Proteomic Characterization of Human Placenta: Insights into Potential Therapeutic Applications for Osteoarthritis.

Biologics have become increasingly prominent as therapeutics in recent years due to their innate immune-privileged nature, biocompatibility, and high levels of protein biofactors. The aim of the study is to characterise the biologic, lyophilized human placenta (LHP) and explore its therapeutic potential for osteoarthritis (OA). The presence of six bioactive constituents that regulate cell-extracellular matrix interaction was identified by liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS). Metalloproteinase inhibitor 3 (TIMP3), alpha-1 anti-trypsin (a1AT), basic fibroblast growth factor (bFGF), and transforming growth factor ß1 (TGFß1) were detected and quantified using ELISA. The total protein content present in LHP by Bradford assay was found to be 409.35 ± 0.005 µg/ml. The analytical techniques such as Attenuated Total Reflectance-Fourier Transform Infrared spectroscopy (ATR-FTIR), solid state carbon-13 Nuclear Magnetic Resonance (ssC13 NMR) spectroscopy, and Differential Scanning Calorimetry (DSC) revealed the secondary structure and conformational stability of LHP. X-Ray diffraction (XRD) studies showed its amorphous nature. Bioactivity assessment of LHP was performed in human keratinocytes (HaCaT) and human dermal fibroblasts (HDF) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The LHP was highly proliferative against skin cells and non-toxic, based on the findings of the bioactivity assay. LHP has the potential to be used as a therapeutic agent for OA, as its characterisation unveiled its physical stability, significant concentration of bioactive components that are pertinent to cartilage repair and its conformational stability.

Progress towards blue emitting MgO-ZnO-Ga2O3 nanocomposites synthesized by bio mediated route.

MgO-ZnO-Ga2O3 nanocomposites are synthesized by solution combustion method using Aloe Vera gel as a reducing agent to increase the efficiency of blue emission. The appearance of Bragg reflections corresponding to MgO, ZnO and Ga2O3 clearly indicates the formation of nanocomposites. The surface morphology consists irregular shape and sized NPs. The Energy dispersive X-ray analysis confirms the purity of the sample. The band energy gap was tuned to 3.1 eV. The Photoluminescence excitation and emission spectra was discussed and compared it with emission spectra of individual oxides as well as with other reported blue emitted nanophosphors. Further, the chromaticity coordinates and Color correlated temperature coordinates clearly confirms their warm blue emission. Further, the powder dusting method was employed to collect the latent fingerprints on the pores and non-pores surfaces. The synthesized MgO-ZnO-Ga2O3 nanocomposites exhibits well-resolved ridge patterns that can be used to identify latent finger prints with clarity. From all these results, the present synthesized MgO-ZnO-Ga2O3 nanocomposite might find an application in display technology as a blue nanophosphor material and for latent finger print detection in crime investigation.

X-ray/gamma radiation shielding properties of zinc ferrite nanoparticles synthesised via solution combustion method.

Zinc ferrite (ZnFe2O4) nanoparticles were synthesised via solution combustion method using urea as a fuel. The synthesised samples were characterised with various techniques. The cubic structure with Fd-3 m space group is confirmed by Powder X-ray Diffraction and Bragg's reflection. The crystallite size estimated from Scherrer's method was found to be 40 nm. The agglomerated irregular shape and sized surface morphology was confirmed by Scanning Electron Microscopy image. The direct energy band gap determined from Wood and Tauc's relation was found to be 5.25 eV. Using a NaI (Tl) detector and multi-channel analyser, the described sample was examined for X-ray and gamma ray shielding characteristics in the energy range of 0.081-1.332 MeV. The measured shielding values are in good agreement with the theory, however below 356 keV, there is a little variation of up to 10%. The current work offers up new possibilities for using this simple, affordable, effective and low temperature approach to create nanomaterials for X-ray and gamma ray shielding.

Red emission of copper aluminate synthesized via chemical and bio-mediated routes.

For the first time, copper aluminate nanoparticles (NPs) are synthesized by a combustion method using urea as a fuel (CAOU) and Ocimum sanctum (tulsi) extract as a reducing agent (CAOT). The Bragg reflections of the as-formed product confirm the formation of a cubic phase with Fd3̄m space group. The crystallite size, crystallinity and other structural parameters are discussed. The surface morphology of CAOU is agglomerated in nature whereas that of CAOT is hexagonal in shape. The smaller crystallite size CAOT NPs show a higher energy band gap. The photoluminescence (PL) analysis excited at 302 nm shows that the CIE coordinates fall in the red region. The oxygen defects are mainly responsible for the PL emission. The CCT coordinates confirm that both CAOU and CAOT NPs can find an application in warm light emitting diodes.

Effect of Eu3+ doping on structural and optical properties of zirconium titanate.

The Europium activated (1-9 mol %) Zirconium Titanate nanoparticles (NPs) have been synthesized by the green solution combustion method using Aloe Vera gel extract as a reducing agent, followed by the calcination at 720 °C for 3hrs. All the synthesized samples crystallize in a pure orthorhombic crystal structure with the space group of Pbcn. The surface and bulk morphology were analyzed. The crystallite size increases, whereas the direct energy band gap was found to decrease with an increase in dopant concentration. Further, the effect of dopant concentration on the photoluminescence properties was studied. The presence of Eu3+ ion in the trivalent state in the host lattice was confirmed by its characteristic emission at 610 nm due to 5D0→7F2 (λex = 464 nm). The CIE coordinates were found in the red region of the CIE 1931 diagram. The CCT coordinates lie in the range 6288-7125 K. The Judd-Ofelt parameters and derived quantities were analyzed. This theory confirms the high symmetry of Eu3+ ions in the host lattice. These findings imply that ZTO:Eu3+ can be employed as a nanopowder material in a red-emitting phosphor material.

Photoluminescence, antibacterial, X-ray/gamma ray absorption, supercapacitor and sensor applications of ZrTiO4 nanorods.

In the present communication, ZrTiO4 nanoparticles (NPs) are synthesized by the solution combustion method using urea (ZTOU) and oxalyl dihydrazide (ODH) (ZTODH) as fuel and calcined at 700 °C. The synthesized samples were characterized with different techniques. Powder X-ray diffraction studies show the presence of diffraction peaks corresponding to ZrTiO4. In addition to these peaks, a few additional peaks corresponding to the monoclinic and cubic phases of ZrO2 and the rutile phase of TiO2 are observed. The surface morphology of ZTOU and ZTODH consists of nanorods with different lengths. The TEM and HRTEM images confirm the formation of nanorods along with NPs, and the estimated crystallite size matches well with that of PXRD. The direct energy band gap was calculated using Wood and Tauc's relation and was found to be 2.7 and 3.2 eV for ZTOU and ZTODH respectively. The photoluminescence emission peaks (λ = 350 nm), CIE and CCT of ZTOU and ZTODH clearly confirm that the present nanophosphor might be a good nanophosphor material for blue or aqua green light emitting diodes. Furthermore, antibacterial activity and a viability test were conducted on two food borne pathogens. The X-ray/gamma ray absorption properties are also studied, which clearly show the ZrTiO4 might be a good absorbing material. Furthermore, cyclic voltammetry (CV) analysis of ZTOU nanorods shows very good redox peaks compared to that of ZTODH. From the electrochemical impedance spectroscopy (EIS) measurements, the charge-transfer resistances for prepared nanorods ZTOU and ZTODH are found to be 151.6 Ω, and 184.5 Ω respectively. The modified graphite electrode with ZTOU shows good sensing activity for both paracetamol and ascorbic acid, compared to ZTODH.

The effect of human amniotic epithelial cells on urethral stricture fibroblasts.

BACKGROUND: Urethral stricture disease (USD) is effectively managed by buccal mucosa (BM) urethroplasty. Lack of adequate healthy BM has led to the use of autologous tissue-engineered BM grafts. Such grafts are costly, not easily scalable and recurrence of the stricture is still a problem. Hence, there is a requirement for cost-effective, scalable cells with innate antifibrotic properties which seem to be fulfilled by human amniotic epithelial cells (HAMECs). The effect of HAMECs on USD is unknown. AIM: To study the effect of HAMECs-CM on human urethral stricture fibroblast (USF) cells by using in-vitro migration assay and molecular techniques. MATERIALS AND METHODS: USF cells were derived from six patients undergoing urethroplasty. HAMECs were derived from one placenta after delivery. The effect of HAMECs-CM on USF cell migration was observed using a standard in vitro scratch assay over a period of 3 days. The effect of HAMECs-CM on the expression levels of markers alpha-smooth muscle actin (α-SMA) and tissue inhibitor of metalloproteinases (TIMP-1) in USF cells was also examined. RESULTS: The HAMECs-CM suppressed the migration of USF cells in in vitro scratch assay. The HAMECs-CM consistently downregulated α-SMA, but not TIMP-1. CONCLUSIONS: HAMECs have shown antifibrotic activity on USF cells in this in vitro study. RELEVANCE FOR PATIENTS: HAMECs could serve as an alternative cell source for tissue-engineered urethroplasty.

Large-scale Isolation, Expansion and Characterization of Human Amniotic Epithelial Cells.

BACKGROUND AND OBJECTIVES: The human Amniotic epithelial cells (AME) derived from amniotic membrane of placenta have been considered as the potential fetal stem cell source with minimal or no ethical concerns and are important therapeutic tool for anti-fibrotic and regenerative therapies. METHODS AND RESULTS: Here, we evaluated the isolation, media screening, scale-up and characterization of AME cells. The isolation, expansion of AMEs were performed by sequential passaging and growth kinetics studies. The AMEs were characterized using immunocytochemistry, immunophenotyping, In-vitro differentiation, and anti-fibrotic assays. The growth kinetics study revealed that the AME cultured in Ultraculture (UC) and DMEM knockout (DMEM-KO) have prominently higher growth rate compared to others. Overall, the AMEs cultured from 5 different media retained basic morphological characteristics and the functional characteristics. CONCLUSIONS: Our result suggests that the AMEs can be successfully cultured in UC based complete media without losing its epithelial cell characteristics even after passaging for passage 2 (P2). However, a careful and methodical pre-clinical and clinical translation studies need to be conducted to show its safety and efficacy.

Gamma, X-ray and neutron shielding parameters for the Al-based glassy alloys.

The X-ray and gamma radiation shielding parameters (mass attenuation coefficient, mean free path, half value layer, tenth value layer, effective atomic numbers, electron density, exposure buildup factors, relative dose, dose rate and specific gamma ray constant) have been studied for the Al-based glassy alloys Al86Y7Ni5Co1Fe0.5Pd0.5, Al85Y8Ni5Co1Fe0.5Pd0.5, Al84Y9Ni4Co1.5Fe0.5Pd1, Al80Y13Ni5Co1Fe0.5Pd0.5, Al70Y23Ni5Co1Fe0.5Pd0.5 and Al60Y33Ni5Co1Fe0.5Pd0.5. For the same alloys, the neutron shielding parameters (coherent neutron scattering length, incoherent neutron scattering lengths, coherent neutron scattering cross section, incoherent neutron scattering cross sections, total neutron scattering cross section and neutron absorption cross sections) have also been explored. Al60Y33Ni5Co1Fe0.5Pd0.5 was found to be a good shielding material for the X-ray/gamma radiation, while Al86Y7Ni5Co1Fe0.5Pd0.5 is a good shielding material for neutrons.

Comparative study of isolation, expansion and characterization of epithelial cells.

BACKGROUND AIMS: The human epithelial cells (EPCs) have been identified as the essential element for the regeneration of skin construct for burns, wounds and various tissue engineer-based products. METHODS: In this study, the isolation, expansion and characterization of EPCs from various sources such as juvenile foreskin (JSK), buccal mucosa (BM), penile skin (PS) and urothelium (UR) in serum-free and xeno-free EpiLife media were evaluated. RESULTS: The growth kinetics study revealed that EPCs from JSK and BM had notably higher growth rates compared with the others. Overall, the EPCs from all sources retained basic morphological characteristics and the functional characteristics such as Pan Cytokeratin (AE1/AE3). In addition, the cryopreservation stability of EPCs was accessed for post-thaw viability and found to be greater than 80% at 1 year of storage, but demonstrated reduced cell recovery (51%) at the second year in fetal bovine serum-free cryopreservation media. CONCLUSIONS: Our result suggests that the EPCs from four cell sources can be grown in feeder-free, serum-free and xeno-free systems using commercially available EpiLife medium without losing epithelial cell characteristics even after passage 4. However, its suitability for clinical application must be accessed by preclinical and clinical studies.

Absorption of glucose from urinary conduits in diabetics and non-diabetics.

The ability of urinary conduits to absorb glucose was investigated. A solution containing 1% glucose and the inert marker phenol red was put into conduits in diabetic and non-diabetic patients. Samples were withdrawn at intervals for analysis and the fall in concentrations plotted against time. The loops in the patients without diabetes did not absorb glucose, but glucose was absorbed in one of the three diabetics. Ileal loops in patients with diabetes may absorb urinary glucose. This should be checked in any diabetic with urinary diversion before advice is given on long term management. Routine testing of conduit urine for glucose may not be an adequate screen for diabetes in patients with urinary diversions.

Prostatic infiltration in leukaemia and lymphoma.

12 cases are reported in which the prostate was the site of a leukaemic or lymphomatous infiltration. Hodgkin's disease, not previously reported as involving the prostate, was seen in one instance. In all cases the diagnosis was made histologically. Although 5 patients were known to have leukaemia or lymphoma prior to surgery, the preoperative clinical diagnosis in 3 cases was benign prostatic hypertrophy and in 3 instances prostatic carcinoma. In 6 patients, the initial diagnosis of leukaemia or lymphoma was made on the prostatic specimen and in one instance infiltration was discovered at postmortem. The management of leukaemic and lymphomatous infiltration is discussed.

Ruptured renal artery aneurysm in a solitary kidney.

The incidence of renal artery aneurysm in a solitary kidney has not been previously documented in the literature. A case of aneurysmal rupture into a pelvicalyceal system of a solitary kidney, successfully treated by primary vascular reconstruction under the cover of inosine and synchronous pelviureteric repair, is reported.

Percutaneous nephroscopy: a review of the physical properties, suitability and cost of the instruments.

The physical properties, suitability and cost of six instruments used for elective percutaneous renal endoscopy have been evaluated. Three were rigid and three were flexible. The optics, flow rate and accessories of the rigid instruments have been demonstrated to be superior to the flexible instruments. Flexible instruments, however, were used in 15% of our 70 elective percutaneous operations. The 21-Charr cystoscope was adequate for extracting smaller calculi, but it was necessary to use the Storz and Wolf rigid nephroscopes when ultrasonic disruption was required. Based on our data, improvements and modifications to both flexible and rigid instrument design are suggested.