and prospective VP37P inhibitors (VP37PIs) for Mpox. The non-patent literature was collected from PubMed, and the patent literature was gathered from free patent databases. Very little work has been carried out on developing VP37PIs. One VP37PI (tecovirimat) has already been approved in Europe to treat Mpox, while another drug, NIOCH-14, is under clinical trial. Developing tecovirimat/NIOCH-14-based combination therapies with clinically used drugs demonstrating activity against Mpox or other OPV infections (mitoxantrone, ofloxacin, enrofloxacin, novobiocin, cidofovir, brincidofovir, idoxuridine, trifluridine, vidarabine, fialuridine, adefovir, imatinib, and rifampicin), immunity boosters (vitamin C, zinc, thymoquinone, quercetin, ginseng, etc.), and vaccines may appear a promising strategy to fight against Mpox and other OPV infections. Drug repurposing is also a good approach for identifying clinically useful VP37PIs. The dearth in the discovery process of VP37PIs makes it an interesting area for further research. The development of the tecovirimat/NIOCH-14-based hybrid molecules with certain chemotherapeutic agents looks fruitful and can be explored to obtain new VP37PI. It would be interesting and challenging to develop an ideal VP37PI concerning its specificity, safety, and efficacy

Targeting Cytochrome P450 Enzymes in Ovarian Cancers: New Approaches to Tumor-Selective Intervention

Over the past decade, there have been significant developments in treatment for ovarian cancer, yet the lack of targeted therapy with few side effects still represents a major issue. The cytochrome P450 (CYP) enzyme family plays a vital role in the tumorigenesis process and metabolism of drugs and has a negative impact on therapy outcomes. Gaining more insight into CYP expression is crucial to understanding the pathophysiology of ovarian cancer since many isoforms are essential to the metabolism of xenobiotics and steroid hormones, which drive the disease’s development. To the best of our knowledge, no review articles have documented the intratumoral expression of CYPs and their implications in ovarian cancer. Therefore, the purpose of this review is to provide a clear understanding of differential CYP expression in ovarian cancer and its implications for the prognosis of ovarian cancer patients, together with the effects of CYP polymorphisms on chemotherapy metabolism. Finally, we discuss opportunities to exploit metabolic CYP expression for the development of novel therapeutic methods to treat ovarian cancer

Cytochrome 4Z1 Expression Connotes Unfavorable Prognosis in Ovarian Cancers

Background and Objective: Ovarian cancer is a leading cause of death in females. Since its treatment is challenging and causes severe side effects, novel therapies are urgently needed. One of the potential enzymes implicated in the progression of cancers is Cytochrome 4Z1 (CYP4Z1). Its expression in ovarian cancer remains unknown. Therefore, the current study aims to assess CYP4Z1 expression in different subtypes of ovarian cancers. Materials and Methods: Immunohistochemistry was used to characterize CYP4Z1 expression in 192 cases of ovarian cancers along with eight normal ovarian tissues. The enzyme’s association with various clinicopathological characteristics and survival was determined. Results: CYP4Z1 was strongly expressed in 79% of ovarian cancers, compared to negative expression in normal ovarian samples. Importantly, significantly high CYP4Z1 expres-sion was determined in patients with advanced-stage cancer and a high depth of invasion (p < 0.05). Surprisingly, CYP4Z1 expression was significantly associated with a low patient survival rate. Univariate analysis revealed that patient survival was strongly associated with CYP4Z1 expression, tumor stage, depth of invasion, and lymph node metastasis (p < 0.05). Multivariate analysis showed that only CYP4Z1 expression was significantly associated with patient survival (p < 0.05). Conclusions: CYP4Z1 expression is correlated with shorter patient survival and has been identified as an independent indicator of a poor prognosis for ovarian cancer patients