Publications by authors named "Adnan Haider"

29 Publications

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Phytochemical and Biological Screening of Leaf, Bark and Fruit Extracts from Wall.

Life (Basel) 2021 Aug 16;11(8). Epub 2021 Aug 16.

Department of Botany, Islamia College University, Peshawar 25120, Khyber Pakhtunkhwa, Pakistan.

The Aquifoliaceae is an important family and has been used traditionally for some time. One of the members of this family is the wall, which itself possesses a potential medicinal importance. This plant is traditionally used for the treatment of various ailments including pain, swelling, burns, and fever. The current study was designed to screen out the antioxidant and analgesic potential of this plant and to verify its traditional uses, along with its phytochemical profile. Extracts were subjected to antioxidant, analgesic, and phytochemical analysis using DPPH, chemical-induced (acetic acid and formalin) nociception models and GC-MS analysis, respectively. The leaf, bark, and fruit extracts showed significant antioxidant activity compared to that of standard. Likewise, all the extracts demonstrated significant ( < 0.01) analgesic activity in a mice model. In acetic acid induced analgesia, the leaf, bark, and fruit extracts caused 51.64, 56.13 and 59.52% inhibition, respectively at a dose of 100 mg/kg while at 200 mg/kg it showed 83.01, 71.69 and 75.47% inhibition, respectively. In Formalin-induced paw-licking assay, fruit extract showed 59.42 and 64.19% inhibition at 200 mg/kg dose in the first and second phase, respectively. The GC-MS analysis revealed the presence of cathinone, phenylpropanolamine, dl-phenylephrine, amphetamine, myristic acid, and palmitic acid. Results of the study suggest that crude extracts from different parts of this plant may be a useful source for the development of novel analgesics. However, further investigation in terms of isolation of bioactive compounds and their toxicological evaluations are needed to validate the observed results.
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http://dx.doi.org/10.3390/life11080837DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8401985PMC
August 2021

Preparation and Physicochemical Characterization of a Diclofenac Sodium-Dual Layer Polyvinyl Alcohol Patch.

Polymers (Basel) 2021 Jul 27;13(15). Epub 2021 Jul 27.

Department of Biological Sciences, National University of Medical Sciences, Rawalpindi 46000, Pakistan.

The aim of this study is to prepare a dual layer polyvinyl (PVA) patch using a combination of electrospinning techniques and cryogelation (freeze-thaw process) then subsequently to investigate the effect of freeze-thaw cycles, nanofiber thickness, and diclofenac sodium (DS) loading on the physicochemical and mechanical properties and formulation of dual layer PVA patches composed of electrospun PVA nanofibers and PVA cryogel. After the successful preparation of the dual layer PVA patch, the prepared patch was subjected to investigation to assess the effect of freeze-thaw cycles, nanofiber thickness and percentages of DS loading on the morphology, physiochemical and mechanical properties. Various spectroscopic techniques such as scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), water contact angle, and tensile tests were used to evaluate the physicochemical and mechanical properties of prepared dual layer PVA patches. The morphological structures of the dual layer PVA patch demonstrated the effectiveness of both techniques. The effect of freeze-thaw cycles, nanofiber thickness, and DS percentage loading on the crystallinity of a dual layer PVA patch was investigated using XRD analysis. The presence of a distinct DS peak in the FTIR spectrum indicates the compatibility of DS in a dual layer PVA patch through in-situ loading. All prepared patches were considered highly hydrophilic because the data obtained was less than 90°. The increasing saturation of DS within the PVA matrix increases the tensile strength of prepared patches, however decreased its elasticity. Evidently, the increasing of electrospun PVA nanofibers thickness, freeze-thaw cycles, and the DS saturation has improved the physicochemical and mechanical properties of the DS medicated dual layer PVA patches, making them a promising biomaterial for transdermal drug delivery applications.
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http://dx.doi.org/10.3390/polym13152459DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8347342PMC
July 2021

A distributed optimisation framework combining natural gradient with Hessian-free for discriminative sequence training.

Neural Netw 2021 May 29;143:537-549. Epub 2021 May 29.

Cambridge University Engineering Department, Trumpington Street, Cambridge, CB2 1PZ, UK. Electronic address:

This paper presents a novel natural gradient and Hessian-free (NGHF) optimisation framework for neural network training that can operate efficiently in a distributed manner. It relies on the linear conjugate gradient (CG) algorithm to combine the natural gradient (NG) method with local curvature information from Hessian-free (HF). A solution to a numerical issue in CG allows effective parameter updates to be generated with far fewer CG iterations than usually used (e.g. 5-8 instead of 200). This work also presents a novel preconditioning approach to improve the progress made by individual CG iterations for models with shared parameters. Although applicable to other training losses and model structures, NGHF is investigated in this paper for lattice-based discriminative sequence training for hybrid hidden Markov model acoustic models using a standard recurrent neural network, long short-term memory, and time delay neural network models for output probability calculation. Automatic speech recognition experiments are reported on the multi-genre broadcast data set for a range of different acoustic model types. These experiments show that NGHF achieves larger word error rate reductions than standard stochastic gradient descent or Adam, while requiring orders of magnitude fewer parameter updates.
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http://dx.doi.org/10.1016/j.neunet.2021.05.011DOI Listing
May 2021

Accurate Segmentation of Nuclear Regions with Multi-Organ Histopathology Images Using Artificial Intelligence for Cancer Diagnosis in Personalized Medicine.

J Pers Med 2021 Jun 4;11(6). Epub 2021 Jun 4.

Division of Electronics and Electrical Engineering, Dongguk University, 30 Pildong-ro 1-gil, Jung-gu, Seoul 04620, Korea.

Accurate nuclear segmentation in histopathology images plays a key role in digital pathology. It is considered a prerequisite for the determination of cell phenotype, nuclear morphometrics, cell classification, and the grading and prognosis of cancer. However, it is a very challenging task because of the different types of nuclei, large intraclass variations, and diverse cell morphologies. Consequently, the manual inspection of such images under high-resolution microscopes is tedious and time-consuming. Alternatively, artificial intelligence (AI)-based automated techniques, which are fast and robust, and require less human effort, can be used. Recently, several AI-based nuclear segmentation techniques have been proposed. They have shown a significant performance improvement for this task, but there is room for further improvement. Thus, we propose an AI-based nuclear segmentation technique in which we adopt a new nuclear segmentation network empowered by residual skip connections to address this issue. Experiments were performed on two publicly available datasets: (1) The Cancer Genome Atlas (TCGA), and (2) Triple-Negative Breast Cancer (TNBC). The results show that our proposed technique achieves an aggregated Jaccard index (AJI) of 0.6794, Dice coefficient of 0.8084, and F1-measure of 0.8547 on TCGA dataset, and an AJI of 0.7332, Dice coefficient of 0.8441, precision of 0.8352, recall of 0.8306, and F1-measure of 0.8329 on the TNBC dataset. These values are higher than those of the state-of-the-art methods.
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http://dx.doi.org/10.3390/jpm11060515DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8226747PMC
June 2021

2-Methoxy-6-Acetyl-7-Methyljuglone: A Bioactive Phytochemical with Potential Pharmacological Activities.

Anticancer Agents Med Chem 2021 Jun 22. Epub 2021 Jun 22.

Department of Chemistry, Islamia College University, Peshawar, Pakistan.

Natural products have been the focus of biomedical and pharmaceutical research to develop new therapies in recent years. 2-methoxy-6-acetyl-7-methyljuglone (2-methoxystypandrone, MAM), a natural bioactive juglone derivative, is known to have various levels of pharmacotherapeutic efficacies as an anti-inflammatory, anticancer, antioxidant, antimicrobial, and anti-HIV activity. MAM fights cancer progression by inducing apoptosis, necroptosis, and deregulating signaling pathways through H2O2-induced JNK/iNOS/NO and MAPK, ERK1/2 pathways, JNK activation, and the RIP1/RIP3 complex. In this review, we summarize the pharmacological importance of MAM in the field of drug discovery. Furthermore, this review not only emphasizes the medicinal properties of MAM but also discusses its potential efficacy in future medicinal products.
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http://dx.doi.org/10.2174/1871520621666210623095636DOI Listing
June 2021

Outside the Thorax: Doege-Potter Syndrome Presenting as a Retroperitoneal Abdominal Mass.

Case Rep Endocrinol 2021 19;2021:9919321. Epub 2021 May 19.

Department of Endocrinology, Department of Medicine, West Virginia University, Morgantown, WV, USA.

Objective: We present a case of refractory hypoglycemia, weight loss, and retroperitoneal solitary fibrous tumor. . A 68-year-old female presented with symptomatic hypoglycemia, weight loss, and abdominal mass identified on CT scan of the abdomen. Blood work during symptomatic hypoglycemia was consistent with an IGF-2-producing tumor. The abdominal mass pathology was consistent with solitary fibrous tumor surrounding the adrenal gland, and resection resulted in complete resolution of hypoglycemia. . Understanding the biochemical mechanisms behind glucose regulation is necessary to diagnose and adequately treat Doege-Potter syndrome, a paraneoplastic syndrome observed in patients with solitary fibrous tumors. Solitary fibrous tumors can be characterized by specific histologic and immunohistochemical studies.

Conclusion: This report describes the clinical workup of a patient presenting with hypoglycemia and a retroperitoneal tumor. This case is unique because of its presentation with severe, refractory hypoglycemia and the tumor's location in the retroperitoneum, given the majority of solitary fibrous tumors are found in the lungs originating from the pleura.
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http://dx.doi.org/10.1155/2021/9919321DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8159632PMC
May 2021

Artificial Intelligence-Based Recognition of Different Types of Shoulder Implants in X-ray Scans Based on Dense Residual Ensemble-Network for Personalized Medicine.

J Pers Med 2021 May 27;11(6). Epub 2021 May 27.

Division of Electronics and Electrical Engineering, Dongguk University, 30 Pildong-ro 1-gil, Jung-gu, Seoul 04620, Korea.

Re-operations and revisions are often performed in patients who have undergone total shoulder arthroplasty (TSA) and reverse total shoulder arthroplasty (RTSA). This necessitates an accurate recognition of the implant model and manufacturer to set the correct apparatus and procedure according to the patient's anatomy as personalized medicine. Owing to unavailability and ambiguity in the medical data of a patient, expert surgeons identify the implants through a visual comparison of X-ray images. False steps cause heedlessness, morbidity, extra monetary weight, and a waste of time. Despite significant advancements in pattern recognition and deep learning in the medical field, extremely limited research has been conducted on classifying shoulder implants. To overcome these problems, we propose a robust deep learning-based framework comprised of an ensemble of convolutional neural networks (CNNs) to classify shoulder implants in X-ray images of different patients. Through our rotational invariant augmentation, the size of the training dataset is increased 36-fold. The modified ResNet and DenseNet are then combined deeply to form a dense residual ensemble-network (DRE-Net). To evaluate DRE-Net, experiments were executed on a 10-fold cross-validation on the openly available shoulder implant X-ray dataset. The experimental results showed that DRE-Net achieved an accuracy, F1-score, precision, and recall of 85.92%, 84.69%, 85.33%, and 84.11%, respectively, which were higher than those of the state-of-the-art methods. Moreover, we confirmed the generalization capability of our network by testing it in an open-world configuration, and the effectiveness of rotational invariant augmentation.
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http://dx.doi.org/10.3390/jpm11060482DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229063PMC
May 2021

Development of Biopolymeric Hybrid Scaffold-Based on AAc/GO/nHAp/TiO Nanocomposite for Bone Tissue Engineering: In-Vitro Analysis.

Nanomaterials (Basel) 2021 May 17;11(5). Epub 2021 May 17.

Department of Biology, College of Sciences, University of Hafr Al Batin, Hafr Al Batin 39524, Saudi Arabia.

Bone tissue engineering is an advanced field for treatment of fractured bones to restore/regulate biological functions. Biopolymeric/bioceramic-based hybrid nanocomposite scaffolds are potential biomaterials for bone tissue because of biodegradable and biocompatible characteristics. We report synthesis of nanocomposite based on acrylic acid (AAc)/guar gum (GG), nano-hydroxyapatite (HAp NPs), titanium nanoparticles (TiO NPs), and optimum graphene oxide (GO) amount via free radical polymerization method. Porous scaffolds were fabricated through freeze-drying technique and coated with silver sulphadiazine. Different techniques were used to investigate functional group, crystal structural properties, morphology/elemental properties, porosity, and mechanical properties of fabricated scaffolds. Results show that increasing amount of TiO in combination with optimized GO has improved physicochemical and microstructural properties, mechanical properties (compressive strength (2.96 to 13.31 MPa) and Young's modulus (39.56 to 300.81 MPa)), and porous properties (pore size (256.11 to 107.42 μm) and porosity (79.97 to 44.32%)). After 150 min, silver sulfadiazine release was found to be ~94.1%. In vitro assay of scaffolds also exhibited promising results against mouse pre-osteoblast () cell lines. Hence, these fabricated scaffolds would be potential biomaterials for bone tissue engineering in biomedical engineering.
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http://dx.doi.org/10.3390/nano11051319DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8156135PMC
May 2021

Light-weighted ensemble network with multilevel activation visualization for robust diagnosis of COVID19 pneumonia from large-scale chest radiographic database.

Appl Soft Comput 2021 Sep 7;108:107490. Epub 2021 May 7.

Division of Electronics and Electrical Engineering, Dongguk University, 30 Pildong-ro 1-gil, Jung-gu, Seoul 04620, Republic of Korea.

Currently, the coronavirus disease 2019 (COVID19) pandemic has killed more than one million people worldwide. In the present outbreak, radiological imaging modalities such as computed tomography (CT) and X-rays are being used to diagnose this disease, particularly in the early stage. However, the assessment of radiographic images includes a subjective evaluation that is time-consuming and requires substantial clinical skills. Nevertheless, the recent evolution in artificial intelligence (AI) has further strengthened the ability of computer-aided diagnosis tools and supported medical professionals in making effective diagnostic decisions. Therefore, in this study, the strength of various AI algorithms was analyzed to diagnose COVID19 infection from large-scale radiographic datasets. Based on this analysis, a light-weighted deep network is proposed, which is the first ensemble design (based on MobileNet, ShuffleNet, and FCNet) in medical domain (particularly for COVID19 diagnosis) that encompasses the reduced number of trainable parameters (a total of 3.16 million parameters) and outperforms the various existing models. Moreover, the addition of a multilevel activation visualization layer in the proposed network further visualizes the lesion patterns as multilevel class activation maps (ML-CAMs) along with the diagnostic result (either COVID19 positive or negative). Such additional output as ML-CAMs provides a visual insight of the computer decision and may assist radiologists in validating it, particularly in uncertain situations Additionally, a novel hierarchical training procedure was adopted to perform the training of the proposed network. It proceeds the network training by the adaptive number of epochs based on the validation dataset rather than using the fixed number of epochs. The quantitative results show the better performance of the proposed training method over the conventional end-to-end training procedure. A large collection of CT-scan and X-ray datasets (based on six publicly available datasets) was used to evaluate the performance of the proposed model and other baseline methods. The experimental results of the proposed network exhibit a promising performance in terms of diagnostic decision. An average F1 score (F1) of 94.60% and 95.94% and area under the curve (AUC) of 97.50% and 97.99% are achieved for the CT-scan and X-ray datasets, respectively. Finally, the detailed comparative analysis reveals that the proposed model outperforms the various state-of-the-art methods in terms of both quantitative and computational performance.
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http://dx.doi.org/10.1016/j.asoc.2021.107490DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8103783PMC
September 2021

Multilevel Deep-Aggregated Boosted Network to Recognize COVID-19 Infection from Large-Scale Heterogeneous Radiographic Data.

IEEE J Biomed Health Inform 2021 06 3;25(6):1881-1891. Epub 2021 Jun 3.

In the present epidemic of the coronavirus disease 2019 (COVID-19), radiological imaging modalities, such as X-ray and computed tomography (CT), have been identified as effective diagnostic tools. However, the subjective assessment of radiographic examination is a time-consuming task and demands expert radiologists. Recent advancements in artificial intelligence have enhanced the diagnostic power of computer-aided diagnosis (CAD) tools and assisted medical specialists in making efficient diagnostic decisions. In this work, we propose an optimal multilevel deep-aggregated boosted network to recognize COVID-19 infection from heterogeneous radiographic data, including X-ray and CT images. Our method leverages multilevel deep-aggregated features and multistage training via a mutually beneficial approach to maximize the overall CAD performance. To improve the interpretation of CAD predictions, these multilevel deep features are visualized as additional outputs that can assist radiologists in validating the CAD results. A total of six publicly available datasets were fused to build a single large-scale heterogeneous radiographic collection that was used to analyze the performance of the proposed technique and other baseline methods. To preserve generality of our method, we selected different patient data for training, validation, and testing, and consequently, the data of same patient were not included in training, validation, and testing subsets. In addition, fivefold cross-validation was performed in all the experiments for a fair evaluation. Our method exhibits promising performance values of 95.38%, 95.57%, 92.53%, 98.14%, 93.16%, and 98.55% in terms of average accuracy, F-measure, specificity, sensitivity, precision, and area under the curve, respectively and outperforms various state-of-the-art methods.
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http://dx.doi.org/10.1109/JBHI.2021.3072076DOI Listing
June 2021

Desperate Times Call for Desperate Measures: Use of Continuous Subcutaneous 1-34 PTH Infusion for Postsurgical Hypoparathyroidism.

Case Rep Endocrinol 2021 9;2021:5593653. Epub 2021 Mar 9.

Department of Medicine, West Virginia University, Morgantown, WV, USA.

Objective: This case highlights use of 1-34 PTH continuous infusion in a patient with postsurgical hypoparathyroidism.

Method: Clinical presentation and biochemical profile were monitored before and after 1-34 PTH infusion, with notable reduction in pill burden in a patient with postsurgical hypoparathyroidism.

Results: We present a case of postsurgical hypoparathyroidism following thyroidectomy for Graves disease. The patient was requiring a total of 34 pills daily and, despite medication compliance, her clinical and biochemical control was unsatisfactory. Following initiation of 1-34 PTH in the form of a subcutaneous pump, we were able to stop all calcium supplementation and reduce calcitriol to 0.5 mcg daily. Her current biochemical control as well as quality of life improved significantly on CSPI, calcitriol, and a daily serving of dietary calcium.

Conclusion: This case highlights the use of 1-34 PTH either as twice-daily dosing or continuous subcutaneous infusion for adult patients with hypoparathyroidism.
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http://dx.doi.org/10.1155/2021/5593653DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7969095PMC
March 2021

Antidepressant-like effect of ethanol in mice forced swimming test is mediated via inhibition of NMDA/nitric oxide/cGMP signaling pathway.

Alcohol 2021 05 11;92:53-63. Epub 2021 Feb 11.

Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran; Brain and Spinal Cord Injury Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran. Electronic address:

There is evidence for a dramatic relationship between depression and alcohol consumption. Depressed patients may abuse ethanol because this agent reduces the symptoms of depression. In the current study, we aimed to investigate the NMDA/nitric oxide/cGMP pathway in the antidepressant-like effect of ethanol in an animal model of behavioral despair. Animals were subjected to locomotor activity in an open-field test separately, followed by a forced swimming test. During the forced swimming test (FST), ethanol (2 and 2.5 g/kg) significantly decreased the immobility time without altering the locomotor activity of animals. The antidepressant-like effect of ethanol (2.5 g/kg) was reversed by co-administration of N-methyl-D-aspartate (NMDA, 75 mg/kg), L-arginine (750 mg/kg), or sildenafil (5 mg/kg). In contrast, co-administration of MK-801 (0.05 mg/kg), ketamine (1 mg/kg), and ifenprodil (0.5 mg/kg) as antagonists of NMDAR, and NG-nitro-L-arginine methyl ester (L-NAME, 10 mg/kg), 7-nitroindazole (7-NI, 30 mg/kg), and methylene blue (10 mg/kg) as inhibitors of nitric oxide synthase (NOS), or 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ) (20 mg/kg), a nitric oxide/cyclic-guanosine monophosphate (NO-cGMP) inhibitor, with a subeffective dose of ethanol (1.5 g/kg), significantly decreased the immobility time in the FST. Furthermore, injection of ethanol 2.5 g/kg alone or 1.5 g/kg with a 7-NI subeffective dose, significantly decreased the nitrite levels in the hippocampus and prefrontal cortex. Hence, it is concluded that blockade of NMDA receptors and the nitric oxide/cyclic-guanosine monophosphate (NO-cGMP) pathway might be involved in the antidepressant-like effect of ethanol in mice.
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http://dx.doi.org/10.1016/j.alcohol.2021.01.005DOI Listing
May 2021

Arabinoxylan/graphene-oxide/nHAp-NPs/PVA bionano composite scaffolds for fractured bone healing.

J Tissue Eng Regen Med 2021 04 9;15(4):322-335. Epub 2021 Mar 9.

Sustainable Energy Technologies Center, College of Engineering, King Saud University, Riyadh, Saudi Arabia.

The importance of bone scaffolds has increased many folds in the last few years; however, during bone implantation, bacterial infections compromise the implantation and tissue regeneration. This work is focused on this issue while not compromising on the properties of a scaffold for bone regeneration. Biocomposite scaffolds (BS) were fabricated via the freeze-drying technique. The samples were characterized for structural changes, surface morphology, porosity, and mechanical properties through spectroscopic (Fourier transform-infrared [FT-IR]), microscopic (scanning electron microscope [SEM]), X-ray (powder X-ray diffraction and energy-dispersive X-ray), and other analytical (Brunauer-Emmett-Teller, universal testing machine Instron) techniques. Antibacterial, cellular, and hemocompatibility assays were performed using standard protocols. FT-IR confirmed the interactions of all the components. SEM illustrated porous and interconnected porous morphology. The percentage porosity was in the range of 49.75%-67.28%, and the pore size was 215.65-470.87 µm. The pore size was perfect for cellular penetration. Thus, cells showed significant proliferation onto these scaffolds. X-ray studies confirmed the presence of nanohydroxyapatite and graphene oxide (GO). The cell viability was 85%-98% (BS1-BS3), which shows no significant toxicity of the biocomposite. Furthermore, the biocomposites exhibited better antibacterial activity, no effect on the blood clotting (normal in vitro blood clotting), and less than 5% hemolysis. The ultimate compression strength for the biocomposites increased from 4.05 to 7.94 with an increase in the GO content. These exciting results revealed that this material has the potential for possible application in bone tissue engineering.
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http://dx.doi.org/10.1002/term.3168DOI Listing
April 2021

Identification of potential inhibitors of Zika virus NS5 RNA-dependent RNA polymerase through virtual screening and molecular dynamic simulations.

Saudi Pharm J 2020 Dec 21;28(12):1580-1591. Epub 2020 Oct 21.

Department of Chemical Engineering, College of Engineering, King Saud University, Riyadh, Saudi Arabia.

Zika virus (ZIKV) is one of the mosquito borne flavivirus with several outbreaks in past few years in tropical and subtropical regions. The non-structural proteins of are suitable active targets for inhibitory drugs due to their role in pathogenicity. In ZIKV, the non-structural protein 5 (NS5) RNA-Dependent RNA polymerase replicates its genome. Here we have performed virtual screening to identify suitable ligands that can potentially halt the ZIKV NS5 RNA dependent RNA polymerase (RdRp). During this process, we searched and screened a library of ligands against ZIKV NS5 RdRp. The selected ligands with significant binding energy and ligand-receptor interactions were further processed. Among the selected docked conformations, top five was further optimized at atomic level using molecular dynamic simulations followed by binding free energy calculations. The interactions of ligands with the target structure of ZIKV RdRp revealed that they form strong bonds within the active sites of the receptor molecule. The efficacy of these drugs against ZIKV can be further analyzed through and studies.
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http://dx.doi.org/10.1016/j.jsps.2020.10.005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7783101PMC
December 2020

Novel functional antimicrobial and biocompatible arabinoxylan/guar gum hydrogel for skin wound dressing applications.

J Tissue Eng Regen Med 2020 10 17;14(10):1488-1501. Epub 2020 Aug 17.

School of Biological Sciences, University of the Punjab, Lahore, Pakistan.

It is a challenging task to develop active biomacromolecular wound dressing materials that are biocompatible and possesses antibacterial properties against the bacterial strains that cause severe skin disease. This work is focused on the preparation of a biocompatible and degradable hydrogel for wound dressing application using arabinoxylan (ARX) and guar gum (GG) natural polymers. Fourier transform infrared spectroscopy (FT-IR) confirmed that both ARX and GG interacted well with each other, and their interactions further increased with the addition of crosslinker tetraethyl orthosilicate. Scanning electron microscope (SEM) micrographs showed uniform porous morphologies of the hydrogels. The porous morphologies and uniform interconnected pores are attributed to the increased crosslinking of the hydrogel. Elastic modulus, tensile strength, and fracture strain of the hydrogels significantly improved (from ATG-1 to ATG-4) with crosslinking. Degradability tests showed that hydrogels lost maximum weight in 7 days. All the samples showed variation in swelling with pH. Maximum swelling was observed at pH 7. The hydrogel samples showed good antibacterial activity against Pseudomonas aeruginosa (Gram-negative) and Staphylococcus aureus (Gram-positive) in PBS, good drug release profile (92% drug release), and nontoxic cellular behavior. The cells not only retained their cylindrical morphologies onto the hydrogel but were also performing their normal activities. It is, therefore, believed that as-developed hydrogel could be a potential material for wound dressing application.
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http://dx.doi.org/10.1002/term.3115DOI Listing
October 2020

Arabinoxylan-co-AA/HAp/TiO nanocomposite scaffold a potential material for bone tissue engineering: An in vitro study.

Int J Biol Macromol 2020 May 17;151:584-594. Epub 2020 Feb 17.

Department of Biological Sciences, National University of Medical Sciences, Rawalpindi, Punjab, Pakistan. Electronic address:

Arabinoxylan (AX) is a natural biological macromolecule with several potential biomedical applications. In this research, AX, nano-hydroxyapatite (n-HAp) and titanium dioxide (TiO) based polymeric nanocomposite scaffolds were fabricated by the freeze-drying method. The physicochemical characterizations of these polymeric nanocomposite scaffolds were performed for surface morphology, porosity, swelling, biodegradability, mechanical, and biological properties. The scaffolds exhibited good porosity and rough surface morphology, which were efficiently controlled by TiO concentrations. MC3T3-E1 cells were employed to conduct the biocompatibility of these scaffolds. Scaffolds showed unique biocompatibility in vitro and was favorable for cell attachment and growth. PNS3 proved more biocompatible, showed interconnected porosity and substantial mechanical strength compared to PNS1, PNS2 and PNS4. Furthermore, it has also showed more affinity to cells and cell growth. The results illustrated that the bioactive nanocomposite scaffold has the potential to find applications in the tissue engineering field.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.02.142DOI Listing
May 2020

Biocompatible Polymers and their Potential Biomedical Applications: A Review.

Curr Pharm Des 2019 ;25(34):3608-3619

Department of Polymer Science and Engineering, Kyungpook National University, Daegu, South Korea.

Background: Biocompatible polymers are gaining great interest in the field of biomedical applications. The term biocompatibility refers to the suitability of a polymer to body and body fluids exposure. Biocompatible polymers are both synthetic (man-made) and natural and aid in the close vicinity of a living system or work in intimacy with living cells. These are used to gauge, treat, boost, or substitute any tissue, organ or function of the body. A biocompatible polymer improves body functions without altering its normal functioning and triggering allergies or other side effects. It encompasses advances in tissue culture, tissue scaffolds, implantation, artificial grafts, wound fabrication, controlled drug delivery, bone filler material, etc.

Objectives: This review provides an insight into the remarkable contribution made by some well-known biopolymers such as polylactic-co-glycolic acid, poly(ε-caprolactone) (PCL), polyLactic Acid, poly(3- hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), Chitosan and Cellulose in the therapeutic measure for many biomedical applications.

Methods: Various techniques and methods have made biopolymers more significant in the biomedical fields such as augmentation (replaced petroleum based polymers), film processing, injection modeling, blow molding techniques, controlled / implantable drug delivery devices, biological grafting, nano technology, tissue engineering etc.

Results: The fore mentioned techniques and other advanced techniques have resulted in improved biocompatibility, nontoxicity, renewability, mild processing conditions, health condition, reduced immunological reactions and minimized side effects that would occur if synthetic polymers are used in a host cell.

Conclusion: Biopolymers have brought effective and attainable targets in pharmaceutics and therapeutics. There are huge numbers of biopolymers reported in the literature that has been used effectively and extensively.
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http://dx.doi.org/10.2174/1381612825999191011105148DOI Listing
June 2020

Intranasal Insulin Administration Does Not Affect LH Concentrations in Men with Diabetes.

Int J Endocrinol 2018 31;2018:6170154. Epub 2018 Oct 31.

Division of Endocrinology, Diabetes and Metabolism, State University of New York, Buffalo and Kaleida Health 462 Grider Street, Buffalo NY-14215, USA.

A quarter of men with obesity or type 2 diabetes have hypogonadotropic hypogonadism. Animal studies and in vitro data have shown that insulin action and insulin responsiveness in the brain are necessary for the maintenance of the functional integrity of the hypothalamo-hypophyseal-gonadal axis. We conducted a randomized, placebo-controlled trial to evaluate the effect of one dose of intranasal insulin (40 IU of regular insulin) or saline on LH concentrations in 14 men (8 with type 2 diabetes and 6 healthy lean men). Insulin or saline was administered intranasally on two different occasions, at least one week apart. Blood samples were collected to measure LH concentrations every 15 minutes for 5 hours. Study drug was administered intranasally after a 2-hour baseline sampling period. Patients remained fasting throughout the procedure. The primary endpoint of the study was to compare the change in LH concentrations after intranasal insulin as compared to placebo (intranasal saline). Change was defined as the difference between baseline LH concentrations (average of the 9 samples collected in two hours prior to drug administration) and average LH concentrations following drug administration (average of the 12 samples collected in 3 hours). There was no change in LH concentrations following insulin administration as compared to placebo in men with diabetes or in lean men. We conclude that one dose of 40 IU of regular insulin administered intranasally does not change LH concentrations acutely in men.
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http://dx.doi.org/10.1155/2018/6170154DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6234437PMC
October 2018

Novel route for amine grafting to chitosan electrospun nanofibers membrane for the removal of copper and lead ions from aqueous medium.

Carbohydr Polym 2018 Nov 10;199:406-414. Epub 2018 Jul 10.

Department of Chemical Engineering, College of Engineering, King Saud University, P.O. Box 800, Riyadh, 11421 Saudi Arabia.

A novel step wise synthetic route was developed to prepare amine grafted nanofibers (AGNFs) affinity membrane. The chemical structure of the nanofibers (NFs) after grafting was studied by acquiring Fourier Transform Infrared (FT-IR) spectra and Carbon, Hydrogen and Nitrogen (CHN) data. The morphology of the NFs before and after grafting was studied by Field Emission Scanning Electron Microscope (FE-SEM). FT-IR and CHN data confirmed the introduction of new functional groups into the primary structure of chitosan (CH). FE-SEM showed denser membrane with no deterioration of the NFs morphology after grafting. The aqueous stability of the membranes was studied in distilled water. The AGNFs membranes showed good aqueous stabilities (with only ∼ 6% loss in weight until 24 h and remained stable thereafter) which was less than the weight loss by glutaraldehyde treated nanofibers (GNFs) (∼44% loss in weight until 24 h) and pristine NFs (100% loss in weight as soon as the NFs were immersed in distilled water). The maximum adsorption (q) capacity of AGNFs for Cu (II) and Pb (II) was observed to be 166.67 mg.g and 94.34 mg.g. The adsorption capacity of the present systems was much higher for Cu (II) when compared to the already existing conventional and chitosan adsorbents. This increased might be related not just to the size, but more potentially to the increase in the number of nitrogen binding sites (chelating sites). Nitrogen donates lone-pair of electrons for chelation. The combination of processing into nano size and amine grafting (AG) has significantly increased the adsorption capacity of CH NFs membrane.
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http://dx.doi.org/10.1016/j.carbpol.2018.07.026DOI Listing
November 2018

A novel use of cellulose based filter paper containing silver nanoparticles for its potential application as wound dressing agent.

Int J Biol Macromol 2018 Mar 6;108:455-461. Epub 2017 Dec 6.

Department of Nano, Medical and Polymer Materials, College of Engineering Yeungnam University, 280 Daehak-Ro, Gyeongsan 712-749, South Korea; School of Chemical Engineering, Yeungnam University, 280-Daehak-ro, Gyeongsan 712-749, South Korea. Electronic address:

The frequent use of antibiotics against microbial infections may lead to the emergence of antibiotic resistant microbial strains. To overcome these microbial strains, we need to fabricate alternative materials which can handle them. It is for this reason, we have fabricated cellulose (CE) based filter paper (FP) composite scaffolds comprising of adsorbed chitosan (CS) and sliver (Ag) nanoparticles (NPs). The AgNPs are incorporated in the CS layer of the composite scaffold. Prior to evaluate the efficacy of the scaffolds against gram positive and gram negative bacterial strains, the scaffolds were characterized for the presence of the Ag NPs with field emission scanning electron microscope (FE-SEM), fourier transform infrared (FTIR) spectroscopy and x-ray diffractometer (XRD). These techniques confirmed the presence of Ag NPs in the composite scaffold. The biocompatibility of the scaffolds was assessed by subjecting pristine FP, CS adsorbed FP (CS-FP) and Ag loaded CS-FP (Ag-CS-FP) composite scaffolds to in vitro studies. From the data obtained, it was observed that NIH3T3 fibroblastic cells adhered and proliferated onto all the scaffolds. Furthermore, the scaffolds exhibited good antibacterial activity against both strains of bacteria. It is, therefore, concluded that these scaffolds could find potential application in biomedical field, particularly as a wound dressing agent.
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http://dx.doi.org/10.1016/j.ijbiomac.2017.12.022DOI Listing
March 2018

Postprandial Hypoglycemia: Complication of Peptic Ulcer Surgery.

Am J Med 2017 12 12;130(12):e527-e529. Epub 2017 Jul 12.

Texas Tech University Health Science Center, Odessa.

Objective: Billroth II procedures (gastrojejunostomy with vagotomy) are seldom performed now but were popular before the advent of histamine 2 receptor blockers and proton pump inhibitors. Such procedures can be a cause of late postprandial hypoglycemia.

Methods: We performed a formal evaluation and discussion of postprandial hypoglycemia.

Results: We present a case of an 85-year-old man who presented to the endocrinology clinic with symptoms of "fainting spells after large meals." The patient previously had extensive evaluations by a neurologist and a cardiologist. Because of the history of Billroth II surgery and the description of his fainting spells, a 5-hour glucose tolerance test was performed in the clinic using the foods that uniformly resulted in symptoms. This confirmed that the late dumping syndrome with associated hypoglycemia was the cause of his spells.

Conclusions: Late dumping syndrome manifesting with hypoglycemia should be considered in the workup of patients with a history of gastric surgery and unusual postprandial symptoms. This case highlights the importance of an appropriate workup that can lead to avoidance of unnecessary testing in such patients.
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http://dx.doi.org/10.1016/j.amjmed.2017.06.010DOI Listing
December 2017

Pamidronic acid-grafted nHA/PLGA hybrid nanofiber scaffolds suppress osteoclastic cell viability and enhance osteoblastic cell activity.

J Mater Chem B 2016 Dec 14;4(47):7596-7604. Epub 2016 Nov 14.

Department of Polymer Science & Engineering, Kyungpook National University, Daegu 702-701, Republic of Korea.

Osteoclasts have the capability to resorb bone. When osteoclastic activity is excessively high, bones generally become weakened and more prone to fracture. In order to treat excessive osteoclastic cell activity, maintain the balance between bone formation and resorption, and enhance osseointegration, pamidronic acid-grafted nanorod hydroxyapatite/poly(lactide-co-glycolide) (P-g-nHA/PLGA) scaffolds were fabricated via an electrospinning technique. Various spectroscopic techniques were used for the structural and morphological characterization of pristine PLGA, nHA/PLGA, and P-g-nHA/PLGA hybrid nanofiber scaffolds. The potential of the P-g-nHA/PLGA hybrid nanofiber scaffold as an implantable material was assessed through in vitro studies. The results showed that the viability of osteoclastic cells on the P-g-nHA/PLGA nanofiber scaffold was significantly suppressed due to the presence of pamidronic acid. Osteoblastic cells adhered and proliferated on all scaffolds tested; however, increased osteoblastic cell proliferation was observed on the P-g-nHA/PLGA hybrid and nHA/PLGA nanofiber scaffolds compared to the pristine PLGA nanofiber scaffolds. Therefore, these types of dual function P-g-nHA/PLGA hybrid nanofiber scaffolds could certainly be used in therapeutic bone implantation.
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http://dx.doi.org/10.1039/c6tb02083fDOI Listing
December 2016

Effect of crosslinking functionality on microstructure, mechanical properties, and in vitro cytocompatibility of cellulose nanocrystals reinforced poly (vinyl alcohol)/sodium alginate hybrid scaffolds.

Int J Biol Macromol 2017 Feb 25;95:962-973. Epub 2016 Oct 25.

School of Chemical Engineering, Yeungnam University, 280 Daehak-Ro, Gyeongsan 712 749, South Korea; Department of Nano, Medical and Polymer Materials, College of Engineering, Yeungnam University, 280 Daehak-Ro, Gyeongsan 712 749, South Korea. Electronic address:

Cellulose nanocrystals reinforced poly (vinyl alcohol)/sodium alginate hybrid scaffolds were fabricated by using freeze casting and freeze drying method. In this study, the effect of crosslinking agents such as calcium chloride, orthophosphoric acid, and borax on morphological, structural, thermal, mechanical, and cytocompatibility (cell adhesion and proliferation) properties was investigated. The results showed that the change in type of crosslinking agent significantly changed the properties of the hybrid scaffolds. Based on this study, borax-crosslinked hybrid scaffold showed good fibrous porous structure with high porosity (95.2%), highest water uptake capacity, good thermal stability, mechanical stability (storage modulus), and in vitro cell adhesion and proliferation with fibroblast (NIH3T3) cells. This primarily research study explores the way for further use of this crosslinking agent to design and fabricate scaffolds for tissue engineering applications.
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http://dx.doi.org/10.1016/j.ijbiomac.2016.10.085DOI Listing
February 2017

Micro/Nano Multilayered Scaffolds of PLGA and Collagen by Alternately Electrospinning for Bone Tissue Engineering.

Nanoscale Res Lett 2016 Dec 4;11(1):323. Epub 2016 Jul 4.

Department of Polymer Science and Engineering, Kyungpook National University, 80, Daehak-ro, Buk-gu, Daegu, 702-701, South Korea.

The dual extrusion electrospinning technique was used to fabricate multilayered 3D scaffolds by stacking microfibrous meshes of poly(lactic acid-co-glycolic acid) (PLGA) in alternate fashion to micro/nano mixed fibrous meshes of PLGA and collagen. To fabricate the multilayered scaffold, 35 wt% solution of PLGA in THF-DMF binary solvent (3:1) and 5 wt% solution of collagen in hexafluoroisopropanol (HFIP) with and without hydroxyapatite nanorods (nHA) were used. The dual and individual electrospinning of PLGA and collagen were carried out at flow rates of 1.0 and 0.5 mL/h, respectively, at an applied voltage of 20 kV. The density of collagen fibers in multilayered scaffolds has controlled the adhesion, proliferation, and osteogenic differentiation of MC3T3-E1 cells. The homogeneous dispersion of glutamic acid-modified hydroxyapatite nanorods (nHA-GA) in collagen solution has improved the osteogenic properties of fabricated multilayered scaffolds. The fabricated multilayered scaffolds were characterized using FT-IR, X-ray photoelectron spectroscopy, and transmission electron microscopy (TEM). The scanning electron microscopy (FE-SEM) was used to evaluate the adhesion and spreads of MC3T3-E1 cells on multilayered scaffolds. The activity of MC3T3-E1 cells on the multilayered scaffolds was evaluated by applying MTT, alkaline phosphatase, Alizarin Red, von Kossa, and cytoskeleton F-actin assaying protocols. The micro/nano fibrous PLGA-Col-HA scaffolds were found to be highly bioactive in comparison to pristine microfibrous PLGA and micro/nano mixed fibrous PLGA and Col scaffolds.
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http://dx.doi.org/10.1186/s11671-016-1532-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4932007PMC
December 2016

BMP-2 Grafted nHA/PLGA Hybrid Nanofiber Scaffold Stimulates Osteoblastic Cells Growth.

Biomed Res Int 2015 11;2015:281909. Epub 2015 Oct 11.

Department of Polymer Science and Engineering, Kyungpook National University, Daegu 702-701, Republic of Korea.

Biomaterials play a pivotal role in regenerative medicine, which aims to regenerate and replace lost/degenerated tissues or organs. Natural bone is a hierarchical structure, comprised of various cells having specific functions that are regulated by sophisticated mechanisms. However, the regulation of the normal functions in damaged or injured cells is disrupted. In order to address this problem, we attempted to artificially generate a scaffold for mimicking the characteristics of the extracellular matrix at the nanoscale level to trigger osteoblastic cell growth. For this purpose, we have chemically grafted bone morphogenetic protein (BMP-2) onto the surface of L-glutamic acid modified hydroxyapatite incorporated into the PLGA nanofiber matrix. After extensive characterization using various spectroscopic techniques, the BMP-g-nHA/PLGA hybrid nanofiber scaffolds were subjected to various in vitro cytocompatibility tests. The results indicated that BMP-2 on BMP-g-nHA/PLGA hybrid nanofiber scaffolds greatly stimulated osteoblastic cells growth, contrary to the nHA/PLGA and pristine PLGA nanofiber scaffold, which are used as control. These results suggest that BMP-g-nHA/PLGA hybrid nanofiber scaffold can be used as a nanodrug carrier for the controlled and targeted delivery of BMP-2, which will open new possibilities for enhancing bone tissue regeneration and will help in the treatment of various bone-related diseases in the future.
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http://dx.doi.org/10.1155/2015/281909DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4619782PMC
August 2016

Photoinduced development of antibacterial materials derived from isosorbide moiety.

Biomacromolecules 2015 Mar 10;16(3):683-94. Epub 2015 Feb 10.

Institut de Chimie et des Matériaux Paris-Est, Equipe Systèmes Polymères Complexes, UMR 7182, CNRS-Université Paris-Est Créteil (UPEC) 2-8 rue Henri Dunant, 94320 Thiais, France.

A straightforward method for immobilizing in situ generated silver nanoparticles on the surface of a photoactivable isosorbide-derived monomer is developed with the objective to design a functional material having antibacterial properties. The photoinduced thiol-ene mechanism involved in these syntheses is described by the electron spin resonance/spin trapping technique. The resulting materials with or without silver nanoparticles (Ag NPs) were used as films or as coatings on glass substrate. The surface of the synthesized materials was characterized by X-ray photoelectron spectroscopy and scanning electron microscopy, and their thermal and mechanical properties were evaluated by dynamic-mechanical thermal tests, differential scanning calorimetry, thermogravimetric analyses, along with pencil hardness, nanoindentation, and scratch resistance tests. The photoinduced formation of Ag NPs is also confirmed by UV spectrophotometry. Finally, a primary investigation demonstrates the antibacterial properties of the isosorbide-derived material against Staphylococcus aureus and Escherichia coli, as well as its cytocompatibility toward NIH 3T3 fibroblastic cells.
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http://dx.doi.org/10.1021/bm501755rDOI Listing
March 2015

PLGA/nHA hybrid nanofiber scaffold as a nanocargo carrier of insulin for accelerating bone tissue regeneration.

Nanoscale Res Lett 2014 25;9(1):314. Epub 2014 Jun 25.

Department of Polymer Science and Engineering, School of Applied Chemical Engineering Kyungpook, National University, Daegu 702-701, South Korea.

The development of tissue engineering in the field of orthopedic surgery is booming. Two fields of research in particular have emerged: approaches for tailoring the surface properties of implantable materials with osteoinductive factors as well as evaluation of the response of osteogenic cells to these fabricated implanted materials (hybrid material). In the present study, we chemically grafted insulin onto the surface of hydroxyapatite nanorods (nHA). The insulin-grafted nHAs (nHA-I) were dispersed into poly(lactide-co-glycolide) (PLGA) polymer solution, which was electrospun to prepare PLGA/nHA-I composite nanofiber scaffolds. The morphology of the electrospun nanofiber scaffolds was assessed by field emission scanning electron microscopy (FESEM). After extensive characterization of the PLGA/nHA-I and PLGA/nHA composite nanofiber scaffolds by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction spectroscopy (XRD), X-ray photoelectron spectroscopy (XPS), energy-dispersive X-ray spectrometry (EDS), and transmission electron microscopy (TEM), the PLGA/nHA-I and PLGA/nHA (used as control) composite nanofiber scaffolds were subjected to cell studies. The results obtained from cell adhesion, alizarin red staining, and Von Kossa assay suggested that the PLGA/nHA-I composite nanofiber scaffold has enhanced osteoblastic cell growth, as more cells were proliferated and differentiated. The fact that insulin enhanced osteoblastic cell proliferation will open new possibilities for the development of artificial scaffolds for bone tissue regeneration.
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http://dx.doi.org/10.1186/1556-276X-9-314DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4082289PMC
July 2014

Morphological effects of HA on the cell compatibility of electrospun HA/PLGA composite nanofiber scaffolds.

Biomed Res Int 2014 26;2014:308306. Epub 2014 Feb 26.

Department of Polymer Science and Engineering, School of Applied Chemical Engineering, Kyungpook National University, Daegu 702-701, Republic of Korea.

Tissue engineering is faced with an uphill challenge to design a platform with appropriate topography and suitable surface chemistry, which could encourage desired cellular activities and guide bone tissue regeneration. To develop such scaffolds, composite nanofiber scaffolds of nHA and sHA with PLGA were fabricated using electrospinning technique. nHA was synthesized using precipitation method, whereas sHA was purchased. The nHA and sHA were suspended in PLGA solution separately and electrospun at optimized electrospinning parameters. The composite nanofiber scaffolds were characterized by FE-SEM, EDX analysis, TEM, XRD analysis, FTIR, and X-ray photoelectron. The potential of the HA/PLGA composite nanofiber as bone scaffolds in terms of their bioactivity and biocompatibility was assessed by culturing the osteoblastic cells onto the composite nanofiber scaffolds. The results from in vitro studies revealed that the nHA/PLGA composite nanofiber scaffolds showed higher cellular adhesion, proliferation, and enhanced osteogenesis performance, along with increased Ca(+2) ions release compared to the sHA/PLGA composite nanofiber scaffolds and pristine PLGA nanofiber scaffold. The results show that the structural dependent property of HA might affect its potential as bone scaffold and implantable materials in regenerative medicine and clinical tissue engineering.
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http://dx.doi.org/10.1155/2014/308306DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3955660PMC
December 2014

Nanofibrous scaffolds in biomedical applications.

Biomater Res 2014 13;18. Epub 2014 Jun 13.

Department of Polymer Science and Engineering, Kyungpook National University, Daegu, 702-701 South Korea.

Nanofibrous scaffolds are artificial extracellular matrices which provide natural environment for tissue formation. In comparison to other forms of scaffolds, the nanofibrous scaffolds promote cell adhesion, proliferation and differentiation more efficiently due to having high surface to volume ratio. Although scaffolds for tissue engineering have been fabricated by various techniques but electrospun nanofibrous scaffolds have shown great potential in the fields of tissue engineering and regeneration. This review highlights the applications and importance of electrospun nanofibrous scaffolds in various fields of biomedical applications ranging from drug delivery to wound healing. Attempts have also been made to highlights the advantages and disadvantages of nanofirbous scaffolds fabricated for biomedical applications using technique of electrospinning. The role of various factors controlling drug distribution in electrospun nanofibrous scaffolds is also discussed to increase the therapeutic efficiency of nanofibrous scaffolds in wound healing and drug delivery applications.
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http://dx.doi.org/10.1186/2055-7124-18-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4549138PMC
September 2015
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