Shreyas Kuddannaya

Surface Chemical Modification of Poly(dimethylsiloxane) for the Enhanced Adhesion and Proliferation of Mesenchymal Stem Cells

The surface chemistry of materials has an interactive influence on cell behavior. The optimal adhesion of mammalian cells is critical in determining the cell viability and proliferation on substrate surfaces. Because of the inherent high hydrophobicity of a poly(dimethylsiloxane) (PDMS) surface, cell culture on these surfaces is unfavorable, causing cells to eventually dislodge from the surface. Although physically adsorbed matrix proteins can promote initial cell adhesion, this effect is usually short-lived. Here, (3-aminopropyl)triethoxy silane (APTES) and cross-linker glutaraldehyde (GA) chemistry was employed to immobilize either fibronectin (FN) or collagen type 1 (C1) on PDMS. The efficiency of these surfaces to support the adhesion and viability of mesenchymal stem cells (MSCs) was analyzed. The hydrophobicity of the native PDMS decreased significantly with the mentioned surface functionalization. The adhesion of MSCs was mostly favorable on chemically modified PDMS surfaces with APTES + GA + protein. Additionally, the spreading area of MSCs was significantly higher on APTES + GA + C1 surfaces than on other unmodified/modified PDMS surfaces with C1 adsorption. However, there were no significant differences in the MSC spreading area on the unmodified/modified PDMS surfaces with FN adsorption. Furthermore, there was a significant increase in cell proliferation on the PDMS surface with APTES + GA + protein functionalization as compared to the PDMS surface with protein adsorption only. Therefore, the covalent surface chemical modification of PDMS with APTES + GA + protein could offer a more biocompatible platform for the enhanced adhesion and proliferation of MSCs. Similar strategies can be applied for other substrates and cell lines by appropriate combinations of self-assembly monolayers (SAMs) and extracellular matrix proteins.

Fabrication, Characterization, and Biocompatibility of Polymer Cored Reduced Graphene Oxide Nanofibers

Graphene nanofibers have shown a promising potential across a wide spectrum of areas, including biology, energy, and the environment. However, fabrication of graphene nanofibers remains a challenging issue due to the broad size distribution and extremely poor solubility of graphene. Herein, we report a facile yet efficient approach for fabricating a novel class of polymer core-reduced graphene oxide shell nanofiber mat (RGO-CSNFM) by direct heat-driven self-assembly of graphene oxide sheets onto the surface of electrospun polymeric nanofibers without any requirement of surface treatment. Thus-prepared RGO-CSNFM demonstrated excellent mechanical, electrical, and biocompatible properties. RGO-CSNFM also promoted a higher cell anchorage and proliferation of human bone marrow mesenchymal stem cells (hMSCs) compared to the free-standing RGO film without the nanoscale fibrous structure. Further, cell viability of hMSCs was comparable to that on the tissue culture plates (TCPs) with a distinctive healthy morphology, indicating that the nanoscale fibrous architecture plays a critically constructive role in supporting cellular activities. In addition, the RGO-CSNFM exhibited excellent electrical conductivity, making them an ideal candidate for conductive cell culture, biosensing, and tissue engineering applications. These findings could provide a new benchmark for preparing well-defined graphene-based nanomaterial configurations and interfaces for biomedical applications.

Enhanced In Vitro Biocompatibility of Chemically Modified Poly(dimethylsiloxane) Surfaces for Stable Adhesion and Long-term Investigation of Brain Cerebral Cortex Cells.

Studies on the mammalian brain cerebral cortex have gained increasing importance due to the relevance of the region in controlling critical higher brain functions. Interactions between the cortical cells and surface extracellular matrix (ECM) proteins play a pivotal role in promoting stable cell adhesion, growth, and function. Poly(dimethylsiloxane) (PDMS) based platforms have been increasingly used for on-chip in vitro cellular system analysis. However, the inherent hydrophobicity of the PDMS surface has been unfavorable for any long-term cell system investigations due to transitory physical adsorption of ECM proteins on PDMS surfaces followed by eventual cell dislodgement due to poor anchorage and viability. To address this critical issue, we employed the (3-aminopropyl)triethoxysilane (APTES) based cross-linking strategy to stabilize ECM protein immobilization on PDMS. The efficiency of surface modification in supporting adhesion and long-term viability of neuronal and glial cells was analyzed. The chemically modified surfaces showed a relatively higher cell survival with an increased neurite length and neurite branching. These changes were understood in terms of an increase in surface hydrophilicity, protein stability, and cell-ECM protein interactions. The modification strategy could be successfully applied for stable cortical cell culture on the PDMS microchip for up to 3 weeks in vitro.

Biocompatible, Free-Standing Film Composed of Bacterial Cellulose Nanofibers–Graphene Composite

In recent years, graphene films have been used in a series of wide applications in the biomedical area, because of several advantageous characteristics. Currently, these films are derived from graphene oxide (GO) via chemical or physical reduction methods, which results in a significant decrease in surface hydrophilicity, although the electrical property could be greatly improved, because of the reduction process. Hence, the comprehensive performance of the graphene films showed practical limitations in the biomedical field, because of incompatibility of highly hydrophobic surfaces to support cell adhesion and growth. In this work, we present a novel fabrication of bacterial cellulose nanofibers/reduced graphene oxide (BC-RGO) film, using a bacterial reduction method. Thus-prepared BC-RGO films maintained excellent hydrophilicity, while electrical properties were improved by bacterial reduction of GO films in culture. Human marrow mesenchymal stem cells (hMSCs) cultured on these surfaces showed improved cellular response with higher cell proliferation on the BC-RGO film, compared to free-standing reduced graphene oxide film without the nanoscale fibrous structure. Furthermore, the cellular adhesion and proliferation were even comparable to that on the tissue culture plate, indicating that the bacterial cellulose nanofibers play a critically contructive role in supporting cellular activities. The novel fabrication method greatly enhanced the biochemical activity of the cells on the surface, which could aid in realizing several potential applications of graphene film in biomedical area, such as tissue engineering, bacterial devices, etc.

Synergistic effects of a novel free-standing reduced graphene oxide film and surface coating fibronectin on morphology, adhesion and proliferation of mesenchymal stem cells

Graphene films have broad use in engineering, energy and biomedical applications. The cost-effective, eco-friendly and easy to scale-up fabrication methods of graphene films are always highly desired. In this work, we develop a novel fabrication method of free-standing reduced graphene oxide (RGO) films by vacuum filtration of graphene oxide aqueous solution through a nanofiber membrane in combination with chemical reduction. Instead of the smooth surface, the generated RGO films have nanoscale patterns transferred from the nanofiber membrane and controlled in a large range by varying the parameters of the electrospinning process. The cellular culture results of the human marrow mesenchymal stem cells (hMSCs) show that the fibronectin modified RGO films could exhibit excellent biocompatibility, which could be attributed to the synergistic effects of the RGO films including both surface morphology and fibronectin modification. The novel fabrication method greatly enhances the fabrication capability and the potential of graphene films for application in cell culture, tissue engineering as well as in other engineering and biomedical applications.

Fabrication and Characterization of Three-Dimensional (3D) Core–Shell Structure Nanofibers Designed for 3D Dynamic Cell Culture

Three-dimensional elastic nanofibers (3D eNFs) can offer a suitable 3D dynamic microenvironment and sufficient flexibility to regulate cellular behavior and functional protein expression. In this study, we report a novel approach to prepare 3D nanofibers with excellent mechanical properties by solution-assisted electrospinning technology and in situ polymerization. The obtained 3D eNFs demonstrated excellent biocompatible properties to meet cell culture requirements under a dynamic environment in vitro. Moreover, these 3D eNFs also promoted human bone marrow mesenchymal stem cells (hMSCs) adhesion and collagen expression under biomechanical stimulation. The results demonstrated that this dynamic cell culture system could positively impact cellular collagen but has no significant effect on the proliferation of hMSCs grown in the 3D eNFs. This work may give rise to a new approach for constructing a 3D cell culture for tissue engineering.

Anti-bacterial properties of collagen-coated glass and polydimethylsiloxane substrates

Attachments of three bacterial species, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa on chemically modified glass slides and polydimethylsiloxane substrates were investigated with an attempt to find anti-bacterial materials which can prevent bacterial infections. The results showed that the attachment of the first two species was largely reduced on surfaces treated with self-assembled monolayers plus collagen type 1, whereas attachment of Pseudomonas aeruginosa was not affected on all the treated/untreated surfaces. Gentamicin protection assay showed that the fewest Pseudomonas aeruginosa were engulfed by macrophages when the substrates were coated with (3-aminopropyl)triethoxysilane/(3-aminopropyl)trimethoxysilane plus glutaraldehyde plus collagen type 1. Considering that both Pseudomonas aeruginosa and macrophage adhesion were not influenced very much by the chemical modifications, the decreased engulfment of Pseudomonas aeruginosa was attributed to its decreased ability to invade macrophage cells on the two coated substrates. The results indicate that by employing appropriate chemical modifications, bacterial adhesion could be weakened with a decreased bacterial engulfment response of macrophages to the coated substrates, which shows an interesting and promising direction for applicability of this surface modification strategy for future biomedical research on anti-inflammatory/anti-microbial cell-material interactions.