Maria Adelaide Continenza

Ultrastructure of isolated mouse ovarian follicles cultured in vitro.

BackgroundIn vitro maturation of ovarian follicles, in combination with cryopreservation, might be a valuable method for preserving and/or restoring fertility in mammals with impaired reproductive function. Several culture systems capable of sustaining mammalian follicle growth in vitro have been developed and many studies exist on factors influencing the development of in vitro grown oocytes. However, a very few reports concern the ultrastructural morphology of in vitro grown follicles.MethodsThe present study was designed to evaluate, by transmission and scanning electron microscopy, the ultrastructural features of isolated mouse preantral follicles cultured in vitro for 6 days in a standard medium containing fetal calf serum (FCS). The culture was supplemented or not with FSH.ResultsThe follicles cultured in FCS alone, without FSH supplementation (FCS follicles), did not form the antral cavity. They displayed low differentiation (juxta-nuclear aggregates of organelles in the ooplasm, a variable amount of microvilli on the oolemma, numerous granulosa cell-oolemma contacts, signs of degeneration in granulosa cell compartment). Eighty (80)% of FSH-treated follicles formed the antral cavity (FSH antral follicles). These follicles showed various ultrastructural markers of maturity (spreading of organelles in ooplasm, abundant microvilli on the oolemma, scarce granulosa cell-oolemma contacts, granulosa cell proliferation). Areas of detachment of the innermost granulosa cell layer from the oocyte were also found, along with a diffuse granulosa cell loosening compatible with the antral formation. Theca cells showed an immature morphology for the stage reached. Twenty (20)% of FSH-treated follicles did not develop the antral cavity (FSH non-antral follicles) and displayed morphological differentiation features intermediate between those shown by FCS and FSH antral follicles (spreading of organelles in the ooplasm, variable amount of microvilli, scattered granulosa cell-oolemma contacts, signs of degeneration in granulosa cell compartment).ConclusionsIt is concluded that FSH supports the in vitro growth of follicles, but the presence of a diffuse structural granulosa cell-oocyte uncoupling and the absence of theca development unveil the incomplete efficiency of the system. The present study contributes to explain, from a morphological point of view, the effects of culture conditions on the development of mouse in vitro grown follicles and to highlight the necessity of maintaining efficient intercellular communications to obtain large numbers of fully-grown mature germ cells.

Effectiveness of platelet-rich plasma as an adjunctive material to bone graft: a systematic review and meta-analysis of randomized controlled clinical trials

The use of platelet-rich plasma (PRP) has become a strategic therapy in tissue regeneration medicine. PRP represents a good source of growth factors. Due to this property, it has been considered a reliable adjunctive material in bone augmentation procedures, such as the sinus lift technique. The aim of this review was to assess the scientific evidence on the effectiveness of PRP as an adjunctive material in the sinus floor elevation technique. The following databases were searched for relevant published studies: Medline, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, CINAHL, Science Direct, ISI Web of Knowledge, and SCOPUS. Only randomized controlled clinical trials comparing a group receiving PRP as an adjunctive material to a control group without PRP, involving adult human subjects (age >18 years) with no systemic disease, were included. Of the studies identified, only one reported a significant difference in bone augmentation in favour of the adjunctive use of PRP, while four studies did not find any significant difference. None of the studies included reported a significant difference in the implant survival rate. Further randomized clinical trials are needed to clarify the effectiveness of adjunctive PRP.

Microleakage of bacteria in different implant-abutment assemblies: an in vitro study

Purpose The aim of the present in vitro study was to evaluate the leakage observed for 2 different microbial species at the level of the implant–abutment (I-A) interface, and the marginal fit and size of microgap at the I-A interface in 2 different implant connections. Methods Ten specimens of each group were tested. The inner parts of 5 implants per group were inoculated with 0.1 μL of a viable Enterococcus faecalis suspension and 5 implants per group with Aggregatibacter actinomycetemcomitans. All of the vials containing the control specimens were incubated at 37°C under aerobic condition for E. faecalis and 37°C in presence of 5% CO2 for A. actinomycetemcomitans. They were maintained for 14 days, and the possible penetration of bacterial suspension into the surrounding solution was determined by the observation of turbidity of the broth. The I-A interface was evaluated for size of microgap and measured under SEM. Five implants of each group were evaluated. The marginal fit between implant and abutment was measured at 8 random locations in each assembly, under different magnifications at the interface. Results No leakages through the I-A interface were demonstrated for either type of connection evaluated. The microgap values of all I-A interfaces ranged from 0.008 to 2.009 μm; the differences between the 2 systems were statistically significant. Conclusions The present study demonstrated that a good marginal fit of implant components seemed to be able to prevent bacterial leakage.

Anterior mandibular lingual foramina: an in vivo investigation.

In descriptions of surgical procedures in mandible, often there is no mention of an anatomical variance, the genial spinal foramina, where nerves and vessels go through. Aim of this study is to investigate frequency, shape, and dimensions of these foramina. 56 computed tomography dentascans were analyzed with an implant planning software. The considered parameters were frequency, number, position, diameters, and length of canals; the collected data were inserted in a spreadsheet and statistically analyzed; therefore, they were compared with those found in the literature. The measurements agree with the ones found in earlier studies, except for the length of the inferior spinal canals, which resulted lesser than that found in the literature. The frequency of the inferior spinal foramina, the data related to the inferior spinal foramina diameter (cross scan), and the measurements related to the superior spinal foramina diameter (axial scan) resulted to be major compared to those reported in literature. These obtained results are clinically interesting because an implant planning software has been employed, daily used by operators, and that permits in vivo investigations. Furthermore, due to the possibility of hemorrhagic accidents in this mandibular region, these data are particularly interesting for all of the operators who make interventions in this area.

Frequency and anatomical features of the mandibular lingual foramina: systematic review and meta-analysis

Lingual mandibular foramina are peculiar anatomical variations often present on the bone surface of the inferior jaw. These structures host vascular and nervous anastomosis deriving from the branches of the submental artery, lingual artery, and the mylohyoid nerve. Reports pointed out intra-operative massive bleeding accidents during implant interventions occurred in this area. The aim of this systematic review and meta-analysis was to summarize quantitatively the features of the lingual foramina on the midline of the inferior jaw. We performed a systematic review and meta-analysis complying the PRISMA statement and registering it in PROSPERO database. The frequency rate of the foramina resulted to have a point estimate of 0.965. The means of the diameters showed to have a point estimate of 0.840 with a standard error of 0.06. The results showed a significant high frequency reported in the literature of this variation, with a quite important diameter, sign of significant caliber of the related vessels. Even if this variation is underreported in modern textbooks of oral anatomy, we suggest an accurate screening via CT scan pre-operatively. In addition, a proper risk management is necessary to minimize injure hazard and properly managing the vascular plexus of the anterior floor of oral cavity.

Development of a new protocol: a macroscopic study of the tongue dorsal surface

The presence on the dorsal tongue of additional irregularities such as fissures, grooves and the distribution of papillae constitutes a retention area for harbouring bacteria, particularly those that produce sulphur compounds. A protocol to study the morphology of the tongue in a macroscopic way was developed, aimed at better investigating on the relationship between the dorsal surface of the tongue, considered as a microbial ecosystem, and the oral halitosis (1). A patient affected by oral malodour was chosen and included in the study. A picture of his lingual dorsum was taken to show the areas where the coating was visible. Then, an impression with alginate was taken obtaining a replication in plaster and, on the base of this one, a sort of impression tray was modelled utilizing an impression material made of silicone putty. After this, a second impression was taken combining the tray in silicone putty with a silicone material having a very low-light density (using the 2-step double-mix impression technique). The impression obtained was divided and cut with a blade in six parts, according to Winkel Tongue Coated Index (2), and their contour was observed with the stereo-microscope. The images were analysed with the Image J programme, and the parameter considered was the depth among the papillae. The mean measure of this parameter, in the parts where the coating was visible, resulted in a range between 0,25 + 0,019 mm and 0,55 + 0,11 mm. The same measures in the parts where the coating was not visible swung instead in a range between 0,14 + 0,08 mm and 0,23 + 0,07 mm. This new protocol can be considered clinically relevant for the suitable diagnosis and for the personalized treatment of halitosis.

In vitro biocompatibility of a new hydrogel with Crocin, powerful antioxidant found in Crocus Sativus L. flowers

Recently, attention has been paid to the identification of natural antioxidants from the petals of Crocus S. flowers that are normally considered waste[1]. The antioxidant activities are mainly attributed to carotenoid compounds, like crocin. Scientific evidences demonstrate that this kind of compounds are among the most important natural plant sources of antioxidant activity in the human diet, protecting the body against damages caused by reactive oxygen species (ROS). This antioxidant was extracted using methanol and ethanol. In this experiment, it was used a new hydrogel consisting of three polymers - Polyvinyl pyrrolidone (PVP), Agar and Polyethylene glycol (PEG) – and Crocin, mixed, reticulated and then sterilized by gamma irradiation at 25 kGy. For the in vitro experimental protocol, it was used a primary culture of fibroblasts taken from the subcutaneous tissue of a newborn mice, seeding the cells on a little square (1cm2 area) of both kinds of hydrogel. As the ISO protocol prescribes, the experiments were repeated 3 times for each kind of hydrogel, stopping the culture at the 3rd, 7th and 14th day after the seeding. For all steps, 3 Petri dishes were used as controls without biomaterials. At fixed deadline, all Petri dishes were stained, using the Wright method for cell counting and morphological evaluations. The microscopic analysis revealed the complete biocompatibility of the hydrogel. The Petri dishes with this kind of new hydrogel has an overproduction of collagen from the fibroblasts. The presence of this natural important compound in saffron petals, now offers new possibilities for the best use of the hole flower. This study is still in progress.

Caracterización Histológica de la Membrana de los Factores de Crecimiento Concentrados Sacco

Along with the emerging needs of the dental patients, numerous techniques for oral tissue stimulation and regeneration were developed to be employed in the modern implant rehabilitation therapies. The Concentrated Growth Factors (CGF) are a relat ively new therapeutic presidium that can be used for this purpose, enhancing the regenerative potential property of blood cells. Alth ough literature presents numerous studies evaluating the CGF for their clinical uses and efficacy, data regarding their biological c haracteristics are very few. The present study evaluates and describes the CGF structural morphology by means of classical histological method s, using haematoxilin-eosin and azan mallory stains. A three layers organization with a fibrin complex network was noted, with blood cor puscular elements entrapped, especially in the most external layer. These descriptions enrich the knowledge about this new type of membr ane, showing the bio-morphological side of the regenerative techniques. These findings will be useful in clinical practice for the c hoice of the most suitable technique in each implant rehabilitation.

Kaempferol, a powerful antioxidant from Crocus sativus L. flowers: an in vitro study

Recently, it has been demonstrated the presence of Kaempferol in the petals of Crocus S. flowers that are normally discarded [1]. Kaempferol is an important antioxidant of the flavonoid family. Numerous reports have shown that kaempferol and/or its glycosides induce cell death in a variety of cancer cells from different tissues, also it can slow skin aging by contrasting enzymes that degrade the extracellular matrix. This antioxidant has been extracted from Crocus S. petals with different alcohols. Isolation and purification was done by flash column chromatography. Fractions were analysed by Thin Layer Chromatography (TLC). Kaempferol was further characterised by infrared (IR), mass spectroscopy (MS) and nuclear magnetic resonance (1H & 13C NMR) spectroscopy. To be sure of the antioxidant property of the extracted Kaempferol, a DPPH test was performed. In this experiment, it was used a new hydrogel consisting of three polymers - Polyvinyl pyrrolidone (PVP), Agar, Polyethylene glycol (PEG) – and Kaempferol, mixed, reticulated and then sterilized by gamma irradiation at 25 kGy. [2]. For the in vitro experimental protocol, it was used a primary culture of fibroblasts taken from the subcutaneous tissue of a newborn mice, seeding the cells on a little square (1cm2 area) of the hydrogel with Kaempferol. As the ISO protocol prescribes, the experiments were repeated 3 times, stopping the culture at the 3rd, 7th and 14th day after the seeding. For all steps, 3 Petri dishes were used as controls without the biomaterial. At fixed deadline, all Petri dishes were stained, using the Wright method for cell counting and morphological evaluations. The microscopic analysis revealed the complete biocompatibility of the hydrogel. The Petri dishes with this kind of new hydrogel showed a production of collagen fibres similar of the Petri dishes without the hydrogel. The presence of this bioactive compound in saffron petals, paves the way of new possibilities for the best use of the Crocus S. hole flower.

New Hydrogels Enriched with Antioxidants from Saffron Crocus Can Find Applications in Wound Treatment and/or Beautification

Saffron extracts have a long history of application as skin protectant, possibly due to their ability to scavenge free radicals. In this work, the performance of a hydrogel enriched with antioxidant compounds isolated from saffron crocus (Crocus sativus L.) petals was tested. These hydrogels could be considered as new drug delivery system. Hydrogels are crosslinked polymer networks that absorb large quantities of water but retain the properties of a solid, thus making ideal dressings for sensitive skin. We tested antioxidant-enriched hydrogels on primary mouse fibroblasts. Hydrogels enriched with kaempferol and crocin extracted from saffron petals showed good biocompatibility with in vitro cultured fibroblasts. These new types of hydrogels may find applications in wound treatment and/or beautification.