Exploring natural anthraquinones as potential MMP2 inhibitors: A computational study.

OBJECTIVE: Matrix metalloproteinase-2 (MMP2) plays a significant role in cleaving extracellular matrix components, leading to many cancer cells' progression and invasion behavior. Therefore, MMP2 inhibition may hold promise for cancer treatment. Anthraquinones have shown antineoplastic effects, some of which have been used in clinical practice as anticancer drugs. This study used a computational drug discovery approach to assess the possible inhibitory effects of selected anthraquinones on MMP2. The results were then compared with that of Captopril, which was considered a standard drug. METHODS: This study used the AutoDock 4.0 tool to evaluate the binding affinity of 21 anthraquinones to the MMP2 catalytic domain. The most favorable scores based on the Gibbs free binding energy scores were given to the highest-ranked ligands. The Discovery Studio Visualizer tool illustrated interactions between MMP2 residues and top-ranked anthraquinones. RESULTS: A total of 12 anthraquinones were identified with ΔGbinding scores less than - 10 kcal/mol. Pulmatin (Chrysophanol-8-glucoside) was the most potent MMP2 inhibitor, with a ΔGbinding score of - 12.91 kcal/mol. This anthraquinone was able to restrict MMP2 activity within a picomolar range. CONCLUSION: MMP2 inhibition by anthraquinones, notably Pulmatin, may be a useful therapeutic approach for cancer treatment.

The bFGF Can Improve Angiogenesis in Oral Mucosa and Accelerate Wound Healing.

Background: The role of the basic fibroblast growth factor (bFGF) has well known in the angiogenesis and ulcer healing. In this study, we aimed to evaluate the effects of bFGF on tissue repair in a rat oral mucosal wound. Methods: Musosal wound induced on the lip mucosa of rats and bFGF was injected along the edge of the mucosal defect immediately after surgery. The tissues were collected on days 3, 7 and 14 after the wound induction. The micro vessel density (MVD) and CD34 expression were done by histochemical studies. Results: The bFGF significantly accelerated granulation tissue formation and MVD was increased three days after ulcer induction but decreased 14 days after surgery. The MVD was significantly higher in the bFGF-treated group. The wound area was decreased in all groups time-dependently and a statistically significant difference (p value?) was observed between the bFGF-treated group and untreated group. The wound area was smaller in the bFGF-treated group compared to the untreated group. Conclusions: Our data demonstrated that bFGF can accelerated and facilitated wound healing.

MicroRNA-Based Markers of Oral Tongue Squamous Cell Carcinoma and Buccal Squamous Cell Carcinoma: A Systems Biology Approach.

Objective: Oral tongue squamous cell carcinoma (OTSCC) and buccal squamous cell carcinoma (BSCC) are the first and second leading causes of oral cancer, respectively. OTSCC and BSCC are associated with poor prognosis in patients with oral cancer. Thus, we aimed to indicate signaling pathways, Gene Ontology terms, and prognostic markers mediating the malignant transformation of the normal oral tissue to OTSCC and BSCC. Methods: The dataset GSE168227 was downloaded and reanalyzed from the GEO database. Orthogonal partial least square (OPLS) analysis identified common differentially expressed miRNAs (DEMs) in OTSCC and BSCC compared to their adjacent normal mucosa. Next, validated targets of DEMs were identified using the TarBase web server. With the use of the STRING database, a protein interaction map (PIM) was created. Using the Cytoscape program, hub genes and clusters within the PIM were shown. Next, gene-set enrichment analysis was carried out using the g:Profiler tool. Using the GEPIA2 web tool, analyses of gene expression and survival analysis were also performed. Results: Two DEMs, including has-miR-136 and has-miR-377, were common in OTSCC and BSCC (p value <0.01; |Log2 FC| > 1). A total of 976 targets were indicated for common DEMs. PIM included 96 hubs, and the upregulation of EIF2S1, CAV1, RAN, ANXA5, CYCS, CFL1, MYC, HSP90AA1, PKM, and HSPA5 was significantly associated with a poor prognosis in the head and neck squamous cell carcinoma (HNSCC), while NTRK2, HNRNPH1, DDX17, and WDR82 overexpression was significantly linked to favorable prognosis in the patients with HNSCC. "Clathrin-mediated endocytosis" was considerably dysregulated in OTSCC and BSCC. Conclusion: The present study suggests that has-miR-136 and has-miR-377 are underexpressed in OTSCC and BSCC than in normal oral mucosa. Moreover, EIF2S1, CAV1, RAN, ANXA5, CYCS, CFL1, MYC, HSP90AA1, PKM, HSPA5, NTRK2, HNRNPH1, DDX17, and WDR82 demonstrated prognostic markers in HNSCC. These findings may benefit the prognosis and management of individuals with OTSCC/BSCC. However, additional experimental verification is required.

3D-Printed Soft Membrane for Periodontal Guided Tissue Regeneration.

OBJECTIVES: The current study aimed to perform an in vivo examination using a critical-size periodontal canine model to investigate the capability of a 3D-printed soft membrane for guided tissue regeneration (GTR). This membrane is made of a specific composition of gelatin, elastin, and sodium hyaluronate that was fine-tuned and fully characterized in vitro in our previous study. The value of this composition is its potential to be employed as a suitable replacement for collagen, which is the main component of conventional GTR membranes, to overcome the cost issue with collagen. METHODS: Critical-size dehiscence defects were surgically created on the buccal surface of the roots of canine bilateral mandibular teeth. GTR treatment was performed with the 3D-printed membrane and two commercially available collagen membranes (Botiss Jason® and Smartbrane-Regedent membranes) and a group without any membrane placement was considered as the control group. The defects were submerged with tension-free closure of the gingival flaps. Histologic and histometric analyses were employed to assess the periodontal healing over an 8-week experimental period. RESULTS: Histometric evaluations confirmed higher levels of new bone formation in the 3D-printed membrane group. Moreover, in all defects treated with the membranes, the formation of periodontal tissues, bone, periodontal ligaments, and cementum was observed after 8 weeks, while in the control group, only connective tissue was found in the defect sites. There was no clinical sign of inflammation or recession of gingiva in any of the groups. SIGNIFICANCE: The 3D-printed gelatin/elastin/sodium hyaluronate membrane can be safe and effective for use in GTR for periodontal tissue regeneration therapies, with better or comparable results to the commercial collagen membranes.

Pathogenesis and prognosis of primary oral squamous cell carcinoma based on microRNAs target genes: a systems biology approach.

Oral squamous cell carcinoma (OSCC) is the most prevalent head and neck malignancy, with frequent cervical lymph-node metastasis, leading to a poor prognosis in OSCC patients. The present study aimed to identify potential markers, including microRNAs (miRNAs) and genes, significantly involved in the etiology of early-stage OSCC. Additionally, the main OSCC's dysregulated Gene Ontology annotations and significant signaling pathways were identified. The dataset GSE45238 underwent multivariate statistical analysis in order to distinguish primary OSCC tissues from healthy oral epithelium. Differentially expressed miRNAs (DEMs) with the criteria of p-value < 0.001 and |Log2 fold change| > 1.585 were identified in the two groups, and subsequently, validated targets of DEMs were identified. A protein interaction map was constructed, hub genes were identified, significant modules within the network were illustrated, and significant pathways and biological processes associated with the clusters were demonstrated. Using the GEPI2 database, the hub genes' predictive function was assessed. Compared to the healthy controls, main OSCC had a total of 23 DEMs. In patients with head and neck squamous cell carcinoma (HNSCC), upregulation of CALM1, CYCS, THBS1, MYC, GATA6, and SPRED3 was strongly associated with a poor prognosis. In HNSCC patients, overexpression of PIK3R3, GIGYF1, and BCL2L11 was substantially correlated with a good prognosis. Besides, "proteoglycans in cancer" was the most significant pathway enriched in the primary OSCC. The present study results revealed more possible mechanisms mediating primary OSCC and may be useful in the prognosis of the patients with early-stage OSCC.

Complementarity of surgical therapy, photobiomodulation, A-PRF and L-PRF for management of medication-related osteonecrosis of the jaw (MRONJ): an animal study.

BACKGROUND: This study aimed to evaluate the complementarity of surgical therapy, photobiomodulation (PBM), advanced platelet-rich fibrin (A-PRF), and Leukocyte and platelet-rich fibrin (L-PRF) for the management of medication-related osteonecrosis of the jaw (MRONJ). METHODS: Sixty rats underwent injection of zoledronate followed by left mandibular first and second molar extractions to induce MRONJ lesions. All rats were examined for the signs of MRONJ 8 weeks post-dental extraction. Forty-nine rats with positive signs of MRONJ were appointed to seven different groups as follows: control (Ctrl); surgery alone (Surg); surgery and PBM (Surg + PBM); surgery and A-PRF insertion (Surg + APRF); surgery and L-PRF insertion (Surg + LPRF); surgery, A-PRF insertion, and PBM (Surg + APRF + PBM); surgery, L-PRF insertion, and PBM (Surg + LPRF + PBM). Euthanasia was carried out 30 days after the last treatment session. The lesions' healing was evaluated clinically, histologically, and radiographically. Data were analyzed using STATA software version 14, and the statistical significance level was set at 5% for all cases. RESULTS: According to the present study, A-PRF and L-PRF treatment resulted in significant improvements in clinical, histological, and radiographical parameters compared to the Ctrl group (P < 0.05). The PBM also decreased wound dimensions and the number of empty lacunae compared to the Ctrl group (P < 0.05). Surg + APRF + PBM and Surg + LPRF + PBM were the only groups that presented a significantly higher mean number of osteocytes (P < 0.05). No significant differences were observed between A-PRF and L-PRF treatment groups (P > 0.05). CONCLUSIONS: Surgical resection followed by applying A-PRF or L-PRF reinforced by PBM showed optimal wound healing and bone regeneration in MRONJ lesions.

3D-printed bi-layered polymer/hydrogel construct for interfacial tissue regeneration in a canine model.

OBJECTIVES: There are complications in applying regenerative strategies at the interface of hard and soft tissues due to the limited designs of constructs that can accommodate different cell types in different sites. The problem originates from the challenges in the adhesion of dissimilar materials, such as polymers and hydrogels, that can be suitable for regenerating different tissues such as bone and soft tissues. This paper presents a design of a new hybrid construct in which a polymer (polycaprolactone (PCL)) membrane firmly adheres to a layer of hydrogen (gelatin). METHODS: PCL membranes with defined size and porosity were fabricated using 3D printing. The gelatin layer was attached to the PCL membranes using the aminolysis procedure. We have examined this construct for the application of Guided Bone Regeneration (GBR) as a typical surgical regenerative procedure of the oral cavity at the interface of bone and soft tissue. Complete in vitro and in vivo investigations on canine tibia bone defects have been performed. Histological analyses for fibrosis morphometric and bone morphometric evaluation, as well as bone-fibrosis histological grading and CBCT imaging, were conducted. RESULTS: Chemical and morphological studies of the membrane proved that gelatin was uniformly attached to the aminolyzed PCL membranes. The in vitro and in vivo studies indicated the membrane's biocompatibility, mechanical stability, and barrier function for the GBR application. Furthermore, in vitro study showed that the membranes could improve osteogenesis and the regeneration of bone defects. The results illustrated that the mean bone density in the membrane groups was about three times more than that of the control group. SIGNIFICANCE: The fabricated 3D-printed hybrid Gelatin/PCL bi-layered membrane can be a good candidate for interfacial tissue engineering and a promising membrane for GBR procedure.

In vivo efficacy of 3D-printed elastin-gelatin-hyaluronic acid scaffolds for regeneration of nasal septal cartilage defects.

Nasal septal cartilage perforations occur due to the different pathologies. Limited healing ability of cartilage results in remaining defects and further complications. This study sought to assess the efficacy of elastin-gelatin-hyaluronic acid (EGH) scaffolds for regeneration of nasal septal cartilage defects in rabbits. Defects (4 × 7 mm) were created in the nasal septal cartilage of 24 New Zealand rabbits. They were randomly divided into four groups: Group 1 was the control group with no further intervention, Group 2 received EGH scaffolds implanted in the defects, Group 3 received EGH scaffolds seeded with autologous auricular chondrocytes implanted in the defects, and Group 4 received EGH scaffolds seeded with homologous auricular chondrocytes implanted in the defects. After a 4-month healing period, computed tomography (CT) and magnetic resonance imaging (MRI) scans were obtained from the nasal septal cartilage, followed by histological evaluations of new tissue formation. Maximum regeneration occurred in Group 2, according to CT, and Group 3, according to both T1 and T2 images with 7.68 ± 1.36, 5.44 ± 2.41, and 8.72 ± 3.02 mm2 defect area respectively after healing. The difference in the defect size was statistically significant after healing between the experimental groups. Group 3 showed significantly greater regeneration according to CT scans and T1 and T2 images. The neocartilage formed over the underlying old cartilage with no distinct margin in histological evaluation. The EGH scaffolds have the capability of regeneration of nasal cartilage defects and are able to integrate with the existing cartilage; yet, they present the best results when pre-seeded with autologous chondrocytes.

Immunohistochemical Comparison of the Expression of CD34 and CD105 in Odontogenic Keratocyst and Dentigerous Cyst.

STATEMENT OF THE PROBLEM: Odontogenic keratocyst (OKC) is a developmental odontogenic cyst with specific histopathological features, high recurrence rate, and aggressive clinical behavior. Angiogenesis might be considered as an important factor for the growth, expansion, and distribution of this lesion. PURPOSE: The aim of the present study was to determine the mean vascular densities (MVD) of OKCs and dentigerous cysts to evaluate their relationship with the biologic behavior of these lesions. MATERIALS AND METHOD: In this cross-sectional analytical study, angiogenesis was assessed in OKC and dentigerous cyst by measuring the MVD. Immunohistochemistry was carried out using CD34 and CD105. The results were analyzed with independent samples t-test. The data were analyzed, setting p value at 0.05. RESULTS: The MVDs with the use of CD34 and CD105 markers were significantly higher in OKC compared to dentigerous cyst (p< 0.05). In addition, MVDs obtained by CD105 in dentigerous cysts and OKC were significantly less than those based on CD34 (p< 0.05). CONCLUSION: Based on the results of the present study, it can be suggested that angiogenesis might be one of the possible mechanisms involved in higher aggressive biologic behavior and greater recurrence rate of OKC compared to dentigerous cysts.

Jaw Intraosseous Lesions Biopsied Extracted From 1998 to 2010 in an Iranian Population.

BACKGROUND: Jaw bones might be potential locations for different lesions. Differences in prevalence and the type of lesions can help in designing and programming prevention procedures in health care centers. OBJECTIVES: The aim of the present study was to evaluate the prevalence of intraosseous lesions in the jaws of patients referred to diagnostic and therapeutic centers in Hamadan during 1990-2010. PATIENTS AND METHODS: This cross-sectional descriptive analytical study was carried out in Hamadan in 2011. Data sheets of the subjects were used to collect all the data of patients with intraosseous lesions, including their age, gender, location of the lesion, the radiographic view of lesions, and their type and histopathological diagnoses. Data were analyzed with SPSS, using means and frequencies. RESULTS: A total of 284 intraosseous lesions were reported in our study. The mean age of the subjects was 28.8 ± 15.2 years. The lesions were distributed in males and females almost similarly. The most prevalent lesions were cystic lesions (54.58%), manifestations of systemic conditions in jaw bones (18.3%), benign tumors (15.5%), malignant lesions (6.7%), and inflammatory lesions (4.92%), in a descending order. The most common cystic lesion was radicular cyst; the most common manifestation of systemic conditions in jaw bones was central giant cell granuloma; the most common benign tumor was ameloblastoma; the most common malignant lesion was osteosarcoma; and the most common inflammatory lesion was periapical granuloma. CONCLUSIONS: Our data provided information on the prevalence and types of intraosseous lesions among an Iranian population. This study provided baseline information to help in designing and programming procedures in health care centers in every community so that preventive therapeutic measures can be adopted.

Immunohistochemical Expression of Mast Cell in Oral Reactive Lesions.

STATEMENT OF THE PROBLEM: Soft tissue reactive lesions are the most common lesions of the oral cavity. Although many studies have shown the interaction of mast cells with fibroblasts and their participation in fibrosis, the role of mast cells in these lesions is not well understood. PURPOSE: The aim of this study was to evaluate the mast cells (MCs) count in oral soft-tissue reactive lesions including peripheral giant cell granuloma (PGCG), peripheral ossifying fibroma (POF), irritation fibroma (IF) and normal oral mucosa. MATERIALS AND METHOD: In this cross-sectional study, 50 samples including IF, PGCG, POF (14 cases for each group) and 8 cases of normal oral mucosa were stained with tryptase antibody through immunohistochemistry. The number of mast cells was counted in 5HPF containing maximum counts for each section stained with tryptase. Statistical analysis including Chi-square test and Tukey test with a significance level of 0.05 were considered. RESULTS: The number of MCs was found to have increased in reactive lesions compared with normal oral mucosa. MCs count in the POF group was higher than the others. CONCLUSION: These findings suggest a possible role of mast cells in the pathogenesis of reactive oral lesions and induction of fibrous tissues. Chemical mediators released from mast cells might influence other cells, especially fibroblasts, to induce fibrosis.

An Immunohistochemical Survey to Evaluate the Expression of CD105 and CD34 in Ameloblastoma and Odontogenic Keratocyst.

UNLABELLED: Statement of the Problem : Ameloblastoma is the most common odontogenic tumor which is slow-growing, locally invasive and exhibit specific biologic behavior and high recurrence rate. Likewise, odontogenic keratocyst is a developmental odontogenic cyst that has a high recurrence rate and aggressive behavior. There are limited studies considering the relationship between the angiogenesis factors and the biologic behavior of these lesions. PURPOSE: the aim of this study was to evaluate the mean density of vessels in odontogenic keratocysts and ameloblastoma and investigate its possible relationship with biological behavior of these lesions. MATERIALS AND METHOD: In this descriptive-analytic cross-sectional study, 40 cases, comprising 10 odontogenic keratocysts and 30 ameloblastomas (10 plexiform, 10 follicular, and 10 unicystic type) were selected and were stained immuno-histochemically with CD34 and CD105. The micro vessel density was assessed and compared in all groups. T- test for the independent samples' One- way Anova, Wilcoxon test and Tukey tests were adopted for statistical analysis. RESULTS: A statistically significant difference was observed in mean vascular density (MVD) between the odontogenic keratocyst and ameloblastoma groups concerning the CD105 and CD34 markers (p= 0.005, p= 0.000, respectively). The MVD was significantly higher in ameloblastomas than odontogenic keratocyst. MVD with CD34 was significantly higher than MVD with CD105 in ameloblastomas (p= 0.00). CONCLUSION: It can be suggested that angiogenesis might be one of the mechanisms that is more possible to contribute the aggressive biological behaviors in ameloblastoma rather than odontogenic keratocyst.

Multiple Calcifying Hyperplastic Dental Follicle (MCHDF): A Case Report.

Evaluation of enlarged follicles with unerupted teeth is always important because some changes may occur. One of the ex-tremely rare conditions seen in dental follicles is multiple calcifying hyperplastic dental follicle. We report a case of multi-ple calcifying hyperplastic dental follicle. Radiographs showed that mandibular third molars had a pericoronal radiolucent zone delineated by a well-defined and sclerotic border. Microscopic examination revealed a combination of fibrous connec-tive and small calcifications. Multiple calcifying hyperplastic dental follicle is a rare condition and its correct diagnosis is necessary to apply appropriate treatment.

Suitability/unsuitability of cell proliferation as an indicator of malignant potential in oral lichen planus: an immunohistochemical study.

BACKGROUND: Oral lichen planus (OLP) is categorized as premalignant lesion although its malignant potential is a matter of controversy. The objective of this study was to investigate Ki67 expression in OLP, oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Expression of Ki67 was evaluated through immunohistochemistry (IHC) in groups of A (17 cases of epithelial hyperplasia), B (16 cases of OLP), C1 (10 cases of mild epithelial dysplasia), C2 (10 cases of severe epithelial dysplasia), D1 (10 cases of well-differentiated OSCC), and D2 (10 cases of poorly-differentiated OSCC). RESULTS: There was a significant difference in Ki67 expression based on quantitative analysis among the six studied groups as well as group B compared bilaterally with groups C2, D1 and D2 (p< 0.0001). However, there was no significant difference between groups B and C1 or between groups D1 and D2 (p> 0.05). CONCLUSIONS: Based on the results of the present study it may not be possible to definitely consider malignant transformation potential for OLP. However, expression of Ki67 was significantly higher in OLP than that of epithelial hyperplasia with no significant difference from that of mild epithelial dysplasia. This should be considered by clinicians to carefully and regularly follow up OLP lesions to detect potential subtle changes at an early stage.

Immunohistochemical comparison of the expression of p53 and MDM2 proteins in ameloblastomas and keratocystic odontogenic tumors.

OBJECTIVE: Ameloblastoma and keratocystic odontogenic tumor (KOT) is characterized by a benign but locally invasive behavior with a high risk of recurrence. MDM2 (murine double minute 2), an amplifier of cell proliferation, and p53, a tumor suppressor gene, are overexpressed in some odontogenic lesions. The aim of this study was to compare the expression of MDM2 and p53 in ameloblastoma and KOT as 2 lesions with similar biologic behavior, by immunohistochemistry. METHODS: The expressions of MDM2 and p53 proteins were determined in 39 ameloblastomas (15 follicular types, 15 plexiform types, and 9 unicystic types) and 15 KOTs. RESULTS: P53 protein was expressed in 100% of KOTs and 77.8% of ameloblastomas, and MDM2 was detected in 74.8% of ameloblastomas and 80% of KOTs. There was no statistical difference between MDM2 and p53 expressions in different subtypes of ameloblastomas and also when KOTs were compared with them (P > 0.05). There was a significant difference between immunohistochemical reactivity of MDM2 among subtypes of ameloblastomas (P < 0.05). MDM2 and p53 expressions were positively correlated. CONCLUSIONS: Overexpression of p53 and MDM2 is associated with the pathogenesis and oncogenesis of ameloblastomas and KOT. Overexpression of these markers can contribute to similar biologic behavior of these lesions.

Odontogenic keratocysts in Nevoid basal cell carcinoma syndrome: a case report.

Nevoid basal cell carcinoma syndrome, a rare autosomal dominant disorder, comprises a number of abnormalities such as multiple nevoid basal cell carcinomas, skeletal abnormalities and multiple odontogenic keratocysts. Considering the rarity of this syndrome, we present a 12-year-old boy affected by this syndrome. He had multiple okcs, calcification of falx cerebri, bifid ribs, frontal bossing and hypertelorism. Characteristic cutaneous manifestation (nevoid basal cell carcinoma) was not present in this patient. The jaw cysts were treated with marsupialization then enucleation. The dental clinician may be the first to encounter and identify this syndrome, when the multiple cystlike radiolucencies are discovered on panoramic view.

Epithelioid hemangioendothelioma of the oral cavity: a case report.

Epithelioid hemangioendothelioma is a rare vascular neoplasm which exhibits the potential for malignancy and recurrence as well as the ability to metastasize. Although numerous sites of involvement are possible, these tumors most commonly arise in soft tissues, lung, liver, bone, and lymph nodes. In this report, we describe a case of oral epithelioid hemangioendothelioma in a child. This tumor appeared as exophytic ulcerated painless masses in the maxillary and mandibular gingiva. Histologically, the tumor was composed of a proliferation of tumor cells arranged in nests, cords, and short strands. Epithelioid cells exhibited abundant eosinophilic cytoplasm with nuclear and cellular pleomorphism and intra-cytoplasmic vacuoles.