Pectin-based injectable biomaterials for bone tissue engineering

A variety of natural polymers and proteins are considered to be 3D cell culture structures able to mimic the extracellular matrix (ECM) to promote bone tissue regeneration. Pectin, a natural polysaccharide extracted from the plant cell walls and having a chemical structure similar to alginate, provides interesting properties as artificial ECM. In this work, for the first time, pectin, modified with an RGD-containing oligopeptide or not, is used as an ECM alternative to immobilize cells for bone tissue regeneration. The viability, metabolic activity, morphology, and osteogenic differentiation of immobilized MC3T3-E1 preosteoblats demonstrate the potential of this polysaccharide to keep immobilized cells viable and differentiating. Preosteoblasts immobilized in both types of pectin microspheres maintained a constant viability up to 29 days and were able to differentiate. The grafting of the RGD peptide on pectin backbone induced improved cell adhesion and proliferation within the microspheres. Furthermore, not only did cells grow inside but also they were able to spread out from the microspheres and to organize themselves in 3D structures producing a mineralized extracellular matrix. These promising results suggest that pectin can be proposed as an injectable cell vehicle for bone tissue regeneration.     

Peripheral Surgical Wounding and Age-Dependent Neuroinflammation in Mice

Post-operative cognitive dysfunction is associated with morbidity and mortality. However, its neuropathogenesis remains largely to be determined. Neuroinflammation and accumulation of β-amyloid (Aβ) have been reported to contribute to cognitive dysfunction in humans and cognitive impairment in animals. Our recent studies have established a pre-clinical model in mice, and have found that the peripheral surgical wounding without the influence of general anesthesia induces an age-dependent Aβ accumulation and cognitive impairment in mice. We therefore set out to assess the effects of peripheral surgical wounding, in the absence of general anesthesia, on neuroinflammation in mice with different ages. Abdominal surgery under local anesthesia was established in 9 and 18 month-old mice. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), Iba1 positive cells (the marker of microglia activation), CD33, and cognitive function in mice were determined. The peripheral surgical wounding increased the levels of TNF-α, IL-6, and Iba1 positive cells in the hippocampus of both 9 and 18 month-old mice, and age potentiated these effects. The peripheral surgical wounding increased the levels of CD33 in the hippocampus of 18, but not 9, month-old mice. Finally, anti-inflammatory drug ibuprofen ameliorated the peripheral surgical wounding-induced cognitive impairment in 18 month-old mice. These data suggested that the peripheral surgical wounding could induce an age-dependent neuroinflammation and elevation of CD33 levels in the hippocampus of mice, which could lead to cognitive impairment in aged mice. Pending further studies, anti-inflammatory therapies may reduce the risk of postoperative cognitive dysfunction in elderly patients.     

Trace metal contamination initiates the apparent auto-aggregation,amyloidosis, and oligomerization of Alzheimer’s Aβ peptides

Nucleation-dependent protein aggregation (seeding) and amyloid fibril-free formation of soluble SDS-resistant oligomers (oligomerization) by hydrophobic interaction is an in vitro model thought to propagate -amyloid (A) deposition, accumulation, and incur neurotoxicity and synaptotoxicity in Alzheimers disease (AD), and other amyloid-associated neurodegenerative diseases. However, A is a high-affinity metalloprotein that aggregates in the presence of biometals (zinc, copper, and iron), and neocortical A deposition is abolished by genetic ablation of synaptic zinc in transgenic mice. We now present in vitro evidence that trace (0.8 µM) levels of zinc, copper, and iron, present as common contaminants of laboratory buffers and culture media, are the actual initiators of the classic A1–42-mediated seeding process and A oligomerization. Replicating the experimental conditions of earlier workers, we found that the in vitro precipitation and amyloidosis of A1–40 (20 µM) initiated by A1–42 (2 µM) were abolished by chelation of trace metal contaminants. Further, metal chelation attenuated formation of soluble A oligomers from a cell-free culture medium. These data suggest that protein self-assembly and oligomerization are not spontaneous in this system as previously thought, and that there may be an obligatory role for metal ions in initiating A amyloidosis and oligomerization.     

Maturation involves suppression of voltage-gated currents in the frog oocyte

Voltage- and time-dependent currents having slow kinetics have been studied in plasma membranes of immature oocytes of the european frog, Rana esculenta. IK, corresponding to an outward flow of K+, is activated at potentials more positive than about -40 mV, and subserves outward rectification; Iir, corresponding to an outward flow of Cl-, is activated at potentials more negative than about -80 mV and subserves inward rectification. Such currents can act as negative feedback mechanisms in the control of membrane potential in the immature oocyte and limit to a somewhat restricted range its possible deviations from resting values. Besides IK, membrane depolarizations to potentials more positive than about +30 mV are capable of activating INa, corresponding to outflow of Na+. By contrast, the frog mature egg-cell has a single voltage- and time-dependent current, IM, activated at potentials more positive than +30 mV, with properties similar to INa. The disappearance of IK and Iir along with remarkable reduction in leakage lowers impedance in the egg membrane. It seems reasonable to suggest that the observed changes in membrane permeability reflect changes which have taken place along the maturation process and are of importance for successful fertilization.     

Use of Victoria blue in the detection of intrahepatocyte HBs Ag: comparison with other methods

The authors tested Victoria blue staining in intrahepatocytic HBs Ag detection and compared the results with those obtained by the following methods: direct immunofluorescence, orcein and peroxidase-antiperoxidase (PAP). Although PAP appears to be the best technique for diagnostic research, Victoria blue is a sound method for routine diagnosis in hepatology.     

Advances in biomedical applications of pectin gels

Pectin, due to its simple and cytocompatible gelling mechanism, has been recently exploited for different biomedical applications including drug delivery, gene delivery, wound healing and tissue engineering. Recent studies involving pectin for the biomedical field are reviewed, with the aim to capture the state of art on current research about pectin gels for biomedical applications, moving outside the traditional fields of application such as the food industry or pharmaceutics. Pectin structure, sources and extraction procedures have been discussed focussing on the properties of the polysaccharide that can be tuned to optimize the gels for a desired application and possess a fundamental role in application of pectin in the biomedical field.     

Genotoxicity of N-acryloyl-N'-phenylpiperazine, a redox activator for acrylic resin polymerization.

N-Acryloyl-N'-phenylpiperazine is a promoter of redox reactions synthesized recently, and proposed as an activator for the polymerization of acrylic resins for biomedical use. The chemical was analyzed for different genotoxicity endpoints, to obtain both information on its possible mutagenic/carcinogenic potential and a model analysis of a tertiary arylamine, which belongs to a class of chemicals commonly used as polymerization accelerators in the biomaterial field. The genotoxicity endpoints considered were: gene mutation in the Salmonella test; structural and numerical chromosome alterations in Chinese hamster V79 cells, evaluated by the micronucleus test together with an immunofluorescent staining specific for kinetochore proteins; in vitro and in vivo DNA damage, evaluated in V79 cells and in mouse liver by the alkaline DNA elution technique. On the whole, the results indicate that N-acryloyl-N'-phenylpiperazine is to be regarded not so much as a DNA-damaging agent, but as a genomic mutagen. Indeed, it was not mutagenic in Salmonella (though its toxicity did not allow testing concentrations over 70 micrograms/plate), and it was weakly positive in inducing chromosomal fragmentation in vitro (one positive, not dose-related, result out of five different doses tested) and in vivo DNA damage (increases in DNA elution rate never doubling control values). The chemical was, however, clearly positive (with dose-dependent effects up to about 25 times the control value) in causing numerical chromosome alterations, at the maximal non-toxic doses.