Preparation and characterization of TiO2-coated polymerization of methyl methacrylate (PMMA) for biomedical applications: In vitro study.

Low surface energy and hydrophobicity of polymethyl methactylate (PMMA) are the main disadvantages of this biomaterial. The aim of this study was to investigate the effects of a new coating process on the surface characteristics and properties of PMMA. A combination of temperature and pressure was used for deposition of titanium dioxide (TiO2) on the surface of PMMA. The PMMA coated with TiO2 thin films and prepared by sputtering and non-coated PMMA were considered as control groups. The surface wettability, functional group, and roughness were determined by contact angle measurement, Fourier transform Infrared spectroscopy (FTIR), and 3D laser scanning digital microscopy, respectively. The flexural strength of coated and non-coated samples was measured using three-point bending test. The cell proliferation, attachment, and viability were determined using 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl tetrazolium bromide, live and dead assay, scanning electron microscope (SEM), and DAPI (4',6-diamidino-2-phenylindole) staining. The antifungal activity of TiO2 was also determined by examining the biofilm attachment of Candida albicans. The obtained results showed that TiO2 was successfully coated on PMMA. The contact angle measurement shows a significant increase of hydrophilicity in TiO2-coated PMMA. FTIR and roughness analysis revealed no loss of TiO2 from coated specimens following sonication. The cell viability after 7 days culturing on TiO2-coated specimens was more than the cell viability on the control groups. SEM images and DAPI staining showed that the total number of the cells increased after 7 days of seeding on TiO2-coated group, whereas it decreased gradually in both control groups. C. albicans attachment also decreased by 63% to 77% on the coated PMMA surface. Overall, this research suggested a new way for developing surface energy of PMMAs for biomedical applications.

3D-printed bi-layered polymer/hydrogel construct for interfacial tissue regeneration in a canine model.

OBJECTIVES: There are complications in applying regenerative strategies at the interface of hard and soft tissues due to the limited designs of constructs that can accommodate different cell types in different sites. The problem originates from the challenges in the adhesion of dissimilar materials, such as polymers and hydrogels, that can be suitable for regenerating different tissues such as bone and soft tissues. This paper presents a design of a new hybrid construct in which a polymer (polycaprolactone (PCL)) membrane firmly adheres to a layer of hydrogen (gelatin). METHODS: PCL membranes with defined size and porosity were fabricated using 3D printing. The gelatin layer was attached to the PCL membranes using the aminolysis procedure. We have examined this construct for the application of Guided Bone Regeneration (GBR) as a typical surgical regenerative procedure of the oral cavity at the interface of bone and soft tissue. Complete in vitro and in vivo investigations on canine tibia bone defects have been performed. Histological analyses for fibrosis morphometric and bone morphometric evaluation, as well as bone-fibrosis histological grading and CBCT imaging, were conducted. RESULTS: Chemical and morphological studies of the membrane proved that gelatin was uniformly attached to the aminolyzed PCL membranes. The in vitro and in vivo studies indicated the membrane's biocompatibility, mechanical stability, and barrier function for the GBR application. Furthermore, in vitro study showed that the membranes could improve osteogenesis and the regeneration of bone defects. The results illustrated that the mean bone density in the membrane groups was about three times more than that of the control group. SIGNIFICANCE: The fabricated 3D-printed hybrid Gelatin/PCL bi-layered membrane can be a good candidate for interfacial tissue engineering and a promising membrane for GBR procedure.

In vivo efficacy of 3D-printed elastin-gelatin-hyaluronic acid scaffolds for regeneration of nasal septal cartilage defects.

Nasal septal cartilage perforations occur due to the different pathologies. Limited healing ability of cartilage results in remaining defects and further complications. This study sought to assess the efficacy of elastin-gelatin-hyaluronic acid (EGH) scaffolds for regeneration of nasal septal cartilage defects in rabbits. Defects (4 × 7 mm) were created in the nasal septal cartilage of 24 New Zealand rabbits. They were randomly divided into four groups: Group 1 was the control group with no further intervention, Group 2 received EGH scaffolds implanted in the defects, Group 3 received EGH scaffolds seeded with autologous auricular chondrocytes implanted in the defects, and Group 4 received EGH scaffolds seeded with homologous auricular chondrocytes implanted in the defects. After a 4-month healing period, computed tomography (CT) and magnetic resonance imaging (MRI) scans were obtained from the nasal septal cartilage, followed by histological evaluations of new tissue formation. Maximum regeneration occurred in Group 2, according to CT, and Group 3, according to both T1 and T2 images with 7.68 ± 1.36, 5.44 ± 2.41, and 8.72 ± 3.02 mm2 defect area respectively after healing. The difference in the defect size was statistically significant after healing between the experimental groups. Group 3 showed significantly greater regeneration according to CT scans and T1 and T2 images. The neocartilage formed over the underlying old cartilage with no distinct margin in histological evaluation. The EGH scaffolds have the capability of regeneration of nasal cartilage defects and are able to integrate with the existing cartilage; yet, they present the best results when pre-seeded with autologous chondrocytes.

Bilayer Scaffolds for Interface Tissue Engineering and Regenerative Medicine: A Systematic Reviews.

PURPOSE: This systematic review focus on the application of bilayer scaffolds as an engaging structure for the engineering of multilayered tissues, including vascular and osteochondral tissues, skin, nerve, and urinary bladder. This article provides a concise literature review of different types of bilayer scaffolds to understand their efficacy in targeted tissue engineering. METHODS: To this aim, electronic search in the English language was performed in PMC, NBCI, and PubMed from April 2008 to December 2019 based on the PRISMA guidelines. Animal studies, including the "bilayer scaffold" and at least one of the following items were examined: osteochondral tissue, bone, skin, neural tissue, urinary bladder, vascular system. The articles which didn't include "tissue engineering" and just in vitro studies were excluded. RESULTS: Totally, 600 articles were evaluated; related articles were 145, and 35 full-text English articles met all the criteria. Fifteen articles in soft tissue engineering and twenty items in hard tissue engineering were the results of this exploration. Based on selected papers, it was revealed that the bilayer scaffolds were used in the regeneration of the multilayered tissues. The highest multilayered tissue regeneration has been achieved when bilayer scaffolds were used with mesenchymal stem cells and differentiation medium before implanting. Among the studies being reported in this review, bone marrow mesenchymal stem cells are the most studied mesenchymal stem cells. Among different kinds of multilayer tissue, the bilayer scaffold has been most used in osteochondral tissue engineering in which collagen and PLGA have been the most frequently used biomaterials. After osteochondral tissue engineering, bilayer scaffolds were widely used in skin tissue engineering. CONCLUSION: The current review aimed to manifest the researcher and surgeons to use a more sophisticated bilayer scaffold in combinations of appropriate stem cells, and different can improve multilayer tissue regeneration. This systematic review can pave a way to design a suitable bilayer scaffold for a specific target tissue and conjunction with proper stem cells.

Poloxamer: A versatile tri-block copolymer for biomedical applications.

Poloxamers, also called Pluronic, belong to a unique class of synthetic tri-block copolymers containing central hydrophobic chains of poly(propylene oxide) sandwiched between two hydrophilic chains of poly(ethylene oxide). Some chemical characteristics of poloxamers such as temperature-dependent self-assembly and thermo-reversible behavior along with biocompatibility and physiochemical properties make poloxamer-based biomaterials promising candidates for biomedical application such as tissue engineering and drug delivery. The microstructure, bioactivity, and mechanical properties of poloxamers can be tailored to mimic the behavior of various types of tissues. Moreover, their amphiphilic nature and the potential to self-assemble into the micelles make them promising drug carriers with the ability to improve the drug availability to make cancer cells more vulnerable to drugs. Poloxamers are also used for the modification of hydrophobic tissue-engineered constructs. This article collects the recent advances in design and application of poloxamer-based biomaterials in tissue engineering, drug/gene delivery, theranostic devices, and bioinks for 3D printing. STATEMENT OF SIGNIFICANCE: Poloxamers, also called Pluronic, belong to a unique class of synthetic tri-block copolymers containing central hydrophobic chains of poly(propylene oxide) sandwiched between two hydrophilic chains of poly(ethylene oxide). The microstructure, bioactivity, and mechanical properties of poloxamers can be tailored to mimic the behavior of various types of tissues. Moreover, their amphiphilic nature and the potential to self-assemble into the micelles make them promising drug carriers with the ability to improve the drug availability to make cancer cells more vulnerable to drugs. However, no reports have systematically reviewed the critical role of poloxamer for biomedical applications. Research on poloxamers is growing today opening new scenarios that expand the potential of these biomaterials from "traditional" treatments to a new era of tissue engineering. To the best of our knowledge, this is the first review article in which such issue is systematically reviewed and critically discussed in the light of the existing literature.

Fabrication and characterization of novel bilayer scaffold from nanocellulose based aerogel for skin tissue engineering applications.

The aim of this study was to fabricate a novel bilayer scaffold containing cellulose nanofiber/poly (vinyl) alcohol (CNF/PVA) to evaluate its potential use in skin tissue engineering. Here, the scaffolds were fabricated using a novel one-step freeze-drying technique with two different concentrations of the aforementioned polymers. FE-SEM analysis indicated that the fabricated scaffolds had interconnected pores with two defined pore size in each layer of the bilayer scaffolds that can recapitulate the two layers of the dermis and epidermis of the skin. Lower concentration of polymers causes higher porosity with larger pore size and increased water uptake and decreased mechanical strength. FTIR proved the presence of functional groups and strong hydrogen bonding between the molecules of CNF/PVA and the efficient crosslinking. The MTT assay showed that these nanofibrous scaffolds meet the requirement as a biocompatible material for skin repair. Here, for the first time, we fabricated bilayer scaffold using a novel one-step freeze-drying technique only by controlling the polymer concentration with spending less time and energy.

Influence of hydrodynamic pressure on chondrogenic differentiation of human bone marrow mesenchymal stem cells cultured in perfusion system.

The natural conditions of chondrocytes in native cartilage including mechanical forces and surface topology could be simulated to enhance chondrogenesis. A perfusion system recapitulating the hydrodynamic pressure of cartilage tissue is designed. Mesenchymal stem cells (MSCs) are isolated and seeded on aligned nanofibrous PCL/PLGA scaffolds that mimic the structure of superficial zone of articular cartilage. The cell-seeded scaffolds are placed into the perfusion bioreactor and exposed to chondrogenic differentiating medium. The chondrogenesis is then investigated by histological analysis and real time PCR for cartilage-specific genes. The highest expression levels of aggrecan and type II collagen are observed in the cells cultured in the presence of differentiating medium and mechanical stimulation. The expression level of type II collagen is higher than aggrecan in presence of differentiating medium and absence of mechanical stimulation. On the contrary, the expression ratio of aggrecan is higher than type II collagen in presence of mechanical stimulation and absence of differentiating medium. These results show the dominant role of mechanical stimulation and differentiating medium on upregulated expression of aggrecan and type II collagen, respectively. The application of mechanical stimulation upon cells-seeded scaffolds could mimic superficial zone of articular cartilage tissue and increase derivation of chondrocytes from MSCs.

Pancreatic Endoderm-Derived From Diabetic Patient-Specific Induced Pluripotent Stem Cell Generates Glucose-Responsive Insulin-Secreting Cells.

Human-induced pluripotent stem cells (hiPSCs) can potentially serve as an invaluable source for cell replacement therapy and allow the creation of patient- and disease-specific stem cells without the controversial use of embryos and avoids any immunological incompatibility. The generation of insulin-producing pancreatic ß-cells from pluripotent stem cells in vitro provides an unprecedented cell source for personal drug discovery and cell transplantation therapy in diabetes. A new five-step protocol was introduced in this study, effectively induced hiPSCs to differentiate into glucose-responsive insulin-producing cells. This process mimics in vivo pancreatic organogenesis by directing cells through stages resembling definitive endoderm, primitive gut-tube endoderm, posterior foregut, pancreatic endoderm, and endocrine precursor. Each stage of differentiation were characterized by stage-specific markers. The produced cells exhibited many properties of functional ß-cells, including expression of critical ß-cells transcription factors, the potency to secrete C-peptide in response to high levels of glucose and the presence of mature endocrine secretory granules. This high efficient differentiation protocol, established in this study, yielded 79.18% insulin-secreting cells which were responsive to glucose five times higher than the basal level. These hiPSCs-derived glucose-responsive insulin-secreting cells might provide a promising approach for the treatment of type I diabetes mellitus. J. Cell. Physiol. 232: 2616-2625, 2017. © 2016 Wiley Periodicals, Inc.

Investigation of 5-HT2A gene expression in PBMCs of patients with allergic asthma.

BACKGROUND: Asthma is an inflammatory airway disorder in which different immune cells in the blood and lungs play a fundamental role. In asthma condition, the airway inflammation accompanied by bronchial smooth muscle spasm cause airway obstruction. A study showed that high concentration of blood serotonin is associated with the intensity and exacerbation of asthma disease. Other studies showed that a subtype of serotonin receptor called 5-Hydroxytriptamine 2A receptor (5- HT2A) can enhance T-cell blastogenesis and production of pro-inflammatory cytokines such as IFNγ. OBJECTIVE: The objective of this study was to assess the level of 5-HT2A in peripheral blood mononuclear cells (PBMCs) of asthmatic patients. METHODS: PBMCs were extracted from blood of 30 patients with asthma and 30 normal people. After synthesizing cDNAs from total mRNAs, real-time PCR was performed to amplify 5-HT2A and ß-actin (as an internal control). The expression ratios were analyzed in patients with asthma in comparison with normal group. RESULTS: The results indicated that gene expression is significantly increased in peripheral blood mononuclear cells (PBMCs) of asthma patients in comparison with normal group (P = 0.003). CONCLUSION: The results of this study can suggest designing a protocol by using of the 5-HT2A receptor expression in PBMCs as a biomarker of asthma, but this requires further studies on a larger number of patients. In addition, the potential role of this receptor in bronchoconstriction can lead us to use its antagonists as a new treatment in asthma.

Investigation of immunomodulatory properties of human Wharton's Jelly-derived mesenchymal stem cells after lentiviral transduction.

Human Wharton's Jelly-derived Mesenchymal Stem Cells (hWJ-MSCs) are considered as an alternative for bone-marrow-derived MSCs. These cells have immunosuppressive properties. It was unclear whether the WJ-MSCs would sustain their immunomodulatory characteristics after lentiviral transduction or not. In this study, we evaluated immunomodulatory properties of WJ-MSCs after lentiviral transduction. HWJ-MSCs were transduced with lentiviral particles. Expression of transduced and un-transduced hWJ-MSCs surface molecules and secretion of IL-10, HGF, VEGF and TGF-ß was analyzed. Cell proliferation and frequency of CD4(+)CD25(+) CD127(low/neg) Foxp3(+) T regulatory cells was measured. There was no difference between the surface markers and secretion of IL-10, HGF, VEGF and TGF-ß in transduced and un-transduced hWJ-MSCs. Both cells inhibited the proliferation of PHA stimulated PBMCs, and improved the frequency of T regulatory cells. These findings suggest that lentiviral transduction does not alter the immunomodulatory function of hWJ-MSCs. However, lentiviral transduction may have a wide range of applications in gene therapy.

Investigation of 5-HT3A receptor gene expression in peripheral blood mononuclear cells of individuals who had been exposed to air pollution.

The role of air pollution in exacerbation of allergic symptoms is well known. Several studies have shown the effect of air pollution on serotonergic system. The changes in serotonergic system could trigger several allergic symptoms. 5-HT(3A) is among serotonin receptors on the peripheral Blood Mononuclear Cells (PBMCs) as well as other cells. In the present study we compared the 5-HT(3A) gene expression in PBMCs of the asthmatic patients as well as individuals who had been exposed to the air pollution. Normal individuals were also included in the study as control for comparison of 5-HT(3A) gene expression. Following the synthesis of the cDNA using mRNA extracted from PBMCs the level of 5- HT(3A) gene expression was measured using real-time PCR. The results showed t a significant increase in the relative expression level of 5-HT(3A) receptor in PBMCs from asthmatic patients and individuals exposed to the air pollutants compared to normal controls. Our result indicates that significant increase in 5-HT(3A) receptor may contribute to the pathogenesis as well as allergic symptoms which resulted from air pollution.