Mesenchymal stromal cells loaded with Paclitaxel (PacliMES) a potential new therapeutic approach on mesothelioma.

Malignant pleural mesothelioma is an asbestos-related tumor originating in mesothelial cells of the pleura that poorly responds to chemotherapeutic approaches. Adult mesenchymal stromal cells derived either from bone marrow or from adipose tissue may be considered a good model for cell-based therapy, a treatment which has experienced significant interest in recent years. The present study confirms that Paclitaxel is effective on mesothelioma cell proliferation in 2D and 3D in vitro cultures, and that 80,000 mesenchymal stromal cells loaded with Paclitaxel inhibit tumor growth at a higher extent than Paclitaxel alone. An in vivo approach to treat in situ mesothelioma xenografts using a minimal amount of 106 mesenchymal stromal cells loaded with Paclitaxel showed the same efficacy of a systemic administration of 10 mg/kg of Paclitaxel. These data strongly support drug delivery system by mesenchymal stromal cells as a useful approach against many solid tumors. We look with interest at the favourable opinion recently expressed by the Italian Drug Agency on the procedure for the preparation of mesenchymal stromal cells loaded with Paclitaxel in large-scale bioreactor systems and their storage until clinical use. This new Advanced Medicinal Therapy Product, already approved for a Phase I clinical trial on mesothelioma patients, could pave the way for mesenchymal stromal cells use as drug delivery system on other solid tumors for adjuvant therapy associated with surgery and radiotherapy.

Regeneration of Critical-Sized Mandibular Defects Using 3D-Printed Composite Scaffolds: A Quantitative Evaluation of New Bone Formation in In Vivo Studies.

Mandibular tissue engineering aims to develop synthetic substitutes for the regeneration of critical size defects (CSD) caused by a variety of events, including tumor surgery and post-traumatic resections. Currently, the gold standard clinical treatment of mandibular resections (i.e., autologous fibular flap) has many drawbacks, driving research efforts toward scaffold design and fabrication by additive manufacturing (AM) techniques. Once implanted, the scaffold acts as a support for native tissue and facilitates processes that contribute to its regeneration, such as cells infiltration, matrix deposition and angiogenesis. However, to fulfil these functions, scaffolds must provide bioactivity by mimicking natural properties of the mandible in terms of structure, composition and mechanical behavior. This review aims to present the state of the art of scaffolds made with AM techniques that are specifically employed in mandibular tissue engineering applications. Biomaterials chemical composition and scaffold structural properties are deeply discussed, along with strategies to promote osteogenesis (i.e., delivery of biomolecules, incorporation of stem cells, and approaches to induce vascularization in the constructs). Finally, a comparison of in vivo studies is made by taking into consideration the amount of new bone formation (NB), the CSD dimensions, and the animal model.

Conditioned Medium of Mesenchymal Stromal Cells Loaded with Paclitaxel Is Effective in Preclinical Models of Triple-Negative Breast Cancer (TNBC).

Triple-negative breast cancer (TNBC) is a very aggressive disease even in its early stages and is characterized by a severe prognosis. Neoadjuvant chemotherapy is one of the milestones of treatment, and paclitaxel (PTX) is among the most active drugs used in this setting. However, despite its efficacy, peripheral neuropathy occurs in approximately 20-25% of cases and represents the dose-limiting toxicity of this drug. New deliverable strategies to ameliorate drug delivery and reduce side effects are keenly awaited to improve patients' outcomes. Mesenchymal stromal cells (MSCs) have recently been demonstrated as promising drug delivery vectors for cancer treatment. The aim of the present preclinical study is to explore the possibility of a cell therapy approach based on the use of MSCs loaded with PTX to treat TNBC-affected patients. For this purpose, we in vitro evaluated the viability, migration and colony formation of two TNBC cell lines, namely, MDA-MB-231 and BT549, treated with MSC-PTX conditioned medium (MSC-CM PTX) in comparison with both CM of MSCs not loaded with PTX (CTRL) and free PTX. We observed stronger inhibitory effects on survival, migration and tumorigenicity for MSC-CM PTX than for CTRL and free PTX in TNBC cell lines. Further studies will provide more information about activity and potentially open the possibility of using this new drug delivery vector in the context of a clinical study.

Micro-fragmented Fat Inhibits the Progression of Human Mesothelioma Xenografts in Mice.

BACKGROUND: Malignant pleural mesothelioma is a pathology with no effective therapy and a poor prognosis. Our previous study demonstrated an in vitro inhibitory effect on mesothelioma cell lines of both the lysate and secretome of adipose tissue-derived Mesenchymal Stromal Cells. The inhibitory activity on tumor growth has been demonstrated also in vivo: five million Mesenchymal Stromal Cells, injected "in situ", produced a significant therapeutic efficacy against MSTO-211H xenograft equivalent to that observed after the systemic administration of paclitaxel. OBJECTIVE: The objective of this study is to evaluate the efficacy of low amount (half a million) Mesenchymal Stromal Cells and micro-fragmented adipose tissues (the biological tissue from which the Mesenchymal Stromal Cells were isolated) on mesothelioma cells growth. METHODS: Tumor cells growth inhibition was evaluated in vitro and in a xenograft model of mesothelioma. RESULTS: The inhibitory effect of micro-fragmented fat from adipose-tissue has been firstly confirmed in vitro on MSTO-211H cell growth. Then the efficacy against the growth of mesothelioma xenografts in mice of both micro-fragmented fat and low amount of Mesenchymal Stromal Cells has been evaluated. Our results confirmed that both Mesenchymal Stromal Cells and micro-fragmented fat, injected "in situ", did not stimulate mesothelioma cell growth. By contrast, micro-fragmented fat produced a significant inhibition of tumor growth and progression, comparable to that observed by the treatment with paclitaxel. Low amount of Mesenchymal Stromal Cells exerted only a little anticancer activity. CONCLUSION: Micro-fragmented fat inhibited mesothelioma cell proliferation in vitro and exerted a significant control of the mesothelioma xenograft growth in vivo.

Significant association between FGFR1 mutation frequency and age in central giant cell granuloma.

Central giant cell granulomas (CGCG) are rare intraosseous osteolytic lesions of uncertain aetiology. Despite the benign nature of this neoplasia, the lesions can rapidly grow and become large, painful, invasive, and destructive. The identification of molecular drivers could help in the selection of targeted therapies for specific cases. TRPV4, KRAS and FGFR1 mutations have been associated with these lesions but no correlation between the mutations and patient features was observed so far. In this study, we analysed 17 CGCG cases of an Italian cohort and identified an interesting and significant (p=0.0021) correlation between FGFR1 mutations and age. In detail, FGFR1 mutations were observed frequently and exclusively in CGCG from young (<18 years old) patients (4/5 lesions, 80%). Furthermore, the combination between ours and previously published data confirmed a significant difference in the frequency of FGFR1 mutations in CGCG from patients younger than 18 years at the time of diagnosis (9/23 lesions, 39%) when compared to older patients (1/31 lesions, 0.03%; p=0.0011), thus corroborating our observation in a cohort of 54 patients. FGFR1 variants in young CGCG patients could favour fast lesion growth, implying that they seek medical attention earlier. Our observation might help prioritise candidates for FGFR1 testing, thus opening treatment options with FGFR inhibitors.

Poly-D,L-lactic acid polymer and cochlear implantation.

This case report describes a peculiar and innovative fixing procedure with a Poly-D,L-lactic acid (PDLLA) polymer in the unusual case of magnet dislodgment and rupture of the cochlear implant (CI) silicone sheath holding the magnet.

Radiographic protrusion of dental implants in the maxillary sinus and nasal fossae: A multidisciplinary consensus utilising the modified Delphi method.

The aim of the present study was to generate an international and multidisciplinary consensus on the clinical management of implant protrusion into the maxillary sinuses and nasal fossae. A total of 31 experts participated, 23 of whom were experts in implantology (periodontologists, maxillofacial surgeons and implantologists), 6 were otolaryngologists and 2 were radiologists. All the participants were informed of the current scientific knowledge on the topic based on a systematic search of the literature. A list of statements was created and divided into three surveys: one for all participants, one for implant providers and radiologists and one for otolaryngologists and radiologists. A consensus was reached on 15 out of 17 statements. According to the participants, osseointegrated implants protruding radiographically into the maxillary sinus or nasal fossae require as much monitoring and maintenance as implants fully covered by bone. In the event of symptoms of sinusitis, collaboration between implant providers and otolaryngologists is required. Implant removal should be considered only after pharmacological and surgical management of sinusitis have failed.

Effects on Muscular Activity after Surgically Assisted Rapid Palatal Expansion: A Prospective Observational Study.

The study aims to investigate the modifications in the temporalis and the masseter activity in adult patients before and after SARPE (Surgically Assisted Rapid Palatal Expansion) by measuring electromyographic and electrokinesographic activity. 24 adult patients with unilateral posterior crossbite on the right side were selected from the Orthodontic Department of the University of Milan. Three electromyographic and electrokinesographic surface readings were taken respectively before surgery (T0) and 8 months after surgery (T1). The electromyographic data of both right and left masseter and anterior temporalis muscles were recorded during multiple tests: standardized maximum voluntary contraction (MVC)s, after transcutaneous electrical nerve stimulation (TENS) and at rest. T0 and T1 values were compared with paired Student's t-test (p < 0.05). Results: Significant differences were found in the activity of right masseter (p = 0.03) and right temporalis (p = 0.02) during clench, in the evaluation of right masseter at rest (p = 0.03), also the muscular activity of masseters at rest after TENS from T0 to T1 (pr = 0.04, pl = 0.04). No significant differences were found in the activity of left masseter (p = 0.41) and left temporalis (p = 0.39) during clench and MVC, in the evaluation of left masseter at rest (p = 0.57) and in the activity during MVC of right masseter (p = 0.41), left masseter (p = 0.34), right temporalis (p = 0.51) and left temporalis (p = 0.77). Results showed that the activity of the masseter and temporalis muscles increased significantly after SARPE during rest and clenching on the side where the cross-bite was treated.

Medication-Related Osteonecrosis of the Jaw in Dental Practice: A Retrospective Analysis of Data from the Milan Cohort.

A retrospective analysis was performed with the aim of understanding whether the risk factors showed in the literature for medication-related osteonecrosis of the jaws (MRONJ) in cancer patients are also relevant in osteoporotic patients taking antiresorptive drugs (ARDs). Data were retrospectively pooled from health records of patients on ARDs who requested a dental visit between January 2006 and April 2020 in the Dental Unit at Fondazione Ca' Granda IRCCS Ospedale Maggiore Policlinico, University of Milan. A total of 434 patients were included. The following variables were collected: sex, age, smoking habit, type of ARD, duration of treatment, route of administration, therapeutic indication, concurrent systemic therapies and pathologies. Statistical analysis confirmed the relevance of chemotherapy, smoking, and immunosuppressive drugs as risk factors. In addition, a higher frequency of MRONJ in osteoporotic patients was reported in our cohort in association with an immunodeficiency disorder of variable origin. In conclusion, the identification of individual risk-profile before dental treatments is crucial for prevention. Anamnesis should include main risk factors, such as immunosuppression, dental extractions, smoking, trauma, and poor dental health. Nevertheless, our suggestion for dental professionals is to conduct a complete medical history of patients who mention long-term per oral therapies with ARDs for osteoporosis. Osteoporotic, as well as cancer patients, may also benefit from periodic monitoring of the ARDs therapy in order to prevent MRONJ.

Post-Operative Patients' Satisfaction and Quality of Life Assessment in Adult Patients with Obstructive Sleep Apnea Syndrome (OSAS).

BACKGROUND: The treatment for severe OSAS includes maxillomandibular advancement surgical option in selected cases. The aim of this study was to evaluate the post-operative impact of bimaxillary surgery on satisfaction and consequently the quality of life of these patients. METHODS: This study included 18 patients with severe OSAS who received maxillomandibular advancement surgery. Patients were divided into Group A (operated by CAD/CAM) and Group B (conventional surgery). The impact of bimaxillary surgery on satisfaction and quality of life of these patients was evaluated by utilizing post-operative life quality and Rustemeyer's patient-satisfaction-based survey. RESULTS: A total of 18 adult OSAS patients (Group A: 11 patients, Group B: 7 patients) with a mean age of 44.39 years (SD ± 9.43) were included. Mean follow-up period was 32.64 months (SD ± 21.91). No intra-operative complications were seen in any patients. Post-operative complication was seen in one patient and the mandible did not integrate. According to the results, overall post-operative satisfaction score was 79.72% (SD ± 9.96). There was no significant difference among those in Group A and Group B. CONCLUSIONS: Maxillomandibular advancement surgery seems to be beneficial in terms of patients' satisfaction in severe adult OSAS patients and can be considered as a valuable option in selected cases.

Dental Trauma at a University Dental Clinic in Milan including the SARS-CoV-2 Period.

The aim of this study i.e., is to present the distribution of traumatic dental injuries (TDI) in 306 patients registered at the Unit of Dental Emergencies at a University Dental Clinic in Milan, Italy, between June 2019 and May 2021. This time frame includes the beginning of the SARS-CoV-2 pandemic. Information regarding age, gender, number and type of injured teeth, type of traumatic injury, and data on where or how the injury happened were recorded. Seventy-nine percent of patients can be classified as pediatric (under 14 years old), and in all age groups, male patients were found to be more susceptible (1.6:1). A total of 480 teeth were involved, 59% of which were deciduous, and 41% permanent. The most affected teeth in both dentitions were upper central incisors. In deciduous teeth, periodontal lesions were more common, whereas in permanent dentitions, dental fractures were diagnosed more often. Most data found in this study confirms the results found in the literature. The biggest difference, due to changes in daily routine during the SARS-CoV-2 pandemic, can be found by analyzing the incidence and etiology. As a matter of fact, there was a decrease in school accidents, whereas domestic falls remained constant.

In Vitro Activity of Monofunctional Pt-II Complex Based on 8-Aminoquinoline against Human Glioblastoma.

A new cationic Pt(II) complex bearing 8-aminoquinoline as chelating ligand (called Pt-8AQ) was evaluated against two human carcinomas, one mesothelioma, and three glioblastoma cell lines. The in vitro comparison to the clinically approved CisPt showed a minor activity of Pt-8AQ against carcinoma and mesothelioma, whereas a significant activity of Pt-8AQ was observed on the proliferation of the three glioblastoma cell lines (U87-MG IC50 = 3.68 ± 0.69 µM; U373-MG IC50 = 11.53 ± 0.16 µM; U138-MG IC50 = 8.05 ± 0.23 µM) that was higher than that observed with the clinically approved CisPt (U87-MG IC50 = 7.27 + 1.80 µM; U373-MG IC50 = 22.69 ± 0.05 µM; U138-MG IC50 = 32.1 ± 4.44 µM). Cell cycle analysis proved that Pt-8AQ significantly affected the cell cycle pattern by increasing the apoptotic cells represented by the sub G0/G1 region related with a downregulation of p53 and Bcl-2. Moreover, an NMR investigation of Pt-8AQ interaction with 9-EtG, GSH, and Mets7 excluded DNA as the main target, suggesting a novel mechanism of action. Our study demonstrated the high stability of Pt-8AQ after incubation at 37 °C and a significant antineoplastic activity on glioblastomas. These features also make Pt-8AQ a good candidate for developing a new selective advanced cell chemotherapy approach in combination with MSCs.

Inhibition of Human Malignant Pleural Mesothelioma Growth by Mesenchymal Stromal Cells.

BACKGROUND: Malignant Pleural Mesothelioma (MPM) is an aggressive tumor that has a significant incidence related to asbestos exposure with no effective therapy and poor prognosis. The role of mesenchymal stromal cells (MSCs) in cancer is controversial due to their opposite effects on tumor growth and in particular, only a few data are reported on MSCs and MPM. METHODS: We investigated the in vitro efficacy of adipose tissue-derived MSCs, their lysates and secretome against different MPM cell lines. After large-scale production of MSCs in a bioreactor, their efficacy was also evaluated on a human MPM xenograft in mice. RESULTS: MSCs, their lysate and secretome inhibited MPM cell proliferation in vitro with S or G0/G1 arrest of the cell cycle, respectively. MSC lysate induced cell death by apoptosis. The efficacy of MSC was confirmed in vivo by a significant inhibition of tumor growth, similar to that produced by systemic administration of paclitaxel. Interestingly, no tumor progression was observed after the last MSC treatment, while tumors started to grow again after stopping chemotherapeutic treatment. CONCLUSIONS: These data demonstrated for the first time that MSCs, both through paracrine and cell-to-cell interaction mechanisms, induced a significant inhibition of human mesothelioma growth. Since the prognosis for MPM patients is poor and the options of care are limited to chemotherapy, MSCs could provide a potential new therapeutic approach for this malignancy.

Comparing prognostic utility between the 8th edition of TNM staging system and the lymph node ratio for oral squamous cell carcinoma.

BACKGROUND: Lymph node metastasis in oral squamous cell carcinoma (OSCC) is associated with poor prognosis. The 8th edition of TNM has implemented new nodal staging criteria. We assess the prognostic utility of the lymph node ratio (LNR) and compare it to that of pN in the TNM 8th edition. METHODS: One hundred and forty-two patients with OSCC were retrospectively studied. Nodal staging was performed using the TMN 8th edition and the prognostic value of the LNR in terms of overall survival (OS) and disease-free survival (DFS) was evaluated. RESULTS: Fifty-seven patients were eligible for inclusion. The LNR was independently prognostic of OS (p = 0.02). Instead N classification was not significantly predictive of OS (p = 0.10). High LNRs resulted in decreases in OS of approximately 40% within 6 months after surgery. CONCLUSIONS: The LNR identifies patients with poor outcomes better than N classification. The lack of reliable LNR cutoffs compromises its utility in clinical practice.

Pros and Cons of CAD/CAM Technology for Infection Prevention in Dental Settings during COVID-19 Outbreak.

The purpose of this commentary is to update the evidence reported in our previous review on the advantages and limitations of computer-aided design/computer-aided manufacturing technology in the promotion of dental business, as well as to guarantee patient and occupational safety. The COVID-19 pandemic led to an unprecedented focus on infection prevention; however, waves of COVID-19 follow one another, asymptomatic cases are nearly impossible to identify by triage in a dental setting, and the effectiveness of long-lasting immune protection through vaccination remains largely unknown. Different national laws and international guidelines (mainly USA-CDC, ECDC) have often brought about dissimilar awareness and operational choices, and in general, there has been very limited attention to this technology. Here, we discuss its advantages and limitations in light of: (a) presence of SARS-CoV-2 in the oral cavity, saliva, and dental biofilm and activation of dormant microbial infections; (b) the prevention of SARS-CoV-2 transmission by aerosol and fomite contamination; (c) the detection of various oral manifestations of COVID-19; (d) specific information for the reprocessing of the scanner tip and the ward from the manufacturers.

Accuracy of splint vs splintless technique for virtually planned orthognathic surgery: A voxel-based three-dimensional analysis.

PURPOSE: This study compared two transferring methods for virtually planned orthognathic surgery - the CAD/CAM intermediate splint and the customized surgical guide with fixation plates. METHODS: This was a prospective clinical study in which participants were consecutively recruited and underwent bimaxillary orthognathic surgery. They were divided into two groups based on the transferring method used. The pre- and postoperative CBCTs were aligned using voxel-based landmark-free registration, and the discrepancies for selected points were compared with the planned displacement of the virtually planned surgery. The maxilla and mandible were analyzed separately, and translation and rotation movements were considered. RESULTS: A total of 16 patients, divided into two groups of eight patients each, were included in this study. The splintless group was significantly more accurate for the translation movement along the x-axes for points A (p = 0.008; mean absolute error 0.527 ± 0.387 for the splint group and 0.137 ± 0.067 for the splintless group) and Ans (p = 0.045; mean absolute error 0.535 ± 0.446 for the splint group and 0.156 ± 0.002 for the splintless group). For the mandible there was a significant difference in accuracy along the x-axes for points B (p = 0.049; mean absolute errors 1.728 ± 1.181 and 0.697 ± 0.519 for the splint and splintless groups, respectively), LL3 (p = 0.049; mean absolute error 1.629 ± 0.912 and 0.851 ± 0.797 for the splint and splintless groups, respectively), LR3 (p = 0.049; mean absolute error 1.711 ± 0.906 and 0.844 ± 0.780 for the splint and splintless groups, respectively), with the splintless group being more accurate. For the rotation the splintless group was significantly more accurate along the y-axes (p = 0.04; mean absolute error 1.62 ± 0.78 and 0.49 ± 0.31 for the splint and splintless groups, respectively) and z-axes (p = 0.04; mean absolute error 0.63 ± 0.45 and 0.17 ± 0.05 for the splint and splintless groups, respectively) for the maxilla, while no significant difference was found for the mandible. CONCLUSIONS: Overall, the customized fixation plate system is more accurate than the intermediate CAD/CAM splint for transferring the virtual plan into the operation room.

Paclitaxel Priming of TRAIL Expressing Mesenchymal Stromal Cells (MSCs-TRAIL) Increases Antitumor Efficacy of Their Secretome.

BACKGROUND: Adipose tissue derived MSCs engineered with the tumor necrosis factor-related apoptosis-inducing ligand protein (MSCs-TRAIL) have a significant anticancer activity. MSCs, without any genetic modifications, exposed to high doses of chemotherapeutic agents are able to uptake the drug and release it in amount affecting tumor proliferation. The purpose of this study was to verify the ability of MSCs-TRAIL to uptake and release paclitaxel (PTX) by providing an increased antitumor efficacy. METHODS: MSCs and MSCs-TRAIL were tested for their sensitivity to Paclitaxel (PTX) by MTT assay and the cells were loaded with PTX according to a standardized procedure. The secretome was analysed by HPLC for the presence of PTX, microarray assay for soluble TRAIL (s-TRAIL) and tested for in vitro anticancer activity. RESULTS: MSCs-TRAIL were resistant to PTX and able to incorporate and then release the drug. The secretion of s-TRAIL by PTX loaded MSCs-TRAIL was not inhibited and the PTX delivery together with s-TRAIL secretion resulted into an increased antitumor efficacy of cell secretoma as tested in vitro on human pancreatic carcinoma (CFPAC-1) and glioblastoma (U87-MG). CONCLUSIONS: Our result is the first demonstration of the possible merging of two new MSCs therapy approaches based on genetic manipulation and drug delivery. If confirmed in vivo, this could potentiate the efficacy of MSCs-TRAIL and strongly contribute to reduce the toxicity due to the systemic treatment of PTX.