Non-cytotoxic Material Research Articles

Development and in vivo response of hydroxyapatite/whitlockite from chicken bones as bone substitute using a chitosan membrane for guided bone regeneration

Biomaterials that meet the requirements to stimulate bone tissue formation play a vital role in orthopedics and dentistry. In this work, chitosan and a biphasic, non-cytotoxic material hydroxyapatite/whitlockite were obtained from natural sources, which are available as organic waste. The osteogenic activity was assessed using a rabbit model animal with a chitosan barrier membrane in combination with a bone-filling graft substitute composed of hydroxyapatite/whitlockite. FT-IR results showed the typical absorption bands of the chitosan and hydroxyapatite. Moreover, the X-ray diffraction pattern revealed a typical hexagonal phase of hydroxyapatite and rhombohedral structures related to whitlockite. Masson's trichrome stain showed an early formation of extracellular matrix mineralized, in accord with the surface morphology of a cortical mature bond observed by Scanning Electron Microscopy. The immunocytochemistry results showed a significant increase of positive immunoreactive cells to osteonectin in the treated defects in comparison with the control defects 6 and 8 weeks postoperatively. Overall, the results confirm that the use of this low-cost and versatile biomaterial as a barrier membrane and a bone substitute graft are useful for bone tissue engineering.

Promising effects of silver tungstate microcrystals on fibroblast human cells and three dimensional collagen matrix models: A novel non-cytotoxic material to fight oral disease

Silver tungstate (α-Ag2WO4) microcrystals have shown encouraging results regarding their antimicrobial activity. However, in addition to the promising outcomes in fighting oral disease, cytotoxic tests are mandatory for screening new materials for biological applications. Here, we developed a better understanding of the effects of microcrystals on the behavior of both human gingival fibroblast (HGF) cells and three-dimensional (3D) collagen matrices. To perform these experiments, the lowest concentration of α-Ag2WO4 capable of preventing the visible growth of Candida albicans (C. albicans) planktonic cells was defined as the test concentration, and it ranged from 0.781 (C1) to 7.81 (C2) to 78.1 (C3) μg/mL. Complete medium and lysis buffer (LB) served as negative (C-) and positive (C+) controls, respectively. The effect of the microcrystal concentration on the morphology, remodeling and proliferation of HGF cells was evaluated by different approaches. Quantitative and qualitative assessments demonstrated that α-Ag2WO4 did not affect the mitochondrial enzymatic activity of HGF cells cultured in a monolayer or the cell viability within 3D collagen matrices. These experiments showed that α-Ag2WO4 at the C2 concentration did not damage the genomic DNA. The development of new materials is attractive for the possible treatment of diseases and for avoiding indiscriminate prescribing of antibiotics. These findings provide information on the effect of α-Ag2WO4 on cell behavior and reveal that these microcrystals are non-cytotoxic against human gingival cells over a sufficient period to measure the hazard potential.

Carbon quantum dot tailored calcium alginate hydrogel for pH responsive controlled delivery of vancomycin

Herein, we demonstrate the preparation of highly luminescent carbon quantum dots (CQDs) from Aloe vera leaf gel; in just 2h at 250°C through carbonization pathway. The prepared CQDs are structurally characterized with high resolution transmission electron microscopy (HRTEM), hydrodynamic diameter, surface polarity, Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), Raman, UV–visible absorption spectrophotometry and fluorescence spectroscopy. The functional carbon nanoparticles are observed as non-cytotoxic materials. The biocompatibility, less cytotoxicity and high aqueous dispersibility of as-synthesized CQDs are motivated to design carbon quantum dot (CQD) tailored calcium alginate (CA) hydrogel films with an aim to controlled delivery of glycopeptides antibiotic vancomycin in the gastrointestinal tract (GI). With CQD, the drug loading capacity of CA/CQD film is increased to 89% from 38% (CA film), whereas; with β-cyclodextrin (β-CD) the vancomycin uptake capacity is increased more, 96%. The release of vancomycin through CA/CQD film is more pronounced at pH1.5, close to the pH of the stomach and it is found that in pH1.5 with β-CD, the release rate of vancomycin is lowered, 56% in 120h. The high drug uptake capacity (96%) and lower release rate (56% in 120h) of CA/CQD hydrogel film in pH1.5 with β-CD can be used for its applicability as drug delivery vehicle for controlled release of vancomycin into the stomach region and therefore it can offer a potential option for oral administration of vancomycin.

Microfluidic Platform for the Long-Term On-Chip Cultivation of Mammalian Cells for Lab-On-A-Chip Applications.

Lab-on-a-Chip (LoC) applications for the long-term analysis of mammalian cells are still very rare due to the lack of convenient cell cultivation devices. The difficulties are the integration of suitable supply structures, the need of expensive equipment like an incubator and sophisticated pumps as well as the choice of material. The presented device is made out of hard, but non-cytotoxic materials (silicon and glass) and contains two vertical arranged membranes out of hydrogel. The porous membranes are used to separate the culture chamber from two supply channels for gases and nutrients. The cells are fed continuously by diffusion through the membranes without the need of an incubator and low requirements on the supply of medium to the assembly. The diffusion of oxygen is modelled in order to find the optimal dimensions of the chamber. The chip is connected via 3D-printed holders to the macroscopic world. The holders are coated with Parlyene C to ensure that only biocompatible materials are in contact with the culture medium. The experiments with MDCK-cells show the successful seeding inside the chip, culturing and passaging. Consequently, the presented platform is a step towards Lab-on-a-Chip applications that require long-term cultivation of mammalian cells.

Electrospun polycaprolactone/chitosan scaffolds for nerve tissue engineering: physicochemical characterization and Schwann cell biocompatibility

Electrospun polycaprolactone (PCL)/chitosan (CH) blend scaffolds with different CH weight ratios were prepared to study the effect of scaffold composition on its physicochemical and biological properties. Scanning electron microscopy showed bead-free homogeneous randomly arranged nanofibers whose average diameter decreased from 240 to 110 nm with increasing CH content. The infrared spectra of the PCL/CH blends were very similar to the neat PCL scaffold. Energy-dispersive x-ray spectroscopy analysis confirmed the presence of carbon, oxygen and nitrogen in the scaffolds, although fluorine—from chemicals used as solvent—was also detected. The water contact angle decreased from 113° (for PCL) to 52° with increasing chitosan content. The biocompatibility was evaluated using fibroblasts and Schwann cell (SC) cultures. Cytotoxicity assays using fibroblasts demonstrated that electrospun scaffolds could be considered as non-cytotoxic material. Biocompatibility tests also revealed that the SCs adhered to scaffolds with different CH content, although the formulation containing CH at 5 wt% exhibited the highest proliferation on days 1 and 3. A better cell distribution was observed in the CH/PCL blends than in the neat PCL or CH scaffolds, where the cells were clustered. Immunochemistry analysis confirmed that SCs expressed the specific p75 cell marker on the scaffolds, suggesting that PCL/CH scaffolds would be good candidates for peripheral nerve tissue engineering.

Evaluation of the Cytotoxicity of NiFe2O4/SiO2 Hybrid Nanoparticles Aiming its Application as Drug Carriers

Magnetic nanoparticles have potential application in biomedicine since their features allow a wide variety of applications, such as drug carriers, destruction of tumor cells and magnetic separation of cells and proteins. Overlooking that, the proposal is to obtain the hybrid NiFe2O4/SiO2 from the surface modifying with the 3-aminopropyltriethoxysilane, and evaluate the structure, morphology and cytotoxicity, to obtain a biocompatible hybrid for biological applications, such as, e.g., drug carrier. The samples were analyzed by XRD, FTIR, SEM, magnetic measurements and cytotoxicity. The results showed the formation of single phase of the NiFe2O4 spinel with crystallite size of 35 and 32 nm referring to the samples before and after the surface modification, and presence of characteristic absorption bands of the spinel and of the silanol group from the silane agent, confirming the hybrid formation. The presence of the silane agent kept the ferrimagnetic characteristic and increased the cell viability, making it a non-cytotoxic material.

Nanopigmented Acrylic Resin Cured Indistinctively by Water Bath or Microwave Energy for Dentures

The highlight of this study was the synthesis of nanopigmented poly(methyl methacrylate) nanoparticles that were further processed using a water bath and/or microwave energy for dentures. The experimental acrylic resins were physicochemically characterized, and the adherence of Candida albicans and biocompatibility were assessed. A nanopigmented acrylic resin cured by a water bath or by microwave energy was obtained. The acrylic specimens possess similar properties to commercial acrylic resins, but the transverse strength and porosity were slightly improved. The acrylic resins cured with microwave energy exhibited reduced C. albicans adherence. These results demonstrate an improved noncytotoxic material for the manufacturing of denture bases in dentistry.

Evaluation of the In Vitro Cytotoxicity of Cross-Linked Biomaterials

This study evaluated the in vitro cytotoxicity of poly(propylene fumarate) (PPF). PPF is an aliphatic biodegradable polymer that has been well characterized for use in bone tissue engineering scaffolds. Four different cell types, human mesenchymal stem cells (hMSC), fibroblasts (L929), preosteoblasts (MC3T3), and canine mesenchymal stem cells (cMSC), were used to evaluate the cytotoxicity of PPF. These cell types represent the tissues that PPF would interact with in vivo as a bone tissue scaffold. The sol fraction of the PPF films was measured and then utilized to estimate cross-linking density. Cytotoxicity was evaluated using XTT assay and fluorescence imaging. Results showed that PPF supported similar cell metabolic activities of hMSC, L929, MC3T3, and cMSC compared to the noncytotoxic control, high-density polyethylene (HDPE) and were statistically different than those cultured with the cytotoxic control, a polyurethane film containing 0.1% zinc diethyldithiocarbamate (ZCF). Results showed differing cellular responses to ZCF, the cytotoxic control. The L929 cells had the lowest cell metabolic activity levels after exposure to ZCF compared to the cell metabolic activity levels of the MC3T3, hMSC, or cMSC cells. Qualitative verification of the results using fluorescence imaging demonstrated no change in cell morphology, vacuolization, or detachment when cultured with PPF compared to HDPE or blank media cultures. Overall, the cytotoxicity response of the cells to PPF was demonstrated to be similar to the cytotoxic response of cells to known noncytotoxic materials (HDPE).

Development of ultra-hydrophilic and non-cytotoxic dental vinyl polysiloxane impression materials using a non-thermal atmospheric-pressure plasma jet

Dental vinyl polysiloxane (VPS) impression materials are widely used for the replication of intraoral tissue where hydrophilicity is important as the oral tissues are surrounded by wet saliva. Recent attempts to improve the wettability of VPS using a ‘surfactant’, however, have resulted in a high level of cytotoxicity. Hence, in this study, application of a non-thermal atmospheric-pressure plasma jet (NTAPPJ) on VPS and its effects in terms of both hydrophilicity and cytotoxicity were investigated. The results showed that the application of the plasma jet resulted in significant improvement of hydrophilicity of VPS that had no surfactant, whereby the results were similar to commercially available products with the surfactant. The surface chemical analysis results indicated that this was due to the oxidation and decreased amount of hydrocarbon on the surface following NTAPPJ exposure. Meanwhile, an NTAPPJ-treated sample was shown to be non-cytotoxic. Therefore, the use of dental VPS impression materials without any surfactant, in conjunction with an NTAPPJ treatment, is a promising method for ultra-hydrophilic but yet non-cytotoxic materials.

A biodegradable and biocompatible PVA–citric acid polyester with potential applications as matrix for vascular tissue engineering

Unique elastomeric and biocompatible scaffolds were produced by the polyesterification of poly(vinyl alcohol) (PVA) and citric acid via a simple polycondensation reaction. The physicochemical characterization of the materials was done by Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), Thermogravimetric Analysis (TGA), mechanical and surface property analyses. The materials are hydrophilic and have viscoelastic nature. Biodegradable, non-cytotoxic materials that can be tailored into 3D scaffolds could be prepared in an inexpensive manner. This polyester has potential implications in vascular tissue engineering application as a biodegradable elastomeric scaffold.

Bone growth around silicon nitride implants—An evaluation by scanning electron microscopy

Silicon nitride has demonstrated to be a potential candidate for clinical applications because it is a non-cytotoxic material and has satisfactory fracture toughness, high wear resistance and low friction coefficient. In this paper, samples of silicon nitride, which were kept into rabbits' tibias for 8 weeks, and the adjacent bone tissue were analysed by scanning electron microscopy in order to verify the bone growth around the implants and the interaction between the implant and the bone. Bone growth occurred mainly in the cortical areas, although it has been observed that the newly bone tends to grow toward the marrow cavity. Differences were observed between the implants installed into distal and proximal regions. In the first region, where the distance between the implant and the cortical bone is greater than in the proximal region, the osteoconduction process was evidenced by the presence of a bridge bone formation toward the implant surface. The results showed that silicon nitride can be used as biomaterial since the newly bone grew around the implants.

Correlation of visual in vitro cytotoxicity ratings of biomaterials with quantitative in vitro cell viability measurements

Medical devices and implanted biomaterials are often assessed for biological reactivity using visual scores of cell-material interactions. In such testing, biomaterials are assigned cytotoxicity ratings based on visual evidence of morphological cellular changes, including cell lysis, rounding, spreading, and proliferation. For example, ISO 10993 cytotoxicity testing of medical devices allows the use of a visual grading scale. The present study compared visual in vitro cytotoxicity ratings to quantitative in vitro cytotoxicity measurements for biomaterials to determine the level of correlation between visual scoring and a quantitative cell viability assay. Biomaterials representing a spectrum of biological reactivity levels were evaluated, including organo-tin polyvinylchloride (PVC; a known cytotoxic material), ultra-high molecular weight polyethylene (a known non-cytotoxic material), and implantable tissue adhesives. Each material was incubated in direct contact with mouse 3T3 fibroblast cell cultures for 24 h. Visual scores were assigned to the materials using a 5-point rating scale; the scorer was blinded to the material identities. Quantitative measurements of cell viability were performed using a 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay; again, the assay operator was blinded to material identities. The investigation revealed a high degree of correlation between visual cytotoxicity ratings and quantitative cell viability measurements; a Pearson's correlation gave a correlation coefficient of 0.90 between the visual cytotoxicity score and the percent viable cells. An equation relating the visual cytotoxicity score and the percent viable cells was derived. The results of this study are significant for the design and interpretation of in vitro cytotoxicity studies of novel biomaterials.