Aligned Fibroporous Matrix Generated from a Silver Ion and Graphene Oxide-Incorporated Ethylene Vinyl Alcohol Copolymer as a Potential Biomaterial for Peripheral Nerve Repair.

Developing an ideal nerve conduit for proper nerve regeneration still faces several challenges. The attempts to fabricate aligned substrates for neuronal growth have enhanced the hope of successful nerve regeneration. In this wok, we have attempted to generate an electrospun matrix with aligned fibers from a silver and graphene oxide-incorporated ethylene vinyl alcohol copolymer (EVAL). The presence of silver was analyzed using UV-visible spectra, XPS spectra, and ICP. Raman spectra and FTIR spectra confirmed the presence of GO. The complexation of Ag+ with - OH of EVAL enabled the generation of aligned fibers. The fiber diameter (>1 µm) provided sufficient space for forming focal adhesion by the neurites and filopodia of N2a and C6 cells, respectively. The fiber diameter enabled the neurites and filopodia of the cells to align on the fibers. The incorporation of GO has contributed to the cell-material interactions. The morphological and mechanical properties of fibers obtained in the study ensure that the EVAL-Ag-GO-0.01 matrix is a potential substrate for developing a nerve guidance conduit/nerve wrap (NGC/W).

Evaluation of the efficiency of estrus synchronization protocols combined with natural service and ultrasonography on ewe reproductive performance during non-breeding season.

Background: Estrus synchronization is an important assisted reproductive technology to improve the reproductive performance in ewes. Various protocols have been used with variable success rates, however; literature regarding field applicable estrus synchronization is meagre. Aims: The present study was designed with the aim to evaluate the estrus synchronization protocols on reproductive performance in ewes using different hormones. Methods: Experimental ewes were divided randomly into three groups (n=15). Ewes of all groups received intravaginal sponge for 12 days. Subsequently, NP4-GnRH and NP4-eCG groups received 8 µg of buserelin acetate or 200 IU of eCG intramuscularly, respectively on day 12 whereas NP4-Insulin group received insulin 0.2 IU/kg body weight subcutaneously for three consecutive days started on the day of sponge removal. Estrus detection commenced 24 h after sponge removal in NP4-GnRH and NP4-eCG groups and 24 h following last injection of insulin in NP4-Insulin group. The ewes in estrus were separated and pen mated. The conception rate was determined by ultrasonography. Results: The estrus response and conception rates were 71.43, 92.86 and 53.85%, and 70.00, 84.61, and 71.43%, respectively in NP4-GnRH, NP4-eCG, and NP4-Insulin groups. The lambing rates were the same as the conception rates. The single and multiple birth rates were 71.41, 36.36 and 60.0%, and 28.57, 63.64, and 40.0% whereas prolificacy was 128.57, 190.91, and 140.00%, respectively in NP4-GnRH, NP4-eCG, and NP4-Insulin groups. Conclusion: In conclusion, the estrus synchronization protocol including intravaginal progesterone sponge and eCG was found to be more effective under field conditions.

Evaluation of allylated gelatin as a bioink supporting spontaneous spheroid formation of HepG2 cells.

The spheroid culture system has gained significant attention as an effective in vitro model to mimic the in vivo microenvironment. Even though numerous studies were focused on developing spheroids, the structural organization of encapsulated cells within hydrogels remains a challenge. Allylated gelatin or GelAGE is used as a bioink due to its excellent physicochemical properties. In this study, GelAGE was evaluated for its capacity to induce spontaneous spheroid formation in encapsulated HepG2 cells. GelAGE was synthesized and characterized using 1HNMR spectroscopy and ninhydrin assay. Then the physicochemical and biological attributes of GelAGE hydrogel was examined. The results demonstrate that GelAGE has remarkable ability to induce the encapsulated cells to self-organize into spheroids.

Silymarin enriched gelatin methacrylamide bioink imparts hepatoprotectivity to 3D bioprinted liver construct against carbon tetrachloride induced toxicity.

Three-dimensional liver bioprinting is an emerging technology in the field of regenerative medicine that aids in the creation of functional tissue constructs that can be used as transplantable organ substitutes. During transplantation, the bioprinted donor liver must be protected from the oxidative stress environment created by various factors during the transplantation procedure, as well as from drug-induced damage from medications taken as part of the post-surgery medication regimen following the procedure. In this study, Silymarin, a flavonoid with the hepatoprotective properties were introduced into the GelMA bioink formulation to protect the bioprinted liver against hepatotoxicity. The concentration of silymarin to be added in GelMA was optimised, bioink properties were evaluated, and HepG2 cells were used to bioprint liver tissue. Carbon tetrachloride (CCl4) was used to induce hepatotoxicity in bioprinted liver, and the effect of this chemical on the metabolic activities of HepG2 cells was studied. The results showed that Silymarin helps with albumin synthesis and shields liver tissue from the damaging effects of CCl4. According to gene expression analysis, CCl4 treatment increased TNF-α and the antioxidant enzyme SOD expression in HepG2 cells while the presence of silymarin protected the bioprinted construct from CCl4-induced damage. Thus, the outcomes demonstrate that the addition of silymarin in GelMA formulation protects liver function in toxic environments.

In vitro Cytotoxicity Evaluation of Flowable Hyaluronic Acid-Acellular Stromal Vascular Fraction (HA-aSVF) Mixture for Tissue Engineering Applications.

Background: The stromal vascular fraction (SVF) is an aqueous fraction isolated from the adipose tissue that constitutes different kinds of cells and extracellular matrix components. Hyaluronic acid (HA) is a linear polysaccharide in vertebrate tissues and is considered a potential tissue engineering scaffold due to its biocompatible nature. In this study, we have evaluated the cytotoxicity of xenofree HA in combination with an acellular component of adipose SVF (HA-aSVF) to propose it as a candidate biomaterial for future applications. Materials and Methods: 3-(4,5-dimethyl thiazolyl-2)-2,5-diphenyltetrazolium bromide assay of L-929 cells treated with HA-aSVF was used in our study. Data were normalized to cell control (untreated) and extracts of copper and ultra-high molecular weight polyethylene were used as positive (PC) and negative controls (NC). Results: Fibroblast cells retained the morphology after 24 h of treatment with HA-aSVF mixture and exhibited a similar percentage of cell activity compared to NC. PC showed a positive cytotoxic response as expected. The cells incubated with HA-aSVF showed a linear increase in cell activity indicating proliferation. Conclusion: The mixture of HA and acellular SVF in its flowable form is non-cytotoxic and showed improved cell proliferation. Hence the mixture can be proposed as a biomaterial and can be further explored for specific tissue engineering applications.

Non-linear dose response of DNA double strand breaks in response to chronic low dose radiation in individuals from high level natural radiation areas of Kerala coast.

BACKGROUND: The human population living in high level natural radiation areas (HLNRAs) of Kerala coast provide unique opportunities to study the biological effects of low dose and low dose rate ionizing radiation below 100 mGy. The level of radiation in this area varies from < 1.0 to 45 mGy/year. The areas with ≤ 1.50 mGy/year are considered as normal level natural radiation areas (NLNRA) and > 1.50 mGy/year, as high level natural radiation areas (HLNRA). The present study evaluated dose response relationship between DNA double strand breaks (DSBs) and background radiation dose in individuals residing in Kerala coast. Venous blood samples were collected from 200 individuals belonging to NLNRA (n = 50) and four dose groups of HLNRA; 1.51-5.0 mGy/year (n = 50), 5.01-10.0 mGy/year (n = 30), 10.01-15.0 mGy/year (n = 33), > 15.0 mGy/year (n = 37) with written informed consent. The mean dose of NLNRA and four HLNRA dose groups studied are 1.21 ± 0.21 (range: 0.57-1.49), 3.02 ± 0.95 (range: 1.57-4.93), 7.43 ± 1.48 (range: 5.01-9.75), 12.22 ± 1.47 (range: 10.21-14.99), 21.64 ± 6.28 (range: 15.26-39.88) mGy/year, respectively. DNA DSBs were quantified using γH2AX as a marker, where foci were counted per cell using fluorescence microscopy. RESULTS: Our results revealed that the frequency of γH2AX foci per cell was 0.090 ± 0.051 and 0.096 ± 0.051, respectively in NLNRA and HLNRA individuals, which were not significantly different (t198 = 0.33; P = 0.739). The frequency of γH2AX foci was observed to be 0.090 ± 0.051, 0.096 ± 0.051, 0.076 ± 0.036, 0.087 ± 0.042, 0.108 ± 0.046 per cell, respectively in different dose groups of ≤ 1.50, 1.51-5.0, 5.01-10.0, 10.01-15.0, > 15.0mGy/year (ANOVA, F4,195 = 2.18, P = 0.072) and suggested non-linearity in dose response. The frequency of γH2AX foci was observed to be 0.098 ± 0.042, 0.078 ± 0.037, 0.084 ± 0.042, 0.099 ± 0.058, 0.097 ± 0.06 and 0.114 ± 0.033 per cell in the age groups of ≤ 29, 30-34, 35-39, 40-44, 45-49 and ≥ 50 years, respectively (ANOVA, F5,194 = 2.17, P = 0.059), which suggested marginal influence of age on the baseline of DSBs. Personal habits such as smoking (No v/s Yes: 0.092 ± 0.047 v/s 0.093 ± 0.048, t198 = 0.13; P = 0.895) and drinking alcohol (No v/s Yes: 0.096 ± 0.052 v/s 0.091 ± 0.045, t198 = 0.62; P = 0.538) did not show any influence on DSBs in the population. CONCLUSION: The present study did not show any increase in DSBs in different dose groups of HLNRA compared to NLNRA, however, it suggested a non-linear dose response between DNA DSBs and chronic low dose radiation.

Phytochemical and biological review of Aegle marmelos Linn.

India has one of the most expanded plant-origin medical traditions in the world. Researchers have evaluated molecules obtained from plants to treat a variety of ailments. Literature review shows that fundamental parts of the plant are used to treat different diseases. The related data is retrieved from Google scholar, PubMed, Science Direct and Scopus. The keywords include Bael, A. marmelos, Vilvam, and Marmelosin. Extensive studies show that A. marmelos has antidiarrhoeal, antimicrobial, antiviral, anticancer, chemopreventive, antipyretic, ulcer healing, antigenotoxic, diuretic, antifertility, and anti-inflammatory properties. In this work, an updated literature review is presented to clarify the current state of research on A. marmelos elucidating its constituents and their most relevant biological activities.

India has one of the most expanded plant-origin medical traditions in the world. A. marmelos Linn, also familiar as bael, belongs to Rutaceae and is widely grown worldwide. A. marmelos is a fruit with various medicinal advantages. We searched various databases, studied elaborately, and understood the importance of this fruit. Thus, its constituents can help mitigate various diseases.

Energy system digitization in the era of AI: A three-layered approach toward carbon neutrality.

The transition toward carbon-neutral electricity is one of the biggest game changers in addressing climate change since it addresses the dual challenges of removing carbon emissions from the two largest sectors of emitters: electricity and transportation. The transition to a carbon-neutral electric grid poses significant challenges to conventional paradigms of modern grid planning and operation. Much of the challenge arises from the scale of the decision-making and the uncertainty associated with the energy supply and demand. Artificial intelligence (AI) could potentially have a transformative impact on accelerating the speed and scale of carbon-neutral transition, as many decision-making processes in the power grid can be cast as classic, though challenging, machine-learning tasks. We point out that to amplify AI's impact on carbon-neutral transition of the electric energy systems, the AI algorithms originally developed for other applications should be tailored in three layers of technology, markets, and policy.

Quality of Life and Depression Assessment in Patients with Acute Coronary Syndrome: A Cross-Sectional Study.

PURPOSE: Acute Coronary Syndrome (ACS) is currently the leading cause of death in industrialized countries. Morbidity after ACS includes physical and mental disorders affecting the patient's whole life situation and Quality of Life (QoL). The main aim of the study was to assess QoL and depression among post-ACS patients. METHODS: This was a cross-sectional observational study. A total of 112 patients who fulfilled the inclusion criteria were included in this study. A semi-structured questionnaire was administered to the patients to collect data from the patients. In this study, men and women aged 18 - 80 with ACS; patients diagnosed with Non-ST Segment Elevated Myocardial Infarction (NSTEMI) or STEMI or Angina Pectoris were included. Patients with severe mental, and physical illness and dementia were excluded from the study. QoL and depression assessment was done by RAND 36-Item Health Survey and Hamilton Depression Rating scale, respectively. RESULTS: Among 112 post-ACS patients, 78 patients were males, and 38 patients were females. The mean age of the study population was 64.25 ± 9.029 and with most individuals in the category of 61 - 70 years. The majority of study populations were married (71.5%) and lived with their families (92.9%). In this study group, 42.9 % of the population reported at least high school education (SSLC) and 54.5% were full-time employees. Most of the patients (91.1%) were taking the medication regularly, while 55.4% of patients reported regular compliance with the follow-up. This study identified that, among various factors, older age, female gender, lower income, unemployment, low education status, poor compliance with medication, and depressive symptoms led to poor QoL. CONCLUSION: This study confirms a negative correlation between depressive symptoms and QoL. This study's results reveal the magnitude of depression that is prevalent in the primary health care clinic that goes undiagnosed and unmanaged. Hence, it is recommended to properly screen depressive symptoms in ACS patients. Therefore, concurrently, better QoL can be achieved by managing both depression and ACS.

Anticancer potential of rhizome extract and a labdane diterpenoid from Curcuma mutabilis plant endemic to Western Ghats of India.

Zingiberaceae plants are well known for their use in ethnomedicine. Curcuma mutabilis Skornick., M. Sabu & Prasanthk., is an endemic Zingiberaceae species from Western Ghats of Kerala, India. Here, we report for the first time, the anticancer potential of petroleum ether extract from C. mutabilis rhizome (CMRP) and a novel labdane diterpenoid, (E)-14, 15-epoxylabda-8(17), 12-dien-16-al (Cm epoxide) isolated from it. CMRP was found to be a mixture of potent bioactive compounds including Cm epoxide. Both the extract and the compound displayed superior antiproliferative activity against several human cancer cell lines, without any display of cytotoxicity towards normal human cells such as peripheral blood derived lymphocytes and erythrocytes. CMRP treatment resulted in phosphatidylserine externalization, increase in the levels of intracellular ROS, Ca2+, loss of mitochondrial membrane potential as well as fragmentation of genomic DNA. Analyses of transcript profiling and immunostained western blots of extract-treated cancer cells confirmed induction of apoptosis by both intrinsic and extrinsic pathways. The purified compound, Cm epoxide, was also found to induce apoptosis in many human cancer cell types tested. Both CMRP and the Cm epoxide were found to be pharmacologically safe in terms of acute toxicity assessment using Swiss albino mice model. Further, molecular docking interactions of Cm epoxide with selected proteins involved in cell survival and death were also indicative of its druggability. Overall, our findings reveal that the endemic C. mutabilis rhizome extract and the compound Cm epoxide isolated from it are potential candidates for development of future cancer chemotherapeutics.

Spondias pinnata (L.f.) Kurz Leaf Extract Derived Zinc Oxide Nanoparticles Induce Dual Modes of Apoptotic-Necrotic Death in HCT 116 and K562 Cells.

This study evaluates the effects of phyto-derived zinc oxide nanoparticles (ZnONPs) on human cancer cells, colon carcinoma HCT 116, and chronic myelogenous leukemic K562, along with normal lymphocytes/erythrocytes. The commercial, chemically synthesized ZnONPs (cZnONPs) were also assessed in parallel. Using an eco-friendly approach devoid of harmful chemicals, biogenic nanoparticles were synthesized from aqueous leaf extract of Spondias pinnata (SpLZnONPs) by a sol-gel method. Optical, structural, and elemental characterization of both particle types were carried out deploying UV-Vis/photoluminescence spectroscopy, FTIR, XRD, FESEM, HRTEM, and EDX. Both SpLZnONPs and cZnONPs displayed hexagonal wurtzite structure with particle sizes averaging 30 and 48.5 nm, respectively. SpLZnONPs were found to be cytotoxic to both cancer cell types while cZnONPs exhibited toxicity only against HCT 116 cells. Interestingly, the cytomorphological changes and analysis of DNA laddering pattern observed in these treated cells were indicative of simultaneous induction of dual modes of death involving apoptosis and necrosis. Flow cytometric analysis of cell-cycle distribution, clonogenic, wound healing, and comet assays provided evidences of the antiproliferative potential of the tested nanoparticles. Apoptosis induction via oxidative stress-mediated Ca2+ release, ROS generation, loss of mitochondrial membrane potential, and externalization of phosphatidylserine was also determined biochemically. Relative expression of apoptotic genes was quantified using RT-qPCR and Western blot analysis. Mitotic index analysis, MTT, and hemolytic assays on lymphocytes and erythrocytes clearly revealed the absence of any deleterious effect(s) of SpLZnONPs in these cells compared with the toxicity of the chemically derived cZnONPs, thereby attesting to the biocompatibility and selective action of the biogenic nanoparticles.

Peripheral Blood As a Source of Stem Cells for Regenerative Medicine: Emphasis Towards Corneal Epithelial Reconstruction-An In Vitro Study.

BACKGROUND: Mesenchymal stem cell-based treatments are now emerging as a therapy for corneal epithelial damage. Although bone marrow, adipose tissue and umbilical cord blood are the main sources of mesenchymal stem cells (MSCs), other tissues like the peripheral blood also harbor mesenchymal-like stem cells called peripheral blood-derived mononuclear cells (PBMNCs). These blood derived stem cells gained a lot of attention due to its minimally invasive collection and ease of isolation. In this study, the feasibility of using PBMNCs as an alternative cell source to corneal limbal stem cells envisaging corneal epithelial regeneration was evaluated. METHODS: Rabbit PBMNCs were isolated using density gradient centrifugation and was evaluated for mesenchymal cell properties including stemness. PBMNCs were differentiated to corneal epithelial lineage using rabbit limbal explant conditioned media and was evaluated by immuno-cytochemistry and gene expression analysis. Further, the differentiated PBMNCs were engineered into a cell sheet using an in-house developed thermo-responsive polymer. RESULTS: These blood derived cells were demonstrated to have similar properties to mesenchymal stem cells. Corneal epithelial lineage commitment of PBMNCs was confirmed by the positive expression of CK3/12 marker thereby demonstrating the aptness as an alternative to limbal stem cells. These differentiated cells effectively generated an in vitro cell sheet that was then demonstrated for cell sheet transfer on an ex vivo excised rabbit eye. CONCLUSION: PBMNCs as an alternative autologous cell source for limbal stem cells is envisaged as an effective therapeutic strategy for corneal surface reconstruction especially for patients with bilateral limbal stem cell deficiency.

Evaluation of the influence of chronic low-dose radiation on DNA repair gene polymorphisms [XRCC1, XRCC3, PRKDC (XRCC7), LIG1, NEIL1] in individuals from normal and high level natural radiation areas of Kerala Coast.

Background: Single Nucleotide Polymorphisms (SNPs) at DNA repair genes are considered as potential biomarkers of radio-sensitivity. The coastal belt of Kerala in south west India has a patchy distribution of monazite in its beach sand that contains Th-232 and its decay products. Thus, radiation levels in this area vary from <1.0mGy to 45.0mGy/year. The areas with external gamma radiation dose >1.5mGy/year are considered as High-Level Natural Radiation Areas (HLNRA) and ≤ 1.5mGy/year are Normal Level Natural Radiation Area (NLNRA).Objective: In the present study, an attempt was made to evaluate the influence of chronic low dose radiation exposure on DNA repair gene polymorphisms in NLNRA and HLNRA population of Kerala coast.Materials and methods: Genomic DNA was isolated from venous blood samples of 246 random, healthy individuals (NLNRA, N = 104; HLNRA, N = 142) and genotyping of five SNPs such as X-ray repair cross complementing 1(XRCC1 Arg399Gln), X-ray repair cross complementing 3 (XRCC3 Thr241Met], Protein kinase, DNA-activated, catalytic subunit (PRKDC) (X-ray repair cross-complementing group 7, XRCC7 G/T), nei like DNA glycosylase 1 (NEIL1 G/T) and DNA ligase 1 (LIG1 A/C) was carried out using PCR based restriction fragment length polymorphism (PCR-RFLP) followed by silver staining.Results: Our results showed no significant difference in genotype frequencies in HLNRA vs NLNRA at three of the five SNPs studied i.e. XRCC1 Arg399Gln (χ2(2) = 5.85, p = .054), XRCC3 Thr241Met (χ2(1) = 0.71, p = .339), PRKDC (XRCC7 G/T) (χ2(2) = 3.72, p = .156), whereas significant difference was observed at NEIL1 G/T (χ2(2) =8.71, p = .013) and LIG1 A/C (χ2(2) = 7.66, p = .022). The odds of heterozygote to homozygote genotypes in HLNRA relative to NLNRA at XRCC1 Arg399Gln (OR = 1.96, 95% CI: 1.13-3.40), XRCC3 Thr241Met (OR = 0.73, 95% CI: 0.41-1.31), PRKDC (XRCC7 G/T), (OR = 0.81; 95% CI: 0.48-1.38), NEIL1 G/T (OR = 0.54; 95% CI: 0.31-0.96) and LIG1 A/C (OR = 1.62; 95% CI: 0.97-2.69) was also not significantly different in HLNRA vs NLNRA, except at XRCC1 and NEIL1.Conclusion: The genotype frequencies at three of these SNPs i.e. XRCC1 Arg399Gln, XRCC3 Thr241Met and PRKDC (XRCC7 G/T) were similar, whereas NEIL1 G/T and LIG1 A/C showed significant difference between HLNRA and NLNRA population. However, further research using more number of SNPs in a larger cohort is required in this study area.

Premature chromosome condensation assay to study influence of high-level natural radiation on the initial DNA double strand break repair in human G0 lymphocytes.

The inherent capacity of individuals to efficiently repair ionizing radiation induced DNA double strand breaks (DSBs) may be inherited, however, it is influenced by several epigenetic and environmental factors. A pilot study tested whether chronic low dose natural radiation exposure influences the rejoining of initial DNA DSBs induced by a 2 Gy γ-irradiation in 22 individuals from high (>1.5 mGy/year) and normal (≤1.5 mGy/year) level natural radiation areas (H&NLNRA) of Kerala. Rejoining of DSBs (during 1 h at 37 °C, immediately after irradiation) was evaluated at the chromosome level in the presence and absence of wortmannin (a potent inhibitor of DSB repair in normal human cells) using a cell fusion-induced premature chromosome condensation (PCC) assay. The PCC assay quantitates DSBs in the form of excess chromosome fragments in human G0 lymphocytes without the requirement for cell division. A quantitative difference was observed in the early rejoining of DNA DSBs between individuals from HLNRA and NLNRA, with HLNRA individuals showing a higher (P = 0.05) mean initial repair ratio. The results indicate an influence of chronic low dose natural radiation on initial DNA DSB repair in inhabitants of HLNRA of the Kerala coast.

Chemical composition, antioxidant and antiproliferative activities of essential oil from rhizome and leaves of Curcuma mutabilis Skornick., M. Sabu & Prasanthk., endemic to Western Ghats of India.

Hydro-distilled essential oils, from fresh rhizomes and leaves of Curcuma mutabilis Skornick., M.Sabu & Prasanthk., characterized by GC-MS revealed the presence of thirty three and twenty three compounds therein respectively. Whilst estrone methyl ether (3-Methoxyestra-1,3,5(10)-trien-17-one) was the major component in rhizome oil (47.35%), sesquiterpene hydrocarbons predominated as the major group (63.92%) in leaf oil with a higher preponderance of ß-caryophyllene (25.48%), ß-farnesene (19.47%) and α-humulene (11.01%). Weak antioxidant activities observed in these oils determined by DPPH and ABTS methods were apparently influenced both by the oil composition and the assay conditions. Rhizome oil showed higher antiproliferative activity than leaf oil against leukemic K562 (IC50-6.8µg/mL) and colorectal HCT116 (IC50-8.5µg/mL) cancer cell lines. This first report reveals composition and biological activities of essential oils from C. mutabilis.

Bioengineered corneal epithelial cell sheet from mesenchymal stem cells-A functional alternative to limbal stem cells for ocular surface reconstruction.

Limbal stem cell deficiency (LSCD) is the loss of limbal stem cells that reside in the corneoscleral junction resulting in vision loss or blindness. Bilateral LSCD is usually treated by allogeneic corneal transplantation, with instances of tissue rejection or failure in long-term follow-up. This study aims to use adipose mesenchymal stem cells (ASC) as an alternative autologous cell source for treating bilateral limbal deficiency conditions. ASCs derived from rabbit fat tissue were differentiated into corneal epithelial lineage using limbal explant condition media. Apart from transdifferentiation, ASC sheets were developed to facilitate effective delivery of these cells to the damage site. A thermoresponsive polymer N-isopropylacrylamide-co-glycidylmethacrylate (NGMA) was synthesized and characterized to demonstrate ASC sheet formation. Transdifferentiated ASCs showed positive expression of corneal epithelial marker CK3/12 on immunostaining, supported by gene expression studies. in vivo studies by transplanting cell sheet in rabbit models of corneal injury showed clear and smooth cornea in comparison to the sham models. Histology revealed a sheet of cells aligned and integrated on to the injured corneal surface, 1 month posttransplantation. Identifying ASCs as an alternative cell source along with cell sheet technology will be a novel step in the field of corneal surface therapies.

Calcium sulfate-based bioactive cement for periodontal regeneration: An In Vitro study.

BACKGROUND AND OBJECTIVE: Various types of osteoconductive graft materials are used for the management of alveolar bone defects arising out of periodontal disease. Inorganic, self-setting, bioactive bone cements are suggested to be most appropriate because they can conformally fill the bone defect and resorb progressively along with the regeneration of the host site. A new calcium sulfate-based bioactive bone cement (BioCaS) is developed, having simplicity and effectiveness for bone grafting applications. The response of primary human periodontal ligament (hPDL) cells to this material is investigated through in vitro cell culture model so as to qualify it for the repair of periodontal infrabony defects. METHOD: The BioCaS was designed as powder-liquid combination with in-house synthesized high purity calcium sulfate hemihydrate incorporating hydrogen orthophosphate ions. hPDL cells were isolated, cultured and characterized using optimized primary cell culture techniques. The cytotoxicity and cytocompatibility of the BioCaS samples were evaluated using the hPDL cells, with hydroxyapatite ceramic material as control. Osteogenic differentiation of the hPDL cells in presence of BioCaS was also evaluated using Alizarin red staining, Alizarin red assay, Von Kossa staining and Masson's trichrome staining. RESULTS: The primary cell culture techniques yielded a healthy population of periodontal ligament cells, with fibroblast morphology and characteristic marker expressions. The hPDL cells exhibited good viability, adhesion and spreading to the BioCaS cement in comparison to sintered hydroxyapatite. In addition, the cells differentiated to osteogenic lineage in the presence of the BioCaS cement, without extraneous osteogenic supplements, confirming the inherent bioactivity of the cement. CONCLUSION: The new BioCaS cement is a potential candidate for the repair of periodontal defects.

Manilkara zapota (L.) P. Royen Leaf Extract Derived Silver Nanoparticles Induce Apoptosis in Human Colorectal Carcinoma Cells Without Affecting Human Lymphocytes or Erythrocytes.

Plant-derived synthesis of silver nanoparticles (AgNPs) has found wide biomedical applications including cancer cure. This report deals with biosynthesis of silver nanoparticles (MZLAgNPs) employing leaf extracts of Manilkara zapota (L.) under optimized conditions. Characterization of MZLAgNPs using UV-Vis spectroscopy, FTIR, XRD, and FESEM analyses revealed that the particles were predominantly spherical averaging 24 nm in size. Their cellular effects were assessed by MTT assay, fluorescence, and scanning electron microscopy of cells stained with propidium iodide, acridine orange/ethidium bromide, and annexin V-FITC to visualize signs of apoptosis. Evaluation of cell proliferation by clonogenic assay, wound healing ability by scratch assay and cell cycle distribution by flow-cytometry was also carried out. Apoptosis-related gene expressions were analyzed by RTq-PCR and western blot analysis. MZLAgNPs selectively inhibited growth of colorectal carcinoma HCT116, HeLa, and non-small lung carcinoma A549 cells, dose-dependently with IC50 concentrations of 8, 16, and 29 µg/mL respectively, following 72-h treatment, without affecting growth of normal human lymphocytes and erythrocytes. Apoptosis induction was observed by fluorescence and scanning electron microscopy. Overproduction of reactive oxygen species (ROS), reduction of mitochondrial membrane potential, upregulation of apoptotic-related genes - PUMA, cas-3, cas-8, cas-9, and BAX, expression of caspase 3, and occurrence of PARP cleavage were observed in MZLAgNPs/cisplatin treated cells. Taken together, our results clearly demonstrate the therapeutic potential of biogenic MZLAgNPs as an effective agent for killing colorectal carcinoma cells by apoptosis induction.

Self-assembling polymeric dendritic peptide as functional osteogenic matrix for periodontal regeneration scaffolds-an in vitro study.

OBJECTIVE: Regeneration of periodontal defects is challenging as it necessitates the formation of complex tissue structure with cementum, periodontal ligament, and alveolar bone. Rather than the conventional barrier membranes, scaffolds mimicking extracellular matrix (ECM) can achieve faster healing as they promote migration, adhesion, and differentiation of native progenitor cells. This work explores the possibility of a functional osteogenic matrix based on self-assembling peptide appended dendritic polydiacetylene in regenerating diseased periodontia. METHOD: The amino acid lysine was appended onto a diacetylene core, which was converted to a polymeric dendritic lysine matrix (Lys-PDA) through photopolymerization. This bioactive matrix was evaluated in vitro for the viability, adhesion, spreading, and differentiation of cultured human periodontal ligament (hPDL) progenitor cells. Its osteogenic differentiation was analysed by histologic staining and expression of osteogenic markers (alkaline phosphatase and Osteonectin). Electrospun polycaprolactone (PCL) mat, a candidate barrier material, was fabricated and functionalized with Lys-PDA matrix, and the cell viability, adhesion, and spreading of hPDL cells were evaluated. RESULTS: The dendritic Lys-PDA matrix well supported the hPDL cell growth and differentiation. The cells were viable and showed good cytoskeletal organization. Early expression of osteogenic markers and mineralization was noted in vitro in the presence of Lys-PDA matrix. The electrospun PCL mat functionalized with Lys-PDA maintained the viability, morphology, and spreading of the hPDL cells. SIGNIFICANCE: The ECM mimetic dendritic peptide matrices are capable of hosting and differentiating cells which can lead to the regeneration of periodontal tissue architecture. They could be used in conjunction with barrier membranes for better results.

Direct cell imprint lithography in superconductive carbon black polymer composites: process optimization, characterization and in vitro toxicity analysis.

Cell imprint lithography (CIL) or cell replication plays a vital role in fields like biomimetic smart culture substrates, bone tissue engineering, cell guiding, cell adhesion, tissue engineering, cell microenvironments, tissue microenvironments, cell research, drug delivery, diagnostics, therapeutics and many other applications. Herein we report a new formulation of superconductive carbon black photopolymer composite and its characterization towards a CIL process technique. In this article, we demonstrated an approach of using a carbon nanoparticle-polymer composite (CPC) for patterning cells. It is observed that a 0.3 wt % load of carbon nanoparticles (CNPs) in a carbon polymer mixture (CPM) was optimal for cell-imprint replica fabrication. The electrical resistance of the 3-CPC (0.3 wt %) was reduced by 68% when compared to N-CPC (0 wt %). This method successfully replicated the single cell with sub-organelle scale. The shape of microvesicles, grooves, pores, blebs or microvilli on the cellular surface was patterned clearly. This technique delivers a free-standing cell feature substrate. In vitro evaluation of the polymer demonstrated it as an ideal candidate for biomimetic biomaterial applications. This approach also finds its application in study based on morphology, especially for drug delivery applications and for investigations based on molecular pathways.