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1.
ACS Appl Bio Mater ; 7(4): 2389-2401, 2024 04 15.
Article in English | MEDLINE | ID: mdl-38502100

ABSTRACT

Graphene-based nanomaterials, renowned for their outstanding electrical conductivity, have been extensively studied as electroconductive biomaterials (ECBs) for electrically stimulated tissue regeneration. However, using eco-friendly reducing agents like l-ascorbic acid (l-Aa) can result in lower conductive properties in these ECBs, limiting their full potential for smooth charge transfer in living tissues. Moreover, creating a flexible biomaterial scaffold using these materials that accurately mimics a specific tissue microarchitecture, such as nerves, poses additional challenges. To address these issues, this study developed a microfibrous scaffold of Bombyx mori (Bm) silk fibroin uniformly coated with graphene nanoplatelets (GNPs) through a vacuum coating method. The scaffold's electrical conductivity was optimized by varying the reduction period using l-Aa. The research systematically investigated how different reduction periods impact scaffold properties, focusing on electrical conductivity and its significance on electrically stimulated axonal growth in PC12 cells. Results showed that a 48 h reduction significantly increased surface electrical conductivity by 100-1000 times compared to a shorter or no reduction process. l-Aa contributed to stabilizing the reduced GNPs, demonstrated by a slow degradation profile and sustained conductivity even after 60 days in a proteolytic environment. ß (III) tubulin immunostaining of PC12 cells on varied silk:GNP scaffolds under pulsed electrical stimulation (ES, 50 Hz frequency, 1 ms pulse width, and amplitudes of 100 and 300 mV/cm) demonstrates accelerated axonal growth on scaffolds exhibiting higher conductivity. This is supported by upregulated intracellular Ca2+ dynamics immediately after ES on the scaffolds with higher conductivity, subjected to a prolonged reduction period. The study showcases a sustainable reduction approach using l-Aa in combination with natural Bm silk fibroin to create a highly conductive, mechanically robust, and stable silk:GNP-based aligned fibrous scaffold. These scaffolds hold promise for functional regeneration in electrically excitable tissues such as nerves, cardiac tissue, and muscles.


Subject(s)
Bombyx , Fibroins , Graphite , Rats , Animals , Silk , Tissue Scaffolds , Graphite/pharmacology , Fibroins/pharmacology , Biocompatible Materials/pharmacology , Electric Conductivity
2.
Article in English | MEDLINE | ID: mdl-37937564

ABSTRACT

According to the World Health Organization (WHO), diabetes has been increasing steadily over the past few decades. In developing countries, it is the cause of increased morbidity and mortality. Diabetes and its complications are associated with education, occupation, and income across all levels of socioeconomic status. Factors, such as hyperglycemia, social ignorance, lack of proper health knowledge, and late access to medical care, can worsen diabetic complications. Amongst the complications, neuropathic pain and inflammation are considered the most common causes of morbidity for common populations. This review is focused on exploring protein kinase C (PKC)-mediated TGF-ß regulation in diabetic complications with particular emphasis on allodynia. The role of PKC-triggered TGF-ß in diabetic neuropathy is not well explored. This review will provide a better understanding of the PKC-mediated TGF-ß regulation in diabetic neuropathy with several schematic illustrations. Neuroinflammation and associated hyperalgesia and allodynia during microvascular complications in diabetes are scientifically illustrated in this review. It is hoped that this review will facilitate biomedical scientists to better understand the etiology and target drugs effectively to manage diabetes and diabetic neuropathy.

3.
Adv Drug Deliv Rev ; 203: 115120, 2023 12.
Article in English | MEDLINE | ID: mdl-37884128

ABSTRACT

Diabetic foot ulcers (DFUs) are a devastating complication for diabetic patients that have debilitating effects and can ultimately lead to limb amputation. Healthy wounds progress through the phases of healing leading to tissue regeneration and restoration of the barrier function of the skin. In contrast, in diabetic patients dysregulation of these phases leads to chronic, non-healing wounds. In particular, unresolved inflammation in the DFU microenvironment has been identified as a key facet of chronic wounds in hyperglyceamic patients, as DFUs fail to progress beyond the inflammatory phase and towards resolution. Thus, control over and modulation of the inflammatory response is a promising therapeutic avenue for DFU treatment. This review discusses the current state-of-the art regarding control of the inflammatory response in the DFU microenvironment, with a specific focus on the development of biomaterials-based delivery strategies and their cargos to direct tissue regeneration in the DFU microenvironment.


Subject(s)
Diabetes Mellitus , Diabetic Foot , Humans , Diabetic Foot/complications , Diabetic Foot/drug therapy , Wound Healing , Skin , Inflammation
4.
ACS Appl Bio Mater ; 5(7): 3193-3211, 2022 07 18.
Article in English | MEDLINE | ID: mdl-35775198

ABSTRACT

Bioscaffolds having electrically conducting polymers (CPs) have become increasingly relevant in tissue engineering (TE) because of their ability to regulate conductivity and promote biological function. With this in mind, the current study shows a conducting polyaniline nanofibers (PNFs) dispersed chitosan (Ch) nanocomposites scaffold with a simple one-step surface functionalization approach using glutaraldehyde for potential neural regeneration applications. According to the findings, 4 wt % PNFs dispersion in Ch matrix is an optimal concentration for achieving desirable biological functions while maintaining required physicochemical properties as evidenced by SEM, XRD, current-voltage (I-V) measurement, mechanical strength test, and in vitro biodegradability test. Surface chemical compositional analysis using XPS and ATR FT-IR confirms the incorporation of aldehyde functionality after functionalization, which is corroborated by surface energy calculations following the Van Oss-Chaudhury-Good method. Surface functionalization induced enhancement in surface hydrophilicity in terms of the polar component of surface energy (γiAB) from 6.35 to 12.54 mN m-1 along with an increase in surface polarity from 13.61 to 22.54%. Functionalized PNF:Ch scaffolds demonstrated improvement in enzyme activity from 67 to 94% and better enzyme kinetics with a reduction of Michaelis constants (Km) from 21.55 to 13.81 mM, indicating favorable protein-biomaterial interactions and establishing them as biologically perceptible materials. Surface functionalization mediated improved cell-biomaterial interactions led to improved viability, adhesion, and spreading of primary adipose derived mesenchymal stem cells (ADMSCs) as well as improved immunocompatibility. Cytoskeletal architecture assessment under differentiating media containing 10 ng/mL of each basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) revealed significant actin remodeling with neurite-like projections on the functionalized scaffolds after 14 days. Immunocytochemistry results showed that more than 85% of cells expressed early neuron specific ß III tubulin protein on the functionalized scaffolds, whereas glial fibrillary acidic protein (GFAP) expression was limited to approximately 40% of cells. The findings point to the functionalized nanocomposites' potential as a smart scaffold for electrically stimulated neural regeneration, as they are flexible enough to be designed into microchanneled or conduit-like structures that mimic the microstructures and mechanical properties of peripheral nerves.


Subject(s)
Chitosan , Mesenchymal Stem Cells , Nanocomposites , Nanofibers , Aniline Compounds , Biocompatible Materials/metabolism , Chitosan/chemistry , Nanofibers/chemistry , Neurons , Spectroscopy, Fourier Transform Infrared , Urease/metabolism
5.
Biomacromolecules ; 22(2): 594-611, 2021 02 08.
Article in English | MEDLINE | ID: mdl-33448795

ABSTRACT

Strategies involving the inclusion of cell-instructive chemical and topographical cues to smart biomaterials in combination with a suitable physical stimulus may be beneficial to enhance nerve-regeneration rate. In this regard, we investigated the surface functionalization of poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV)-based electroconductive electrospun nanofibers coupled with externally applied electrical stimulus for accelerated neuronal growth potential. In addition, the voltage-dependent conductive mechanism of the nanofibers was studied in depth to interlink intrinsic conductive properties with electrically stimulated neuronal expressions. Surface functionalization was accomplished using 3-aminopropyltriethoxysilane (APTES) and 1,6-hexanediamine (HDA) as an alternative to costly biomolecule coating (e.g., collagen) for cell adhesion. The nanofibers were uniform, porous, electrically conductive, mechanically strong, and stable under physiological conditions. Surface amination boosted biocompatibility, 3T3 cell adhesion, and spreading, while the neuronal model rat PC12 cell line showed better differentiation on surface-functionalized mats compared to nonfunctionalized mats. When coupled with electrical stimulation (ES), these mats showed comparable or faster neurite formation and elongation than the collagen-coated mats with no-ES conditions. The findings indicate that surface amination in combination with ES may provide an improved strategy to faster nerve regeneration using MEH-PPV-based neural scaffolds.


Subject(s)
Nanofibers , Animals , Neurons , PC12 Cells , Rats , Tissue Engineering , Tissue Scaffolds
6.
Prog Biomater ; 9(4): 219-237, 2020 Dec.
Article in English | MEDLINE | ID: mdl-33206319

ABSTRACT

The naturally obtained protein Bombyxmori silk is a biocompatible polymer with excellent mechanical properties and have the potential in controlled drug delivery applications. In this work, we have demonstrated dielectric barrier discharge (DBD) oxygen (O2) plasma surface modified electrospun Bombyxmori silk/Amoxicillin hydrochloride trihydrate (AMOX)/polyvinyl alcohol (PVA) nanofibers for drug release applications with controlled plasma treatment duration (1-10 min). The findings indicate that plasma treated electrospun nanofibers for 1-3 min exhibited significant enhancement in tensile strength, Young's modulus, wettability and surface energy. The plasma treated electrospun nanofibers for 1-5 min showed remarkable increase in AMOX released rate, whereas the electrospun nanofibers treated with plasma irradiation beyond 5 min showed only marginal increase. Moreover, the plasma treated nanofibers also exhibited good antibacterial activity against both E. coli (gram negative) and S. aureus (gram positive) bacteria. The untreated and the plasma treated silk/AMOX/PVA electrospun nanofibers for 1-3 min showed enhanced viability of primary adipose derived mesenchymal stem cells (ADMSCs) growth on them and much less hemolysis activity (< 5%). The in vitro biocompatibility of various electrospun nanofibers were further corroborated by live/dead imaging and cytoskeletal architecture assessment demonstrating enhanced cell adhesion and spreading on the plasma treated nanofibers for 1-3 min. The findings of the present study suggest that the silk/AMOX/PVA electrospun nanofibers with plasma treatment (1-3 min) due to their enhanced drug release ability and biocompatibility can be used as potential wound dressing applications.

7.
Biomater Sci ; 6(9): 2342-2359, 2018 Aug 21.
Article in English | MEDLINE | ID: mdl-30019048

ABSTRACT

The purpose of this study was to prepare an electrically conducting poly[2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene vinylene] (MEH-PPV) based nanofibrous scaffold and to investigate the synergetic effect of nanofibre structure and electrical stimulation on neuronal growth for possible use in nerve repair. Nanofibres were produced by electrospinning of blended MEH-PPV with polycaprolactone (PCL) at a ratio of 20 : 80, 40 : 60, 50 : 50 and 60 : 40 (v/v). A better electrical conductivity was achieved by using core-sheath structured nanofibres of PCL (core) and MEH-PPV (sheath) produced using the coaxial electrospinning technique. The highest electrical conductivity was observed in the core-sheath nanofibres, while it increased with increasing concentration of MEH-PPV for the blended electrospun nanofibres. The biocompatibility of the electrospun nanofibres was confirmed by MTS and live-dead staining assays using 3T3 fibroblasts and a neuronal rat pheochromocytoma (PC12) cell line. Beta (III) tubulin immunochemistry showed that PC12 cells differentiated into sympathetic neurons on these porous and stiffer electrospun nanofibres coated with collagen I. Improved cell morphology and attachment on the collagen I coated electrospun meshes has been confirmed by SEM analysis. Significant enhancement in neurite formation and neurite outgrowth of PC12 cells on the conductive scaffolds under electrical potential of 500 mV cm-1 for 2 h day-1 suggests the potential use of these scaffolds for nerve repair.


Subject(s)
Nanofibers/chemistry , Polyesters/chemistry , Polymers/chemistry , Vinyl Compounds/chemistry , Adrenal Gland Neoplasms/metabolism , Animals , Cell Adhesion/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Drug Stability , Electric Conductivity , Electric Stimulation , Mice , NIH 3T3 Cells , Nanofibers/administration & dosage , PC12 Cells , Pheochromocytoma/metabolism , Polyesters/administration & dosage , Polymers/administration & dosage , Rats , Tubulin/metabolism , Vinyl Compounds/administration & dosage
8.
ACS Biomater Sci Eng ; 4(9): 3327-3346, 2018 Sep 10.
Article in English | MEDLINE | ID: mdl-33435069

ABSTRACT

In the present study, a conducting polymer, poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV) along with a biodegradable polymer poly(ε-caprolactone) (PCL) was used to prepare an electrically conductive, biocompatible, bioactive, and biodegradable nanofibrous scaffold for possible use in neural tissue engineering applications. Core-sheath electrospun nanofibers of PCL as the core and MEH-PPV as the sheath, were surface-functionalized with (3-aminopropyl) triethoxysilane (APTES) and 1,6-hexanediamine to obtain amine-functionalized surface to facilitate cell-biomaterial interactions with the aim of replacing the costly biomolecules such as collagen, fibronectin, laminin, and arginyl-glycyl-aspartic acid (RGD) peptide for surface modification. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) confirmed the formation of core-sheath morphology of the electrospun nanofibers, whereas Fourier-transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) revealed successful incorporation of amine functionality after surface functionalization. Adhesion, spreading, and proliferation of 3T3 fibroblasts were enhanced on the surface-functionalized electrospun meshes, whereas the neuronal model rat pheochromocytoma 12 (PC12) cells also adhered and differentiated into sympathetic neurons on these meshes. Under a constant electric field of 500 mV for 2 h/day for 3 consecutive days, the PC12 cells displayed remarkable improvement in the neurite formation and outgrowth on the surface-functionalized meshes that was comparable to those on the collagen-coated meshes under no electrical signal. Electrical stimulation studies further demonstrated that electrically stimulated PC12 cells cultured on collagen I coated meshes yielded more and longer neurites than those of the unstimulated cells on the same scaffolds. The enhanced neurite growth and differentiation suggest the potential use of these scaffolds for neural tissue engineering applications.

9.
Mater Sci Eng C Mater Biol Appl ; 61: 762-72, 2016 Apr 01.
Article in English | MEDLINE | ID: mdl-26838907

ABSTRACT

Polyaniline nanofibers (PNFs) synthesized by dilute polymerization method have been surface functionalized with glutaraldehyde at their N-terminals in Phosphate Buffered Saline (PBS) at P(H)=7.4 in order to achieve improved interaction of surface functionalized polyaniline nanofibers (SF-PNFs) with aromatic amino acids-Tyrosine, Tryptophan and Phenylalanine through incorporation of aldehyde (-CHO) and hydroxyl (-OH) functionalities. HRTEM reveals nanofibers of average diameter of 35.66 nm. FESEM depicts interconnected networks of nanofibers of polyaniline (PAni). UV-visible absorption and Fluorescence spectroscopy indicate that the PNFs and SF-PNFs are in emeraldine base (EB) form. FT-IR, (1)H NMR spectroscopy suggests covalent interactions of SF-PNFs with aromatic amino acids and possible reaction mechanisms have been proposed based on these results. Remarkable enhancement in fluorescence signals of SF-PNFs in the presence of aromatic amino acids has been observed and the apparent binding constant (KA) and the number of binding sites (n) have been calculated using fluorescence enhancement equation. The KA value is found to be highest for SF-PNFs+Tyrosine and n is two for all the polymer amino acid complexes, which are in agreement with the FT-IR and (1)H NMR results. Fluorescence resonance energy transfer (FRET) efficiency has been found to be highest for SF-PNFs+Tyrosine giving maximum fluorescence enhancement. The study of interaction mechanisms by means of an extremely sensitive technique like fluorescence using SF-PNFs as a substrate may provide a promising analytical tool for detection and monitoring any biochemical reactions involving these three aromatic amino acids.


Subject(s)
Aniline Compounds/chemistry , Glutaral/chemistry , Nanofibers/chemistry , Amino Acids, Aromatic/chemistry , Fluorescence Resonance Energy Transfer , Magnetic Resonance Spectroscopy , Spectrophotometry, Ultraviolet , Spectroscopy, Fourier Transform Infrared , Tyrosine/chemistry
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