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      Development of Biopolymeric Hybrid Scaffold-Based on AAc/GO/nHAp/TiO 2 Nanocomposite for Bone Tissue Engineering: In-Vitro Analysis

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          Abstract

          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 2 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 2 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 ( MC3T3-E1) cell lines. Hence, these fabricated scaffolds would be potential biomaterials for bone tissue engineering in biomedical engineering.

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          Neutral red uptake assay for the estimation of cell viability/cytotoxicity.

          Guillermo Repetto, Ana del Peso, Jorge L Zurita (2008)
          The neutral red uptake assay provides a quantitative estimation of the number of viable cells in a culture. It is one of the most used cytotoxicity tests with many biomedical and environmental applications. It is based on the ability of viable cells to incorporate and bind the supravital dye neutral red in the lysosomes. Most primary cells and cell lines from diverse origin may be successfully used. Cells are seeded in 96-well tissue culture plates and are treated for the appropriate period. The plates are then incubated for 2 h with a medium containing neutral red. The cells are subsequently washed, the dye is extracted in each well and the absorbance is read using a spectrophotometer. The procedure is cheaper and more sensitive than other cytotoxicity tests (tetrazolium salts, enzyme leakage or protein content). Once the cells have been treated, the assay can be completed in <3 h.
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            Biodegradable synthetic polymers for tissue engineering.

            Ajeya Adhikari, P Gunatillake (2003)
            This paper reviews biodegradable synthetic polymers focusing on their potential in tissue engineering applications. The major classes of polymers are briefly discussed with regard to synthesis, properties and biodegradability, and known degradation modes and products are indicated based on studies reported in the literature. A vast majority of biodegradable polymers studied belongs to the polyester family, which includes polyglycolides and polylactides. Some disadvantages of these polymers in tissue engineering applications are their poor biocompatibility, release of acidic degradation products, poor processability and loss of mechanical properties very early during degradation. Other degradable polymers such as polyorthoesters, polyanhydrides, polyphosphazenes, and polyurethanes are also discussed and their advantages and disadvantages summarised. With advancements in tissue engineering it has become necessary to develop polymers that meet more demanding requirements. Recent work has focused on developing injectable polymer compositions based on poly (propylene fumarate) and poly (anhydrides) to meet these requirements in orthopaedic tissue engineering. Polyurethanes have received recent attention for development of degradable polymers because of their great potential in tailoring polymer structure to achieve mechanical properties and biodegradability to suit a variety of applications.
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              Electrospun poly(epsilon-caprolactone) microfiber and multilayer nanofiber/microfiber scaffolds: characterization of scaffolds and measurement of cellular infiltration.

              Upma Sharma, Quynh Pham, Antonios Mikos (2006)
              The physical and spatial architectural geometries of electrospun scaffolds are important to their application in tissue engineering strategies. In this work, poly(epsilon-caprolactone) microfiber scaffolds with average fiber diameters ranging from 2 to 10 microm were individually electrospun to determine the parameters required for reproducibly fabricating scaffolds. As fiber diameter increased, the average pore size of the scaffolds, as measured by mercury porosimetry, increased (values ranging from 20 to 45 microm), while a constant porosity was observed. To capitalize on both the larger pore sizes of the microfiber layers and the nanoscale dimensions of the nanofiber layers, layered scaffolds were fabricated by sequential electrospinning. These scaffolds consisted of alternating layers of poly(epsilon-caprolactone) microfibers and poly(epsilon-caprolactone) nanofibers. By electrospinning the nanofiber layers for different lengths of time, the thickness of the nanofiber layers could be modulated. Bilayered constructs consisting of microfiber scaffolds with varying thicknesses of nanofibers on top were generated and evaluated for their potential to affect rat marrow stromal cell attachment, spreading, and infiltration. Cell attachment after 24 h did not increase with increasing number of nanofibers, but the presence of nanofibers enhanced cell spreading as evidenced by stronger F-actin staining. Additionally, increasing the thickness of the nanofiber layer reduced the infiltration of cells into the scaffolds under both static and flow perfusion culture for the specific conditions tested. The scaffold design presented in this study allows for cellular infiltration into the scaffolds while at the same time providing nanofibers as a physical mimicry of extracellular matrix.
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                Author and article information

                Contributors
                Cristina Martín: Role: Academic Editor
                Journal
                Journal ID (nlm-ta): Nanomaterials (Basel)
                Journal ID (iso-abbrev): Nanomaterials (Basel)
                Journal ID (publisher-id): nanomaterials
                Title: Nanomaterials
                Publisher: MDPI
                ISSN (Electronic): 2079-4991
                Publication date (Electronic): 17 May 2021
                Publication date Collection: May 2021
                Volume: 11
                Issue: 5
                Electronic Location Identifier: 1319
                Affiliations
                [1 ]BioInspired Device and Tissue Engineering Research Group, School of Biomedical Engineering and Health Sciences, Faculty of Engineering, Universiti Teknologi Malaysia, Skudai 81300, Johor, Malaysia; saifulizwan@ 123456utm.my
                [2 ]Department of Polymer Engineering and Technology, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590, Pakistan
                [3 ]Institute for Personalized Medicine, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China
                [4 ]Department of Metallurgy and Materials Engineering, CEET, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590, Pakistan; mohsin.ceet@ 123456pu.edu.pk
                [5 ]Department of Chemistry, College of Science, King Faisal University, Al-Ahsa 31982, Saudi Arabia; walarjan@ 123456kfu.edu.sa
                [6 ]Department of Chemistry, Faculty of Science, University of Jeddah, Jeddah 21589, Saudi Arabia; mbinkadem@ 123456uj.edu.sa
                [7 ]Department of Engineering Management, College of Engineering, Prince Sultan University, Rafha Street, Riyadh 11586, Saudi Arabia; hmehboob@ 123456psu.edu.sa
                [8 ]Department of Biological Sciences, National University of Medical Sciences, Rawalpindi, Punjab 46000, Pakistan; adnan_phd@ 123456outlook.com
                [9 ]Center for Advanced Composite Materials, Universiti Teknologi Malaysia, Skudai 81300, Johor, Malaysia
                [10 ]Biomedical Research Center, Qatar University, Doha 2713, Qatar; hasan.anwarul.mit@ 123456gmail.com
                [11 ]Department of Mechanical and Industrial Engineering, Qatar University, Doha 2713, Qatar
                [12 ]Department of Biology, College of Sciences, University of Hafr Al Batin, Hafr Al Batin 39524, Saudi Arabia; rashida@ 123456uhb.edu.sa
                Author notes
                Author information
                https://orcid.org/0000-0002-2724-1336
                https://orcid.org/0000-0003-1184-1326
                https://orcid.org/0000-0002-2544-4754
                Article
                Publisher ID: nanomaterials-11-01319
                DOI: 10.3390/nano11051319
                PMC ID: 8156135
                PubMed ID: 34067844
                SO-VID: 6dd31840-8f59-415a-ab89-2e9cde078ce7
                Copyright © © 2021 by the authors.
                License:

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( https://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 09 April 2021
                Date accepted : 09 May 2021
                Categories
                Subject: Article

                Keywords: biocompatibility,biodegradation,cytotoxicity,drug delivery,graphene oxide,hybrid scaffolds,nanocomposite,tissue engineering
                Data availability:
                Keywords: biocompatibility, biodegradation, cytotoxicity, drug delivery, graphene oxide, hybrid scaffolds, nanocomposite, tissue engineering

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