Silver Nanoparticles at Biocompatible Dosage Synergistically Increases Bacterial Susceptibility to Antibiotics.

Antibiotics used to treat bacterial infections can become ineffective over time or result in the emergence of antibiotic resistant pathogens. With the advent of nanotechnology, silver nanoparticles (AgNPs) have gained significant attention as a therapeutic agent due to the well-known antimicrobial properties of silver. However, there are concerns and limited literature on the potential cytotoxicity of nanoparticles at effective antimicrobial concentrations. AgNPs prepared from silver nitrate with glucose reduction were characterized by surface plasmon resonance, dynamic light scattering, zeta potential analysis and transmission electron microscopy. The cytotoxicity of AgNPs towards human gingival fibroblasts over 7 days was determined using cell proliferation assays and confocal microscopy. AgNP MIC and antibacterial effects alone and in combination with 11 antibiotics were determined against a panel of nine microbial species including gram-positive and gram-negative bacterial species. AgNPs concentrations ≤ 1 µg/mL were non-cytotoxic but also showed no antibacterial effects. However, when combined with each of eleven antibiotics, the biocompatible concentration of AgNPs (1 µg/mL) resulted in significant inhibition of bacterial growth for multiple bacterial species that were resistant to either the antibiotics or AgNPs alone. This study presents a promising strategy with further testing in vivo, to develop novel antimicrobial agents and strategies to confront emerging antimicrobial resistance.

Anodized anisotropic titanium surfaces for enhanced guidance of gingival fibroblasts.

Ensuring the formation of a robust trans-mucosal soft-tissue seal at the dental abutment surface is crucial towards protecting the underlying dental implant associated tissues from the external microbial-rich oral environment. The ability to mechanically enhance fibroblast functions at the dental abutment-mucosa interface, without the use of bioactive agents, holds great promise towards reducing the ingress of oral pathogens into the dental implant microenvironment. We hereby propose fabrication of unique anisotropic titania nanopores (TNPs) on the surface of titanium (via electrochemical anodization, EA) towards enhancing the soft-tissue integration and wound healing abilities of the conventional abutments. Using optimized EA, mechanically robust TNPs of varied diameters were fabricated on Ti surfaces with preserved underlying substrate micro-features: dual micro-nanostructured surfaces. Next, we evaluated the mechanical stability of such structures and demonstrated the ease of fabrication on commercial abutment geometries. The functions of primary human gingival fibroblasts (GFs) cultured on these surfaces in vitro were evaluated from 1 h to 7 days, and were compared between TNPs and clinically relevant titanium controls: as-received irregular rough Ti (Rough Ti) and mechanically prepared micro-rough Ti (Micro Ti). Improved cell viability was observed on TNPs as compared to controls. Additionally, cellular spreading morphology indicated cell alignment along the direction of the nanopores with strong anchoring evident by enhanced filopodia and stress fibers. RT-PCR showed improved wound healing, cell migration/adhesion and angiogenesis related mRNA, especially for TNPs with large diameters. This study provides a proof-of-concept towards using anodization for improving soft-tissue sealing around dental abutment surfaces, with implications towards reducing implant failure/peri-implantitis and achieving long-term success, especially in compromised patient conditions.

Correction to: Antimicrobial Drugs Encapsulated in Fibrin Nanoparticles for Treating Microbial Infested Wounds.

In the original manuscript, the Figure 2 a-i is inadvertently repeated as Figure 2 a-ii. This mistake has been rectified and the corrected Figure 2 is presented below.

Surface Modification of 3D Printed Polycaprolactone Constructs via a Solvent Treatment: Impact on Physical and Osteogenic Properties.

One promising strategy to reconstruct bone defects relies on 3D printed porous structures. In spite of several studies having been carried out to fabricate controlled, interconnected porous constructs, the control over surface features at, or below, the microscopic scale remains elusive for 3D polymeric scaffolds. In this study, we developed and refined a methodology which can be applied to homogeneously and reproducibly modify the surface of polymeric 3D printed scaffolds. We have demonstrated that the combination of a polymer solvent and the utilization of ultrasound was essential for achieving appropriate surface modification without damaging the structural integrity of the construct. The modification created on the scaffold profoundly affected the macroscopic and microscopic properties of the scaffold with an increased roughness, greater surface area, and reduced hydrophobicity. Furthermore, to assess the performance of such materials in bone tissue engineering, human mesenchymal stem cells (hMSC) were cultured in vitro on the scaffolds for up to 7 days. Our results demonstrate a stronger commitment toward early osteogenic differentiation of hMSC. Finally, we demonstrated that the increased in the specific surface area of the scaffold did not necessarily correlate with improved adsorption of protein and that other factors, such as surface chemistry and hydrophilicity, may also play a major role.

Exploration of alginate hydrogel/nano zinc oxide composite bandages for infected wounds.

Alginate hydrogel/zinc oxide nanoparticles (nZnO) composite bandage was developed by freeze-dry method from the mixture of nZnO and alginate hydrogel. The developed composite bandage was porous with porosity at a range of 60%-70%. The swelling ratios of the bandages decreased with increasing concentrations of nZnO. The composite bandages with nZnO incorporation showed controlled degradation profile and faster blood clotting ability when compared to the KALTOSTAT® and control bandages without nZnO. The prepared composite bandages exhibited excellent antimicrobial activity against Escherichia coli, Staphylococcus aureus, Candida albicans, and methicillin resistant S. aureus (MRSA). Cytocompatibility evaluation of the prepared composite bandages done on human dermal fibroblast cells by Alamar assay and infiltration studies proved that the bandages have a non-toxic nature at lower concentrations of nZnO whereas slight reduction in viability was seen with increasing nZnO concentrations. The qualitative analysis of ex-vivo re-epithelialization on porcine skin revealed keratinocyte infiltration toward wound area for nZnO alginate bandages.

Antibacterial and bioactive alpha- and beta-chitin hydrogel/nanobioactive glass ceramic/nano silver composite scaffolds for periodontal regeneration.

Alveolar bone loss and bone defects are the commonly encountered periodontal problems. Large defects do not heal spontaneously and thus require surgical interventions with bone substitutes. Bone grafts have the disadvantages of eliciting an immunologic response with subsequent graft rejection. The success rate of Guided Tissue Regeneration (GTR) is variable because of high susceptibility to infection. Thus emerged the important role of synthetic biomaterials and hence for this purpose we developed a nanocomposite scaffold, using alpha- and beta-chitin hydrogel with bioactive glass ceramic nanoparticles (nBGC) and silver nanoparticles (nAg) by lyophilization technique (aalpha and beta-chitin hydrogel/nBGC/nAg nanocomposite scaffold). The prepared nanoparticles and nanocomposite scaffolds were characterized. In addition, the porosity, swelling, mechanical properties, antibacterial activity, in vitro degradation and biomineralization, cell viability, cell attachment and cell proliferation ability of the prepared composite scaffolds were also evaluated. The results showed that alpha- and beta-chitin/nBGC/nAg composite scaffolds were porous and have the capacity to absorb fluids and swell. The composite scaffolds also showed enhanced antibacterial activity, bioactivity and controlled degradation in comparison to the control scaffolds. Cell viability studies proved the non-toxic nature of the nanocomposite scaffolds. Cell attachment and cell proliferation studies revealed the attachment and spreading nature of cells. All these studies revealed that, these antibacterial nanocomposite scaffolds could be a promising approach for the management of periodontal defects.

Drug delivery and tissue engineering applications of biocompatible pectin-chitin/nano CaCO3 composite scaffolds.

In this work, we have developed a nanocomposite scaffold using a mixture of pectin, chitin and nano CaCO3 using the technique of lyophilization, with an intended use towards biomedical applications such as tissue engineering and drug delivery. The prepared composite scaffold was characterized using scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). In addition, swelling, degradation and biomineralization capability of the composite scaffold was evaluated. The developed composite scaffold showed controlled swelling and degradation in comparison with the control scaffold. Cytocompatibility evaluation of the scaffold was tested on NIH3T3, L929 and human dermal fibroblast (HDF) cells, showed negligible toxicity towards cells. Cell attachment and proliferation studies were also conducted using these cells, which showed that cells attached onto the scaffolds and started to proliferate after 48 h of incubation. Further, drug delivery through the scaffold was examined using a bisphosphonate called Fosamax. These results suggest that the developed composite scaffold possess the essential requisites for their application in the fields of tissue engineering and drug delivery.

Synthesis and biological evaluation of chitin hydrogel/nano ZnO composite bandage as antibacterial wound dressing.

We developed chitin hydrogel/nano ZnO composite bandages using chitin hydrogel and ZnO nanoparticles (nZnO). The homogenized mixture of chitin hydrogel and nZnO was freeze-dried to obtain micro-porous composite bandages. The prepared nanocomposite bandages were characterized using FT-IR, XRD and SEM. In addition, blood clotting, antibacterial, swelling, cytocompatibility and cell attachment capability of the prepared nanocomposite bandages were evaluated. The nanocomposite bandages showed enhanced swelling, blood clotting and antibacterial activity. The incorporation of nZnO helped to attain antibacterial activity. Cytocompatibility studies were carried out using human dermal fibroblast (HDF) cells proved the non-toxic nature of the composite bandages. HDF cell attachment and infiltration analysis showed that the cells were attached and penetrated into the interior (250 microm) of the nanocomposite bandages. These studies revealed that, this nanocomposite can be used for burn, diabetic and chronic wound defects.

Flexible and microporous chitosan hydrogel/nano ZnO composite bandages for wound dressing: in vitro and in vivo evaluation.

Current wound dressings have disadvantages such as less flexibility, poor mechanical strength, lack of porosity, and a tendency for dressings to adhere onto the wound surface; in addition, a majority of the dressings did not possess antibacterial activity. Hydrogel-based wound dressings would be helpful to provide a cooling sensation and a moisture environment, as well as act as a barrier to microbes. To overcome these hassles, we have developed flexible and microporous chitosan hydrogel/nano zinc oxide composite bandages (CZBs) via the incorporation of zinc oxide nanoparticles (nZnO) into chitosan hydrogel. The prepared nanocomposite bandages were characterized using Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometry (XRD), and scanning electron microscopy (SEM). In addition, swelling, degradation, blood clotting, antibacterial, cytocompatibility, cell attachment on the material, and cell infiltration into the composite bandages were evaluated. The nanocomposite bandage showed enhanced swelling, blood clotting, and antibacterial activity. Cytocompatibility of the composite bandage has been analyzed in normal human dermal fibroblast cells. Cell attachment and infiltration studies showed that the cells were found attached to the nanocomposite bandages and penetrated into the interior. Furthermore, the in vivo evaluations in Sprague-Dawley rats revealed that these nanocomposite bandages enhanced the wound healing and helped for faster re-epithelialization and collagen deposition. The obtained data strongly encourage the use of these composite bandages for burn wounds, chronic wounds, and diabetic foot ulcers.

Synthesis, characterization and cytocompatibility studies of α-chitin hydrogel/nano hydroxyapatite composite scaffolds.

α-chitin hydrogel/nano hydroxyapatite (nHAp) composite scaffold have been synthesized by freeze-drying approach with nHAp and α-chitin hydrogel. The prepared nHAp and nanocomposite scaffolds were characterized using DLS, SEM, FT-IR, XRD and TGA studies. The porosity, swelling, degradation, protein adsorption and biomineralization (calcification) of the prepared nanocomposite scaffolds were evaluated. Cell viability, attachment and proliferation were investigated using MG 63, Vero, NIH 3T3 and nHDF cells to confirm that the nanocomposite scaffolds were cytocompatible and cells were found to attach and spread on the scaffolds. All the results suggested that these scaffolds can be used for bone and wound tissue engineering.

Bioactive and osteoblast cell attachment studies of novel alpha- and beta-chitin membranes for tissue-engineering applications.

Chitin is a novel biopolymer and has excellent biological properties such as biodegradation in the human body and biocompatible, bioabsorable, antibacterial and wound healing activities. In this work, alpha- and beta-chitin membranes were prepared using alpha- and beta-chitin hydrogel. The bioactivity studies were carried out using these chitin membranes with the simulated body fluid solution (SBF) for 7, 14 and 21 days. After 7, 14 and 21 days the membranes were characterized using SEM, EDS and FT-IR. The SEM, EDS and FT-IR studies confirmed the formation of calcium phosphate layer on the surface of the both chitin membranes. These results indicate that the prepared chitin membranes were bioactive. Cell adhesion studies were also carried out using MG-63 osteoblast-like cells. The cells were adhered and spread over the membrane after 24h of incubation. These results indicated that the chitin membranes could be used for tissue-engineering applications.