Cell Proliferations Research Articles

New Innovations in Seed Morphology, and Plant Physiology for Crop Protection

The seed is a mainly reproductive unit of the spermatophyta and links the subsequent generations. Other essential seed activities include survival in arid, cold, or other adverse conditions and dispersal. There is a lot of variety in the internal and external architecture of seeds. OsMKP1, a mitogen-activated protein kinase phosphatase, is encoded by GSN1. Rice glume cells proliferate when GSN1 expression is suppressed, producing larger but fewer rice grains. The GSN1 directly interacts with OsMAPK6 and dephosphorylates it, rendering it inactive. Thus, precise regulation of OsMAPK6 activity via reversible phosphorylation is essential for regulating the rice grain size. Some genes also negatively control the grain width and weight by inhibiting cell proliferation. The embryo, which is made up of cotyledons, hypocotyl, and radicle; the endosperm, which feeds the growing embryo; and the seed coat, which envelops the embryo and the endosperm, are the three main parts of plant seeds, each of which has unique biological functions and outcome. The growing seed can benefit from the ability to predict when a sugar burst would arrive through the phloem, since this would allow the seed to quickly adapt to its storage product synthesis. However, there are still certain obstacles that prevent the use against a variety of insect species. The adoption of various formulation procedures that improve dsRNA persistence and cellular absorption in insects is expected to overcome these issues, which are mostly related to the varying RNAi sensitivity of oral RNAi in insects. The CRISPR/Cas system has become the most popular being shown to be useful for editing the genome of plants, its uses in plants have grown significantly in comparison to other technologies.

Unicentric Castleman Disease: Updates and Novel Insights Into Spindle Cell Proliferations and Aggressive Forms of a Localized Disease.

Castleman Disease (CD) is a rare lymphoproliferative disorder that can be separated into two primary forms: Unicentric Castleman disease (UCD) and multicentric Castleman disease (MCD). UCD is localized, while MCD is systemic. Though UCD generally has a favorable prognosis following surgical resection, more aggressive forms of this disease have been identified, including cases associated with dendritic and spindle cell proliferation. Genetic analysis has deepened our understanding of UCD. Despite advancements in better understanding the pathophysiology of UCD, challenges persist in the diagnosis, management, and treatment due to its rarity and heterogeneity. Here, we review current knowledge on UCD, highlighting the epidemiology, clinical presentation, diagnostic criteria, and treatment options while emphasizing the need for further research and innovation in therapeutic strategies.

Antitumor effects of inhibitors of ERK and Akt pathways in canine histiocytic sarcoma cell lines

Canine histiocytic sarcoma (CHS) is characterized by aggressive biological behavior. In our previous study, ERK and Akt pathways were found to be activated in CHS tissues. Thus, the objective of this study was set to investigate the relationships between the activation status of these pathways and the proliferation of CHS cell lines by examining the effects of single and co-administrations of drugs targeting these pathways. First, we evaluated the changes in cell proliferations and the activations of ERK and Akt pathways after treatments with ERK and Akt-specific inhibitors in CHS cells. Then, these changes after treatments with dasatinib and trametinib were also examined in CHS cells.Inhibitors specific to ERK and Akt pathways successfully inhibited the respective pathways in CHS cell lines. It was also indicated that these pathways were associated with the regulations of proliferations of CHS cells, although the anti-proliferative effect was not necessarily observed by inhibition of Akt pathway alone. Dasatinib and trametinib also showed the inhibitions of Akt and ERK pathway activations, respectively, in CHS cells. However, the anti-proliferative effects of these drugs varied among CHS cell lines, and co-administration showed enhanced anti-proliferative effects in only a part of CHS cell lines. Further studies are needed to investigate the molecular mechanisms associated with the sensitivities to these molecular-targeted drugs in CHS cells.

Future directions in myelodysplastic syndromes/neoplasms and acute myeloid leukaemia classification: from blast counts to biology.

Myelodysplastic syndromes/neoplasms (MDS) and acute myeloid leukaemia (AML) are neoplastic haematopoietic cell proliferations that are diagnosed and classified based on a combination of morphological, clinical and genetic features. Specifically, the percentage of myeloblasts in the blood and bone marrow is a key feature that has historically separated MDS from AML and, together with several other morphological parameters, defines distinct disease entities within MDS. Both MDS and AML have recurrent genetic abnormalities that are increasingly influencing their definitions and subclassification. For example, in 2022, two new MDS entities were recognised based on the presence of SF3B1 mutation or bi-allelic TP53 abnormalities. Genomic information is more objective and reproducible than morphological analyses, which are subject to interobserver variability and arbitrary numeric cut-offs. Nevertheless, the integration of genomic data with traditional morphological features in myeloid neoplasm classification has proved challenging by virtue of its sheer complexity; gene expression and methylation profiling also can provide information regarding disease pathogenesis, adding to the complexity. New machine-learning technologies have the potential to effectively integrate multiple diagnostic modalities and improve on historical classification systems. Going forward, the application of machine learning and advanced statistical methods to large patient cohorts can refine future classifications by advancing unbiased and robust previously unrecognised disease subgroups. Future classifications will probably incorporate these newer technologies and higher-level analyses that emphasise genomic disease entities over traditional morphologically defined entities, thus promoting more accurate diagnosis and patient risk stratification.

Mediastinal Teratoma with Angiosarcomatous Overgrowth in a Background of Vasculogenic Mesenchymal Tumor

Abstract Introduction/Objective Vasculogenic mesenchymal tumor refers to neoplastic reiteration of embryonic vasculogenesis. It is most commonly associated with nonseminomatous germ cell tumors of the mediastinum, in particular yolk sac tumor. In rare instances these lesions progress to angiosarcoma. Herein, we describe a case of angiosarcoma in a background of vasculogenic mesenchymal tumor in the setting of a mediastinal teratoma. Methods/Case Report A 33-year-old male, with a past medical history of anxiety, and right orchiectomy for testicular torsion, presented with shortness of breath, chest pain, and left arm swelling. Imaging studies demonstrated a mediastinal mass. Biopsy of the mass showed a germ cell tumor most compatible with a teratoma in the limited specimen. The patient received two rounds of chemotherapy and additional treatments were discontinued due to poor tolerance. Subsequent imaging showed continued increase in the size along with mass effect causing moderate to severe compression of the superior vena cava. Subsequently, the patient underwent resection of the mass. Grossly the mass was predominantly encapsulated with areas of capsular disruption. Sectioning revealed heterogeneous cut surfaces with fleshy, cystic, and necrotic areas. Morphologically the tumor was composed predominantly of teratomatous elements with areas of anastomosing atypical vascular channels within a hypocellular stroma suggestive of vascular genic mesenchymal as well as areas reminiscent of Masson lesion. Focally within these areas were malignant spindle cell proliferations compatible with angiosarcoma. The patient had an elevated AFP; however, no yolk sac tumor component was identified after thorough sampling. Final diagnosis of teratoma with angiosarcoma in a background of vasculogenic mesenchymal tumor was rendered after expert consultation. Results (if a Case Study enter NA) NA Conclusion Somatic-type malignancy and sarcomatous overgrowth are known to occur in mediastinal germ cell tumors; however, classification of the sarcomatous overgrowth can be challenging. Vasculogenic mesenchymal tumor is an exceedingly rare sarcomatous overgrowth that can rarely progress to angiosarcoma.

9291 Hypothyroidism impairs skeletal muscle regeneration after injury

Abstract Disclosure: P. Aguiari: None. V. Villani: None. K.Y. Liu: None. G.A. Brent: None. L. Perin: None. A. Milanesi: None. Thyroid hormone (TH) signaling plays an essential role in muscle development and function, in the maintenance of muscle mass, and in regeneration after injury. Disruption of TH signaling results in an abnormal skeletal muscle phenotype, impaired regeneration, and sarcopenia with aging, reflecting the myopathic changes described in hypothyroid patients. Muscle stem cells (MuSCs) are the mediators of post-natal skeletal muscle plasticity. Normally quiescent, in response to injury they enter the cell cycle (myoblasts) and proliferate. Myoblasts then exit the cell cycle and differentiate into myocytes that will fuse with existing myofibers to restore tissue architecture and function. Via TH receptor alpha, TH regulates all the stages of the regeneration program and regulates the expression of multiple myogenic genes. Despite all this evidence, to date there are no studies investigating MuSCs behavior in response to injury during hypothyroidism. We characterized injury-induced regeneration of the tibialis anterior muscle (TAM) in euthyroid and hypothyroid mice, fed a low iodine + 0.15% propylthiouracil diet. To investigate the cell cycle dynamics of MuSCs, we generated Pax7-Fucci mice carrying the fluorescent ubiquitination-based cell-cycle indicator (Fucci) system under the Pax7 promoter. Muscle injury was induced via cardiotoxin injection in the TAM of 3-month-old Pax7-Fucci mice. Hypothyroid mice present signs of impaired regeneration compared to euthyroid mice. From histological analysis, at different time points, we observed smaller fiber diameters, myofibers with central nuclei, and a significantly reduced number of fast glycolytic type IIb fibers, the prevalent muscle fiber type in the TA muscle. ScRNAseq analysis shows that at a late phase of regeneration (14 days after injury) hypothyroid MuSCs and myoblasts are still in the activation state and overexpress genes related to DNA replication and cell proliferation, while genes related to myogenic differentiation and cell cycle exit are downregulated. Differentiated myocytes in the hypothyroid fibers present an immature phenotype and are characterized by overexpression of embryonic/temporary and slow myosin genes and downregulation of mature fast myosins. Cell cycle analysis of the Fucci signal through Flow Cytometry confirmed a higher number of activated hypothyroid MuSCs at 4, 7, and 28 days after injury. These cells accumulate in the S phase and are unable to exit the cell cycle and differentiate towards myocytes. These data demonstrate that hypothyroidism impairs muscle tissue regeneration halting MuSCs progression through the cell cycle, myoblast cell cycle exit, and fiber maturation. Investigating muscle stem cell behavior and response to injury during hypothyroidism is crucial to understanding the mechanism behind thyroid hormone-induced myopathies and identifying possible therapeutical targets. Presentation: 6/2/2024

8670 Hypothyroidism impairs skeletal muscle regeneration after injury

Abstract Disclosure: P. Aguiari: None. V. Villani: None. K.Y. Liu: None. G.A. Brent: None. L. Perin: None. A. Milanesi: None. Thyroid hormone (TH) signaling plays an essential role in muscle development and function, in the maintenance of muscle mass, and in regeneration after injury. Disruption of TH signaling results in an abnormal skeletal muscle phenotype, impaired regeneration, and sarcopenia with aging, reflecting the myopathic changes described in hypothyroid patients. Muscle stem cells (MuSCs) are the mediators of post-natal skeletal muscle plasticity. Normally quiescent, in response to injury they enter the cell cycle (myoblasts) and proliferate. Myoblasts then exit the cell cycle and differentiate into myocytes that will fuse with existing myofibers to restore tissue architecture and function. Via TH receptor alpha, TH regulates all the stages of the regeneration program and regulates the expression of multiple myogenic genes. Despite all this evidence, to date there are no studies investigating MuSCs behavior in response to injury during hypothyroidism. We characterized injury-induced regeneration of the tibialis anterior muscle (TAM) in euthyroid and hypothyroid mice, fed a low iodine + 0.15% propylthiouracil diet. To investigate the cell cycle dynamics of MuSCs, we generated Pax7-Fucci mice carrying the fluorescent ubiquitination-based cell-cycle indicator (Fucci) system under the Pax7 promoter. Muscle injury was induced via cardiotoxin injection in the TAM of 3-month-old Pax7-Fucci mice. Hypothyroid mice present signs of impaired regeneration compared to euthyroid mice. From histological analysis, at different time points, we observed smaller fiber diameters, myofibers with central nuclei, and a significantly reduced number of fast glycolytic type IIb fibers, the prevalent muscle fiber type in the TA muscle. ScRNAseq analysis shows that at a late phase of regeneration (14 days after injury) hypothyroid MuSCs and myoblasts are still in the activation state and overexpress genes related to DNA replication and cell proliferation, while genes related to myogenic differentiation and cell cycle exit are downregulated. Differentiated myocytes in the hypothyroid fibers present an immature phenotype and are characterized by overexpression of embryonic/temporary and slow myosin genes and downregulation of mature fast myosins. Cell cycle analysis of the Fucci signal through Flow Cytometry confirmed a higher number of activated hypothyroid MuSCs at 4, 7, and 28 days after injury. These cells accumulate in the S phase and are unable to exit the cell cycle and differentiate towards myocytes. These data demonstrate that hypothyroidism impairs muscle tissue regeneration halting MuSCs progression through the cell cycle, myoblast cell cycle exit, and fiber maturation. Investigating muscle stem cell behavior and response to injury during hypothyroidism is crucial to understanding the mechanism behind thyroid hormone-induced myopathies and identifying possible therapeutical targets. Presentation: 6/2/2024

The Role of SIRT1 in Leukemia.

Leukemia is a type of hematological malignancy (HM) caused by uncontrolled proliferation, apoptosis, and differentiation of hematopoietic stem cells (HSCs). Leukemia cells proliferate greatly in the bone marrow (BM), infiltrate other tissues and organs, and affect the normal hematopoietic function. Although the emergence of new targeted agents and immune agents has improved the prognosis of patients, due to the complex pathogenic factors and heterogeneity of leukemia, there are still some patients with poor prognosis. Recent studies have shown that silent information regulator 1 (SIRT1) is involved in the proliferation, apoptosis, metabolism, and senescence of leukemia cells. As a double-edged sword in leukemia cells, SIRT1 can both promote and inhibit the growth of leukemia cells. Since its mechanism of action has not been elucidated, it is urgent to explore the regulatory mechanism of SIRT1 in leukemia. In this review, we discussed the mechanisms of SIRT1 in different aspects of leukemia, providing a theoretical basis for the treatment of patients with leukemia.

When Do Tumours Develop? Neoplastic Processes Across Different Timescales: Age, Season and Round the Circadian Clock.

While it is recognised that most, if not all, multicellular organisms harbour neoplastic processes within their bodies, the timing of when these undesirable cell proliferations are most likely to occur and progress throughout the organism's lifetime remains only partially documented. Due to the different mechanisms implicated in tumourigenesis, it is highly unlikely that this probability remains constant at all times and stages of life. In this article, we summarise what is known about this variation, considering the roles of age, season and circadian rhythm. While most studies requiring that level of detail be done on humans, we also review available evidence in other animal species. For each of these timescales, we identify mechanisms or biological functions shaping the variation. When possible, we show that evolutionary processes likely played a role, either directly to regulate the cancer risk or indirectly through trade-offs. We find that neoplastic risk varies with age in a more complex way than predicted by early epidemiological models: rather than resulting from mutations alone, tumour development is dictated by tissue- and age-specific processes. Similarly, the seasonal cycle can be associated with risk variation in some species with life-history events such as sexual competition or mating being timed according to the season. Lastly, we show that the circadian cycle influences tumourigenesis in physiological, pathological and therapeutic contexts. We also highlight two biological functions at the core of these variations across our three timescales: immunity and metabolism. Finally, we show that our understanding of the entanglement between tumourigenic processes and biological cycles is constrained by the limited number of species for which we have extensive data. Improving our knowledge of the periods of vulnerability to the onset and/or progression of (malignant) tumours is a key issue that deserves further investigation, as it is key to successful cancer prevention strategies.

Development of a P(L-D,L)LA Foam as a Dura Substitute and Its In Vitro Evaluation

Dura substitutes are used to reduce the risk of postoperative complications following neurosurgical interventions, and to facilitate the healing of dura damages or defects caused by injuries. Traditional tissue transplants have limitations like limited tissue availability, potential risk of immune rejection and disease transmission. The use of biomaterials composed of synthetic polymers as dura substitutes offers a promising approach to overcome these limitations to replace and treat damaged dura mater. Potential biocompatible porous scaffolds still need to be developed to minimize the risks of immune response and disease transmission, while also ensuring effective cell migration and cell ingrowth in three dimension. The aim of the present study was to develop a poly(L-lactide-co-D,L-lactide) (P(L-D,L)LA) foam with an optimal pore size for dura mater substitution, investigate its morphological characteristics, and evaluate its potential for dura mater regeneration by assessing the spreading and growth of meningeal cells within it through in vitro studies. Foams were produced by lyophilization using different concentrations of P(L-D,L)LA solution. A GMP-grade P(L-D,L)LA, suitable for medical device applications, was used in this study. Morphological analysis was performed using scanning electron microscopy, and porosity of the foams was studied with mercury porosimetry. In in vitro studies, meningeal cells were seeded onto the polymeric foams, and their behavior and proliferation in these scaffolds were investigated with cytoskeleton and nucleus staining, and colorimetric cell proliferation assay, respectively. Scanning electron microscopy results showed that the foams prepared with 2.5% and 3% polymer solutions displayed good structural integrity and convenient interconnectivity, with pore sizes ranging from 80 to 150 µm. However, the foams prepared with 2% and 4% polymer solution demonstrated poor structural integrity and low interconnectivity, respectively. In vitro studies showed that the foams prepared with 2.5% and 3% polymer solutions served effectively as scaffolds for meningeal cells, and the cells attached, spread and homogeneously distributed. In addition, the cells proliferated and increased in number over time within these polymeric scaffolds. These findings suggest that the foams produced with 2.5% and especially 3% P(L-D,L)LA polymer solutions could effectively serve as a suitable substitute for the dura mater, providing an appropriate environment for cell ingrowth and tissue integration. This indicates that the developed foam could be a promising treatment for dura mater damage or defects, with the potential approach to promote regeneration in future in vivo and clinical studies.

PSI-3 Effects of a Retinoid X Receptor agonist on primary ovine satellite cells

Abstract Sheep producers are typically paid based on body weight, so understanding the mechanisms of postnatal skeletal muscle growth is vital. Satellite cells are responsible for the capacity for postnatal skeletal muscle growth. Satellite cells proliferate and then differentiate into myoblasts before fusing with existing myotubes. The retinoid X receptor (RXR) is a transcription factor that enhances myogenic differentiation 1 (MyoD) expression, which increases differentiation and fusion of myoblasts in sheep. LG 100268 is an RXR agonist, which activates the RXR. The objective of this project is to measure the effect of LG 100268 on primary ovine satellite cell (OSC) proliferation and protein synthesis rates. Primary OSC were isolated from the hind legs of 9-mo old commercial hair sheep wethers (n = 3). For proliferation, cultures were grown to approximately 70% confluency, followed by treatment with 5% fetal bovine serum (FBS) or 300 nM of LG 100268 in 5% FBS. For protein synthesis, cultures were grown to approximately 80% confluency, at which point cultures were induced to differentiate and treated 48-h later with serum free media (SFM) or 300 nM LG 100268 in SFM. Proliferation rates were measured at 24-h post-treatment, and protein synthesis rates were measured at 6-h post-treatment. Treatment with 300 nM LG 100268 increased (P = 0.004) OSC proliferation rates compared with control cultures. Preliminary protein synthesis results suggest that LG 100268 does not alter (P = 0.22) protein synthesis rates of fused OSC. However, more research is warranted to clarify the role of the RXR in sheep skeletal muscle growth.

Spectra of well-differentiated neuroendocrine lesions in the extrahepatic biliary system: a case series.

Neuroendocrine tumours (NETs) occurring in the extrahepatic biliary system are exceedingly rare. While NETs typically manifest as mass lesions, the occurrence of microscopic neuroendocrine cell proliferation without a distinct mass remains undocumented at this location. This study aims to characterise the clinicopathological features of a series of well-differentiated neuroendocrine lesions involving the extrahepatic biliary tree, including mass forming NETs and microscopic non-mass-forming neuroendocrine cell proliferation, designated neuroendocrine cell micronests (NCMs). Surgical resections of NETs/NCMs involving the extrahepatic bile ducts and gallbladder were identified from electronic pathology databases among seven institutions spanning from January 2011 to September 2023. Clinical and histological findings were recorded. Ten patients (four female, six male: age range = 34-75 years) were included in the study. Histopathological examination revealed visible mass-forming lesions in four cases (1.6-14.0 cm in size), identified in the gallbladder (n = two) or extrahepatic bile duct (n = two), all diagnosed as well-differentiated NETs. The remaining six cases revealed incidental non-mass-forming NCMs in either the cystic duct (n = two), common bile duct (n = three) or gallbladder (n = one), ranging from < 0.1 to 0.4 cm; four were associated with biliary lithiasis. No evidence of metastasis or recurrence was seen in the follow-up period (range = 0.1-11.2 years). This study highlights the spectrum of extrahepatic biliary well-differentiated neuroendocrine lesions, ranging from incidental microscopic NCMs to grossly apparent mass-forming NETs, potentially requiring different clinical management. Noteworthy is the frequent association of incidental microscopic neuroendocrine cell proliferations with biliary lithiasis, indicating a potential neuroendocrine metaplastic pathogenesis that merits further exploration.

Sebaceous gland epithelioma with potential malignancy in a dog

Sebaceous gland tumors consist of neoplastic proliferations of sebaceous gland cells located around hair follicles in the dermis. These tumors are subclassified as sebaceous epithelioma, sebaceous adenoma, sebaceous adenocarcinoma and nodular hyperplasia. In this case report, a nodular growth in the sacral region of an eight-year-old male Belgian Malinois dog was presented. Macroscopically, the nodular mass had a slightly soft consistency, grayish-white, and dark red-black appearance. The mass was measured in the dimension of 2x1x1 cm. Histopathologically, the tumor was observed to have a multilobular structure shaped by neoplastic cell islands. The tumor consisted mainly of eosinophilic cells with small cytoplasm resembling epithelial basaloid cells and to a lesser extent differentiated sebocytes. The parenchyma of the tumor consisted of irregular islets containing a small number of mature sebocytes. Their nuclei are oval with one to three small nucleoli. Immunohistochemically, tumor cells for Ki67 antibody showed strongly positive immunoreactivity. Based on the histopathological and immunohistochemical features, the tumor was diagnosed as sebaceous epithelioma having, a potential malignancy. Since no case of sebaceous epithelioma with malignant potential has been reported in our country, we aimed to present the case with histopathological and immunohistochemical features.

Biomonitoring of Heavy Metal Toxicity in Freshwater Canals in Egypt Using Creeping Water Bugs (Ilyocoris cimicoides): Oxidative Stress, Histopathological, and Ultrastructural Investigations.

The abundance of metal pollutants in freshwater habitats poses serious threats to the survival and biodiversity of aquatic organisms and human beings. This study intends for the first time to assess the pernicious influences of heavy metals in Al Marioteya canal freshwater in Egypt, compared to Al Mansoureya canal as a reference site utilizing the creeping water bug (Ilyocoris cimicoides) as an ecotoxicological model. The elemental analysis of the water showed a significantly higher incidence of heavy metals, including cadmium (Cd), cobalt (Co), chromium (Cr), nickel (Ni), and lead (Pb), in addition to the calcium (Ca) element than the World Health Organization's (WHO) permitted levels. The Ca element was measured in the water samples to determine whether exposure to heavy metals-induced oxidative stress engendered Ca deregulation in the midgut tissues of the creeping water bug. Remarkably, increased levels of these heavy metals were linked to an increase in chemical oxygen demand (COD) at the polluted site. Notably, the accumulation of these heavy metals in the midgut tissues resulted in a substantial reduction in antioxidant parameters, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and ascorbate peroxidase (APOX), along with a marked rise in malondialdehyde (MDA), cytochrome P450, and protein carbonyl levels. These results clearly indicate a noticeable disturbance in the antioxidant defense system due to uncontrollable reactive oxygen species (ROS). Notably, the results demonstrated that oxidative stress caused disturbances in Ca levels in the midgut tissue of I. cimicoides from polluted sites. Furthermore, the comet and flow cytometry analyses showed considerable proliferations of comet cells and apoptotic cells in midgut tissues, respectively, exhibiting prominent correlations, with pathophysiological deregulation. Interestingly, histopathological and ultrastructural examinations exposed noticeable anomalies in the midgut, Malpighian tubules, and ovarioles of I. cimicoides, emphasizing our findings. Overall, our findings emphasize the potential use of I. cimicoides as a bioindicator of heavy metal pollution in freshwater to improve sustainable water management in Egypt.

MM-003 The Role of Bone Marrow Galectin-1 in Tumor Progression in Plasma Cell Proliferations

MM-003 The Role of Bone Marrow Galectin-1 in Tumor Progression in Plasma Cell Proliferations

The Role of Bone Marrow Galectin-1 in Tumor Progression in Plasma Cell Proliferations

The Role of Bone Marrow Galectin-1 in Tumor Progression in Plasma Cell Proliferations

Evaluating Osteogenic Cell Differentiation Efficacy in the Presence of Polylactide Samples With Varied Compositions for Bone Grafting: In Vitro Study.

In oral implantology, surgeons often confront the need to improve alveolar bone quality and volume before implantation in patients with bone defects. Whereas guided bone regeneration with titanium meshes is a clinical gold standard for bone augmentation, mesh removal pre-implantation presents a drawback. This study explores biodegradable scaffolds as an alternative. The research investigates the impact of various compositions of customized bone-grafting scaffolds on proliferation and osteogenic differentiation processes in vitro. Plates (10 × 10 × 0.5 mm) were fabricated from polylactide (PLA), PLA with 15% hydroxyapatite nanoparticles (PLA/HA), and polylactide with glycolic acid copolymers (PLGA 60:40 and 85:15). Gingival fibroblasts assessed the influence of experimental samples on proliferation and osteogenic differentiation in a low-glucose medium. Osteogenic differentiation was induced, and alizarin red staining measured extracellular matrix calcification via spectrophotometry. Active proliferation of gingival fibroblasts occurred along scaffold edges during cultivation. Although cells proliferated with experimental samples, rates were lower than control cells. PLA/HA showed higher alizarin red staining intensity, indicating enhanced matrix calcification. Experimental samples (PLA, PLA/HA, PLGA 85:15, PLGA 60:40) supported cell proliferation at lower rates than control. PLA/HA demonstrated increased matrix calcification. Biodegradable membranes were nontoxic, suggesting potential for bone augmentation.

Evolution of Initial Treatment for Desmoid Tumors.

Desmoid tumors (DTs) are rare, fibroblastic cell proliferations that can exhibit locally aggressive behavior but lack metastatic potential. Initial management has traditionally involved upfront resection; however, contemporary guidelines and expert panels have increasingly advocated for prioritizing active surveillance strategies. A single-institution, retrospective chart review identified all patients diagnosed with a primary DT at any site from 2007 to 2020. The primary outcome was the initial management strategy over time. Secondary outcomes included treatment-free survival (TFS) and time to treatment (TTT) for those undergoing active surveillance, as well as recurrence-free survival (RFS) and time to recurrence for those undergoing resection. Overall, 103 patients were included, with 68% female and a median follow-up of 44 months [24-74]. The most common tumor locations included the abdominal wall (27%), intra-abdominal/mesenteric (25%), chest wall (19%), and extremity (10%). Initial management included resection (60%), systemic therapy (20%), active surveillance (18%), and cryoablation (2%). Rates of surgical resection significantly decreased (p<0.001) over time, from 69.6% prior to 2018 to 29.2% after 2018. For those treated with upfront resection, 5-year RFS was 41.2%, and for patients undergoing initial active surveillance, TFS was 66.7% at 2 years, with a median TTT of 4 months [4-10]. This single-institution cohort at a tertiary medical center spanning over a decade demonstrates the transition to active surveillance for initial management of DTs, and highlights salient metrics in the era of surveillance. This trend mirrors recommended treatment strategies by expert panels and consensus guidelines.

The ability of SPEEK to promote the proliferation and osteogenic differentiation of BMSCs on PEEK surfaces

To investigate the ability of sulfonated polyetheretherketone (SPEEK) to promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and compare the effects of different degrees of sulfonation (DS), SPEEK was made with two different DS. The L-SPEEK group had a lower DS, while the H-SPEEK group had a higher DS. The physicochemical properties of both species were evaluated by scanning electron microscopy (SEM), capitilize Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), X-ray diffraction (XRD) and differential scanning calorimetry (DSC). Then, proliferation and osteogenic differentiation between the two groups and with pure polyetheretherketone (PEEK) were compared after surface inoculation of bone marrow mesenchymal stem cells (BMSCs). Scanning electron microscopy (SEM) revealed that the surface of the PEEK substrates could be smooth or coarse, and the degree of roughness increased with increasing sulfonation. FTIR spectroscopy showed that both the L-SPEEK and H-SPEEK samples contained sulfonic acid. TGA and XRD revealed that the components in the two groups were the same, but the intensities were different. After BMSC inoculation, a CCK8 assay revealed that the cells proliferated more on the H-SPEEK surface and little on the L-SPEEK surface compared with the PEEK surface. Then, osteogenic differentiation was verified by immunofluorescence staining for OCN and Runx2, which indicated that H-SPEEK had the greatest effect on improving differentiation. The results of alizarin red staining (ARS) and alkaline phosphatase staining (APS) also revealed this trend. Sulfonation can change the microsurface of PEEK, which can improve both BMSC proliferation and osteogenic differentiation.

P-349 Impact of an Aurora kinase inhibitor on AML cells and mouse ovarian follicles in vitro

Abstract Study question Can an aurora kinase B/C inhibitor suppress AML cell line proliferation while not impacting on mouse follicles in vitro? Summary answer Exposure to an aurora kinase inhibitor at a predetermined cytotoxic concentration suppressed AML cell proliferation but had no impact on subsequent follicle health in vitro. What is known already There is a risk that cryopreserved ovarian tissue from young women with a leukaemia diagnosis harbours leukemic cells, which on grafting could transfer disease. Novel chemotherapy agents, specifically aurora kinase inhibitors [ e.g. GSK 1070916 (GSK) ], target proteins involved in chromosomal alignment and segregation during mitosis and meiosis. It is anticipated that aurora kinase inhibitors would have an impact on the development of all somatic cells, including granulosa cells proliferating during follicle growth, however a previous study using ovarian tissue pieces indicated no impact on follicles. Study design, size, duration Primary and secondary follicles (diameter ≤ 100 µm) were manually dissected from adult mouse ovaries and embedded in an extracellular scaffold together with &amp;gt;1,000 AML cells. Individual follicles and AML cells were cultured for 7 days followed by exposure to GSK for 24 hours. GSK was removed by washing and the culture continued for another two days at which time cells were assessed for normal morphology and survival with live/dead staining (Calcein AM/ Ethidium Homodimer-1). Participants/materials, setting, methods The leukemic cell line OC1-AML-3 (DSMZ) was cultured in RPMI-1640 +10% FCS and routinely passaged as recommended. Cell proliferation and cytotoxicity were assessed using the Alamar Blue assay. 20% DMSO was used as a positive control in the cytotoxicity assays and live/dead evaluation. Follicles were cultured in αMEM, ITS, FSH, ascorbic acid and 10% FCS under oil in 5%CO2 : 95% air. Survival and growth were evaluated in two extracellular scaffolds; UltiMatrix and Hystem™-HP. Main results and the role of chance Follicle survival and growth was better (p &amp;lt; 0.05) in UltiMatrix (84.7%; 61/72) compared to Hystem™-HP (72.0%;77/107) with an average increase of 20 µm in diameter at day 7. When OCI-AML-3 cells (0.7 x105/ml) were exposed to a range of concentrations of GSK (10nM to 10µM) for 24hr in an in vitro cytotoxicity assessment, survival was reduced to 0% at a concentration of 10µM. The 20% DMSO positive control also resulted in 0% survival. However, in the 3- dimensional scaffold culture, exposure to 10 µM GSK for 24hr resulted in a mean survival of OCI-AML-3 cells of 27% (100 cells counted/ well) which was similar to the 20% DMSO positive control (32%) and significantly different to the negative (no GSK) control (95%; p &amp;lt; 0.01). Follicles exposed to GSK appeared alive, emitting a strong green fluorescence for both oocyte and granulosa cells, although some of the surface follicular cells were dead. Live/dead staining of GSK exposed follicles was similar to follicles not exposed to GSK. In contrast, the majority of the follicular cells were dead following exposure to 20% DMSO. Limitations, reasons for caution Due to whole mount assessment of the follicles for live/dead staining, it is difficult to determine whether the oocyte is alive. In scaffold culture some of the free cells present may be of follicular origin, potentially resulting in overestimation of the proportion of AML cells alive after GSK treatment. Wider implications of the findings This 3- dimensional culture system, incorporating a follicle and a leukemic cell line within a scaffold provides a good model to assess the impact of chemotherapy on both cell types and could be applicable for human follicles. Trial registration number Not applicable