Physiomimetic In Vitro Human Models for Viral Infection in the Liver.

Viral hepatitis is a leading cause of liver morbidity and mortality globally. The mechanisms underlying acute infection and clearance, versus the development of chronic infection, are poorly understood. In vitro models of viral hepatitis circumvent the high costs and ethical considerations of animal models, which also translate poorly to studying the human-specific hepatitis viruses. However, significant challenges are associated with modeling long-term infection in vitro. Differentiated hepatocytes are best able to sustain chronic viral hepatitis infection, but standard two-dimensional models are limited because they fail to mimic the architecture and cellular microenvironment of the liver, and cannot maintain a differentiated hepatocyte phenotype over extended periods. Alternatively, physiomimetic models facilitate important interactions between hepatocytes and their microenvironment by incorporating liver-specific environmental factors such as three-dimensional ECM interactions and co-culture with non-parenchymal cells. These physiologically relevant interactions help maintain a functional hepatocyte phenotype that is critical for sustaining viral hepatitis infection. In this review, we provide an overview of distinct, novel, and innovative in vitro liver models and discuss their functionality and relevance in modeling viral hepatitis. These platforms may provide novel insight into mechanisms that regulate viral clearance versus progression to chronic infections that can drive subsequent liver disease.

Signature quality attributes of CD146+ mesenchymal stem/stromal cells correlate with high therapeutic and secretory potency.

CD146+ bone marrow-derived mesenchymal stem/stromal cells (BM-MSCs) play key roles in the perivascular niche, skeletogenesis, and hematopoietic support; however, comprehensive evaluation of therapeutic potency has yet to be determined. In this study, in vitro inflammatory priming to crude human BM-MSCs (n = 8) captured a baseline of signature responses, including enriched CD146+ with coexpression of CD107aHigh , CXCR4High , and LepRHigh , transcriptional profile, enhanced secretory capacity, and robust immunomodulatory secretome and function, including immunopotency assays (IPAs) with stimulated immune cells. These signatures were significantly more pronounced in CD146+ (POS)-sorted subpopulation than in the CD146- (NEG). Mechanistically, POS BM-MSCs showed a markedly higher secretory capacity with significantly greater immunomodulatory and anti-inflammatory protein production upon inflammatory priming compared with the NEG BM-MSCs. Moreover, IPAs with stimulated peripheral blood mononuclear cells and T lymphocytes demonstrated robust immunosuppression mediated by POS BM-MSC while inducing significant frequencies of regulatory T cells. in vivo evidence showed that POS BM-MSC treatment promoted pronounced M1-to-M2 macrophage polarization, ameliorating inflammation/fibrosis of knee synovium and fat pad, unlike treatment with NEG BM-MSCs. These data correlate the expression of CD146 with innately higher immunomodulatory and secretory capacity, and thus therapeutic potency. This high-content, reproducible evidence suggests that the CD146+ (POS) MSC subpopulation are the mediators of the beneficial effects achieved using crude BM-MSCs, leading to translational implications for improving cell therapy and manufacturing.

Engineering anisotropic cardiac monolayers on microelectrode arrays for non-invasive analyses of electrophysiological properties.

A standard culture of cardiac cells as unorganized monolayers on tissue culture plastic or glass does not recapitulate the architectural or the mechanical properties of native myocardium. We investigated the physical and protein cues from the extracellular matrix to engineer anisotropic cardiac tissues as highly aligned monolayers on top of the microelectrode array (MEA). The MEA platform allows non-invasive measurement of beating rate and conduction velocity. The effect of different extracellular proteins was tested by using the most common extracellular matrix proteins in the heart, fibronectin and gelatin, after aligning myocytes using a microcontact (µC) printing technique. Both proteins showed similar electrophysiological results before the monolayer began to delaminate after the sixth day of culture. Additionally, there were no significant differences on day 4 between the two microcontact printed proteins in terms of sarcomere alignment and gap junction expression. To test the effect of substrate stiffness, a micromolded (µM) gelatin hydrogel was fabricated in different concentrations (20% and 2%), corresponding to the elastic moduli of approximately 33 kPa and 0.7 kPa, respectively, to cover both spectra of the in vivo range of myocardium. Cardiac monolayers under micromolded conditions beat in a much more synchronized fashion, and exhibited conduction velocity that was close to the physiological value. Both concentrations of gelatin hydrogel conditions yielded similar sarcomere alignment and gap junction expression on day 4 of culture. Ultimately, the 3D micromolded gelatin hydrogel that recapitulated myocardial stiffness improved the synchronicity and conduction velocity of neonatal rat ventricular myocytes (NRVM) without any stimulation. Identifying such microenvironmental factors will lead to future efforts to design heart on a chip platforms that mimic in vivo environment and predict potential cardiotoxicity when testing new drugs.

Survival of Implants after Indirect Maxillary Sinus Elevation Procedure: A Two Years Longitudinal Study.

AIM: The aim of the study was to evaluate the survival rate of two diverse implant systems with different implant surfaces with the same geometrical design. MATERIALS AND METHODS: One hundred fifty patients were included in the study in which 95 were males and 55 were females and 150 implants were placed using indirect sinus floor elevation technique and only one implant was placed in each subject and they were categorized into two groups of 100 in group A and 50 in group B as per two different implant systems. At review appointments, implants were tested clinically and radiographically and were examined for signs of infection. The patients were examined periodically after placement of the implants, and follow-up was conducted annually. RESULTS: Results of the Chi-square analysis showed no significant association between the type of implant surface and rate of success or failure of the implant. There was no significant difference between the observed and expected frequency of successful implants in group A as well as group B, indicating that the surface type of implant had no significant association with the success of the implant in group A and B. CONCLUSION: To date, there is no consensus in the literature regarding the best surface and even on the macrotopography of the implants for better osseointegration. However, Surface treatments improve the result of osseointegration, especially in the early stages, benefiting bone affixation with qualitative and quantitative enhancements. In the present study, we achieved clinical success with both kinds of implant surfaces however Bioetched implant surface showed promising results comparable to Tiunite surface of Nobel BioCare Implants. In the future, more case-controlled studies with longer follow-up are needed to validate the results of the present findings.

Prevalence of oral squamous cell carcinoma in Bareilly Region: A seven year institutional study.

BACKGROUND: Oral squamous cell carcinoma is a major contributor to disability and death caused by malignant tumors. Variations in social, cultural, and geographic factors affect the tumor behavior and response to treatment. In this study, we undertake a seven years institutional review and analysis of Oral squamous cell carcinoma cases in Bareilly. MATERIALS AND METHODS: A total of 1938 histologically diagnosed cases during the period of seven years i.e. from 2010 to 2016 were extracted from the archives of Department of Oral & Maxillofacial Pathology, Institute of Dental Sciences, Bareilly and evaluated to know the prevalence of Oral squamous cell carcinoma. The details like age, sex, habits and anatomical site were also recorded from the archived patient's case sheets. RESULTS: Out of the total 1938 cases, 318 were found to be Oral squamous cell carcinoma of which 232 were males and 86 were females. Incidence was highest in 40-49 year age group. The most common site was buccal mucosa (190 cases), followed by tongue (68 cases). Most lesions were well differentiated Oral squamous cell carcinoma (230 cases). Patients with poorly differentiated lesions had a comparatively lower mean age than their counterparts with other histological varieties. CONCLUSION: The pattern of Oral squamous cell carcinoma differs from that of previous studies in relation to incidence and age correlation with the grade of carcinoma. The majority of the lesions were well differentiated. There is a need for intensive oral health awareness to encourage early presentation to cancer center as early detection will further enhance prognosis.

Velocity Fluctuations in Kinesin-1 Gliding Motility Assays Originate in Motor Attachment Geometry Variations.

Motor proteins such as myosin and kinesin play a major role in cellular cargo transport, muscle contraction, cell division, and engineered nanodevices. Quantifying the collective behavior of coupled motors is critical to our understanding of these systems. An excellent model system is the gliding motility assay, where hundreds of surface-adhered motors propel one cytoskeletal filament such as an actin filament or a microtubule. The filament motion can be observed using fluorescence microscopy, revealing fluctuations in gliding velocity. These velocity fluctuations have been previously quantified by a motional diffusion coefficient, which Sekimoto and Tawada explained as arising from the addition and removal of motors from the linear array of motors propelling the filament as it advances, assuming that different motors are not equally efficient in their force generation. A computational model of kinesin head diffusion and binding to the microtubule allowed us to quantify the heterogeneity of motor efficiency arising from the combination of anharmonic tail stiffness and varying attachment geometries assuming random motor locations on the surface and an absence of coordination between motors. Knowledge of the heterogeneity allows the calculation of the proportionality constant between the motional diffusion coefficient and the motor density. The calculated value (0.3) is within a standard error of our measurements of the motional diffusion coefficient on surfaces with varying motor densities calibrated by landing rate experiments. This allowed us to quantify the loss in efficiency of coupled molecular motors arising from heterogeneity in the attachment geometry.

Removal of biofilms by intermittent low-intensity ultrasonication triggered bursting of microbubbles.

In this study, a chemical-free cleaning method for biofilms removal is presented, which is based on intermittent low-intensity ultrasonication (US) triggered bursting of microbubbles (MB) in such a sequence that MB were continuously introduced into the reaction vessel for 15 min, while US was activated for 2 s after every 2 min of microbubbling. It was found that the fixed biomass, and the extracellular proteins and polysaccharides of 24-h old biofilms grown on a nylon membrane surface were reduced, respectively, by 75, 79 and 72% after treatment by the US + MB method. Fourier transform infrared (FTIR) analysis further revealed that the chemical composition of the biofilms was not altered by the US + MB treatment, suggesting that biofilms were removed through physical forces due to the generation of a shock wave and a high-speed water jet through US-triggered bursting of the MB. The proposed method can be considered a chemical-free technology for biofilm removal.

LiCoO2 impregnated nano-hierarchical ZSM-5 assisted catalytic upgrading of Kraft lignin-derived liquefaction bio-oil.

In this study, Kraft lignin-derived bio-oil was upgraded with LiCoO2 or Co3O4-impregnated hierarchical nano-ZSM-5 catalysts. The synthesized catalysts were characterized by N2-Ads-Des, XRD, XPS, NH3-TPD, FTIR, FESEM and ICP-OES analyses. Upon incorporation of LiCoO2 and Co3O4 onto the HZSM-5 support, the MFI structure of HZSM-5 remained intact. All the catalysts displayed a combination of Type-I and -IV isotherms. The upgraded bio-oil showed a significant increase in the amounts of alkylated guaiacols owing to the reduction in unsubstituted guaiacols, alkenyl guaiacols, and homovanillic acid. Hydrogenation, alkylation, and deoxygenation were the plausible bio-oil upgrading pathways. With the increase in cobalt content, weak acidity decreased through all the catalysts, while LiCoO2 provided supplementary acid sites that increased the total acidity of LiCoO2/HZSM-5 compared to the Co3O4/HZSM-5 catalyst. LiCoO2/HZSM-5 with a low cobalt content (5% and 10% Co) displayed high selectivity for the production of alkylated guaiacols owing to their strong acidity. The upgraded bio-oils showed an increase in carbon and hydrogen followed by a decrease in oxygen content. The maximum higher heating value (∼29.83 MJ kg-1) was obtained for the 10% Co (LiCoO2)/HZSM-5 catalyst. In general, LiCoO2/HZSM-5 outperformed the Co3O4/HZSM-5 catalyst. XRD of the spent 10% Co (LiCoO2)/HZSM-5 suggested the complete loss of lithium from the catalyst with the retention of the MFI structure of the HZSM-5 support. In this study, it was successfully demonstrated that the main constituent of the cathode material of spent lithium-ion batteries i.e. LiCoO2 could be employed to synthesize a novel and cheap catalyst for bio-oil upgrading while addressing the e-waste management issue in a sustainable manner.

Roadblocks confronting widespread dissemination and deployment of Organs on Chips.

Organ on Chip platforms hold significant promise as alternatives to animal models or traditional cell cultures, both of which poorly recapitulate human pathophysiology and human level responses. Within the last 15 years, we have witnessed seminal scientific developments from academic laboratories, a flurry of startups and investments, and a genuine interest from pharmaceutical industry as well as regulatory authorities to translate these platforms. This Perspective identifies several fundamental design and process features that may act as roadblocks that prevent widespread dissemination and deployment of these systems, and provides a roadmap to help position this technology in mainstream drug discovery.

Comparative Evaluation of Octenidine with Chlorhexidine Mouthwash in Gingivitis and Periodontitis Patients: A Randomized Clinical Trial.

Introduction: Chlorhexidine gluconate (CHX) has been still regarded as the most efficient mouthwash. Due to its recognized negative effects, it can only be used for a short duration. Octenidine dihydrochloride (OCT), an antiseptic substance found by Sterling Winthrop in the 1980s, has proven helpful in preventing the co-aggregation of dental plaque microbial invaders without disrupting the typical, healthy oral flora. However, there is very little research on octenidine's effectiveness as a mouthwash for preventing plaque. Therefore, this study is being conducted to examine the effectiveness of mouthwashes containing 0.1% Octenidine and 0.2% chlorhexidine. Methodology: In this clinical trial, subjects were divided into two groups, 60 patients each with gingivitis and periodontitis. Then, from each group, 30 patients were advised to use octenidine mouthwash, and 30 patients were prescribed chlorhexidine mouthwash as an adjunct to scaling and root planning. Clinical parameters like O'Leary plaque index, Bleeding index, Probing pocket depth, and clinical attachment loss were evaluated at baseline and after 3 months. Result: In both the gingivitis and periodontitis groups, the octenidine group significantly outperformed the chlorhexidine group in all clinical metrics. Conclusion: Octenidine showed better results in comparison to chlorhexidine with respect to all the above-mentioned clinical parameters. Hence, it can be considered a promising mouthwash for future therapeutic and research studies.

Exploring Anthocyanins: Unveiling Sources, Analytical Techniques, and Biological Activities.

Anthocyanins (ANCs) are obtained from pigmented foods like blueberry, strawberry, and red cabbage and are phenolic compounds belonging to the flavonoids family. ANCs have garnered significant attention in recent years due to their diverse biological activities and potential health benefits. This comprehensive review presents a holistic exploration of anthocyanins, spanning from their chemical structure and biosynthesis pathways to the myriad analytical techniques employed for their identification and quantification. Furthermore, the rich tapestry of plant sources yields anthocyanins is delved into, highlighting their incorporation into various pharmaceutical formulations. This review aims to provide a comprehensive synthesis of current knowledge on anthocyanins, spanning from their origins in nature to their multifaceted pharmacological activities and innovative pharmaceutical applications.

SliceChip: a benchtop fluidic platform for organotypic culture and serial assessment of human and rodent pancreatic slices.

Enzymatically isolated pancreatic islets are the most commonly used ex vivo testbeds for diabetes research. Recently, precision-cut living slices of human pancreas are emerging as an exciting alternative because they maintain the complex architecture of the endocrine and exocrine tissues, and do not suffer from the mechanical and chemical stress of enzymatic isolation. We report a fluidic pancreatic SliceChip platform with dynamic environmental controls that generates a warm, oxygenated, and bubble-free fluidic pathway across singular immobilized slices with continuous deliver of fresh media and the ability to perform repeat serial perfusion assessments. A degasser ensures the system remains bubble-free while systemic pressurization with compressed oxygen ensures slice medium remains adequately oxygenated. Computational modeling of perfusion and oxygen dynamics within SliceChip guide the system's physiomimetic culture conditions. Maintenance of the physiological glucose dependent insulin secretion profile across repeat perfusion assessments of individual pancreatic slices kept under physiological oxygen levels demonstrated the culture capacity of our platform. Fluorescent images acquired every 4 hours of transgenic murine pancreatic slices were reliably stable and recoverable over a 5 day period due to the inclusion of a 3D-printed bioinert metallic anchor that maintained slice position within the SliceChip. Our slice on a chip platform has the potential to expand the useability of human pancreatic slices for diabetes pathogenesis and the development of new therapeutic approaches, while also enabling organotypic culture and assessment of other tissue slices such as brain and patient tumors.

Detection of microplastics in the human penis.

The proliferation of microplastics (MPs) represents a burgeoning environmental and health crisis. Measuring less than 5 mm in diameter, MPs have infiltrated atmospheric, freshwater, and terrestrial ecosystems, penetrating commonplace consumables like seafood, sea salt, and bottled beverages. Their size and surface area render them susceptible to chemical interactions with physiological fluids and tissues, raising bioaccumulation and toxicity concerns. Human exposure to MPs occurs through ingestion, inhalation, and dermal contact. To date, there is no direct evidence identifying MPs in penile tissue. The objective of this study was to assess for potential aggregation of MPs in penile tissue. Tissue samples were extracted from six individuals who underwent surgery for a multi-component inflatable penile prosthesis (IPP). Samples were obtained from the corpora using Adson forceps before corporotomy dilation and device implantation and placed into cleaned glassware. A control sample was collected and stored in a McKesson specimen plastic container. The tissue fractions were analyzed using the Agilent 8700 Laser Direct Infrared (LDIR) Chemical Imaging System (Agilent Technologies. Moreover, the morphology of the particles was investigated by a Zeiss Merlin Scanning Electron Microscope (SEM), complementing the detection range of LDIR to below 20 µm. MPs via LDIR were identified in 80% of the samples, ranging in size from 20-500 µm. Smaller particles down to 2 µm were detected via SEM. Seven types of MPs were found in the penile tissue, with polyethylene terephthalate (47.8%) and polypropylene (34.7%) being the most prevalent. The detection of MPs in penile tissue raises inquiries on the ramifications of environmental pollutants on sexual health. Our research adds a key dimension to the discussion on man-made pollutants, focusing on MPs in the male reproductive system.

Micropatterning Alginate Substrates for in vitro Cardiovascular Muscle on a Chip.

Soft hydrogels such as alginate are ideal substrates for building muscle in vitro because they have structural and mechanical properties close to the in vivo extracellular matrix (ECM) network. However, hydrogels are generally not amenable to protein adhesion and patterning. Moreover, muscle structures and their underlying ECM are highly anisotropic, and it is imperative that in vitro models recapitulate the structural anisotropy in reconstructed tissues for in vivo relevance due to the tight coupling between sturcture and function in these systems. We present two techniques to create chemical and structural heterogeneities within soft alginate substrates and employ them to engineer anisotropic muscle monolayers: (i) microcontact printing lines of extracellular matrix proteins on flat alginate substrates to guide cellular processes with chemical cues, and (ii) micromolding of alginate surface into grooves and ridges to guide cellular processes with topographical cues. Neonatal rat ventricular myocytes as well as human umbilical artery vascular smooth muscle cells successfully attach to both these micropatterned substrates leading to subsequent formation of anisotropic striated and smooth muscle tissues. Muscular thin film cantilevers cut from these constructs are then employed for functional characterization of engineered muscular tissues. Thus, micropatterned alginate is an ideal substrate for in vitro models of muscle tissue because it facilitates recapitulation of the anisotropic architecture of muscle, mimics the mechanical properties of the ECM microenvironment, and is amenable to evaluation of functional contractile properties.

Cleaning of biologically fouled membranes with self-collapsing microbubbles.

Increasing global demand for water reclamation has driven the widespread development of membrane processes, where membrane biofouling due to microbial attachment onto membrane surfaces remains the biggest challenge. Given the potential ability of microbubbles (MBs) to generate pressure waves upon collapse, the responses of biofilms of different ages on nylon membrane surfaces towards self-collapsing air MBs was investigated. Changes in the fixed biomass, extracellular polysaccharides and proteins were determined for 3-h, 12-h, 18-h and 24-h old biofilms before and after treatment with MBs. The resistance-in-series model was further applied for analysis of various resistances after treatment of biofilms of different ages with MBs. The results showed substantial flux recovery 1 h after MB treatment for stationary phase biofilms in comparison with initial and exponential growth phase biofilms, which was consistent with the relatively larger percentage reduction in fixed biomass, extracellular polysaccharides and proteins for stationary phase biofilms. However, pore blocking still remained a big challenge even after treatment with MBs, regardless of biofilm age. The experimental data were further supported by confocal laser scanning microscope images. Collapsing MBs appear to be an alternative green chemical-free technology for mitigation of membrane biofouling.

Nanoscale transport enables active self-assembly of millimeter-scale wires.

Active self-assembly processes exploit an energy source to accelerate the movement of building blocks and intermediate structures and modify their interactions. A model system is the assembly of biotinylated microtubules partially coated with streptavidin into linear bundles as they glide on a surface coated with kinesin motor proteins. By tuning the assembly conditions, microtubule bundles with near millimeter length are created, demonstrating that active self-assembly is beneficial if components are too large for diffusive self-assembly but too small for robotic assembly.

Post-COVID mucormycosis in presentation of periodontal abscess.

COVID-19 pandemic has traumatized deep inside in minds and lives of human beings. Those who have survived are at risk of many post-COVID complications; mucormycosis being one of the most common as well as morbid infections. Mucormycosis is also known as black fungus, it is a life-threatening opportunistic fungal infection. Inoculation occurs by inhalation of spores in nose, paranasal air sinuses and in lungs. Mucormycosis is often associated in patients with compromised immunity and it presents with characteristic black necrotic eschar and necrosis. Disease affecting the facial region possess a challenge because it often disseminates with sino-orbital and cranial involvement at the time of diagnosis. Dental practice occasionally encounters unusual and subtle symptoms with non-pathognomonic clinical signs of several fatal diseases which may pretend like a common oral disease. Hence, the key knowledge to oral and primary care physicians and its precise application is necessary for the early diagnosis of such fatal infections to prevent untoward consequences in this COVID era. This article presents a case of mucormycosis with chief complaint of pain and swelling in right front tooth region, which mimicked as periodontal abscess in a patient, leading to delay in the diagnosis possess greater challenges during the treatment.

Spermatogonial Stem Cells and In Vitro Spermatogenesis: How Far Are We from a Human Testis on a Chip?

An in vitro testis model will provide a superior platform for studying the testis microenvironment and molecular mechanisms that affect male fertility. The ultimate aim is to provide reproductive hope for children diagnosed with cancer who were sterilized by aggressive gonadotoxic therapies.

Immunohistochemical expression of paxillin in ameloblastoma and odontogenic keratocyst: An observational study.

Background: Cell adhesion molecules (CAMs) are found on the surface of all cells, where they allow dynamic processes to take place. These include cadherins, integrins, selectins and Immunoglobulin superfamily. Directly associated with ß-integrin tails is a multidomain protein known as paxillin. However, CAMs participate in cell-cell and extracellular matrix-cell interactions during histomorphogenesis in the various phases of odontogenesis. Some tumours or cysts like ameloblastoma (AB) or odontogenic keratocyst (OKC) having odontogenic origin show disturbance in the interaction of these CAMs. Hence, the assessment of paxillin expression in AB and OKC was carried out. Materials and Methods: The present observational study comprised 30 clinically and histologically confirmed cases of AB and OKC. All the slides were stained immunohistochemically using a paxillin antibody. Results: Upon comparison of staining intensity of paxillin among AB and OKC showed statistically significant result, whereas quantitative staining and final summation showed non-significant result. Gender-wise comparison of paxillin staining intensity, quantitative staining and final summation among OKC showed significant result; however, in AB, staining intensity showed non-significant result, whereas quantitative staining and final summation showed significant result. Conclusion: Paxillin has the greatest influence on tissue morphogenesis and development. The regulation of cell mobility is aided by the multiple roles that paxillin plays in a range of cells and tissues. However, further studies using a large sample size, along with other molecular analytical methods, may be essential to draw a definite conclusion about the association of paxillin and its exact function in OKC and AB.

Bioinspired bi-phasic 3D nanoflowers of MgO/Mg(OH)2 coated melamine sponge as a novel bactericidal agent.

By roughly mimicking the surface architectural design of dragonfly wings, novel bi-phasic 3D nanoflowers of MgO/Mg(OH)2 were successfully synthesized via the electrospinning technique. The 3D nanoflowers were coated over a commercial melamine sponge and extensively characterized by SEM, XRD, FTIR, and EDS. The formation of distinct dense 3D nano petals was revealed by SEM images whereby the mean petal thickness and mean distance between the adjacent petals were found to be 36 nm and 121 nm, respectively. The bactericidal activities of synthesized 3D nano-flowers coated melamine sponges were assessed against five different bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa). This study demonstrated significant bactericidal activity of MgO/Mg(OH)2 3D nanoflowers coated MS against Gram-positive and Gram-negative bacteria. Plausible bactericidal mechanisms include envelope deformation, penetration, and induction of oxidative stress. This study introduces novel bioinspired biomaterial with the capacity to reduce the risk associated with pathogenic bacterial infections, especially in medical devices.