The Role of Immunohistochemistry in the Differential Diagnosis between Intrahepatic Cholangiocarcinoma, Hepatocellular Carcinoma and Liver Metastasis, as Well as Its Prognostic Value.

Intrahepatic cholangiocarcinoma (iCCA) is the second most frequent primary hepatic malignant tumor, after hepatocellular carcinoma (HCC). Its incidence has risen worldwide, yet the only potentially curative treatment, surgical resection, is seldom applicable, and the median overall survival remains extremely low. So far, there are no personalized therapy regimens. This study investigated whether routine immunohistochemical stains have diagnostic and/or prognostic value in iCCA. Clinical, imaging, and pathology data were retrospectively gathered for patients diagnosed with iCCA, HCC, or liver metastases assessed using liver needle biopsies. Three study groups with an equal number of cases (n = 65) were formed. In the iCCA group, CK19, CA19-9, CK7, and CEA demonstrated the highest sensitivities (100%, 100%, 93.7%, and 82.6%, respectively). The most relevant stains used for diagnosing HCCs were Glypican 3, CD34 (sinusoidal pattern), and Hep Par 1, with corresponding sensitivities of 100%, 100%, and 98.2%. The immunohistochemical panels for diagnosing metastatic tumors were chosen after correlating the clinical data and morphologic H&E aspects. Moderate/intensely positive CK7 expression and absent/low amount of intratumoral immune cells were favorable prognostic factors and correlated with increased overall survival in both the univariate analysis and the multivariate regression adjusted for age, existence of cirrhosis, number of tumors, and tumor differentiation.

In Vitro and In Vivo Characterisation of a Mucoadhesive Buccal Film Loaded with Doxycycline Hyclate for Topical Application in Periodontitis.

Mucoadhesive films loaded with doxycycline hyclate (Doxy Hyc), consisting of mixtures of hydroxypropylmethyl cellulose (HPMC) E3, K4 and polyacrylic acid (Carbopol 940), were prepared by casting method, aiming to design a formulation intended for application in the oral cavity. The obtained film formulations exhibited a Doxy Hyc content between 7.52 ± 0.42 and 7.83 ± 0.41%, which had adequate mechanical properties for application in the oral cavity and pH values in the tolerance range. The x-ray diffraction studies highlighted the amorphisation of Doxy Hyc in the preparation process and the antibiotic particles present on the surface of the films, identified in the TEM images, which ensured a burst release effect in the first 15 min of the in vitro dissolution studies, after which Doxy Hyc was released by diffusion, the data presenting a good correlation with the Peppas model, n < 0.5. The formulation F1, consisting of HPMC K4 combined with C940 in a ratio of 5:3, the most performing in vitro, was tested in vivo in experimentally-induced periodontitis and demonstrated its effectiveness in improving the clinical parameters and reducing the salivary levels of matrix metalloproteinase-8 (MMP-8). The prepared Doxy Hyc loaded mucoadhesive buccal film could be used as an adjuvant for the local treatment of periodontitis, ensuring prolonged release of the antibiotic after topical application.

Immunomodulatory Effect of Novel Electrospun Nanofibers Loaded with Doxycycline as an Adjuvant Treatment in Periodontitis.

The immunomodulatory effect of a novel biomaterial obtained through electrospinning, based on polylactic acid (PLA) and nano-hydroxyapatite (nano-HAP), loaded with doxycycline (doxy) was evaluated in an animal model. The treatment capabilities as a local non-surgical treatment of periodontitis was investigated on the lower incisors of Wistar rats, after the induction of localized periodontitis using the ligature technique. Following the induction of the disease, the non-surgical treatment of scaling and root planing was applied, in conjunction with the application of the new material. The results of the treatment were evaluated clinically, using the tooth mobility and gingival index scores, as well as histologically. The salivary concentrations of matrix metalloproteinase 8 (MMP-8) and plasmatic concentrations of interleukin 1 (IL-1), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) were also monitored. Two weeks after the ligature application, the periodontal disease was successfully induced in rats. The application of the novel biomaterial obtained through electrospinning was proven to be more effective in improving the clinical parameters, while decreasing the salivary MMP-8 and plasmatic IL-1 and TNF-α concentrations, compared to the simple scaling and root planing. Thus, the novel electrospun biomaterial could be a strong candidate as an adjuvant to the non-surgical periodontal therapy.

In Vitro Antimicrobial Effect of Novel Electrospun Polylactic Acid/Hydroxyapatite Nanofibres Loaded with Doxycycline.

The present study aimed to assess the in vitro antimicrobial effects of a novel biomaterial containing polylactic acid (PLA), nano-hydroxyapatite (nano-HAP) and Doxycycline (Doxy) obtained by electrospinning and designed for the non-surgical periodontal treatment. The antimicrobial activity of two samples (test sample, PLA-HAP-Doxy7: 5% PLA, nano-HAP, 7% Doxy and control sample, PLA-HAP: 5% PLA, nano-HAP) against two periodontal pathogens­Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis­was assessed using the Kirby−Bauer Disk Diffusion Susceptibility Test and compared with the effect of four antibiotics used as adjuvants in periodontal therapy: Amoxicillin, Ampicillin, Doxy and Metronidazole. The test sample (embedded with Doxy) showed higher inhibitory effects than commonly used antibiotics used in the treatment of periodontitis, while the control sample showed no inhibitory effects. Moreover, significant differences were observed between the inhibition zones of the two samples (p < 0.05). The Doxy-loaded PLA nanofibres had an antimicrobial effect against the periodontal pathogens. Based on these results, the novel biomaterial could be a promising candidate as adjuvant for the non-surgical local treatment in periodontitis.

Multiple Faces of the Glioblastoma Microenvironment.

The tumor microenvironment is a highly dynamic accumulation of resident and infiltrating tumor cells, responsible for growth and invasion. The authors focused on the leading-edge concepts regarding the glioblastoma microenvironment. Due to the fact that the modern trend in the research and treatment of glioblastoma is represented by multiple approaches that target not only the primary tumor but also the neighboring tissue, the study of the microenvironment in the peritumoral tissue is an appealing direction for current and future therapies.

High-Frequency Ultrasound Assessment of Skin and Oral Mucosa in Metabolic Syndrome Patients-A Cross-Sectional Study.

BACKGROUND: Exogenous factors (such as sun exposure, smoking habits, and diet) and endogenous (inflammatory status, general diseases) have a direct influence on skin and soft tissue characteristics. The study's objective was to assess the impact of metabolic syndrome (MS) on characteristics of skin layers in sun-exposed and non-exposed maxillofacial tissues evaluated by high-frequency ultrasound (HFU), as a potential diagnosis and monitoring tool for the aging process. MATERIAL AND METHODS: The present study included 102 subjects (24 with MS; 78 without MS). Anthropometric parameters and disease history were recorded, and blood samples were harvested in order to assess biochemical parameters of MS. Sun-exposed skin (zygomatic region) and non-exposed oral mucosa of the lower lip were assessed using HFU (DUB® cutis, Taberna Pro Medicum) with a 22 MHz probe. RESULTS: Patients with cardiac disease had significantly lower values for epidermis density (p = 0.002). Gender was independently linked to the aged dermis depth (p < 0.001), aged dermis no. of px (pixels) (p < 0.001), dermis depth (p < 0.001), dermis no. of px (p < 0.001), and subcutaneous tissue density (p < 0.001). Patients with MS had thinner epidermis (p = 0.008) and thinner aged dermis (p = 0.037) when compared to non-MS subjects. CONCLUSION: Patients with MS had thinner epidermis and a lower epidermis number of pixels in sun-exposed skin. Women had lower epidermis density and thicker dermis in sun-exposed skin. Our study showed that HFU, as a non-invasive investigation approach, is useful to diagnose and monitor the aging process in skin and oral mucosa, correlated with skin phenotype pathological conditions.

Cannabidiol and Vitamin D3 Impact on Osteogenic Differentiation of Human Dental Mesenchymal Stem Cells.

Background and objective: The aim of the present study was to establish a new differentiation protocol using cannabidiol (CBD) and vitamin D3 (Vit. D3) for a better and faster osteogenic differentiation of dental tissue derived mesenchymal stem cells (MSCs). Materials and methods: MSCs were harvested from dental follicle (DFSCs), dental pulp (DPSCs), and apical papilla (APSCs) of an impacted third molar of a 17-year old patient. The stem cells were isolated and characterized using flow cytometry; reverse transcription polymerase chain reaction (RT-PCR); and osteogenic, chondrogenic, and adipogenic differentiation. The effects of CBD and Vit. D3 on osteogenic differentiation of dental-derived stem cell were evaluated in terms of viability/metabolic activity by alamar test, expression of collagen1A, osteopontin (OP), osteocalcin (OC), and osteonectin genes and by quantification of calcium deposits by alizarin red assay. Results: Stem cell characterization revealed more typical stemness characteristics for DFSCs and DPSCs and atypical morphology and markers expression for APSCs, a phenotype that was confirmed by differences in multipotential ability. The RT-PCR quantification of bone matrix proteins expression revealed a different behavior for each cell type, APSCs having the best response for CBD. DPSCs showed the best osteogenic potential when treated with Vit. D3. Cultivation of DFSC in standard stem cell conditions induced the highest expression of osteogenic genes, suggesting the spontaneous differentiation capacity of these cells. Regarding mineralization, alizarin red assay indicated that DFSCs and APSCs were the most responsive to low doses of CBD and Vit. D3. DPSCs had the lowest mineralization levels, with a slightly better response to Vit. D3. Conclusions: This study provides evidence that DFSCs, DPSCs, and APSCs respond differently to osteoinduction stimuli and that CBD and Vit. D3 can enhance osteogenic differentiation of these types of cells under certain conditions and doses.

Osseointegration of titanium scaffolds manufactured by selective laser melting in rabbit femur defect model.

The aim of this study was to assess the osseointegration of two series of titanium (Ti) scaffolds with 0.8 and 1 mm cell size obtained by Selective Laser Melting (SLM) technique. One of the series had the Ti surface unmodified, while the other had the Ti surface coated with silicon-substituted nano-hydroxyapatite (nano-HapSi). The scaffolds were implanted in the femur bone defects of 6 White Californian male rabbits: three animals were implanted with 0.8 mm cell size scaffolds and three animals with 1 mm cell size scaffolds, respectively. The bone fragments and scaffolds harvested at 2, 4 and 6 months were histologically analyzed using conventional light microscopy (LM) and scanning electron microscopy (SEM) for the qualitative evaluation of the bone tissue formed in contact with the scaffold. Both LM and SEM images indicated a better osseointegration for nano-HapSi coated Ti scaffolds. LM revealed that the compact bone formed in the proximity of nano-HapSi-coated scaffolds was better organized than spongy bone associated with unmodified scaffolds. Moreover, Ti scaffolds with meshes of 0.8 mm showed higher osseointegration compared with 1 mm. SEM images at 6 months revealed that the bone developed not only in contact with the scaffolds, but also proliferated inside the meshes. Nano-HapSi-coated Ti implants with 0.8 mm meshes were completely covered and filled with new bone. Ti scaffolds osseointegration depended on the mesh size and the surface properties. Due to the biocompatibility and favorable osseointegration in bone defects, nano-HapSi-coated Ti scaffolds could be useful for anatomical reconstructions.

Temporal bone trauma effects on auditory anatomical structures in mastoid obliteration.

PURPOSE: The risk of temporal bone fractures in head trauma is not negligible, as injuries also depend on the resistance and integrity of head structures. The capacity of mastoid cells to absorb part of the impact kinetic energy of the temporal bone is diminished after open cavity mastoidectomy, even if the surgical procedure is followed by mastoid obliteration. The aim of our study was to evaluate the severity of lesions in auditory anatomical structures after a lateral impact on cadaveric temporal bones in which open cavity mastoidectomy followed by mastoid obliteration was performed, compared to cadaveric temporal bones with preserved mastoids. METHODS: The study was carried out on 20 cadaveric temporal bones, which were randomly assigned to two groups. In the study group, open cavity mastoidectomy followed by mastoid obliteration with heterologous materials was performed. All temporal bones were impacted laterally under the same conditions. Temporal bone fractures were evaluated by CT scan. RESULTS: External auditory canal fractures were six times more seen in the study group. Tympanic bone fractures were present in 80% of the samples in the study group and 10% in the control group (p = .005). Middle ear fractures were found in 70% of the samples in the study group and 10% in the control group (p = .02). Otic capsule violating fractures of the temporal bone were present only in the study group. CONCLUSIONS: Mastoid obliteration with heterologous materials after open cavity mastoidectomy increases the risk of fracture, with the involvement of auditory anatomical structures.