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

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

A Nontoxic and Biocompatible Method for Augmenting Mechanical Strength of Acellular Matrix by Silk Fibroin Impregnation.

Biological scaffolds are plagued by poor biomechanical properties and untimely degradation. These limitations have yet to be addressed without compromising their biocompatibility. It is desirable to avoid inflammation and have degradation with concomitant host collagen deposition or even site-appropriate in situ regeneration for the successful outcome of an implanted biological scaffold. This work aims to achieve this by utilizing a biocompatible method to modify acellular scaffolds by impregnating alkaline-catalyzed citric acid (CA) cross-linking between the extracellular matrix proteins and silk fibroin (SF)/SF-gelatin (SFG) blends. Combinatorial detergent decellularization was employed to prepare a decellularized porcine liver scaffold (DPL). After proving the decellularization efficiency, the scaffold underwent modification by vacuum impregnation with CA containing SF (SF100DPL) and SFG blends (SFG5050DPL and SFG3070DPL) following pre-cross-linking, drying, and post-cross-linking. The subsequent strength augmentation was demonstrated by significant improvement in tensile strength from 2.4 ± 0.4 MPa (DPL) to, 3.8 ± 0.7 MPa (SF100DPL), 3.4 ± 0.7 MPa (SFG5050DPL), and 3.5 ± 0.2 MPa (SFG3070DPL); Young's modulus from 8.7 ± 1.8 MPa (DPL) to 20 ± 1.9 MPa (SF100DPL), 13.3 ± 2.6 MPa (SFG5050DPL), and 16 ± 1.2 MPa (SFG3070DPL); and suture retention strength from 0.9 ± 0.08 MPa (DPL) to 2.3 ± 0.2 MPa (SF100DPL), 2.8 ± 1.2 MPa (SFG5050DPL), and 2.6 ± 0.9 MPa (SFG3070DPL). The degradation resistance of the modified scaffolds was also markedly improved. Being cytocompatible, its ability to incite tolerable inflammatory and immune responses was confirmed by rat subcutaneous implantation for 14, 30, and 90 days, in terms of inflammatory cell infiltration, neoangiogenesis, and in vitro cytokine release to assess B-cell and T-cell activation. Such ECM composite scaffolds with appropriate strength and biocompatibility offer great promise in soft tissue repair applications such as skin grafting.

Design and evaluation of propranolol hydrochloride loaded thiolated Zein/PEO electrospun fibrous matrix for transmucosal drug delivery.

Thiolated polymers have garnered wide attention from researchers on mucoadhesive drug delivery. This work explores the thiolation of zein protein using cysteine amino acid via the EDC crosslinker. The optimization of thiolation and purification have been done and confirmed using Ellman's assay and Raman spectra. The thiolated Zein/PEO polymer blend has been appraised for electrospinning to fabricate fibrous matrices. The extent of thiol modification augmented the mechanical properties and adhesion in rabbit intestinal mucosa. In vitro cytotoxicity evaluations such as direct contact assay, MTT assay, and live dead assay performed in RPMI 2650 cells corroborated the non-cytotoxicity of the fabricated matrices with and without propranolol hydrochloride (PL). Detailed drug release studies were conducted in PBS. Drug release in PBS followed the Korsmeyer Peppas model of release. On treating RPMI 2650 cells with the matrix, F-actin and adherens junctional proteins retained integrity, and consequently, drug permeation would proceed through the transcellular transport mechanism. Transepithelial electrical resistance (TEER) measurement of the RPMI 2650 cell monolayer also supported the transcellular transport mechanism. Ex vivo permeation study through porcine buccal mucosa showed 41.26 ± 0.56% PL permeation within 24 h of study. It validated the competence of the electrospun thiolated Zein/PEO matrix for transmucosal drug delivery.

Electrospun Mucoadhesive Zein/PVP Fibroporous Membrane for Transepithelial Delivery of Propranolol Hydrochloride.

Mucoadhesive drug delivery systems have been extensively studied to effectively reduce the limitations of conventional drug delivery systems. Zein and polyvinyl pyrrolidone (PVP) are appraised for mucoadhesive properties. This study focuses on developing a mechanically stable zein/PVP electrospun membrane for propranolol hydrochloride (PL) transport. Fourier transform infrared, Raman spectra, and swelling studies gave evidence for PVP crosslinking, whereas circular dichroism spectroscopy revealed crosslinking of zein owing to the conformational change from α-helix to ß-sheet. A 10 h thermal treatment of zein/PVP imparted 3.92 ± 0.13 MPa tensile strength to the matrix. Thermally crosslinked electrospun zein/PVP matrix showed 22.1 ± 0.1 g mm work of adhesion in porcine buccal mucosa tissue. Qualitative and quantitative evaluation of cytotoxicity in RPMI 2650 has been carried out. The in vitro drug release profile of PL from thermally crosslinked zein/PVP best fitted with the Korsmeyer-Peppas model. Immunostaining of ß-catenin adherens junctional protein confirmed the absence of paracellular transport through the junctional opening. Still, drug permeation was observed through the porcine buccal mucosa, attributed to the transcellular transport of PL owing to its lipophilicity. The ex vivo permeation of PL through porcine buccal mucosa was also evaluated.

Neuronal cell response on aligned fibroporous electrospun mat generated from silver ion complexed ethylene vinyl alcohol copolymer.

Generating electrospun mats with aligned fibers and obtaining neurite extension in the aligned fiber direction could provide hope for fabricating nerve guidance conduits or wraps through an easy method. The growing interest in generating electrospun mats with aligned fibers for tissue engineering is looking for simple methods to generate the same. Here, in this study, ethylene vinyl alcohol copolymer (EVAL) chains were complexed with silver ions (Ag+ ) to generate aligned fibers during the electrospinning process. The fibers thus produced were subjected to physico-chemical characterization and biological studies to ensure their properties and to examine whether suitable for neuronal cell attachment and neurite extension that may be useful in making nerve guidance conduits or wraps. The presence of silver ions and its complex formation with -OH of EVAL has been confirmed with EDX and XPS analysis respectively. The alignment of fibers was visualized from SEM analysis and confirmed using directionality analysis using Fiji-ImageJ software. Mechanical properties done with dumbbells punched out in longitudinal and transverse directions also substantiated the alignment of fibers. The results obtained from direct contact, MTT, and live/dead assay showed the cells are viable on the material. From the actin staining and immunostaining assays, it was evident that the PC12 cells could attach and extend their neurites in an aligned manner on the fibers. The maximum neurite extension was up to 200 µm in length. These properties of electrospun EVAL-Ag mat with aligned fibers indicated that it could be developed as a biocompatible nerve guidance conduit or wrap.

Recent advances in functionally modified polymers for mucoadhesive drug delivery.

Novel methods for the delivery of drugs other than the conventional method of oral administration have been a thrust area of research for a few decades. Mucoadhesive delivery of drugs opened up a new domain where rapid and patient-friendly delivery of drugs can be achieved. Delivery of drugs through the mucosal sites such as buccal, nasal, ocular, sublingual, rectal and vaginal facilitates bypassing the first-pass metabolism and the drug reaches the systemic circulation directly. This helps to increase the bioavailability of the drug. The study of the chemical characteristics of polymers with mucoadhesive properties and how the molecules or the pharmaceuticals are transported across the mucosa is very much needed for the advancement of research in this field. And at the same time, it is very pertinent to know about the anatomy and the physiology of the mucosal tissue and its variation in different regions of the body. In this review, we try to present a comprehensive understanding of relevant topics of mucoadhesion giving more emphasis on the mechanism of transport of drugs across mucosa, and different possible functional modifications of polymers to enhance the property of mucoadhesion.

UV-Crosslinked Electrospun Zein/PEO Fibroporous Membranes for Wound Dressing.

Electrospun zein membranes are suitable for various biomedical applications. A UV-crosslinked electrospun membrane of a zein/PEO blend for wound healing application was explored in this work. The improvement in mechanical properties of the membrane after UV crosslinking was attributed to the change in protein conformation from an α-helix to a ß-sheet. The circular dichroism (CD) spectra and FTIR spectra confirmed this conformational change. XRD analysis was shown to prove the amorphous nature of polymer blends with specific broad peaks at 2θ = 9° and 20°. The water vapor transmission rate (WVTR) of the membrane was found to be in the range of 1500-2000 g m-2 day-1, which was well suited with that of commercially available wound dressing material. Enough number of available functional groups like thiol, amino, and hydroxyl groups supplement a blood clotting index (BCI) to the matrix, causing 99% BCI within 4 min. A 91% cell viability result in the MTT assay with human dermal fibroblast cells confirmed the noncytotoxicity of the membrane. Tripeptides produced after the thermolysin-based hydrolysis of zein caused inhibition of TGF ß1 expression and thus increased fibroblast and collagen production. The membrane stimulated 54% more collagen production compared to control cells at day 2 and caused 84% wound closure in human dermal fibroblast cells, which were desirable index markers of a potential wound care material.

Accelerated Outgrowth of Neurites on Graphene Oxide-Based Hybrid Electrospun Fibro-Porous Polymeric Substrates.

Fabrication of a surface-engineered electrospun scaffold having biomimetic properties like the extracellular matrix (ECM) is essential for neural tissue engineering. An electroconductive and elastomeric scaffold with aligned fibers acting as a substrate may have a great impact on the directional outgrowth of neurites. In this study, we have electrospun electrically conductive, polyurethane-based elastomeric and topographically aligned fibro-porous neural scaffolds. Adhesive proteins of the ECM are documented to have an important role in controlling neuronal cell behavior, including cell adhesion, proliferation, and neurite outgrowth. These bio-adhesion proteins or nanomaterials mimicking their action, if used for surface modification of neural scaffolds, may have the potential to accelerate the nerve repair process. Thus, electrospun scaffolds fabricated were surface-engineered using a unique and modified single-step electrospraying technique to coat the scaffold surface with an exploratory bio-adhesion agent, a thin layer of graphene oxide (GO) films. The study was then carried out to determine if the GO-coated electrospun electroconductive polycarbonate urethane (PCU) substrate can improve the bio-interface attributes of these scaffolds or may alter the neurite outgrowth of PC-12 cells like any other bio-adhesion proteins. Therefore, the hybrid scaffolds with GO coatings were compared with similar scaffolds coated with poly-l-lysine (PLL) for neural cell adhesion, proliferation, and neurite extension. Neurite outgrowth studies showed that although the average neurite length was comparable on both GO- and PLL-coated surfaces, the length profile of neurites, when categorized based on length, showed an increased number of lengthier neurites on the GO-coated hybrid scaffolds. In particular, the study brings out an innovative surface engineering technique for the coating of GO on polymeric scaffolds. It may be further put together in designing of hybrid surfaces with nanotopographical biophysical cues on three-dimensional neural scaffolds, which in turn may stimulate an accelerated neuronal regeneration via providing an enhanced ECM like milieu.

Pamidronate-Encapsulated Electrospun Polycaprolactone-Based Composite Scaffolds for Osteoporotic Bone Defect Repair.

Bone fractures associated with osteoporosis are a major concern all over the world especially among the elderly population and postmenopausal women. Bisphosphonates (BPs) are widely used clinically for both treatment and prevention of osteoporosis despite their poor oral bioavailability and undesired side effects. Local delivery of BPs from polymeric scaffolds can improve the efficacy and overcome the undesirable side effects associated with oral bisphosphonate therapy. The aim of the present study is to explore the effectiveness of pamidronate (PDS) encapsulated electrospun polycaprolactone/polycaprolactone-polyethyleneglycol-polycaprolactone/nanohydroxyapatite (PCH) scaffolds in healing critical-size calvarial defects in an osteoporotic rat animal model. Prior to implantation studies, the effect of PDS on the fiber architecture, mechanical properties, and in vitro degradation behavior was evaluated. The in vitro release of PDS from PCH scaffolds in phosphate buffer saline (PBS) at 37 °C was monitored for a period of 21 days. An osteoporotic animal model was successfully developed in Wistar rats by bilateral ovariectomy. Results of micro CT (computed tomography) and blood serum analysis confirmed the osteoporotic model induction in rats. Critical-size calvarial defects of 8 mm size were created in osteoporotic rats, and the in vivo osteogenic efficacy of PCH-PDS scaffolds was evaluated by micro CT, histology, and histomorphometry. Micro CT analysis showed improved osseous tissue integration with the use of PDS-loaded PCH scaffolds after 12 week post implantation. Histology, density measurement using micro CT, and histomorphometry further substantiate that PCH-PDS scaffolds have the potential to be used for the repair of osteoporotic bone defects. Our findings revealed that incorporation of PDS onto PCH scaffolds provides a promising biomaterial that could be used for regenerating osteoporosis-related fractures.

Differential Adhesive and Bioactive Properties of the Polymeric Surface Coated with Graphene Oxide Thin Film.

Surface engineering of implantable devices involving polymeric biomaterials has become an essential aspect for medical implants. A surface enhancement technique can provide an array of unique surface properties that improve its biocompatibility and functionality as an implant. Polyurethane-based implants that have found extensively acclaimed usage as an implant in biomedical applications, especially in the area of cardiovascular devices, still lack any mechanism to ward off bacterial or platelet adhesion. To bring out such a defense mechanism we are proposing a surface modification technique. Graphene oxide (GO) in very thin film form was wrapped onto the electrospun fibroporous polycarbonate urethane (PCU) membrane (GOPCU) by a simple method of electrospraying. In the present study, we have developed a simple single-step method for coating a polymeric substrate with a thin GO film and evaluated the novel antiadhesive activity of these films. SEM micrographs after coating showed the presence of very thin GO films over the PCU membrane. On the GOPCU surface, the contact angle was shifted by ∼30°, making the hydrophobic PCU surface slightly hydrophilic, while Raman spectral characterization and mapping showed the presence and distribution of GO over 75% of the membrane. A reduced platelet adhesion on the GOPCU surface was observed; meanwhile, bacterial adhesion also got reduced by 85% for Staphylococcus aureus (Gram positive, cocci) and 64% for Pseudomonas aeruginosa (Gram negative, bacilli). A cell adhesion study conducted using mammalian fibroblast cells projected its proliferation percentage in a MTT assay, with 82% cell survival on PCU and 86% on GOPCU after 24 h culture, while a study for an extended period of 72 h showed 87% of survival on PCU and 88% on GOPCU. This plethora of functionalities by a simple modification technique makes thin GO films a self-sufficient surface engineering material for future biomedical applications.