The efficacy of hyaluronic acid fragments with amino acid in combating facial skin aging: an ultrasound and histological study.

BACKGROUND: Various techniques have been employed in aesthetic medicine to combat skin aging, in particular that of the facial region. Hyaluronic acid is utilized to enhance moisture levels and extracellular matrix molecules. This study aims to histologically assess the effects of low molecular weight hyaluronic acid fragments combined with amino acids (HAAM) on facial skin rejuvenation through intradermal microinjections. METHODS: A total of twenty women, with an average age of 45 and ranging from 35 to 64 years old, participated in the study, including 8 in menopause and 12 in the childbearing age group. Mesotherapy was used to administer HAAM to the patients. Prior to and three months after the treatment, each patient underwent small circular punch biopsies. Ultrasound examinations were conducted using B-mode, capturing 2D images in longitudinal or transverse orientations with frequencies ranging from 5 to 13 Mega-hertz (MY LAB X8, ESAOTE, Genova, Italy). A total of 60 ultrasound examinations were taken, with 30 collected before treatment and 30 after treatment. RESULTS: The histological analysis demonstrates an increase in fibroblast activity resulting in the production of Type III reticular collagen, as well as an increased number of blood vessels and epidermal thickness. However, the analysis of ultrasound data before and after treatment showed no statistical difference in skin thickness in malar area, chin and mandibular angle. CONCLUSIONS: Histological assessments indicate that subcutaneous infiltration of HAAM has a substantial impact on the dermis of facial skin.

Tumor and peritumoral adipose tissue crosstalk: De-differentiated adipocytes influence spread of colon carcinoma cells.

Colorectal cancer is the second leading cause of cancer and often has a fatal course. There are many studies in the literature that have described a close functional relationship between the tumor mass and surrounding tissue, or tumor stroma, which is affected by the continuous metabolic exchange that occurs at the interface between tumor and tissues in contact with it. There is much evidence that the presence of adipose tissue in stroma plays a fundamental role in modulating the tumor microenvironment and promote tumor development, growth, and angiogenesis due to its endocrine characteristics. In this analysis, we have studied the alterations of adipose tissue surrounding colorectal tumors with MRI and optical imaging in vivo techniques to monitor tumor progression and also performed histological and molecular analysis. We detected differences in the principal adipose markers expressed by adipocytes residing around the rectal colon and observed that peritumoral adipose tissue is exposed to a mesenchymal transition process that leads to the acquisition of a less differentiated phenotype of adipocyte that represents the main cellular type present in tumor stroma. The mesenchymal transition correlated with the acquisition of more aggressive tumor phenotype and could represent a valid target for tumor therapy.

Clinical evaluation of efficacy and tolerance of a skin reconditioning compound for anti-aging.

Facial aging involves all facial structures located at different levels: bones soft tissues and skin with a reduction of the extracellular matrix. The aim of the study was to evaluate the efficacy of the injectable solution antiaging complex composed by non-reticulated hyaluronic acid (HA) and amino acids vitamins and antioxidants conveyed with mesotherapy technique in subjects with different expressions of aging. 114 patients with different expressions of aging were enrolled in this study with mean age (49±6). HA and amino acids vitamins and antioxidants complex solution Neofound (Love Cosmedical, Castagneto, Italy) was injected on the dermal plane or superficial subdermal plane. Among the various imperfections, fine roughness surface irregularities skin firmness brightness/discoloration cutaneous hydration were those with the greatest response to therapy. The clinical data showed that the medical device Neofound is effective and safe to treat various skin signs of chrono and photoaging thanks to its ability to protect tissues from oxidative stress and hydrate the skin.

CROCC-mutated rhabdoid colorectal carcinoma showing in intercellular spaces lamellipodia and cellular projections revealed by electron microscopy.

BACKGROUND: Rhabdoid colorectal carcinoma (RC) is a rare lesion localized to the proximal colon of patients with a mean age at diagnosis of around 70 years. This tumor shows an aggressive behavior with an overall survival period shorter than 12 months. The diagnostic hallmark is the presence of rhabdoid cells. Alterations in chromatin remodeling (SMARCB1) and in the centrosome structure (CROCC) are reported in RC usually BRAFmut and MSI-H. RKO intestinal neoplastic cells culture (BRAFmut, SMARCB1wt, MSI-H) with CROCC knockdown exhibit rhabdoid features and develop prominent projections from the edge of the cell. METHODS: Here, we investigated two cases of CROCCmutSMARCB1wt RC by scanning and transmission electron microscopy (SEM, TEM). RESULTS: TEM confirmed the diagnostic presence of intermediate cytoplasmic filaments and nucleolar margination. SEM showed cellular protrusions (lamellipodia) in the intercellular spaces not evident at light microscopy. CONCLUSIONS: These protrusions CROCC-related might represent the pathogenetic mechanism underlying the rhabdoid aggressive behavior, independently of tumor staging. To our knowledge, the SEM technique was applied in the study of this neoplasm for the first time.

Hyaluronic acid fillers in lip augmentation procedure: a clinical and histological study.

Several approaches have been proposed for soft tissue augmentation using injectable materials for lip and soft-tissue augmentation due to their physical and biological properties. Hyaluronic acid represents a natural component of the connective tissues, related to wound healing and skin regeneration. The aim of this research was to investigate the clinical and histological effectiveness of cross-linked hyaluronic acid for lip augmentation. A total of 8 patients were treated for lip augmentation with a clinical follow up of 60 days. After this period a histological evaluation was performed to evaluate the healing of the treated regions. The healing phase reported no inflammatory response, tissue contractions and no local flogistic evidence in the treated areas, where the filling volume appeared maintained. The 60 days histological evaluation showed evidence of filler resorption with few infiltrated inflammatory cells. The clinical and histological findings suggested that cross-linked hyaluronic acid represents a safe and effective tool for lip augmentation.

Evaluation of the effectiveness and safety of peppermint peel (PMP) soft peeling for skin ageing.

Turning to peeling in a dermatological sphere is extensively common and has been used for a long time. From the use of single acids moving on to the so-called compound peelings (associations of more than one substance in the same product) and the combined peelings which take advantage of the action of different substances in a synergistic manner (different products are applied sequentially) in order to best guarantee a greater effectiveness of the treatment for the recommended target. Superficial peelings, combined and not, have led to a drastic reduction in the percentage of incidence of adverse events typical of medium and deep peels. Nevertheless, it has been demonstrated that superficial peels bring about a rejuvenating effect through the mechanical stimulation of the Skin Stress Response System (SSRS), system designated to repairing the damaged tissue and restoring of the normal homeostasis. Clinical trials aims to evaluate the effectiveness and safety of the peppermint peel (PMP) medical device in subjects with different ageing expressions both in qualitative terms (different blemishes such as discolouration, fine wrinkles, elastosis, atony and skin inelasticity, laxity, scarce superficial hydration) and in quantitative terms (degree, extension and number of lesions). A non-controlled multi-centric clinical trial was done in 121 subjects. The use protocol calls for a session every 2 weeks for a total of 4 sessions. Subjects were evaluated before each subsequent session at the first and at 2-4-8 weeks of the fourth and last treatment. During the study there were no adverse events. Only a minimal scurfy flaking and a very slight redness were reported. From an effectiveness point of view, the percentage of therapeutic failure, judged with a score equal to or greater than 4 or 5 in Global Aesthetic Improvement Scale (GAIS) scale was 0%. Best score was obtained in subjects ranging in ages between 38 and 57 (2.02) and in women (2.02) years, while the less satisfactory one was obtained in males (2.14). The study has demonstrated that PMP and the proposed protocol are effective and safe to treat subject with skin signs of chrono and photo ageing, thanks to its capabilities of carrying out a mechanic action indicated as a coadjuvant in the treatment of the dermoepidermic revitalisation through chemical exfoliation and hydration.

Non-surgical treatment of lower eyelid fat pads with an injectable solution acid deoxycholic based.

The use of sodium deoxycholate (DC) in aesthetic medicine for reducing unwanted localized fat deposits is a procedure in use for over 30 years. Lower eyelid fat pads are one of the main imperfections of the middle third of the face. The purpose of the study is to assess the effectiveness and the safety of a second-generation injectable solution containing sodium deoxycholate 1.25% (DB125) for the treatment of lower eyelid fat pads. A multi-centre observational prospective study was carried out between May and October 2017. Patients presented various forms and degrees of lower eyelid fat pads. They were treated montly apart until the clinical result was obtained. The technique was explained. The study treated 120 patients for a total of 306 infiltration sessions (average 2.55 per patient). Patients gave the effectiveness of the treatment an average score of 7.125 (7.28 in men and 7.03 in women). The greatest successes were in patients under the age of 40. The medical evaluation showed therapeutic success in 85.83%. Adverse events were reported. The results of the study are encouraging since they have shown a therapeutic success from both the viewpoint of specialist medical assessment and from the personal, subjective view of the patients treated. This success did not show any significant differences between sex. On the other hand, outcome has been better in the younger age groups. The high degree of effectiveness shown in the study was associated with a minimal occurrence of adverse events. Therefore DB125, used with the right technique and dosage, is effective and safe to treat lower eyelid fat pads.

Histological evaluation of periodontal ligament in human after orthodontic treatment with piezosurgery and monolateral tooth dislocation and ligament distraction technique: a first morphologic and histologic evaluation.

Traditional orthodontic tooth movement is based on the concept that application of a protracted force causes alveolar bone remodelling and adaptive changes in periodontal and dental tissues. Thus, if orthodontic tooth movement is described as a biological bone reaction to orthodontic forces mediated by the periodontal ligament (PDL), this event involves a series of sophisticated signal transduction processes that allows the PDL compression with specific histologic and biomolecular modifications. However, the preservation of the integrity of the PDL is generally difficult to achieve when it is associated with a long duration of orthodontic treatment. A total of 20 Caucasian patients with different dental-skeletal were treated using the Monocortical Tooth Dislocation and Ligament Distraction (MTDLD) technique with Piezosurgery associated with morphologic and histological evaluation of the PDL. The histological results obtained, confirm a good clinical outcome with an improvement of the speed on orthodontic treatment without any signs of tissue injury of PDL fiber without areas of hyalinization. The data suggests that MTDLD with Piezosurgery seems to be a valid alternative to the traditional orthodontic movement in adult patients preserving the anatomy and the integrity of PDL.

Filamentous fungi associated with natural infection of noble rot on withered grapes.

The effects of noble rot infection of grapes on the characteristics of different types of wine, including Italian passito wine, are well known. Nevertheless, there is still little information on filamentous fungi associated with noble-rotten grapes. In this study, withered Garganega grapes for passito wine production, naturally infected by noble rot, were analyzed and compared to sound grapes. Skin morphology and fungal population on berry surfaces were analyzed. Scanning electron microscopy analysis revealed microcracks, germination conidia and branched hyphae on noble-rotten berries. Penicillium, Aureobasidium and Cladosporium were the most frequent genera present. Analysis of single berries displayed higher heterogeneity of epiphytic fungi in those infected by noble-rot than in sound berries. Penicillium adametzoides, Cladosporium cladospoirioides and Coniochaeta polymorpha were recovered. These, to the best of our knowledge, had never been previously isolated from withered grapes and, for C. polymorpha, from grapevine. This study provided novel data on noble rot mycobiota and suggests that fungi that co-habit with B. cinerea could have an important role on grape and wine quality.

Molecules implicated in glucose homeostasis are differentially expressed in the trachea of lean and obese Zucker rats.

Recent studies indicate that the processes mediated by the (T1R2/T1R3) glucose/sugar receptor of gustatory cells in the tongue, and hormones like leptin and ghrelin contribute to the regulation of glucose homeostasis. Altered plasma levels of leptin and ghrelin are associated with obesity both in humans and rodents. In the present study, we evaluated the ultrastructure of the mucosa, and the expression of molecules implicated in the regulation of glucose homeostasis (GLUT2, SGLT1, T1R3, ghrelin and its receptor) in the trachea of an animal model of obesity (Zucker rats). We found that the tracheal epithelium of obese animals was characterized by the presence of poorly differentiated cells. Ciliated and secretory cells were the cell lineages with greatest loss of differentiation. Severe epithelial alterations were associated with marked deposit of extracellular matrix in the lamina propria. The expression pattern of GLUT2 and SGLT1 glucose transporters was similar in the trachea of both the Zucker rat genotypes, whereas that of T1R3 was reduced in ciliated cells of obese rats. A different immunolocalization for ghrelin was also found in the trachea of obese rats. In conclusion, the tracheal morphological alterations in obese animals seem to compromise the expression of molecules involved in the homeostasis of glucose.

In vivo imaging techniques: a new era for histochemical analysis.

In vivo imaging techniques can be integrated with classical histochemistry to create an actual histochemistry of water. In particular, Magnetic Resonance Imaging (MRI), an imaging technique primarily used as diagnostic tool in clinical/preclinical research, has excellent anatomical resolution, unlimited penetration depth and intrinsic soft tissue contrast. Thanks to the technological development, MRI is not only capable to provide morphological information but also and more interestingly functional, biophysical and molecular. In this paper we describe the main features of several advanced imaging techniques, such as MRI microscopy, Magnetic Resonance Spectroscopy, functional MRI, Diffusion Tensor Imaging and MRI with contrast agent as a useful support to classical histochemistry.

Characterization of magnetic nanoparticles from Magnetospirillum Gryphiswaldense as potential theranostics tools.

We investigated the theranostic properties of magnetosomes (MNs) extracted from magnetotactic bacteria, promising for nanomedicine applications. Besides a physico-chemical characterization, their potentiality as mediators for magnetic fluid hyperthermia and contrast agents for magnetic resonance imaging, both in vitro and in vivo, are here singled out. The MNs, constituted by magnetite nanocrystals arranged in chains, show a superparamagnetic behaviour and a clear evidence of Verwey transition, as signature of magnetite presence. The phospholipid membrane provides a good protection against oxidation and the MNs oxidation state is stable over months. Using an alternate magnetic field, the specific absorption rate was measured, resulting among the highest reported in literature. The MRI contrast efficiency was evaluated by means of the acquisition of complete NMRD profiles. The transverse relaxivity resulted as high as the one of a former commercial contrast agent. The MNs were inoculated into an animal model of tumour and their presence was detected by magnetic resonance images two weeks after the injection in the tumour mass.

Chromium-induced diffuse dermatitis with lymph node involvement resulting from Langerhans cell histiocytosis after metal-on-metal hip resurfacing.

Total hip arthroplasty (THA) is a highly effective surgical treatment for severe joint involvement. However, due to the release of metal ions in the blood, the patients who undergo hip replacement with metal-on-metal (MOM) bearings may develop signs of allergic skin disease. We report a case of a 60-year-old man who had received MOM hip resurfacing 5 years earlier for osteoarthritis. He presented with a 3-year history of diffuse dermatitis that did not respond to antihistamines and corticosteroids and also had elevated serum levels of chromium and cobalt. A patch test revealed chromium-sulfate hypersensitivity. A skin biopsy showed nonspecific perivascular lymphocytic infiltrate associated with histiocytes. A biopsy of an inguinal lymph node demonstrated large aggregates of Langerhans cells, suggesting type IV delayed-type hypersensitivity. The prosthesis was replaced using ceramic-on-ceramic bearings and the dermatitis resolved after 3 months. The lymph nodes decreased in volume and the serum chromium levels normalized within 24 months of revision surgery. The high levels of serum ions associated with the metal debris from MOM-THAs may induce sensitization and type IV hypersensitivity reactions. Replacing the prosthesis using alternative coupling surfaces is the only approach that has the capacity to resolve these symptoms. Physicians who are not familiar with this issue may misdiagnose systemic symptoms and provide inadequate treatment.

The post-adipocytic phase of the adipose cell cycle.

Subcutaneous white adipose tissue harvested by liposuction has been studied with the aim to understand how the adipocytes modify their morphology when subjected to the passage in a needle for liposuction and to cryopreservation. The work try to clarify the ultrastructural aspects of adipose tissue, in the conditions described before, examining samples of body fat employed in fat graft procedures, and samples after cryopreservation. Scanning and transmission electron microscopy show that the first event that occur in the adipocytes is a lesion of mild degree detectable early in the samples fixed immediately after liposuction. The sequence of events following the adipocyte stress appeared composed by different phases: plasmatic membrane interruption, loss of lipid charge, formation of cup-like adipocytes and formation of post-adipocytes (i.e. cells that survive to traumatic events and restart to internalize lipid droplets). In conclusion, the study suggests that the loss of lipid charge in adipose cell is an active process that can be due to a small hole in the cytoplasmic membrane with the preservation of a large part of the cytoplasmatic content and that at the end of the process of lipid extrusion the cell can maintain viability.

The biocompatibility of porous vs non-porous bone cements: a new methodological approach.

Composite cements have been shown to be biocompatible, bioactive, with good mechanical properties and capability to bind to the bone. Despite these interesting characteristic, in vivo studies on animal models are still incomplete and ultrastructural data are lacking. The acquisition of new ultrastructural data is hampered by uncertainties in the methods of preparation of histological samples due to the use of resins that melt methacrylate present in bone cement composition. A new porous acrylic cement composed of polymethylmetacrylate (PMMA) and ß-tricalciumphosphate (ß-TCP) was developed and tested on an animal model. The cement was implanted in femurs of 8 New Zealand White rabbits, which were observed for 8 weeks before their sacrifice. Histological samples were prepared with an infiltration process of LR white resin and then the specimens were studied by X-rays, histology and scanning electron microscopy (SEM). As a control, an acrylic standard cement, commonly used in clinical procedures, was chosen. Radiographic ultrastructural and histological exams have allowed finding an excellent biocompatibility of the new porous cement. The high degree of osteointegration was demonstrated by growth of neo-created bone tissue inside the cement sample. Local or systemic toxicity signs were not detected. The present work shows that the proposed procedure for the evaluation of biocompatibility, based on the use of LR white resin allows to make a thorough and objective assessment of the biocompatibility of porous and non-porous bone cements.

3D printing of rat salivary glands: The submandibular-sublingual complex.

The morphology and the functionality of the murid glandular complex, composed of the submandibular and sublingual salivary glands (SSC), were the object of several studies conducted mainly using magnetic resonance imaging (MRI). Using a 4.7 T scanner and a manganese-based contrast agent, we improved the signal-to-noise ratio of the SSC relating to the surrounding anatomical structures allowing to obtain high-contrast 3D images of the SSC. In the last few years, the large development in resin melting techniques opened the way for printing 3D objects starting from a 3D stack of images. Here, we demonstrate the feasibility of the 3D printing technique of soft tissues such as the SSC in the rat with the aim to improve the visualization of the organs. This approach is useful to preserve the real in vivo morphology of the SCC in living animals avoiding the anatomical shape changes due to the lack of relationships with the surrounding organs in case of extraction. It is also harmless, repeatable and can be applied to explore volumetric changes occurring during body growth, excretory duct obstruction, tumorigenesis and regeneration processes. 3D printing allows to obtain a solid object with the same shape of the organ of interest, which can be observed, freely rotated and manipulated. To increase the visibility of the details, it is possible to print the organs with a selected zoom factor, useful as in case of tiny organs in small mammalia. An immediate application of this technique is represented by educational classes.

Lipid droplets characterization in adipocyte differentiated 3T3-L1 cells: size and optical density distribution.

The 3T3-L1 cell line, derived from 3T3 cells, is widely used in biological research on adipose tissue. 3T3-L1 cells have a fibroblast-like morphology, but, under appropriate conditions, they differentiate into an adipocyte-like phenotype. During the differentiation process, 3T3-L1 cells increase the synthesis of triglycerides and acquire the behavior of adipose cells. In particular, triglycerides accumulate in lipid droplets (LDs) embedded in the cytoplasm. The number and the size distribution of the LDs is often correlated with obesity and many other pathologies linked with fat accumulation. The integrated optical density (IOD) of the LDs is related with the amount of triglycerides in the droplets. The aim of this study is the attempt to characterize the size distribution and the IOD of the LDs in 3T3-L1 differentiated cells. The cells were differentiated into adipocytes for 5 days with a standard procedure, stained with Oil Red O and observed with an optical microscope. The diameter, area, optical density of the LDs were measured. We found an asymmetry of the kernel density distribution of the maximum Feret's diameter of the LDs with a tail due to very large LDs. More information regarding the birth of the LDs could help in finding the best mathematical model in order to analyze fat accumulation in adipocytes.