Andrea Cochis

Tissue-engineered skin substitutes: an overview

Extensive skin loss and chronic wounds are still a significant challenge to clinicians: even if injured epidermis is normally able to self-renew, deep injuries can cause negative regulation of the wound healing cascade, leading to chronic wound formation. Skin-autografting surgical procedures are often limited by the poor availability of healthy tissue, whereas the use of non-self-tissues for allografts presents some severe risks. Tissue-engineered skin substitutes have recently become viable as a suitable alternative to auto- and allografts. However, biologists, biochemists, and technical engineers are still struggling to produce complex skin substitutes that can readily be transplanted in large quantities. The ambitious goal is now to construct a dermoepidermal substitute that rapidly vascularizes and optimally supports a stratifying epidermal graft on a biodegradable matrix. This review analyzes these aspects in light of the available literature and the authors’ experience.

Biofilm formation on titanium implants counteracted by grafting gallium and silver ions

Biofilm-associated infections remain the leading cause of implant failure. Thanks to its established biocompatibility and biomechanical properties, titanium has become one of the most widely used materials for bone implants. Engineered surface modifications of titanium able to thwart biofilm formation while endowing a safe anchorage to eukaryotic cells are being progressively developed. Here surfaces of disks of commercial grade 2 titanium for bone implant were grafted with gallium and silver ions by anodic spark deposition. Scanning electron microscopy of the surface morphology and energy dispersive X-ray spectroscopy were used for characterization. Gallium-grafted titanium was evaluated in comparison with silver-grafted titanium for both in vivo and in vitro antibiofilm properties and for in vitro compatibility with human primary gingival fibroblasts. Surface-modified materials showed: (i) homogeneous porous morphology, with pores of micrometric size; (ii) absence of cytotoxic effects; (iii) ability to support in vitro the adhesion and spreading of gingival fibroblasts; and (iv) antibiofilm properties. Although both silver and gallium exhibited in vitro strong antibacterial properties, in vivo gallium was significantly more effective than silver in reducing number and viability of biofilm bacteria colonies. Gallium-based treatments represent promising titanium antibiofilm coatings to develop new bone implantable devices for oral, maxillofacial, and orthopedic applications.

Biosurfactants prevent in vitro Candida albicans biofilm formation on resins and silicon materials for prosthetic devices

OBJECTIVE The aim of this study was to evaluate in vitro the preventive antiadhesion activity of biosurfactants against Candida albicans biofilm. STUDY DESIGN Disks of silicon and acrylic resin for denture prostheses were precoated with increasing concentrations of biosurfactants obtained from endophyte biofilms selected from Robinia pseudoacacia and from Nerium oleander, and afterward infected with C. albicans cells. The number of biofilm cells were detected by colony-forming unit (CFU) counting, cell viability was established by the 2,3-bis(2-methoxy-4-nitro-5-sulphophenyl)-5-[(phenyl amino)carbonyl]-2H-tetrazolium hydroxide (XTT) assay, and biosurfactant cytotoxicity was evaluated by the [3-(4,5-dimethyliazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium] (MTT) assay. Chlorhexidine was used as control. RESULTS Precoating with biosurfactants caused a greater reduction (P < .01) in biofilm cell number and viability than chlorhexidine. The antiadhesion activity of the biosurfactants was observed at low concentrations (78.12 μg/mL and 156.12 μg/mL) which were noncytotoxic. CONCLUSIONS This study demonstrated the preventive antiadhesion activity of biosurfactants against C. albicans biofilm. These agents are amphiphilic, interfere with microbial adhesion, and demonstrate cycompatibility with epithelial cells and fibroblasts.

Biofilm formation on composite resins for dental restorations: an in situ study on the effect of chlorhexidine mouthrinses.

Purpose Biofilm formation on the surface of dental restorative materials by oral bacteria is considered an important step in the development of secondary caries. The aim of this study was to evaluate the in situ effect of a chlorhexidine (CHX)-containing mouthrinse on the biofilm formation occurring on the surface of human enamel and of two resin-based commercially available materials: a silorane-based material (Filtek Silorane®) and a methacrylate-based material (Filtek Supreme XT®). Methods 53 disks were obtained for each of the two composites and 37 disks for enamel. The surface was characterized by determining the surface roughness and the surface free energy of 5 samples for each of the three materials tested, then the remaining samples were mounted on splints worn by 16 volunteers. The participants were randomly divided into two groups: an experimental group that used 0.12% CHX-based mouthrinse and a control group that used a placebo mouthrinse. Biofilm formation on the different surfaces after a 24 h period was assessed using MTT assay. Results The two composites in the group treated with the placebo mouthrinse showed a similar biofilm formation, which was significantly higher than that occurring on enamel surfaces. The CHX-based mouthrinse significantly reduced biofilm formation on the surfaces of the two resin-based materials when compared with the placebo mouthrinse. The reduction was particularly relevant on the Filtek Silorane surfaces. Conclusions The new silorane-based material seems to interact with CHX in a promising way from the point of view of biofilm formation control.

Essential Oil from Berries of Lebanese Juniperus excelsa M. Bieb Displays Similar Antibacterial Activity to Chlorhexidine but Higher Cytocompatibility with Human Oral Primary Cells

Chlorhexidine (CHX), one of the most effective drugs administered for periodontal treatment, presents collateral effects including toxicity when used for prolonged periods; here, we have evaluated the bactericidal potency and the cytocompatibility of Juniperus excelsa M. Bieb essential oil (EO) in comparison with 0.05% CHX. The EO was extracted from berries by hydrodistillation and components identified by gas chromatography and mass spectrometry. Bacterial inhibition halo analysis, quantitative cell viability 2,3-bis(2-methoxy-4-nitro-5-sulphophenyl)-5-[(phenyl amino) carbonyl]-2H-tetrazolium hydroxide assay (XTT), and colony forming unit (CFU) count were evaluated against the two biofilm formers Aggregatibacter actinomycetemcomitans and Streptococcus mutans. Finally, cytocompatibility was assessed with human primary gingival fibroblasts (HGF) and mucosal keratinocytes (HK). The resulting EO was mainly composed of monoterpene hydrocarbons and oxygenated monoterpenes. An inhibition halo test demonstrated that both bacteria were sensitive to the EO; XTT analysis and CFU counts confirmed that 10-fold-diluted EO determined a statistically significant (p < 0.05) reduction in bacteria count and viability towards both biofilm and planktonic forms in a comparable manner to those obtained with CHX. Moreover, EO displayed higher cytocompatibility than CHX (p < 0.05). In conclusion, EO exhibited bactericidal activity similar to CHX, but a superior cytocompatibility, making it a promising antiseptic alternative to CHX.

Composite bone cements loaded with a bioactive and ferrimagnetic glass-ceramic: Leaching, bioactivity and cytocompatibility.

In this work, composite bone cements, based on a commercial polymethylmethacrylate matrix (Palamed®) loaded with ferrimagnetic bioactive glass-ceramic particles (SC45), were produced and characterized in vitro. The ferrimagnetic bioactive glass-ceramic belongs to the system SiO2-Na2O-CaO-P2O5-FeO-Fe2O3 and contains magnetite (Fe3O4) crystals into a residual amorphous bioactive phase. Three different formulations (containing 10, 15 and 20 wt.% of glass-ceramic particles respectively) have been investigated. These materials are intended to be applied as bone fillers for the hyperthermic treatment of bone tumors. The morphological, compositional, calorimetric and mechanical properties of each formulation have been already discussed in a previous paper. The in vitro properties of the composite bone cements described in the present paper are related to iron ion leaching test (by graphite furnace atomic absorption spectrometer), bioactivity (i.e. the ability to stimulate the formation of a hydroxyapatite - HAp - layer on their surface after soaking in simulated body fluid SBF) and cytocompatibility toward human osteosarcoma cells (ATCC CRL-1427, Mg63). Morphological and chemical characterizations by scanning electron microscopy and energy dispersion spectrometry have been performed on the composite samples after each test. The iron release was negligible and all the tested samples showed the growth of HAp on their surface after 28 days of immersion in a simulated body fluid (SBF). Cells showed good viability, morphology, adhesion, density and the ability to develop bridge-like structures on all investigated samples. A synergistic effect between bioactivity and cell mineralization was also evidenced.

Degradable polymers may improve dental practice

The use of biomaterials in dentistry is more widespread than in any other medical field in terms of both amount and variety. Most of them were not originally designed for dental applications but for other medical applications or, sometimes, for no medical purposes. Among these materials, biodegradable materials play an important role, especially in bone regeneration and in periodontal surgery. This paper briefly reviews some degradable polymers developed as tools for the treatment of periodontal and bone diseases. We discuss materials previously applied in other industrials contexts, such as polyesters, methylcellulose, and chitosan and we provide perspectives for their use in periodontal regeneration.

Biomimetic calcium–phosphates produced by an auto-catalytic route on stainless steel 316L and bio-inert polyolefin

A melange of hydroxyapatite and calcium phosphate hydrate coatings have been deposited on the surfaces of 316L type stainless steel and polyolefin (PP-PE), both components of 316L/PP-PE/316L sandwich, by an auto-catalytic route using an acidic bath. Coatings on 316L are made of spherules size ranged from 100 nm to 1 μm while those observed on PP-PE are smaller in the 50–600 nm range. Cell viability is much higher in samples with novel auto-catalytic layer of Ca–P than in the uncoated ones. Furthermore, our results show that the Ca–P coating produced by an auto-catalytic route act as promoter for osteoblasts proliferation. Osteoblasts morphology investigated by immunofluorescence proves that they are attached and well spread confirming the cytocompatibility.

Rutin, a Quercetin Glycoside, Restores Chemosensitivity in Human Breast Cancer Cells

Several studies have documented the ability of flavonoids to sensitize cancer cells to chemotherapeutics and reverse multidrug resistance by inhibition of efflux pumps (adenosine triphosphate‐binding cassette transporters), apoptosis activation, and cell cycle arrest. In this study, the flavonoid rutin (quercetin 3‐O‐β‐d‐rutinoside) was investigated as chemosensitizer towards two different human epithelial breast cancer cell lines: (i) MB‐MDA‐231, selected as representative for triple‐negative breast cancer and (ii) MCF‐7 used as a well‐characterized model of HER2‐negative breast cancer. To assess the cytocompatibility of rutin against non‐cancer cells, primary human mammary fibroblasts were used as control and non‐target cells. In MDA‐MB‐231 cells, 20 μM rutin enhanced cytotoxicity related to cyclophosphamide and methotrexate. Rutin significantly (p < 0.05) increased the anticancer activity of both chemotherapeutics, at 24–48–72 h, and decreased the activity of the adenosine triphosphate‐binding cassette transporters, namely, P‐glycoprotein (P‐gp) and breast cancer resistance protein (BCRP). Flow cytometry analysis showed 20 μM and 50 μM rutin arrested cell cycle at G2/M and G0/G1 phases, respectively, significantly promoting cell apoptosis. Rutin, via non‐selective inhibition of P‐gp and BCRP pumps, efficiently reverses multidrug resistance and restores chemosensitivity to cyclophosphamide and cyclophosphamide of human chemoresistant, triple‐negative breast cancer cells, successfully arresting cell cycle progression. Copyright

Bioceramic Materials Show Reduced Pathological Biofilm Formation

The aim of the present work was to assess the surface ability of three bioceramic materials (A: alumina BIOLOX®forte; B: Si3N4; C: alumina matrix composite BIOLOX®delta) to inhibit bacterial biofilm formation. For this purpose, ceramic disks at standardized roughness (Ra = 0,25 μm) were used as test materials while commercial polystyrene was considered as control. Two biofilm-producing bacterial strains (S. epidermidis ATCC14990, Escherichia coli ATCC25922) were used for experiments. The viable biomass was assessed by the metabolic MTT assay after 24h incubation. Morphological data regarding biofilms structure were obtained by scanning electron microscopy. In general, results revealed that all bioceramics materials were significantly less colonized compared to polystyrene. The degree of biofilm formation onto bioceramics ranged between about 30 to 60% less than the polystyrene control. Moreover, some differences were noticed by comparing the three bioceramics inhibition ratio: bioceramic A showed significanlty less S. epidermidis biofilm formation (p<0.005) compared to B and C that showed similar performance. Conversely, no difference were noted for E. coli biofilm amount for A, B and C. In conclusion, the tested materials showed capability to reduce biofilm formation to a different extent depending on the tested bacterial strains.