Histology and Long-term Clinical Outcome of Crushed Cartilage with Double-layer Gelatin Sponge Membrane for Dorsum Refinement in Primary Rhinoplasty.

This article demonstrates the ability to use autologous crushed cartilage grafts in rhinoplasty with rapid recovery and optimal nasal functionality without any tissue damage and allows its rapid rejuvenation. Eligible patients underwent primary rhinoplasty using autologous crushed cartilage graft followed by microscopy imaging of the grafted tissue after recovery. Tissue and cytological analysis using optical microscopy, transmission electronic microscopy (TEM), and scanning electronic microscopy (SEM) showed complete viability of chondrocytes, formation of new collagen fibers, neo-perichondrium, neo-angiogenesis, and exhibiting optimal aesthetic outcome. The surgical approach is easy to perform, feasible, and less time-consuming, with excellent tissue rejuvenation and rapid recovery.

The Use of Double-Layer Collagen Membrane for the Improvement of Nasal Dorsum Skin Thickness and Texture in Primary Nose Surgery.

OBJECTIVES: To place a collagen membrane containing crushed nasal septal cartilage over the nasal dorsum to see how this graft can improve the results of visible postsurgical irregularities in thin skinned patients. METHODS: Fifty-seven patients were treated between 2006 and 2010 (26 males and 31 females) whose ages ranged between 31 and 55 years old. They were divided into group a, defect <1 mm, group b defect between 1 and 2 mm, group c defect >3 mm. These patients presented intraoperative nasal dorsum irregularities that were corrected with 57 cartilage grafts in conjunction with collagen membrane. RESULTS: All of our patients showed an initial over-correction. A dense network of collagen fiber bundles was observed running parallel to the surface of the membrane at the connective tissue-membrane interface. The clinical percentage of volume reabsorption was about 5% after 6 months, measured with clinical pictures and a clinical follow-up. No infection was noted, only 1 patient of dislocation was observed, and 5 patients required some refinements at the long-term follow-up. CONCLUSIONS: In the authors' experience the use of bilayered combined cartilage and collagen membrane grafts gives the best aesthetic results with balanced tip projection and dorsum fullness, and avoids thickness and texture modification of the skin above the graft. The use of Bio-gide membrane avoids all problems related to the donor site and shortens surgical time.

Stamp Technique: An Explorative SEM Analysis.

BACKGROUND: Achieving correct tooth anatomy and saving time at the dental chair are some of the goals of modern restorative dentistry. Stamp technique has gained acceptance in clinical practice. The aim of this study was to evaluate the effectiveness of this technique in terms of microleakage, voids, overhangs and marginal adaptation of Class I restorations, and to analyse the operative times in comparison with traditional restorative procedures. METHODS: Twenty extracted teeth were divided into 2 groups. Ten teeth in the study group (SG) were Class I prepared and restored using stamp technique, and ten teeth in the control group (CG) were Class I restored traditionally. SEM analysis was performed to evaluate voids, microleakage, overhangs, and marginal adaptation, and operative times were recorded. A statistical analysis was performed. RESULTS: There were no significant differences in microleakage, marginal adaptation and filling defects between the two groups, however, the stamp technique seems to facilitate the formation of large overflowing margins that require a careful finishing phase. CONCLUSIONS: Stamp technique does not seem to have any critical aspects in terms of restoration durability and it can be performed in a short time.

Grafting of large mandibular advancement with a collagen-coated bovine bone (Bio-Oss Collagen) in orthognathic surgery.

Current principles for correction of dentoskeletal deformities ask to satisfy different treatment goals, making large mandibular advancements a common practice in orthognathic surgery. A main consequence of significant mandibular movements is the potential for unfavorable bone healing of osteotomy sites after traditional sagittal split procedures. This drawback, which mainly occurs at the level of inferior mandibular borders, can affect the stability and support of overlying soft tissues. Whereas the role of bone grafting for upper jaw defects after Le Fort I osteotomy is well addressed in the Literature, until now, just a few articles discussed the potential for grafting of mandibular osteotomy sites. The aim of this study is to evaluate the healing of mandibular bone defects because of large advancement (>8 mm) after sagittal spit procedures. In 20 patients treated for correction of class II dentoskeletal deformities, mandibular osteotomies defects have been grafted with a collagen-coated bovine bone substitute. Clinical, radiological, and histological evaluation of grafted sites showed a good healing of grafted area both in terms of recontouring of inferior mandibular borders and in terms of quality of newly formed bone. This confirms how this procedure could help to avoid the drawbacks related to significant mandibular advancement.

Evaluation of biocompatibility, osteointegration and biomechanical properties of the new Calcemex® cement: An in vivo study.

The mixture of polymethylmethacrylate (PMMA) and ß-tricalciumphospate (ß-TCP) is the most widely used bone graft. Common features of bone cement are the biocompatibility, bioactivity, mechanical stability and ability to fuse with the host's bone tissue. However, there are still few studies that have evaluated these characteristics in vivo. Our study aims to acquire these parameters, using an animal model with functional characteristics similar to those of humans. The analyzed cement is Calcemex®, evaluated both in compact and fluid formulation. The chosen animal models were 5 pigs, treated with femoral and tibial implants of Calcemex® samples. After one year, the pigs were sacrificed and the specimens explanted for morphological, histological, ultrastructural and mechanical evaluations. For both formulations, the investigation highlighted the absence of foreign body reactions in the host, the histological integration with the surrounding tissues and the preservation of mechanical compression resistance.

Tissue-Material Integration and Biostimulation Study of Collagen Acellular Matrices.

BACKGROUND: Breast reconstruction after mastectomy using silicone implants is a surgical procedure that occasionally leads to capsular contracture formation. This phenomenon constitutes an important and persistent cause of morbidity, and no successful therapies are available to date. Recently, the use of acellular membranes as a protective material for silicone prostheses has been gaining attention due to their ability to prevent this adverse outcome. For this reason, the evaluation of the tissue-material integration and the induced biostimulation by acellular membranes results crucial. Evaluation of in vivo tissue integration and biostimulation induced by three different natural acellular collagen membranes. METHODS: Scanning electron microscopy was performed to analyse the membrane porosity and cells-biomaterial interaction in vitro, both in dry and wet conditions. Adipose-derived stem cells were cultured in the presence of membranes, and the colonisation capacity and differentiation potential of cells were assessed. In vivo tests and ex vivo analyses have been performed to evaluate dermal integration, absorption degree and biostimulation induced by the evaluated membrane. RESULTS: Analysis performed in vitro on the three different acellular dermal matrices evidenced that porosity and the morphological structure of membranes influence the liquid swelling ratio, affecting the cell mobility and the colonisation capacity. Moreover, the evaluated membranes influenced in different manner the adipose derived stem cells differentiation and their survival. In vivo investigation indicated that the absorption degree and the fluid accumulation surrounding the implant were membrane-dependent. Finally, ex vivo analysis confirmed the membrane-dependent behavior revealing different degree of tissue integration and biostimulation, such as adipogenic stimulation. CONCLUSION: The physico-chemical characteristics of the membranes play a key role in the biostimulation of the cellular environment inducing the development of well-organized adipose tissue.

Biocompatible, photo-responsive layer-by-layer polymer nanocapsules with an oil core: in vitro and in vivo study.

In cancer therapy, stimulus-responsive drug delivery systems are of particular interest for reducing side effects in healthy tissues and improving drug selectivity in the tumoral ones. Here, a strategy for the preparation of a photo-responsive cross-linked trilayer deposited onto an oil-in-water nanoemulsion via a layer-by-layer technique is reported. The system is made of completely biocompatible materials such as soybean oil, egg lecithin and glycol chitosan, with heparin as the polymeric shell. The oil core is pre-loaded with curcumin as a model lipophilic active molecule with anti-tumoral properties. The trilayer cross-linkage is performed via a photoinitiator-free thiol-ene 'click' reaction. In particular, the system is implemented with an o-nitrobenzyl group functionalized with a thiol moiety which can perform both the thiol-ene 'click' reaction and the cleavage meant for controlled drug release at two different wavelengths, respectively. So the preparation and characterization of a photo-responsive natural nanocarrier (PNC) that is stable under physiological conditions owing to the thiol-ene cross-linkage are reported. PNC performance has been assessed in vitro on melanoma cells as well as in vivo on xenograft tumour-induced mice.

Ultrastructural features of the double capsulated connective tissue around silicone prostheses.

The implantation of breast prostheses for both aesthetic and reconstructive purposes has been growing exponentially in the last 20 years. Safety and prosthesis lifespan are majorly debated issues in relation to the correlated long-term complications. Mainly the periprosthetic capsule that develops around the implant is often the cause of complications and particularly for macrotextured silicone breast implants. Some reports have tried to elucidate the mechanism by which macrotextured silicone implants undergo damage and cause double capsule formation. In this study, we investigated the morphological characteristics of double capsule of macrotextured implants surgically removed from patients. With the use of microscopy techniques, this work analyzed the newly formed tissue observed in the interaction between synthetic and biological surfaces.

A novel near-infrared indocyanine dye-polyethylenimine conjugate allows DNA delivery imaging in vivo.

Near-infrared (NIR) fluorescence light has been applied to monitor several biological events in vivo since it penetrates tissues more efficiently than visible light. Dyes exhibiting NIR fluorescence and having large Stokes shift are key elements for this promising optical imaging technology. Here, we report the synthesis of a novel conjugate between a near-infrared indocyanine dye and an organic polyamine polymer (polyethylenimine, PEI) (IR820-PEI) with high chemical stability and good optical properties. IR820-PEI absorbs at 665 nm, emits at 780 nm, and displays a large Stokes shift (115 nm). Moreover, the reported conjugate is able to bind DNA, and the delivery process can be monitored in vivo with noninvasive optical imaging techniques. These characteristics make IR820-PEI one of the most effective and versatile indocyanine dye polymeric-conjugate reported so far.

Post-traumatic taste disorders: a case series.

Since 1800s there are reports of post-traumatic anosmia, but few studies investigated post-traumatic gustatory deficit and adopted validated evaluation tests. Peripheral and/or central mechanisms may be involved in the genesis of post-traumatic gustatory dysfunction. Beyond a reduction/loss (hypogeusia/ageusia) of gustatory function following a trauma, qualitative taste changes (dysgeusia) may occur. Especially when persistent, taste deficits might be particularly relevant for patient's quality of life and activities of daily life, but knowledge on this topic is limited. Fifty-three consecutive patients with previous head trauma were recruited. Every patient underwent a careful history taking and thorough olfactory and gustatory chemosensory testing by Sniffin'Sticks Extended test, Whole Mouth Test and Taste Strips Test. All patients had olfactory abnormalities (hyposmia: n = 10, functional anosmia: n = 43), while 10 of them (19%) showed taste deficits (dysgeusia: n = 3, dysgeusia with hypogeusia: n = 1, hypogeusia: n = 5, ageusia: n = 1). Here, we report clinical and neuroimaging data and detailed description of four meaningful cases representing central and peripheral injury patterns. Chemosensory evaluation might be useful to explore taste disorder, a still neglected and underestimated sequela of head trauma.

Regenerative potential of the Bichat fat pad determined by the quantification of multilineage differentiating stress enduring cells.

Published studies regarding Bichat fat pad focused, quite exclusively, on the implant of this adipose depot for different facial portions reconstruction. The regenerative components of Bichat fat pad were poorly investigated. The present study aimed to describe by an ultrastructural approach the Bichat fat pad, providing novel data at the ultrastructural and cellular level. This data sets improve the knowledge about the usefulness of the Bichat fat pad in regenerative and reconstructive surgery. Bichat fat pads were harvested form eight patients subjected to maxillofacial, dental and aesthetic surgeries. Biopsies were used for the isolation of mesenchymal cell compartment and for ultrastructural analysis. Respectively, Bichat fat pads were either digested and placed in culture for the characterization of mesenchymal stem cells (MSCs) or, were fixed in glutaraldehyde 2% and processed for transmission or scanning electron microscopy. Collected data showed very interesting features regarding the cellular composition of the Bichat fat pad and, in particular, experiments aimed to characterized the MSCs showed the presence of a sub-population of MSCs characterized by the expression of specific markers that allow to classify them as multilineage differentiating stress enduring cells.  This data set allows to collect novel information about regenerative potential of Bichat fat pad that could explain the success of its employment in reconstructive and regenerative medicine.

The biocompatibility of bone cements: progress in methodological approach.

The ideal bone graft substitute should have certain properties and there are many studies dealing with mixture of polymethylmetacrilate (PMMA) and ß-tricalciumphospate (ß-TCP) presenting the best characteristics of both. Scanning Electron Microscopy (SEM), for ultra-structural data, resulted a very reliable in vivo model to better understand the bioactivity of a cement and to properly evaluate its suitability for a particular purpose. The present study aims to further improve the knowledge on osteointegration development, using both parameters obtained with the Environmental Scanning Electron Microscopy (ESEM) and focused histological examination. Two hybrid bone graft substitute were designed among ceramic and polymer-based bone graft substitutes. Based on ß-TCP granules sizes, they were created with theoretical different osteoconductive properties. An acrylic standard cement was chosen as control. Cements were implanted in twelve New Zealand White (NZW) rabbits, which were sacrificed at 1, 2, 3, 6, 9 and 12 months after cement implantation. Histological samples were prepared with an infiltration process of LR white resin and then specimens were studied by X-rays, histology and Environmental Scanning Electron Microscopy (ESEM). Comparing the resulting data, it was possible to follow osteointegration's various developments resulting from different sizes of ß-TCP granules. In this paper, we show that this evaluation process, together with ESEM, provides further important information that allows to follow any osteointegration at every stage of develop.

Hyaluronic Acid (HA) Scaffolds and Multipotent Stromal Cells (MSCs) in Regenerative Medicine.

Traditional methods for tissue regeneration commonly used synthetic scaffolds to regenerate human tissues. However, they had several limitations, such as foreign body reactions and short time duration. In order to overcome these problems, scaffolds made of natural polymers are preferred. One of the most suitable and widely used materials to fabricate these scaffolds is hyaluronic acid. Hyaluronic acid is the primary component of the extracellular matrix of the human connective tissue. It is an ideal material for scaffolds used in tissue regeneration, thanks to its properties of biocompatibility, ease of chemical functionalization and degradability. In the last few years, especially from 2010, scientists have seen that the cell-based engineering of these natural scaffolds allows obtaining even better results in terms of tissue regeneration and the research started to grow in this direction. Multipotent stromal cells, also known as mesenchymal stem cells, plastic-adherent cells isolated from bone marrow and other mesenchymal tissues, with self-renew and multi-potency properties are ideal candidates for this aim. Normally, they are pre-seeded onto these scaffolds before their implantation in vivo. This review discusses the use of hyaluronic acid-based scaffolds together with multipotent stromal cells, as a very promising tool in regenerative medicine.

Might Helicobacter pylori infection be associated with distortion on taste perception?

Helicobacter pylori (H. pylori) has been found in dental plaque, saliva and lingual sites. To date, taste or olfaction disorders related to H. pylori infections have never been reported. In a review of the literature we found two papers just referring to a sour taste sensation during H. pylori infection. Studies in animal models suggest that changes in taste perception may relate to infections which damage taste buds. We observed an interesting clinical case of a 24-year-old Ghanaian woman with documented H. pylori gastric infection, complaining of cacosmia and cacogeusia. Taste evaluation indicated hypogeusia and highlighted a specific difficulty in discriminating between bitter and acid tastes. Saliva fluid was found positive for the ureA gene (H. pylori ureasi A). On the basis of this report, we hypothesize that taste perception might be correlated with a documented H. pylori infection. So, in a dyspeptic clinical picture in both pre and post diagnostic phase when H. pylori infection is suspected, taste evaluation might be important. Further studies are certainly needed in a large patient population to clarify the possible connection between H. pylori infection and smell-taste distortion.

Mapping molecular agents distributions in whole mice hearts using born-normalized optical projection tomography.

To date there is a lack of tools to map the spatio-temporal dynamics of diverse cells in experimental heart models. Conventional histology is labor intensive with limited coverage, whereas many imaging techniques do not have sufficiently high enough spatial resolution to map cell distributions. We have designed and built a high resolution, dual channel Born-normalized near-infrared fluorescence optical projection tomography system to quantitatively and spatially resolve molecular agents distribution within whole murine heart. We validated the use of the system in a mouse model of monocytes/macrophages recruitment during myocardial infarction. While acquired, data were processed and reconstructed in real time. Tomographic analysis and visualization of the key inflammatory components were obtained via a mathematical formalism based on left ventricular modeling. We observed extensive monocyte recruitment within and around the infarcted areas and discovered that monocytes were also extensively recruited into non-ischemic myocardium, beyond that of injured tissue, such as the septum.

Design, synthesis, and evaluation of gadolinium cationic lipids as tools for biodistribution studies of gene delivery complexes.

Gadolinium-chelating cationic lipids have been synthesized to obtain lipoplexes with MRI contrast properties. These compounds were designed to follow the biodistribution of synthetic DNA for gene delivery by nuclear magnetic resonance imaging. The lipid MCO-I-68 was synthesized, and chelate complexes with gadolinium were formed and characterized in terms of physicochemical and DNA binding properties. The transfection activity of MCO-I-68-Gd/DNA complexes was assayed in vitro on NIH 3T3. Different formulations of the product were tested. When up to 5% of the gadolinium lipid complexes were co-formulated with the cationic lipid RPR120535 used as a reference, the transfection levels were maintained as compared to RPR120535 alone. To date, only a liposomal formulation of a gadolinium-cationic lipid chelate without DNA had been observed using magnetic resonance imaging. In vivo intratumoral administration of MCO-I-68-Gd/DNA lipoplexes to tumor model led to an important increase of the NMR signal. It was demonstrated that the new complexes also acted as transfection carriers when they were formulated from liposomes.