Collagen has been used extensively in tissue engineering applications. However, the source of collagen has been primarily bovine and porcine. In view of the potential risk of zoonotic diseases and religious constraints associated with bovine and porcine collagen, fish collagen was examined as an alternative. The aim of this study is to use tilapia fish collagen to develop a cross-linked electrospun membrane to be used as a barrier membrane in guided bone regeneration. As there is limited data available on the cytotoxicity and immunogenicity of cross-linked tilapia collagen, in vitro and in vivo tests were performed to evaluate this in comparison to the commercially available Bio-Gide membrane. In this study, collagen was extracted and purified from tilapia skin and electrospun into a nanofibrous membrane. The resultant membrane was cross-linked to obtain a cross-linked electrospun tilapia collagen (CETC) membrane, which was characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), degradation studies, cytotoxicity studies, and cell proliferation studies. The membranes were also implanted subcutaneously in rats and the host immune responses were examined. The DSC data showed that cross-linking increased the denaturation temperature of tilapia collagen to 58.3°C ± 1.4°C. The in vitro tests showed that CETC exhibited no cytotoxicity toward murine fibroblast L929 cells, and culture of murine preosteoblast MC3T3-E1 cells demonstrated better proliferation on CETC as compared to Bio-Gide. When implanted in rats, CETC caused a higher production of interleukin IL-6 at early time points as compared to Bio-Gide, but there was no long-term inflammatory responses after the acute inflammation phase. This finding was supported with histology data, which clearly illustrated that CETC has a decreased inflammatory response comparable to the benchmark control group. In all, this study demonstrated the viability for the use of CETC as a tissue engineering scaffold and provides an insight on the in vivo immune responses toward xenogenic collagen scaffolds.
Bone transplants are used to treat fractures and increase new tissue development in bone tissue engineering. Grafting of massive implantations showing slow curing rate and results in cell death for poor vascularization. The potentials of biocomposite scaffolds to mimic extracellular matrix (ECM) and including new biomaterials could produce a better substitute for new bone tissue formation. A purpose of this study is to analyze polycaprolactone/silk fibroin/hyaluronic acid/minocycline hydrochloride (PCL/SF/HA/MH) nanoparticles initiate human mesenchymal stem cells (MSCs) proliferation and differentiation into osteogenesis. Electrospraying technique was used to develop PCL, PCL/SF, PCL/SF/HA and PCL/SF/HA/MH hybrid biocomposite nanoparticles and characterization was analyzed by field emission scanning electron microscope (FESEM), contact angle and Fourier transform infrared spectroscopy (FT-IR). The obtained results proved that the particle diameter and water contact angle obtained around 0.54 ± 0.12 to 3.2 ± 0.18 µm and 43.93 ± 10.8° to 133.1 ± 12.4° respectively. The cell proliferation and cell-nanoparticle interactions analyzed using (3-(4,5-dimethyl thiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt) MTS assay (Promega, Madison, WI, USA), FESEM for cell morphology and 5-Chloromethylfluorescein diacetate (CMFDA) dye for imaging live cells. Osteogenic differentiation was proved by expression of osteocalcin, alkaline phosphatase activity (ALP) and mineralization was confirmed by using alizarin red (ARS). The quantity of cells was considerably increased in PCL/SF/HA/MH nanoparticles when compare to all other biocomposite nanoparticles and the cell interaction was observed more on PCL/SF/HA/MH nanoparticles. The electrosprayed PCL/SF/HA/MH biocomposite nanoparticle significantly initiated increased cell proliferation, osteogenic differentiation and mineralization, which provide huge potential for bone tissue engineering.
Hybrid biocomposite nanofibrous structures that mimics native extracellular matrix have been extensively applied for bone tissue engineering (BTE) due to their potential in efficiently inducing cellular response for the secretion of extracellular matrix (ECM). This study performed fabrication of uniform porous polycaprolactone (PCL), polycaprolactone/ silk fibroin (PCL/SF), polycaprolactone/silk fibroin/ minocycline hydrochloride (PCL/SF/MH), polycaprolactone/ collagen (PCL/COL), and polycaprolactone/collagen/ minocycline hydrochloride (PCL/COL/MH) biocomposites nanofibrous scaffolds by electrospinning, for comparing their properties to use in bone tissue regeneration. Field emission scanning electron microscopy (FESEM) images of fabricated nanofibrous scaffolds revealed porous, beadless, uniform fibers of diameter in the range of 147.13 ± 28.02 to 176.53 ± 22.34 nm and porosity around 82-93 %. Adipose-derived stem cells (ADSCs) considered as the novel cell therapeutics were cultured on these electrospun fibrous scaffolds to undergo osteogenic differentiation for BTE. The cell morphology, proliferation, and interactions were analyzed by CMFDA dye extrusion, MTS assay, and FESEM analysis, respectively. Differentiation of ADSCs into osteogenesis was determined by alkaline phosphatase activity, mineralization by alizarin red staining, and osteogenic protein expression by immunofluorescence analysis. The results demonstrated that the addition of SF and MH to PCL-based scaffolds improved the mechanical stability, interconnected pores, and surface roughness of the scaffolds initiating heightened biological functions such as ADSCs adhesion, proliferation, differentiation, and mineralization into osteogenesis for bone tissue regeneration. Lay Summary Globally, the rate of bone defects or trauma has been trending upwards and is predicted to rapidly increase by 2020. This is mainly due to the lack of physical activity, age, and increased obese populations. A basic understanding of the morphological characteristics and biological functions of mineralized bone tissue substrates is required for effective clinical treatments. The engineered bone tissue substrates have been potentially used as an alternative to the traditional bone grafts. However using these engineered bone substrates in clinical practices has certain limitations. These limitations can be overcome by the inclusion of natural polymers, growth factors, and stem cells. The current study demonstrates the addition of silk fibroin and minocycline hydrochloride to polycaprolactone-based scaffolds potentially enhanced adipose-derived stem cells differentiation into osteogenesis.
A new strategy for supercapacitor formation was carried out in the study using electrodes made of graphene oxide (GO) and manganese dioxide (MnO2) nanocomposites. To the present knowledge, only a few investigations have been carried out concerning the synthesis of GO-MnO2-based nanocomposites and their electrochemical properties, with varying mass ratios, as well as the change in electrochemical properties of their components as MnO2 and GO were tested individually for the enhanced stability and performances. A synthetic method was performed successfully to manufacture MnO2/GO nanocomposites. The findings of the present study show that the composites have a lot of potential as an effective conduction property. A composite of graphene oxide supported manganese dioxide nanocomposites fabricated with the simple soft chemical route. As-prepared nanocomposites can be improved in performance by the interactions between GO and MnO2.
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