Platelet rich fibrin in the management of medication-related osteonecrosis of the jaw: a clinical and histopathological evaluation.

Bisphosphonates are antiresorptive pharmacological agents used in the treatment of osteoporosis. Recently, osteonecrosis of the jaw has been recognized as a potential side effect in patients on long-term bisphosphonate therapy. This condition, popularly called bisphosphonate-related osteonecrosis of the jaw (BRONJ), has been rechristened as MRONJ (medication-related osteonecrosis of the jaw) to accommodate the increasing number of cases of osteonecrosis of jaws associated with various other antiresorptive and antigiogenic pharmacological therapies. The aim of the present study was to assess the outcome of using platelet-rich fibrin (PRF) for the treatment of MRONJ in a single study group. Twenty-three consecutive patients (15 females and 8 males; aged 52-73 years) with MRONJ were enrolled in this study. These patients presented a history of bisphosphonate medication of varying duration, presence of exposed bone in the maxillofacial region for more than eight weeks, and no history of radiation therapy to the jaws. These patients were managed by surgical curettage and application of platelet rich fibrin (PRF). The outcomes were assessed using clinical and histopathological methods. On the basis of the present findings, we can conclude that PRF can act as an effective barrier membrane between the alveolar bone and the oral cavity and may offer a fast, easy and effective alternative method for the closure of bone exposure in MRONJ patients.

Lights and shadows of dental implants: focus on mucositis and perimplantitis and their biological markers.

The increase in oral rehabilitation by means of dental implants has required an evolution of the related managing protocols and correct updating of the skills of dental professionals. Postsurgical management of the clinical case is aimed to stabilize the obtained results and preserve them from adverse conditions: a healthy implant prosthesis is maintained thanks to the huge number of consolidated protocols of oral hygiene. This practice plays a decisive role in the prevention of perimplant pathologies, forming a strong basis to ensure long implant life and avoid unnecessary and painful new surgical procedures. Furthermore, dental companies, in order to satisfy the new needs of professionals in oral hygiene, have produced new instrumentations and targeted drugs, in agreement to the cutting-edge scientific literature, thus creating a new market attracting huge interests in healthcare. The purpose of this topical review is to briefly comment on the state of the art of post-surgical dental implant management. This research is aimed to report the current protocols available to reduce the risk of oral diseases and prevent the progression of perimplant complications. Special focus has been dedicated to the most effective surgical and non-surgical protocols for treating mucositis and perimplantitis.

Anti-plaque and antimicrobial efficiency of different oral rinses in a 3-day plaque accumulation model.

The idea of incorporating a mouthrinse with normal tooth brushing could be a useful adjunct to oral hygiene. Despite the principle nature of the toothpaste vehicle, most alcohol-based chemical plaque-control agents have been evaluated and later formulated in the mouthrinse vehicle. The current study was aimed to investigate the persistence of antimicrobial action and plaque inhibitory properties of a new alcohol-free mouthrinse when compared with positive control, chlorhexidine 0.12% and placebo control, physiologic saline solution mouthrinses. The evaluation of the antimicrobial activity was performed by saliva samples collected during the 3 days of usage. The results of this study indicate that this new oral rinse has an equivalent plaque inhibitory action to chlorhexidine, and the plaque inhibitory action of the rinse appears to be derived from a persistence of antimicrobial action in the mouth. Furthermore, no side effects were reported during the study, and the additional benefit of no alcohol presence in the rinse solution.

DENTAL PULP STEM CELLS AND HUMAN PERIAPICAL CYST MESENCHYMAL STEM CELLS IN BONE TISSUE REGENERATION: COMPARISON OF BASAL AND OSTEOGENIC DIFFERENTIATED GENE EXPRESSION OF A NEWLY DISCOVERED MESENCHYMAL STEM CELL LINEAGE.

Bone regeneration is an interesting field of biomedicine. The most recent studies are aimed to achieve a bone regeneration using mesenchymal stem cells (MSCs) taken from more accessible sites: oral and dental tissues have been widely investigated as a rich accessible source of MSCs. Dental Pulp Stem Cells (DPSCs) and human Periapical Cysts Mesenchymal Stem Cells (hPCy-MSCs) represent the new generation MSCs. The aim of this study is to compare the gene expression of these two innovative cell types to highlight the advantages of their use in bone regeneration. The harvesting, culturing and differentiating of cells isolated from dental pulp as well as from periapical cystic tissue were carried out as described in previously published reports. qRT-PCR analyses were performed on osteogenic genes in undifferentiated and osteogenic differentiated cells of DPSC and hPCy-MSC lineage. Real-time RT-PCR data suggested that both DPSCs and hPCy-MSCs cultured in osteogenic media are able to differentiate into osteoblast/odontoblast-like cells: however, some differences indicated that DPSCs seem to be directed more towards dentinogenesis, while hPCy-MSCs seem to be directed more towards osteogenesis.

Immediately loaded dental implants bioactivated with platelet-rich plasma (PRP) placed in maxillary and mandibular region.

BACKGROUND AND AIM: Dental implants have today deeply changed the dental field. In fact, after almost 50 years, the permanent placement of a titanium screw with an attached tooth have, step by step, changed the society's standard toward a permanently replacement of missed or severely damaged teeth. In fact, the host of benefits born from dental implants have affected both patients and dental professionals. The aim of the present study is to report the outcomes of an implant therapy protocol supporting fixed prostheses implanted soon after extractions and loaded with flapless guided surgery by a 3D software planning. MATERIALS AND METHODS: 37 patients, requiring rehabilitation of booth dental arches with a one-to-one technique, were enrolled in a follow-up study plan which established clinical and radiological examinations on the day after surgery, seven months and one year later. Platelet Rich Plasma (PRP), in combination with autogenous bone, organic bone material and organic bone substitutes, was used before implant-prosthetic rehabilitation with an its prompt loading just before the implant insertion phase. RESULTS: Definitive restorations were delivered at 6-12 months after surgical procedure. One year after loading there were no dropouts and no failure of the definitive prosthesis. Furthermore, the feedback from patients resulted positive. CONCLUSIONS: Patients resulted satisfied both aesthetically and functionally regarding these types of prosthetic reconstructions made at the time.

Human periapical cyst-mesenchymal stem cells differentiate into neuronal cells.

It was recently reported that human periapical cysts (hPCys), a commonly occurring odontogenic cystic lesion of inflammatory origin, contain mesenchymal stem cells (MSCs) with the capacity for self-renewal and multilineage differentiation. In this study, periapical inflammatory cysts were compared with dental pulp to determine whether this tissue may be an alternative accessible tissue source of MSCs that retain the potential for neurogenic differentiation. Flow cytometry and immunofluorescence analysis indicated that hPCy-MSCs and dental pulp stem cells spontaneously expressed the neuron-specific protein ß-III tubulin and the neural stem-/astrocyte-specific protein glial fibrillary acidic protein (GFAP) in their basal state before differentiation occurs. Furthermore, undifferentiated hPCy-MSCs showed a higher expression of transcripts for neuronal markers (ß-III tubulin, NF-M, MAP2) and neural-related transcription factors (MSX-1, Foxa2, En-1) as compared with dental pulp stem cells. After exposure to neurogenic differentiation conditions (neural media containing epidermal growth factor [EGF], basic fibroblast growth factor [bFGF], and retinoic acid), the hPCy-MSCs showed enhanced expression of ß-III tubulin and GFAP proteins, as well as increased expression of neurofilaments medium, neurofilaments heavy, and neuron-specific enolase at the transcript level. In addition, neurally differentiated hPCy-MSCs showed upregulated expression of the neural transcription factors Pitx3, Foxa2, Nurr1, and the dopamine-related genes tyrosine hydroxylase and dopamine transporter. The present study demonstrated for the first time that hPCy-MSCs have a predisposition toward the neural phenotype that is increased when exposed to neural differentiation cues, based on upregulation of a comprehensive set of proteins and genes that define neuronal cells. In conclusion, these results provide evidence that hPCy-MSCs might be another optimal source of neural/glial cells for cell-based therapies to treat neurologic diseases.

Diagnosis and treatment of a rare case of adenomatoid odontogenic tumor in a young patient affected by attenuated familial adenomatosis polyposis (aFAP): case report and 5 year follow-up.

BACKGROUND: The adenomatoid odontogenic tumor (AOT) is a quite rare odontogenic tumor, with an incidence rate of approximately 12 cases/year worldwide. Attenuated familial adenomatous polyposis (aFAP) is a syndrome characterized by a significant risk to develop colon cancer. The aim of the paper is to describe a case never reported before in the literature: an AOT developed in a patient with aFAP; moreover, we want to show how it appears 5 years after surgery and after the regeneration of the eroded bone tissue, using the Platelet-Rich Fibrin (PRF) as filling material. CASE PRESENTATION: We report the case of a female 18 years old patient, affected by aFAP; she comes to us with a swelling on the right hemi-face. We performed several radiological exams, and they showed a neoformation approximately 2 cm in diameter: this neoformation packed the upper right canine, therefore, we hypothesized a dentigerous cyst. We decided to proceed to open biopsy and enucleation of the lesion. An intra-operative endodontic treatment on the adjacent partially resorbed teeth was also performed. Finally, we performed a reconstruction of eroded bone tissue, by use of Platelet-Rich Fibrin as filling material. The samples fixed and embedded in paraffin have led to the diagnosis of AOT. After 5 years from the surgery, we did not find any clear sign of relapse, in addition, the use of PRF has favored an optimal osteogenesis at the surgical site. CONCLUSIONS: Undoubtedly, a correct diagnosis of AOT allows to have a more performing clinical and surgical approach. Furthermore, this case could document a new manifestation of aFAP in extra-intestinal site. The onset of an AOT is quite rare in the general population, and this rarity could represent a critical point for its diagnosis; AOT onset in a patient with aFAP is a finding that could represent a new element of diagnosis and, therefore, the starting point to perform a more effective therapy.

Applications of natural compounds in the photodynamic therapy of skin cancer.

Despite significant advances in early diagnosis and treatment, skin cancer is one of the leading causes of death. Photodynamic therapy (PDT) is a new therapeutic modality that is emerging as an important resource against malignant tumors. This strategy is based on the action of photosensitizers, i.e. of molecules which may accumulate preferentially inside tumor cells where they exert a cytotoxic effect after excitation by light at appropriate wavelengths. Some forms of skin cancers and also some non-tumor pathologies are now treated with PDT. Several compounds with photosensitizing activity have been identified, and some of these molecules are commercially available. Many photoactive principles are natural compounds. Numerous reviews in the last decade have focused on photodynamic therapy, its effects and applications, but less attention has been paid to plant extracts or molecules of natural origin studied for their phototoxic activity to date.This review critically examines the potential role of various plant extracts and naturally occurring compounds in the treatment of skin cancer. Both in vitro and in vivo effects of these agents, together with their known related cellular and molecular mechanisms, are presented and discussed.

Influence of PRF in the healing of bone and gingival tissues. Clinical and histological evaluations.

INTRODUCTION: The healing of bone tissues around dental implants is based primarily on a correct osseointegration. BACKGROUND: Typically, implants stability and peri-implant tissues health are anticipated to decrease during the early weeks of healing; this is followed by an increase in stability. AIM: The aim of the present study is to assess a predictable protocol in order to increase the peri-implant tissues maintenance around post-extractive dental implants, thanks to the use of Platelet-Rich Fibrin (PRF) membrane. MATERIALS AND METHODS: This is a retrospective observational study of 127 tapered dental implants placed in the immediate post-extraction sites of maxillary bone of 59 patients. Atraumatic dental extraction and flapless implant surgery was performed in all reported cases. The cortical bone position relative to the implant reference point was evaluated at implant placement and 10 to 24 months following implantology. The gap between bone tissue and the implant surface was measured up to a maximum of 3 mm. After placing implants we have filled the surgical site with a PRF gel, so as to fill the gap between bone tissue and the implant surface, and then we have covered the surgical site with a PRF membrane, so as to coat the gap between the alveolar crest and the implant. RESULTS: In all cases, we observed the complete covering of the dental implants, with newly formed soft tissue of variable thickness between 1 and 3 mm. Cortical bone adaptation from the time of implant placement up to 30 months following prosthetic restoration ranged from 0.4 mm to 1.7 mm. CONCLUSIONS: Our study showed a series of successful rehabilitations, with post-extraction implantology technique, in 99.8 percent of cases, despite the success rates in the medium and long-term post-extraction implantology reported in the literature range between 92.7 percent and 98.0 percent. Long-term maintenance of crestal bone and the rapid healing of soft tissue dimension with maintenance of peri-implant papilla were observed as outcomes after post-extractive implants insertion.

Hypericum perforatum L. subsp. perforatum induces inhibition of free radicals and enhanced phototoxicity in human melanoma cells under ultraviolet light.

OBJECTIVES: Our interest continues in discovering phytocomplexes from medicinal plants with phototoxic activity against human melanoma cells; thus the aim of the present study was to assess antioxidant, anti-inflammatory and phototoxic activity of Hypericum perforatum L. subsp. perforatum, and relate these properties to the plant's chemical composition. MATERIALS AND METHODS: Components of H. perforatum subsp. perforatum were extracted by hydroalcoholic solution and chemical profiles of preparations (HyTE-3) performed by HPTLC. Linoleic acid peroxidation and DPPH tests were used to assess antioxidant activity, while MTT assay allowed evaluation of anti-proliferative activity with respect to A375 human melanoma cells after irradiation with UVA dose, 1.8 J/cm(2) . Inhibition of nitric oxide production of macrophages was also investigated. RESULTS: HyTE-3 indicated better antioxidant activity with ß-carotene bleaching test in comparison to DPPH assay (IC50 = 0.89 µg/ml); significant phototoxicity in A375 cells at 78 µg/ml concentration resulted in cell destruction of 50%. HyTE-3 caused significant dose-related inhibition of nitric oxide production in murine monocytic macrophage cell line RAW 264.7 with IC50 value of 342 µg/ml. CONCLUSIONS: The H. perforatum subsp. perforatum-derived product was able to suppress proliferation of human malignant melanoma A375 cells; extract together with UVA irradiation enhanced phototoxicity. This biological activity of antioxidant effects was combined with inhibition of nitric oxide production.

Regenerative surgery performed with platelet-rich plasma used in sinus lift elevation before dental implant surgery: an useful aid in healing and regeneration of bone tissue.

OBJECTIVES: Aim of this work is to show the effectiveness of a protocol involving the use of platelet-rich plasma (PRP) as a grafting material in bone regeneration before dental implant rehabilitation. MATERIALS AND METHODS: 127 patients, requiring maxillary sinus lift, were enrolled in a follow-up study plan, which established clinical and radiological examinations on the day after surgery and six months later. PRP, in combination with autogenous bone, an organic bone material and organic bone substitutes, was used before implant-prosthetic rehabilitation. RESULTS AND CONCLUSIONS: After implant placement, 63 patients, previously treated with PRP, reached a statistically significant improvement in implant-prosthetic rehabilitation, established by primary stability and radiographic integration criteria, in comparison with the other 64 patients receiving implant-prosthetic rehabilitation without PRP treatment.

Cachrys pungens Jan inhibits human melanoma cell proliferation through photo-induced cytotoxic activity.

OBJECTIVE: To date, plants belonging to the genus Cachrys have not been amply studied. In the present study, aerial components of Cachrys pungens Jan from Italy, were examined to assess their free radical-scavenging and antioxidant activity, and their phototoxicity on A375 melanoma cells. In view of potential pharmaceutical applications, a relationship between antioxidant, phototoxic activities and polyphenolic composition has also been investigated. MATERIALS AND METHODS: Content of sterols, terpenes, fatty acids and coumarins was assessed by gas chromatography-mass spectrometry and GC. Total phenolic content was also determined. Antioxidant activity of the methanol extract and fractions of C. pungens Jan was assessed using DPPH scavenging assay and ß-carotene bleaching test. Plant phototoxicity was also investigated in this human tumour cell line (amelanotic melanoma). RESULTS: Analysis of the chloroform extract was particularly interesting, as it led to identification of many coumarins, of which five were linear and one angular furanocoumarins. Methanol and ethyl acetate fractions exhibited substantial antioxidant activity. Moreover, chloroform extract and isolated coumarin fraction had strong phototoxic activity on UVA-induced A375 cells after irradiation at UVA dose of 1.08 J/cm. CONCLUSIONS: Plant-derived natural compounds are an important source for development of cancer-fighting drugs. This study has demonstrated strong phototoxic activity of the coumarin fraction of C. pungens, a plant which, to our knowledge, has never been studied before. This investigation offers a new perspective for developing other formulations potentially useful in photodynamic therapy for treatment of non-melanoma skin cancers as well as melanomas.

Biological potential and structure-activity relationships of most recently developed vascular disrupting agents: an overview of new derivatives of natural combretastatin a-4.

Tumor blood vessels are an important emerging target for anti-cancer therapy. The antimitotic agent combretastatin A-4 (CA-4), a cis-stilbene natural product isolated from the South African tree Combretum caffrum Kuntze, is the lead compound of a new class of anti-cancer drugs that target tumor vasculature. CA-4 inhibits tubulin polymerization by interacting at the colchicine binding site on tubulin. This alters the morphology of endothelial cells and causes vascular shutdown and regression of tumor vasculature. Some tubulin-binding vascular-disrupting agents (VDAs) are currently in clinical trials for cancer therapy. As a consequence of the potential favorable applications of these compounds, several analogs projected to induce rapid and selective vascular shutdown in tumors have been synthesized during the last few years. Many of these molecules have already been tested for their effects on tubulin polymerization as well as for their antiproliferative activity and other biological properties, and possible mechanisms of action have been investigated. The aim of the present review is to offer an overview of most recently developed combretastatin derivatives, focusing on biological effects exerted by these compounds. The published data about new analogs are presented and compared, and a detailed investigation of structure-activity relationships is described.

Clinical trial with bromelain in third molar exodontia.

BACKGROUND AND OBJECTIVE: Bromelain is a proteolytic enzyme, particularly effective in the treatment of soft tissue inflammations and traumas, in localized inflammations, especially in presence of hydropsy and also in postoperative tissue reactions. The aim of the present study is to assess the efficacy of bromelain in controlling the edema and its related pain in the inflamed area after upper third molar exodontia. MATERIALS AND METHODS: The effectiveness of our protocol was evaluated by a clinical assessment of the profile of the hemiface corresponding to the treated area: indeed, the linear values of the trago-pogonion distances were measured. Algogens were determined by VAS (Visual Analogue Scale) (Figure 1) with integers ranging from 1 (no pain) to 8 (maximum pain) up to a maximum of 10 (paroxysmal and unbearable pain). RESULTS AND CONCLUSIONS: The obtained results clearly demonstrate the effectiveness of bromelain in treating postoperative edema after third molar surgery.

Antiproliferative activity against human tumor cell lines and toxicity test on Mediterranean dietary plants.

Sixteen edible plants from Southern Italy were evaluated for their in vitro antiproliferative properties, using the sulforodamine B (SRB) assay, on four human cancer cell lines: breast cancer MCF-7, prostate cancer LNCaP, amelanotic melanoma C32 and renal adenocarcinoma ACHN. After 48 h of incubation the most antiproliferative plant extract was Cynara cardunculus ssp. cardunculus on C32 and ACHN cell lines with IC(50) of 21 and 18 microg/ml, respectively. Mentha aquatica showed a selective antiproliferative activity on breast cancer while significant activity was exerted by Cichorium intybus on melanoma. These species contained the highest amount of phenolics. The acute toxicity of the hydroalcohol extracts from all the plants were evaluated by using the Microtox acute toxicity test. This bacterial test measures the decrease in light emission from the marine luminescent Vibrio fischeri bacteria when exposed to organic extracts. This inhibition test was revealed to be highly sensitive, cost effective and easy to operate, requiring just 15 min to predict the sample toxicity. All the extracts analyzed resulted to give values very less than a limit of 20% value, demonstrating so an irrelevant toxicity for the human health. In contrast, Echium vulgare and Malva sylvestris showed bioluminescence inhibition values of 19.42% and 17.32%, respectively, just under the established limit.

Antioxidant and cytotoxic activities of methanolic extract and fractions from Senecio gibbosus subsp. gibbosus (GUSS) DC.

The aim of this study was to evaluate the in vitro antioxidant and cytotoxic activities of the methanolic extract and fractions of Senecio gibbosus subsp. gibbosus aerial parts. The antioxidant activity was assessed by means of two different tests: (1) bleaching of the stable 2,2-diphenyl-1-picrylhydrazil (DPPH) radical; and (2) lipid peroxidation of liposomes which were prepared from bovine brain extract. In both tests used methanolic extract and AcOEt fraction showed a significant antioxidant effect. The cytotoxic activity of the methanolic extract and fractions was carried out using the SRB assay. The methanolic extract demonstrated a good cytotoxic activity against human breast cancer cell line (MCF-7) and human prostate cancer cell line (LNCaP). Dichloromethane and AcOEt fractions showed the greatest cytotoxic activity, particularly on LNCaP cell line.

Analysis of salivary gland proteins of the mosquito Anopheles darlingi (Diptera: Culicidae).

The salivary proteins of Anopheles darlingi Root, the principal vector of malaria in the Amazon Region, Brazil, were analyzed. Comparison of the protein profiles between adult males and females revealed that most of the polypeptides are present in both sexes, but female-specific polypeptides also were observed. SDS-PAGE analysis of sugar-fed female mosquitoes with ages varying from 1 to 10 d after adult emergence indicated that the proteins start to be accumulated in the first day of life and are present throughout the period analyzed. Analysis of blood-fed mosquitoes showed no differences in salivary proteins when compared with sugar fed ones, suggesting that there is no specific protein induced by blood. The protein profiles of the salivary glands dissected from wild-caught female mosquitoes from different geographical regions of Brazil were compared and some differences were observed.

Seasonal variation of anti-Plasmodium falciparum antibodies directed against a repetitive peptide of gametocyte antigen pfs2400 in inhabitants in the State of Amapá, Brazil.

Antibodies to the Pfs2400 gametocyte antigen have been shown to inhibit the development of Plasmodium falciparum in anophelines and therefore this antigen is a candidate for a transmission-blocking vaccine. To test seasonal variation of these antibodies under field conditions, sera from 72 individuals were collected twice, first during the long-rains season with low malaria transmission and then, 6 months later, during the short-rains season, when transmission is high. This study was conducted in several localities in the State of Amapá, Brazil. All but three individuals had a positive indirect fluorescent antibody test (IFAT) with asexual forms of P. falciparum. Most of them did not report malaria attacks during the period between the first and second sampling. Their sera were tested by IFAT with P. falciparum gametocytes. The overall positivity of this test did not vary between seasons, and was 47.2 (34/72) and 48.6% (35/72), respectively. The sera were also tested by ELISA with the Pfs2400 repeat peptide. The positivity rate dropped from 29.2 (21/72) to 15.3% (11/72) and the mean absorbancies from 0.623 to 0.354, when we compared the results of the long-rains and short-rains seasons. Fifteen out of the 21 ELISA positive sera turned negative, with no change of geometric mean of titres (GMT) of asexual IFAT, while five negatives became ELISA positive on second sampling, with increase of GMT. Soon after the second sampling a malaria outbreak was reported in one of the localities. These results point toward a relatively short persistence of anti-Pfs2400 repeat peptide antibodies, under natural field conditions. A gametocyte antigen booster before a high transmission period might contribute towards lowering malaria incidence by eliciting a partially effective antibody response.