Protective Role of Nanoceria-Infused Nanofibrous Scaffold toward Bone Tissue Regeneration with Senescent Cells.

The presence of oxidative stress in bone defects leads to delayed regeneration, especially in the aged population and patients receiving cancer treatment. This delay is attributed to the increased levels of reactive oxygen species (ROS) in these populations due to the accumulation of senescent cells. Tissue-engineered scaffolds are emerging as an alternative method to treat bone defects. In this study, we engineered tissue scaffolds tailored to modulate the adverse effects of oxidative stress and promote bone regeneration. We used polycaprolactone to fabricate nanofibrous mats by using electrospinning. We exploited the ROS-scavenging properties of cerium oxide nanoparticles to alleviate the high oxidative stress microenvironment caused by the presence of senescent cells. We characterized the nanofibers for their physical and mechanical properties and utilized an ionization-radiation-based model to induce senescence in bone cells. We demonstrate that the presence of ceria can modulate ROS levels, thereby reducing the level of senescence and promoting osteogenesis. Overall, this study demonstrates that ceria-infused nanofibrous scaffolds can be used for augmenting the osteogenic activity of senescent progenitor cells, which has important implications for engineering bone tissue scaffolds for patients with low regeneration capabilities.

Cellular Senescence Program is Sensitive to Physical Differences in Polymeric Tissue Scaffolds.

A typical cellular senescence program involves exposing cells to DNA-damaging agents such as ionization radiation or chemotherapeutic drugs, which cause multipronged changes, including increased cell size and volume, the onset of enhanced oxidative stress, and inflammation. In the present study, we examined if the senescence onset decision is sensitive to the design, porosity, and architecture of the substrate. To address this, we generated a library of polymeric scaffolds widely used in tissue engineering of varied stiffness, architecture, and porosity. Using irradiated A549 lung cancer cells, we examined the differences between cellular responses in these 3D scaffold systems and observed that senescence onset is equally diminished. When compared to the two-dimensional (2D) culture formats, there were profound changes in cell size and senescence induction in three-dimensional (3D) scaffolds. We further establish that these observed differences in the senescence state can be attributed to the altered cell spreading and cellular interactions on these substrates. This study elucidates the role of scaffold architecture in the cellular senescence program.

A study to estimate the serum IgA and salivary IgA levels in patients with oral leukoplakia and oral squamous cell carcinoma.

Context: The increasing death rate because of oral cancer is mainly due to its late diagnosis. Tumour markers are often detected in abnormal amounts in blood, urine or saliva of patients with certain types of cancer. Diagnosing cancer through human saliva has advantages such as low invasiveness, minimum cost and easy sample collection. We have used serum immunoglobulin A (IgA) and salivary IgA for our present study. Aims: The aim of present study was to estimate serum and salivary IgA levels in oral leukoplakia and oral squamous cell carcinoma (OSCC) patients. Settings and Design: The study included 40 patients; 10 in the control group, 15 cases with oral leukoplakia and 15 cases with OSCC. Methods and Material: The blood samples and saliva were taken from clinically diagnosed oral leukoplakia and OSCC patients and were tested for IgA levels. Statistical analysis used: The data were analysed using SPSS 16.0. The mean values were compared between the groups by using analysis of variance (ANOVA) followed by post-hoc test for group-wise comparison. P value ≤ 0.05 was considered significant. Results: It was observed that the comparison of levels of serum IgA in control and leukoplakia group; control and OSCC group; leukoplakia and OSCC group were found to be statistically significant. Also, comparison between the levels of salivary IgA in control and OSCC group was found to be statistically significant. Conclusion: It is suggested that the serum and salivary IgA levels could be a better adjuvant diagnostic marker along with routine markers in patients with premalignant and malignant lesions.

A Study of the Degree of Gall Bladder Wall Thickness and Its Impact on Patients Undergoing Laparoscopic Cholecystectomy.

Background The gold standard management for symptomatic gallstone disease is elective laparoscopic cholecystectomy, which has replaced open cholecystectomy. The wall thickness of the gallbladder is an indicator of cholecystitis in patients who have presented with symptoms of gallstone disease. The aim of this study was to evaluate preoperative gall bladder wall thickness by ultrasonography and assess its impact on the outcome of laparoscopic cholecystectomy, including conversion rate, complications, operative time, and postoperative hospital stay. Method This prospective study was conducted on 350 patients with symptomatic gallstone disease, those who had undergone laparoscopic cholecystectomy in Dr. Sampurnanand Medical College, Jodhpur, and attached hospitals from July 2019 to November 2021. On the basis of ultrasonography findings of gallbladder wall thickness, patients were divided into four groups: normal thickness - up to 2 mm, mild thickness - 3-4mm, moderate thickness - 5-6mm, and severe wall thickness - more than 6mm). Up to 2 millimeters thickness was considered as normal. Results The incidence of conversion rate, as well as intra or postoperative complications, were higher in moderate and severe wall thickness groups. The maximum incidence of complication rate is seen in moderately thickened group (33.33%). In severely thickened group, complication was seen in 100% of patients. Operative time, as well as postoperative hospital stay, were more in higher thickness groups. There was a statistically significant correlation between gallbladder wall thickness and conversion rate, complications operative time, and postoperative length of stay. Conclusion Increased gallbladder wall thickness causes increased intra as well as postoperative complications, more conversion to open procedure rate, increased operative time, and enhanced postoperative hospital stay. Among the total study population, 29.71% of patients had increased gallbladder wall thickness. In our study, a positive correlation was seen among gallbladder wall thickness, complication rate, conversion rate, intraoperative time, and postoperative hospital stay.

Light-based 3D bioprinting of bone tissue scaffolds with tunable mechanical properties and architecture from photocurable silk fibroin.

Three-dimensional (3D) bioprinting based on digital light processing (DLP) offers unique opportunities to prepare scaffolds that mimic the architecture and biomechanical properties of human tissues. Limited availability of biocompatible and biodegradable bioinks amenable for DLP-bioprinting is an impediment in this field. This study presents a bioink prepared from silk fibroin (SF) tailored for DLP bioprinting. Photocurable methacrylated-SF (SF-MA) was synthesized with 67.3% of methacrylation. Physical characterization of rheological and mechanical properties revealed that the 3D printed hydrogels of SF-MA (spanning from 10 to 25 wt%) exhibit bone tissue-like viscoelastic behavior and compressive modulus ranging from ≈12 kPa to ≈96 kPa. The gels exhibited favorable degradation (≈48 to 91% in 21 days). This SF-MA bioink afforded the printing of complex structures, with high precision. Pre-osteoblasts were successfully encapsulated in 3D bioprinted SF-MA hydrogels with high viability. 15% SF-MA DLP bioprinted hydrogels efficiently supported cell proliferation with favorable cell morphology and cytoskeletal organization. A progressive increase in cell-mediated calcium deposition up to 14 days confirmed the ability of the gels to drive osteogenesis, which was further augmented by soluble induction factors. This work demonstrates the potential of silk fibroin-derived bioinks for DLP-based 3D bioprinting of scaffolds for tissue engineering.

Senescent cells in 3D culture show suppressed senescence signatures.

Cellular senescence, an irreversible proliferation arrested but viable cellular state, has been implicated in the progression of several age-associated pathologies. A vast amount of information about senescence has been acquired in cultured cells; however, senescence in living organisms (in vivo) remains poorly understood, mainly because of technical limitations. Furthermore, it is now widely recognized that three-dimensional (3D) culture systems are a better mimic of the in vivo physiology. Herein, senescence was induced in HeLa cells by irradiation. Non-senescent or senescent cells were cultured in soft 3D polymer scaffolds and compared with cells in conventional two-dimensional (2D) culture. This work shows that the morphology of the senescent cells markedly varies between substrates/culture platforms, driving the differences in the cytoskeletal organization, cellular division, and nanomechanical properties. One characteristic feature of senescent cells on 2D culture systems is the enlarged and flattened morphology; however, such drastic changes are not seen in vivo. This is an artificial effect of the substrate, which renders such non-physiological morphology to senescent cells. In the 3D scaffolds, this artifact is reduced. Hence, it serves as a better mimic of tissues, leading to reduced expression of senescence-associated genes, implying that the 3D scaffolds suppress the senescence in cells.

Computational approach to study the synthesis of noscapine and potential of stereoisomers against nsP3 protease of CHIKV.

Chikungunya fever is a major public health issue in India affecting millions of people and occurs due to Chikungunya. Chikungunya virus (CHIKV) is a single stranded RNA virus from the family of Togaviridae and genus alpha virus. It contain three structural proteins: glycosylated E1 and E2, embedded in the viral envelope, and a non-glycosylated nucleocapsid protein. Till date, researchers are working on inhibition of CHIKV but till now no cheap and effective medicine is available in the market. Therefore, the authors of this work thought of isoquinoline based noscapine to inhibit the nsP3 protease of CHIKV. The aim of the work is to understand the mechanism for the synthesis of noscapine theoretically using DFT. Further study the potential of all four isomers of noscapines {(13 (S,R), 14 (R,R), 15 (R,S) and 16 (S,S)} against nsP3 protease of CHIKV with the help of docking and MD simulation. The integrated e-pharmacophore binding affinity based virtual screening, docking and molecular dynamics simulation recognized four hits isomers as inhibition nsP3 protease of CHIKV. The docking energies of all the isomers of noscapine (13-16) with nsP3 protease CHIKV was found out to be more negative than baicalin (-8.06 kcal/mol) on selected sites. Amongst the isomers of noscapine, CMPD 13 possessed best binding affinity with four hydrogen bonding interactions. Further, ADME properties and blood-brain barrier permeability properties have been calculated. DFT studies of all the isomers of noscapine was investigated.

Development of optically sensitive liver cells.

Optogenetics is a new and emerging field that involves techniques of optics and genetic engineering to influence cellular functionality. In this work, we have successfully incorporated a non-selective cationic channel channelrhodopsin-2 (ChR2) into human hepatocellular carcinoma (HepG2) cells. A plasmid construct AAV-CAG-ChR2-GFP was used for liposomal transfection into the cells. ChR2 is a light sensitive membrane channel that opens upon illumination with blue light. Plasmid DNA isolation from E. coli XL 10 gold bacteria by alkaline lysis method resulted in a DNA concentration of 1150 µg/mL. A significant difference (p < 0.05) was observed between the fluorescent intensities of transfected cells and the control. The percentage of transfected cells was estimated to be 41.26%. Overall, the study delivers an optimized methodology to produce the transfected HepG2 cells that can be controlled with the light stimulation. Although ChR2 has mostly been associated with excitable cells, we anticipate that its presence into HepG2 cells may also result changes in biological functionalities by modulating the concentration of cations inside the cell. Furthermore, the transfected HepG2 cells can be co-cultured with fibroblasts such as NIH 3T3 to form liver spheroids that can serve as models for toxicological and pharmacological studies.

The impact of Bakri balloon tamponade on the rate of postpartum hysterectomy for uterine atony.

OBJECTIVE: Our objective was to evaluate the impact of uterine tamponade with a Bakri balloon on the rate of postpartum hysterectomy due to uterine atony. METHODS: We performed a retrospective cohort study of all deliveries >20 weeks gestation from January 2002 to March 2013 at Baystate Medical Center. Charts were reviewed to determine incidence of postpartum hysterectomy, Bakri balloon placement, uterine artery embolization (UAE) and the B-Lynch procedure. Patients with evidence of placenta accreta were excluded. The primary outcome was the change in rates of postpartum hysterectomy for uterine atony before and after the introduction of Bakri balloon tamponade, using chi-square testing. RESULTS: There were 48 767 deliveries during the study period, with 17 950 before and 30 817 after the introduction of the Bakri balloon. A total of 43 Bakri balloons were placed during the study period and 21 hysterectomies were performed for postpartum hemorrhage secondary to uterine atony, 14 before and 7 after the introduction of the Bakri balloon. This was consistent with a decrease in the rate of postpartum hysterectomy from 7.8/10 000 deliveries to 2.3/10 000 deliveries (p = 0.01). CONCLUSION: Our findings show that utilization of the Bakri balloon is associated with a decreased rate of postpartum hysterectomy.

Minimally invasive diagnosis and treatment of endometrial cancer after LeFort colpocleisis.

BACKGROUND: Endometrial carcinoma is rare after LeFort colpocleisis. Standards for its diagnosis and treatment have not been established. CASE: A 74-year-old woman presented with postmenopausal bleeding 14 months after LeFort colpocleisis. Here, we describe the use of the colpocleisis channels in our novel 2-stage approach. In the first stage, endometrial carcinoma was diagnosed with vaginohysteroscopy and dilatation and curettage via the channels. In the second stage, the cancer was optimally treated with total robotic hysterectomy, bilateral salpingo-oophorectomy, and lymph node dissection. Assistance and specimen retrieval were achieved through the vaginal channels. The patient recovered without compromise to the pelvic floor. CONCLUSIONS: Endometrial cancer after LeFort colpocleisis can be diagnosed and treated with minimally invasive approaches without disrupting the colpocleisis or the pelvic floor support.

Imaging of trauma in a pregnant patient.

Trauma is the number one nonobstetric cause of maternal death. This chapter presents the latest consensus from the literature on the best approach to radiographic imaging of these patients. The central issues of discussion include the rationale and protocols for screening for pregnancy in trauma setting; the effects of radiation and its risks to the fetus; obtaining informed consent; how to estimate fetal dose; and the role of ultrasound, computed tomography, and magnetic resonance imaging, including the intravenous contrast agents used for the assessment of abdominal trauma. The team approach to the management of these patients is also highlighted.