Multiparametric correlation of laboratory biomarkers to multiorgan failure outcome in hospitalized COVID-19 patients: a retrospective observational study.

OBJECTIVE: COVID-19 is an extremely contagious illness caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), that will keep broadly circulating and evolving. Collected evidence revealed the clinical profile of COVID-19 patients as a potential predictor of their outcome. The aim of this study was to investigate the causal relationship between poor outcomes and laboratory parameters in hospitalized COVID-19 patients, in this sense observing how SARS-CoV-2 infection affects other organs. PATIENTS AND METHODS: We retrospectively evaluated a cohort of 133 patients, positive for SARS-CoV-2, aged between 30 to 94 years, between January 12th and April 25th, 2021. Discharge from the hospital, transferral to the ordinary ward or nursing home, intensive care unit (ICU) admission, and in-hospital mortality were recorded, along with demographic, laboratory and clinical parameters. The whole sample was summarized by median (interquartile range) for quantitative data, and absolute and relative percentage frequencies for qualitative variables. Univariable logistic regression models were performed to assess the association between all the parameters of interest and COVID-19 adverse outcomes, single (in-hospital mortality) and composite (in-hospital mortality and ICU admission). Hence, a multivariable model was fitted to identify potential independent predictors of the composite outcome. The accuracy of the model was assessed through appropriate fitting indices, such as the C-statistic and Hosmer-Lemeshow test. Moreover, to detect multicollinearity, the variance inflation factor (VIF) was used. RESULTS: Our study sample had a median age of 72 years old (59.0-83.0). The most common comorbidities were hypertension (63.7%), cardiovascular disease (41.9%), diabetes (33.6%), and cerebrovascular disease (21.5%); while as the most common symptoms, we observed dry cough (32.5%), dyspnoea (50.8%), and fatigue (29.8%). Totally, 18 patients died during hospitalization (13.5%), 10 required ICU admission (7.5%), 78 (58.6%) were discharged from the hospital, and 27 (20.3%) were transferred to either ordinary wards or nursing homes. We disclosed an association of older age with both composite [OR 1.06, 95% CI 1.02-1.09; p=0.003] and single outcome [OR 1.10, 95% CI 1.04-1.16; p=0.001]. A higher oxygen saturation (SpO2) was associated with a better outcome [OR 0.75, 95% CI 0.60-0.93; p=0.009 and OR 0.76, 95% CI 0.61-0.95, p=0.009]. Among laboratory parameters, higher levels of neutrophils increased the risk of a poor outcome [OR 1.05, 95% CI 1.00-1.10; p=0.043]; while higher levels of lymphocytes seem associated with a better outcome [OR 0.94, 95% CI 0.88-0.99; p=0.043]. Higher levels of creatinine were associated with a higher risk of both adverse outcomes [OR 6.20, 95% CI 2.16-17.81; p<0.001 and OR 19.90, 95% CI 5.07-78.06; p<0.001, respectively]. Higher levels of sodium (Na) were associated with a higher risk of adverse events [OR 1.15, 95% CI 1.03-1.28; p=0.014 and OR 1.14, 95% CI 1.01-1.27]. Similar findings were also observed for C-reactive protein (CRP) levels [OR 1.01, 95% CI 1.00-1.02; p=0.010 and OR 1.01, 95% CI 1.00-1.02; p=0.024]. Conversely, being positive to IgM and IgG decreases the risk of adverse outcomes [IgM: OR 0.33, 95% CI 0.14-0.77; p=0.011 and OR 0.23, 95% CI 0.08-0.66; p=0.006. IgG: OR 0.30 95% CI 0.13-0.72; p=0.007 and OR 0.22 95% CI 0.07-0.66; p=0.007]. Hence, a multivariable model was fitted to identify potential independent laboratory predictors of the composite outcome, with laboratory parameters that showed an association with composite outcome. The model can be considered accurate according to LH-Test and C-statistic [p>0.83, C-stat=0.90]. CONCLUSIONS: Our findings confirm that COVID-19 is a multiorgan disease. In fact, the analysis of laboratory parameters has revealed a strong relationship between poorer outcomes and multiple organ dysfunction, particularly established by higher levels of neutrophils, creatinine, sodium, and CRP. Alongside, cerebrovascular diseases, chronic kidney disease and older age supported this finding. Of note, higher levels of SpO2, and lymphocytes, as well as positivity to IgM and IgG were associated with a lower risk of a poor outcome.

Oral bisphosphonate-induced osteonecrosis complications in patients undergoing tooth extraction: a systematic review and literature updates.

OBJECTIVE: Bisphosphonates, the most common anti-resorptive medications, are internalized by osteoclasts, where they inhibit the macrophage colony-stimulating factor (M-CSF) pathway, preventing their differentiation, inhibiting anchorage to the cell membrane, and inducing apoptosis. In patients undergoing oral bisphosphonate therapy, oral surgery involves a high risk of developing drug-related osteonecrosis of the jaws (BRONJ/MRONJ), among the possible complications. MATERIALS AND METHODS: A systematic search was carried out on the PubMed, Scopus and Cochrane Library search engines, using the keywords "oral bisphosphonates AND tooth extraction", "third molar extraction AND oral bisphosphonates". In addition, we manually evaluated the articles included in references from other sources and an analysis of the Gray Literature was performed. A secondary outcome was to evaluate the assessment of pharmacological (antibiotics) use in the BRONJ/MRONJ management. The revision protocol followed the indications of the Cochrane Handbook, and was registered in the INPLASY database, while the drafting of the manuscript was based on PRISMA. RESULTS: The results of the systematic review, after the study identification and selection process, included a total of 7 studies: 4 retrospective studies, 2 prospective studies and 1 case report. The main complication was represented by osteonecrosis of the jaws, which appears to be related to the duration of treatment with bisphosphonates; in addition, data regarding the anatomical location of post-extraction sites, the sex and age of patients, comorbidities and various systemic risk factors were extrapolated. The most frequent post-extraction complication in patients treated with oral bisphosphonates is osteonecrosis of the jaws, with a significant prevalence in the posterior region of the mandible. In some cases, delayed healing of the surgical wound was also found; moreover, the duration of exposure to oral bisphosphonates influences the onset of complications. CONCLUSIONS: Ongoing studies continue to unravel the role of the oral environment response in alveolar bone homeostasis and how it might contribute to the induction of BRONJ/MRONJ. Approaching the problem from this perspective could provide new directions for the prevention of BRONJ/MRONJ and expand our understanding of the unique oral microenvironment.

The human microbiota key role in the bone metabolism activity.

Data collection has suggested a complex correlation between the gut microbiota (GM) and bone homeostasis involving host-microbiota crosstalk. Although the GM is known to affect bone metabolism, the mechanisms linked with these effects remain unclear. The aim of this review is to current insight advances regarding how gut-derived hormones regulate bone homeostasis in humans, emphasizing gut-bone axis and bone regeneration. The GM may be engaged in bone metabolism and fracture risk. Additional investigations of the fundamental microbiota-related pathways in bone metabolism may uncover treatment strategies and enable the prevention of osteoporosis. A better knowledge of gut hormones' action on bone homeostasis may lead to new strategies for preventing and treating skeletal frailty related to age.

Gender medicine: the impact of probiotics on male patients.

PURPOSE: Numerous studies in the literature confirm the importance of the use of probiotics in inflammatory states of the prostate in humans. Our pilot study aimed to test probiotics strains ability to improve urinary bacterial load in male subjects. METHODS: Twenty healthy men aged 55-65 years, with recurrent urinatory infections were enrolled and randomized into 2 groups. Urine culturing to detect Enterobacteriaceae (Escherichia Coli and Enterococcus faecalis) was performed at baseline and at 12 weeks after the starting of the study. RESULTS: We found that taking the probiotic reduces the bacterial load of E. coli and E. faecalis in urine cultures. The use of the probiotic in these inflammatory states does not affect the change in weight in the tested subjects. CONCLUSIONS: Our data confirm and improve the main results reported in the scientific literature on the importance of taking probiotics in prostatitis.

The role of PIAS3, p-STAT3 and ALR in colorectal cancer: new translational molecular features for an old disease.

OBJECTIVE: Human colorectal cancer (CRC) is characterized by a sequence of biological events that determine its induction and progression. Gut microbiota has an important role in this multistep model of carcinogenesis, as well as constitutive activation of Signal Transducer and Activator Factors 3 (p-STAT3) and Protein Inhibitor of Activated STAT3 (PIAS3), which negatively controls STAT3. It has been reported that a liver growth factor, the Augmenter of Liver Regeneration (ALR), an anti-apoptotic, anti-metastatic factor, exerts protective/cell survival and anti-metastatic activities and has been detected highly expressed in neoplastic cells. PATIENTS AND METHODS: To evaluate, by immunohistochemistry, p-STAT3, PIAS3 and ALR expression in neoplastic human tissues from CRC patients, grouping the data in accordance with the histological alterations (G1, G2 and G3) and metastasis presence. Western blot (WB) analysis of ALR was also determined in neoplastic and surrounding tissues. Finally, cell proliferation (Ki-67) and apoptosis (Bcl-2) were determined. RESULTS: Colon cancer tissue samples showed: (1) ALR and p-STAT3 strongly over-expression in 100% of G1 tissue samples, reducing in G2 and G3 tissue samples; (2) PIAS3 immunological determination was poorly expressed in G1 tissue samples and highly expressed in the 100% of colorectal tissues from group G2 and G3. Ki-67 progressively increases with the importance of the anatomic-pathological alterations and Bcl-2 resulted higher in G3 tissue samples compared to G1 neoplastic tissues. WB data evidenced, in neoplastic tissues, compared to the tumour-surrounding tissues, ALR over-expressed in G1 neoplastic tissues and down-expressed in G3 neoplastic tissues. CONCLUSIONS: Our data demonstrate a different dynamism of the investigated factors in relation to the severity of CRC histological findings. We hypothesize that the positive expression of ALR and p-STAT3 in the neoplastic tissue samples from CRC G1 group, associated to the absence of PIAS3, could be useful marker to identify an early stage of the disease. Based on these data and on our previous studies on gut microbiota in precancerous intestinal lesions, we are confident that, after microbial priming, a cascade of molecular events is started. So, the detectable molecules acting in these initial steps should be considered for the study of CRC progression and therapy.

Rapid and sensitive diagnostic procedure for multiple detection of pandemic Coronaviridae family members SARS-CoV-2, SARS-CoV, MERS-CoV and HCoV: a translational research and cooperation between the Phan Chau Trinh University in Vietnam and University of Bari "Aldo Moro" in Italy.

OBJECTIVE: A new pandemic coronavirus causing coronavirus disease-2019 (COVID-19), initially called 2019-nCoV and successively named Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The COVID-19 refers to the disease while the SARS-CoV-2 refers to the virus and is characterized by a rapid contagious capacity able to spread worldwide in a very short time. The rise in the number of infected patients and deaths is of great concern especially because symptoms are vague and similar to other forms of flu infection and corona syndrome infections characterized by fever, fatigue, dry cough, and dyspnea. According to the latest guidelines published by the World Health Organization (WHO), the diagnosis of COVID-19 must be confirmed by quantitative reverse transcription polymerase chain reaction (rRT-PCR) or gene sequencing of specimen obtained from throat, sputum and blood samples. However, the limitations due to logistics, as well as low sensitivity and specificity diagnostic tools currently available have been reported as the main cause of high incidence of either false-negative or positive results. PATIENTS AND METHODS: The purpose of the present translational research protocol is to discuss and present the original findings from our research team on new diagnostic technique to detect four Coronaviridae family members (SARS-CoV-2, SARS-CoV, HCoV and MERS-CoV), highlighting the methodology, the procedure and the possible advantages. Moreover, the authors review the current epidemiology, precautions and safety measures for health personnel to manage patients with known or suspected COVID-19 infection. RESULTS: Implementation of an effective and rapid plan of diagnosing, screening and checking is a key factor to reduce and prevent further transmission. This procedure based on rRT-PCR could be of great help to decisively validate the results obtained from more conventional diagnostic procedures such as chest computed tomography (CT) imaging and chest ultrasound. CONCLUSIONS: This translational diagnostic tool will assist emergency and primary care clinicians, as well as out-of-hospital providers, in effectively managing people with suspected or confirmed SARS-CoV-2.

The alteration of stress-related physiological parameters after probiotics administration in oral surgeons with different degrees of surgical experience.

PURPOSE: Stress is a multifactorial and complex pathway, gaining growing attention from the healthcare community. Surgeons are subjected to higher levels of stress, due to surgical procedures that are demanding and repetitive; unfortunately, high-stress levels may also cause side-effects, as surgical mistakes. This study aimed to evaluate the efficacy of specific probiotics strains formula on stress levels in oral and maxillofacial surgeons, to improve their quality of life. METHODS: We have investigated the hormonal (salivary Cortisol; sC), immune (salivary Immunoglobulin A; sIgA) and cardiovascular (Heart rate, HR, and systolic blood pressure, SBP) responses induced by stress conditions in 40 oral surgeons, randomly selected and allocated, according to their experience level, in three categories: senior, expert, and junior. RESULTS: The results described how the number of heartbeats/ minute and SBP are slightly raised in all surgeons at different timepoints. Such data allow us to assess that work-related stress can induce an increase in cardiovascular parameters, even if they are not significantly modified by the use of probiotics. On the other hand, our data indicate that 10 weeks of probiotic integration may induce the improvement of other stress-related physiological parameters in oral surgeons with different degrees of surgical experience, such as the salivary cortisol levels, even under stress conditions. Moreover, in the test group (probiotics administration), the immunoglobulin levels were higher than the control (placebo administration) group: this happens as a consequence of the regular use of probiotics, which may induce an increased number of IgA producing cells. DISCUSSION: Our data indicated that 10 weeks of probiotics-enriched diet modify some stress-related physiological parameters in oral surgeons with different degrees of surgical experience, but it does not impact on the overall cardiovascular risk.

In vitro effects of concentrated growth factors (CGF) on human SH-SY5Y neuronal cells.

OBJECTIVE: The aim of this study was to test the in vitro differentiation effects of concentrated growth factors (CGF), a platelet rich preparation, using SH-SY5Y cells, derived from human neuroblastoma. MATERIALS AND METHODS: SH-SY5Y cells were cultured in presence of CGF or retinoic acid (RA). After 72 h of treatment, different parameters were investigated: cell proliferation by an automated cell counter; cell viability by thiazolyl blue tetrazolium bromide (MTT) assay; cell differentiation markers, i.e., neuronal nuclear antigen (NeuN), synaptophysin (SYP) and ß3-tubulin, by immunocytochemistry and Western blotting techniques; release of nerve growth factor (NGF) and brain-derived growth factor (BDNF) by enzyme-linked immunosorbent assay (ELISA) and neurite outgrowth by a dedicated image software. RESULTS: In presence of CGF, the cell proliferation rate and viability decreased, as expected for differentiated SH-SY5Y cells. On the contrary, the cellular differentiation markers increased their expression together with the release of growth factors. Moreover, the neurite outgrowth was improved. CONCLUSIONS: The data suggest that CGF treatment positively affects the cell differentiation, regulating the expression of neuronal markers, the release of growth factors and the neurite length. Taken together these results seem to be promising in the development of new approaches for neural regeneration.

Effects of sea salt rinses on subjects undergone to oral surgery: a single blinded randomized controlled trial.

PURPOSE: It has been customary to explain the dentally beneficial effects of xylitol and certain other natural compound as lysozyme and seas salt in terms of microbiological effects only. Several studies have tested the use of natural ingredients, alcohol and fluoride free, in mouthwashes. The purpose of this study was to evaluate a combined mouthwash formulation containing natural antibiofilm agents in oral care wound healing after routinely oral surgery (extraction) procedures. METHODS: Patients were assigned following a blinded randomized controlled trial and divided into two groups, an experimental group (I = 15) and a control (placebo) group (II = 15). Any infectious complications, wound healing, plaque accumulation in the stitches, and presence of trismus and inflammation were evaluated at ten and thirty days after extraction procedure. Pain and swelling were evaluated using the well-known visual analogue scale (VAS) scale throughout study period following extraction. The mean difference in Pre and Post values were compared among the groups. The change in pre-post score was analyzed using the paired t test. RESULTS: An appreciable wound healing was seen in the experimental group when compared to the control sites, with no reported adverse effects. Four weeks postoperative patient's satisfaction level, to subjective and objective outcome measurements in documenting the result of a mouthwash treatment showed an interesting difference between groups. CONCLUSIONS: Since combined mouthwash formulations, containing natural/bioactive substances, could provide a cheap, safe and acceptable alternative in oral care, further studies will also be required to study these effects and their mechanism of action in detail.

Steroids and growth factors in oral squamous cell carcinoma: useful source of dental-derived stem cells to develop a steroidogenic model in new clinical strategies.

OBJECTIVE: Head and neck region is involved in a high percentage of malignant lesions, and oral squamous cell carcinoma (OSCC) is undoubtedly the most frequently found, accounting for over 90% of malignant tumors. Hormone receptor overexpression, like Estrogen Receptor (ER), Progesterone Receptor (PR) and Endothelial Growth Factor Receptor (EGFR), and signaling have been related to the pathogenesis of OSCC. For metastasis of OSCC, Cancer Stem Cells (CSCs) undergo epithelial to mesenchymal transition (EMT) under the influence of growth factors, cytokines, and regulation of cadherins from the tumor's microenvironment. In this context, the stem cells may become a potential therapeutic target for OSCC through modulation of cytokines and RAS pathway, which is involved in intracell signal transduction. The objective of this study was to suggest an experimental steroidogenic model for OSCC in translational research. PATIENTS AND METHODS: Dental-derived Stem Cells (D-dSCs) have been obtained from apical papilla tissue that surrounds the developing tooth of healthy donors and cultured in vitro. The cells have been exposed to different concentrations of Estradiol (E2 - 10 nM and 40 nM) in order to verify their response. The number of cells and cell viability has been evaluated up to 96 hours of treatment. RESULTS: The results showed that cell growth was increased under estradiol treatments compared with cells maintained without estradiol. Moreover, no significant difference in cell death levels was detected among treatments. CONCLUSIONS: This work underlines as D-dSCs could represent a useful steroidogenic model for the development of the target and gene therapies in OSCC.

Induction of mitochondrial dysfunction in patients under cardiopulmonary by-pass: preliminary results.

OBJECTIVE: Cardiac surgery is often performed by cardiopulmonary by-pass (CPB), generally associated with organ dysfunction. The aim of this work was to determine if and how this phenomenon is related to mitochondrial damage. To this purpose, the effect of the addition of serum from CPB patients to human fibroblasts cultures on mitochondrial respiratory chain and oxidative phosphorylation (OXPHOS) activities was investigated. PATIENTS AND METHODS: Serum samples of five patients were obtained before (pre-CPB) and after 6 h CPB weaning (CPB). Mitochondrial OXPHOS activities were examined by polarographic and spectrophotometric assays, and reactive oxygen species (ROS) production was measured by a spectrofluorimeter. RESULTS: Addition of CPB serum to fibroblasts determined a decrease of mitochondrial oxygen consumption due to an inhibition of mitochondrial respiratory chain and some OXPHOS enzymes activities. This inhibition seems to be mainly related to a reduced activity of complex I. CONCLUSIONS: Our data represent the first translational research evidence showing that CPB determines mitochondrial dysfunction which leads to impairment of OXPHOX activities and to an increase in ROS production, compromising tissue bioenergetic efficiency.

Effect of probiotics on the occurrence of nutrition absorption capacities in healthy children: a randomized double-blinded placebo-controlled pilot study.

OBJECTIVE: Recent advances in the translational research showed that dietary nutrients have critical importance to the microbioma balance in the gastrointestinal tract. Therefore, the alteration of the intestinal microbiota in order to achieve, restore, and maintain favorable balance in the ecosystem, and the activity of microorganisms present in the gastrointestinal tract is necessary for the improved health condition of the host. The objective of this translational study was to evaluate, in a pediatric population, the efficacy and safety of prophylactic probiotics for a better nutritional absorption capacity in the view to enhance their overall health and immunity. PATIENTS AND METHODS: A total of 40 pediatric patients between the ages of 14 and 18 years were enrolled in the study and divided under two categories (treated/active group and placebo group). Three-time points clinical evaluations were performed: a baseline assessment (Time 0), a second evaluation at 5 weeks after the start of probiotic use (Time 1), and a final evaluation at the timeline after 10 weeks (Time 2). In the initial phase of the study, the recruited subjects underwent a panel of initial T0 clinical tests. For each of the patients, a blood sample was taken in order to evaluate the following biochemical measurements: Vitamin D, Vitamin A, Calcium, Zinc, and Iron. Moreover, an initial nutritional evaluation was carried out through which the nutritionist estimated the body composition of the subject (weight and body mass index), the caloric needs and dietary behaviour of each recruited patient. RESULTS: Eligible participants were randomized into placebo (n = 20) or treated/active (n = 20) treatment conditions by random allocation using a computerized random number generator, ensuring all investigators remained blind to the treatment distribution. The data were compared within and between groups using statistical methods. The results confirmed that the probiotic supplementation was effective in increasing the overall blood biomarkers levels of vitamins, calcium, and mineral absorption from baseline to 10 weeks of treatment, compared with the placebo. CONCLUSIONS: Probiotics may be suggested as supplements to improve biomarkers serum concentration if administered for a period of at least ≥ 5 weeks. However, further studies are required for optimal recommendations in patient treatment.

Analysis of lip pigmentations by reflectance confocal microscopy: report of two cases.

Oral mucosa pigmentations belong to a heterogeneous variety of lesions, which are usually divided into two groups: exogenous or endogenous pigmentations. The pigmented lesions most frequently found in the oral mucosa are the amalgam tattoo, the melanotic macula and the nevus. All these lesions may affect every part of the oral mucosa, and they may represent a hard diagnostic challenge for the clinician; the clinical objective examination is not sufficient to make a correct diagnosis. Reflectance Confocal Microscopy provides a real-time microscopic evaluation of tissue layers, and is widely considered a useful auxiliary tool in monitoring skin and mucosa lesions. In this context, Reflectance Confocal Microscopy imaging is a valid aid in the management of oral mucosa pigmented lesions, to corroborate and support the diagnostic process.

Prevalence of caries and dental malocclusions in the apulian paediatric population: an epidemiological study.

AIM: The purpose of this epidemiological study was to assess the prevalence of malocclusion, its association with caries experience, and level of oral hygiene in the Apulian population. MATERIALS AND METHODS: Study design: A total of 530 paediatric patients (267 girls, 263 boys), aged 8-10 years (±SD 1.2) were randomly selected from primary schools in Apulia (Italy). The Decayed, Missing and Filled Teeth (DMFT/dmtf) index, the Dental Aesthetic Index (DAI), recorded according to the WHO criteria, were used by two calibrated examiners to diagnose dental caries and malocclusion, respectively. RESULTS: Except for the presence of dental calculus, Class II malocclusion, open bite and dental crowding, (p>0.05), all variables in the negative binomial regression showed a significant relationship with the incidence rate of caries in deciduous teeth. STATISTICS: Children were clinically examined in a community dental office. Statistical analysis was carried out using R version 3.5.1. CONCLUSION: The study outcomes underline the need for preventive care programmes to improve oral health conditions as well as to decrease oral pathology risk factors in the Apulia region.

Anti-calculus efficacy of Periogen® oral rinse in gingivitis patients.

Dental biofilm is a complex, organized microbial community that is the primary etiologic factor for plaque, the most frequently occurring oral contamination for gingivitis, dental caries and periodontal diseases. In dentistry, calculus is a form of hardened dental plaque, caused by precipitation of calcium (Ca) and phosphorous (P) ions deposited from saliva and gingival crevicular fluid. Although the dental biofilm cannot be eliminated, it can be controlled with comprehensive mechanical and chemotherapeutic oral hygiene practices. Chemotherapeutic agents have difficulty penetrating the polysaccharide matrix to reach and affect the microorganisms. The purpose of the present study was to evaluate the anti-calculus efficiency of a mouthwash containing an association of sodium tripolyphosphate, tetrapotassium pyrophosphate, sodium bicarbonate and citric acid in patients affected with gingivitis.

Stemness genes expression in naïve vs. osteodifferentiated human dental-derived stem cells.

OBJECTIVE: Mesenchymal stem cells (MSCs) have been deeply investigated in regenerative medicine because of their crucial role in tissue healing, such as tissue regeneration. Dental-derived stem cells (d-DSCs) are easily available from dental tissues, which can be isolated from all age patients with minimal discomfort. PATIENTS AND METHODS: Normal unerupted third molars tooth buds were collected from adolescents' patients underwent to extractions for orthodontic reasons. The expression of the genes Kruppel-like factor 4 (Klf-4), octamer-binding transcription factor 4 (Oct-4), homeobox transcription factor Nanog (NANOG) was investigated in d-DSCs obtained from dental bud (DBSCs), differentiated toward osteoblastic phenotype and not. RESULTS: Our results showed that DBSCs expressed Oct-4, Nanog, and Klf-4 in undifferentiated conditions and interestingly the expression of such genes increased when the cells were kept in osteogenic medium. CONCLUSIONS: These attractive stemness properties, together with the effortlessly isolation, during common oral and maxillofacial surgical procedures, from undifferentiated tissues such as dental bud, make this kind of d-DSCs a promising tool in regenerative medicine, having the potential for clinical applications, and reinforcing the present challenge to develop new preventive and healing strategies in tissue regeneration.

A pilot study of human mesenchymal stem cells from visceral and sub-cutaneous fat tissue and their differentiation to osteogenic phenotype.

OBJECTIVE: To evaluate the different behavior of two different human adult adipocytes derived stem cells (hASCs) during proliferation and osteogenic differentiation. PATIENTS AND METHODS: Human adult adipocytes stem cells (hAT-SCs) from visceral (hAV-SCs) and subcutaneous (hAS-SCs) sites were obtained after surgery procedures of seven patients. All samples were fully investigated and the different proliferation rates were evaluated. All MSCs clusters were cultured with an osteogenic and adipogenic differentiation medium. Homogeneous pools of Mesenchymal Stem Cells (MSCs) were confirmed by Flow-Cytometry Analysis (FACS) and Spectrophotometric Assay. The differentiated cells were eventually assessed for the expression of Alkaline Phosphatase (ALP), Alizarin Red (AR) and Oil Red-O (OR-O) detection, and analyzed by the Spectrophotometric Assay. After osteogenic differentiation, the cell clusters were incubated and analyzed with Real Time-Polymerase Chain Reaction (qRT-PCR) and fluorescence microscopy. RESULTS: The FACS analysis performed on hAT-SCs confirmed the homogenous presence of MSCs in all samples. The ALP, AR stain confirmed the osteogenic differentiation capacity of MSCs towards osteoblast-like-cells. The colorimetric cell metabolic activity (MTS) assay showed an increase in the proliferation rate with different values in both sets hAS-SCs vs. hAV-SCs. CONCLUSIONS: These in vitro findings of both hAS-SCs and hAV-SCs suggested an important role of these stem cells for future clinical use in bone regeneration. Indeed, the final outcomes suggested a better performance of cells coming from subcutaneous adipose tissue vs. those from visceral fat tissue.