Generative Adversarial Networks in Digital Histopathology: Current Applications, Limitations, Ethical Considerations, and Future Directions.

Generative adversarial networks (GANs) have gained significant attention in the field of image synthesis, particularly in computer vision. GANs consist of a generative model and a discriminative model trained in an adversarial setting to generate realistic and novel data. In the context of image synthesis, the generator produces synthetic images, whereas the discriminator determines their authenticity by comparing them with real examples. Through iterative training, the generator allows the creation of images that are indistinguishable from real ones, leading to high-quality image generation. Considering their success in computer vision, GANs hold great potential for medical diagnostic applications. In the medical field, GANs can generate images of rare diseases, aid in learning, and be used as visualization tools. GANs can leverage unlabeled medical images, which are large in size, numerous in quantity, and challenging to annotate manually. GANs have demonstrated remarkable capabilities in image synthesis and have the potential to significantly impact digital histopathology. This review article focuses on the emerging use of GANs in digital histopathology, examining their applications and potential challenges. Histopathology plays a crucial role in disease diagnosis, and GANs can contribute by generating realistic microscopic images. However, ethical considerations arise because of the reliance on synthetic or pseudogenerated images. Therefore, the manuscript also explores the current limitations and highlights the ethical considerations associated with the use of this technology. In conclusion, digital histopathology has seen an emerging use of GANs for image enhancement, such as color (stain) normalization, virtual staining, and ink/marker removal. GANs offer significant potential in transforming digital pathology when applied to specific and narrow tasks (preprocessing enhancements). Evaluating data quality, addressing biases, protecting privacy, ensuring accountability and transparency, and developing regulation are imperative to ensure the ethical application of GANs.

Oral epithelial dysplasia with lymphocytic immune response: clinicopathological characterisation of 44 cases.

AIMS: Oral epithelial dysplasia (OED) often exhibits a lymphocytic/lichenoid immune response (LIR), imparting histological resemblance to lichenoid mucositis and rendering diagnosis challenging. The clinical appearances of OED and lichenoid inflammatory processes are generally divergent, presenting as well-demarcated hyperkeratotic plaques and diffuse white and/or red mucosal change with variably prominent Wickham striae, respectively. To date, clinicopathological characterisation of OED with LIR, including clinical/gross appearance, has not been depicted. METHODS AND RESULTS: Cases of solitary OED with LIR for which a clinical photograph was available were identified in the authors' institutional files. Clinical and histological features were documented. In 44 identified cases, dysplasia was mild (19 of 44, 43.2%), moderate (19 of 44, 43.2%) and severe (six of 44, 13.6%). Clinically/grossly, all 44 cases (100.0%), presented as well-demarcated hyperkeratotic plaques lacking diffuse white-and-red mucosal change or Wickham striae. Histologically, OED with LIR exhibited numerous 'lichenoid' features beyond the lymphocytic band in the superficial lamina propria, including: leucocyte transmigration (38 of 44, 86.4%), spongiosis (37 of 44, 84.1%), Civatte/colloid bodies (36 of 44, 81.8%), basal cell degeneration (29 of 45, 65.9%), sawtooth rete ridges (11 of 44, 25.0%) and subepithelial clefting (7 of 44, 15.9%). CONCLUSIONS: Virtually any lichenoid histological feature may be seen in OED with LIR, representing a significant diagnostic pitfall. The typical clinical appearance of OED with LIR is of a well-demarcated hyperkeratotic plaque, characteristic of keratinising dysplasia and devoid of lichenoid features. This suggests that pathologist access to clinical photographs during diagnostic interpretation of biopsied white lesions, which represents opportunity to perform gross examination of the disease process, may reduce interobserver variability and improve diagnostic accuracy in this challenging differential diagnosis.

Deep learning for temporomandibular joint arthropathies: A systematic review and meta-analysis.

BACKGROUND AND OBJECTIVE: The accurate diagnosis of temporomandibular disorders continues to be a challenge, despite the existence of internationally agreed-upon diagnostic criteria. The purpose of this study is to review applications of deep learning models in the diagnosis of temporomandibular joint arthropathies. MATERIALS AND METHODS: An electronic search was conducted on PubMed, Scopus, Embase, Google Scholar, IEEE, arXiv, and medRxiv up to June 2023. Studies that reported the efficacy (outcome) of prediction, object detection or classification of TMJ arthropathies by deep learning models (intervention) of human joint-based or arthrogenous TMDs (population) in comparison to reference standard (comparison) were included. To evaluate the risk of bias, included studies were critically analysed using the quality assessment of diagnostic accuracy studies (QUADAS-2). Diagnostic odds ratios (DOR) were calculated. Forrest plot and funnel plot were created using STATA 17 and MetaDiSc. RESULTS: Full text review was performed on 46 out of the 1056 identified studies and 21 studies met the eligibility criteria and were included in the systematic review. Four studies were graded as having a low risk of bias for all domains of QUADAS-2. The accuracy of all included studies ranged from 74% to 100%. Sensitivity ranged from 54% to 100%, specificity: 85%-100%, Dice coefficient: 85%-98%, and AUC: 77%-99%. The datasets were then pooled based on the sensitivity, specificity, and dataset size of seven studies that qualified for meta-analysis. The pooled sensitivity was 95% (85%-99%), specificity: 92% (86%-96%), and AUC: 97% (96%-98%). DORs were 232 (74-729). According to Deek's funnel plot and statistical evaluation (p =.49), publication bias was not present. CONCLUSION: Deep learning models can detect TMJ arthropathies high sensitivity and specificity. Clinicians, and especially those not specialized in orofacial pain, may benefit from this methodology for assessing TMD as it facilitates a rigorous and evidence-based framework, objective measurements, and advanced analysis techniques, ultimately enhancing diagnostic accuracy.

Oral manifestations of mpox (monkeypox).

The zoonotic infectious disease mpox (previously known as monkeypox) is caused by the monkeypox virus (MPXV) from the Poxviridae family. Presently, mpox is receiving worldwide attention because of its emergence in countries that have never previously documented the illness, resulting in a public health emergency. MPXV is transmitted via human-to-human contact, and sexual contact is especially implicated in spread of the disease. Affected individuals experience fever, headache, malaise, and early lymphadenopathy, followed by a secondary mucotaneous rash. Oral ulcers and perioral papules may be the first evidence of the disease. Although there are numerous articles in medical publications documenting the cutaneous presentations of mpox, there is limited information in the dental literature regarding oral lesions. The objective of this article is to review the oral manifestations of mpox and strategies for management of the disease.

Dental implant failure possibly attributed to embedded pencil graphite: first clinical report.

Implant failures have been associated with a diversity of etiologic processes, predominately arising from bone loss (peri-implantitis) due to inadequate maintenance of oral hygiene or excess luting agents. The aim of this article is to report a novel case of the apparent failure of a dental implant to undergo osseointegration in the presence of submerged pencil graphite. Practitioners are advised to carefully evaluate the clinical and radiographic site of a proposed implant for occult foreign substances. Embedded pencil graphite in the jawbone may promote a foreign body reaction and should be considered in the list of possible contributing factors to dental implant complications.

Spindle cell lipoma of the buccal mucosa: Uncommon finding in an older adult.

OBJECTIVE: This article reports a case of a longstanding, slowly enlarging intraoral spindle cell lipoma (SCL) that had become increasingly painful during mastication. BACKGROUND: The SCL represents an uncommon variant of the conventional lipoma. There is limited information regarding this lesion in the gerodontologic literature. MATERIALS AND METHODS: A 68-year-old patient underwent an excisional biopsy of a 9-mm slightly yellow papule along the buccal mucosa. RESULTS: The surgical specimen was composed of mature adipocytes with abundant spindle cell populations and was diagnosed as a SCL. CONCLUSIONS: Timely removal of the SCL may reduce the incidence of clinical and surgical complications, particularly in older adults. The management of a SCL is complete excision, and recurrence is rare. Lesions must be carefully distinguished microscopically from its malignant counterpart, the spindle cell liposarcoma.

Idiopathic bone cavity mimicking a botryoid odontogenic cyst: A rare radiographic presentation in an older adult.

OBJECTIVES: This report describes an unusual case of a multilocular idiopathic bone cavity (IBC) that presented as a botryoid odontogenic cyst situated between the mandibular lateral incisor and canine in an older adult. BACKGROUND: The IBC represents an intraosseous concavity that appears radiographically as a unilocular or multilocular radiolucent lesion found in various skeletal sites, including the jaw. Atypical cases of gnathic IBC have not been appreciated in the gerodontologic literature. MATERIALS AND METHODS: The teeth adjacent to the bony lesion had normal pulpal responses to cold. A full-thickness flap was elevated and provided a direct entry into a bony concavity, which was devoid of an epithelial lining and fluid. RESULTS: The lack of a cystic lining within the empty osseous lesion following surgical entry, concomitant with the vital pulpal status of the proximal teeth, led to a diagnosis of an IBC. The bony walls underwent curettage and copious irrigation prior to primary closure. A 10-month follow-up revealed partial evidence of osseous repair. The patient will continue to be monitored. CONCLUSION: Timely surgical intervention of central lesions of the jaws may improve clinical outcomes. Variants of the IBC should be included in the differential diagnosis of multilocular lesions, particularly in the geriatric population.

Prosthodontic implications of saliva and salivary gland dysfunction.

PURPOSE: To provide a detailed overview of the fundamentals of saliva constituents and production. The review outlines the clinical manifestations as a consequence of salivary gland dysfunction and management strategies for patients with salivary gland dysfunction. Prosthodontic implications of saliva and salivary gland dysfunction are presented. MATERIALS AND METHODS: English-language literature relating to saliva constituents, physiologic saliva production, clinical manifestations secondary to salivary gland dysfunction, salivary biomarkers, and management strategies were retrieved via electronic search. Relevant articles were summarized for this manuscript with a view toward providing pragmatic information. RESULTS: Saliva is produced by three pairs of major and minor salivary glands. The major salivary glands, namely, the parotid, submandibular, and sublingual glands, contribute approximately 90% of saliva production. Saliva contains serous and mucinous secretions produced by different types of cells within salivary glands. Parasympathetic and sympathetic fibers innervate the major salivary glands, and upon stimulation, the parasympathetic innervation increases serous secretions, while the sympathetic innervation increases protein secretion. Stimulated saliva is mainly derived from the parotid glands which are composed of serous acini, while unstimulated saliva is mainly derived from the submandibular glands which are composed of mixed seromucous acini. As major salivary glands contribute the most to salivary flow, local or systemic factors influencing those glands can disrupt saliva production resulting in clinically significant oral manifestations. CONCLUSION: This review provides a fundamental overview of saliva production. In addition, the review highlights the various clinical manifestations secondary to salivary gland dysfunction, explores salivary biomarkers for screening of systemic diseases, discusses management strategies for patients with salivary gland dysfunction, and outlines the prosthodontic implications of saliva and salivary gland dysfunction.

Denture stomatitis-An interdisciplinary clinical review.

PURPOSE: An interdisciplinary clinical review on denture stomatitis formulated by experts from prosthodontics, oral medicine, and oral microbiology is presented. The review outlines the etiopathogenesis, clinical features, and management strategies of denture stomatitis from an interdisciplinary perspective. MATERIALS AND METHODS: English-language articles relating to the definition, incidence, gender distribution, geographical distribution, etiology, pathogenesis, symptoms, signs, treatment, and prognosis of denture stomatitis were retrieved via electronic literature search. Relevant articles were summarized for this manuscript with a view toward providing pragmatic information. RESULTS: Denture stomatitis represents a very common, multifactorial infectious, inflammatory, and hyperplastic condition which is primarily caused by poor oral hygiene, poor denture hygiene, and full-time; mainly night-time denture wear, bringing about the emergence of advanced Candida-containing polymicrobial biofilms in close proximity to the host's mucosal tissues. Denture stomatitis shares clinically relevant associations with dental caries, periodontitis, median rhomboid glossitis, angular cheilitis, aspirational pneumonia, and associated mortality. CONCLUSIONS: Effective, long-term treatment of denture stomatitis relies upon sustained patient-driven behavioral change which should focus on daily prosthesis-level cleaning and disinfection, removal of dentures at night, every night, engagement with professional denture maintenance, and when required, denture replacement. Antifungal medications offer limited benefits outside of short-term use due to the emergence of antifungal resistance. For frail, older, medically compromised, and nursing home populations, treatment of denture stomatitis lowers the risk of aspirational pneumonia and associated mortality.

Rise of the natural red pigment 'prodigiosin' as an immunomodulator in cancer.

Cancer is a heterogeneous disease with multifaceted drug resistance mechanisms (e.g., tumour microenvironment [TME], tumour heterogeneity, and immune evasion). Natural products are interesting repository of bioactive molecules, especially those with anticancer activities. Prodigiosin, a red pigment produced by Serratia marcescens, possesses inherent anticancer characteristics, showing interesting antitumour activities in different cancers (e.g., breast, gastric) with low or without harmful effects on normal cells. The present review discusses the potential role of prodigiosin in modulating and reprogramming the metabolism of the various immune cells in the TME, such as T and B lymphocytes, tumour-associated macrophages (TAMs), natural killer (NK) cells, and tumour-associated dendritic cells (TADCs), and myeloid-derived suppressor cells (MDSCs) which in turn might introduce as an immunomodulator in cancer therapy.

Association of TNF-α G-308 a Promoter Polymorphism with the Course and Outcome of COVID-19 Patients.

BACKGROUND: Tumor necrosis factor-ɑ (TNF-ɑ) is one of the most important cytokines that manage the host defense mechanism, which may play a role in the pathogenesis of COVID-19 patients. The work aims to study the association of TNF-ɑ G-308 A gene polymorphism with the course and outcome of COVID-19 patients in Mansoura University Hospital. METHODS: 900 patients with COVID-19 infection and 184 controls were tested for TNF-ɑ G-308 A promoter polymorphism. Different genotypes of TNF-ɑ G-308 A were compared as regards the severity and prognosis of the disease. RESULTS: No statistically significant difference was found between patients and controls as regards the demographic data. The AA genotype of TNF-ɑ showed a higher incidence of the disease in comparison to the other genotypes. As regards the demographic and laboratory characters, no statistically significant difference was found between the different genotypes except for age, lymphopenia, CRP, and serum ferritin levels. In 336(80.0%) cases of the AA genotype, the disease was severe in comparison to 90(41.7%) cases in the GA genotype and no cases in the GG genotype with P = .001. CONCLUSION: People who carry the A allele of TNF-ɑ polymorphism are more prone to COVID-19 infection. The AA genotype of TNF-ɑ is associated with a more aggressive pattern of the disease. In those patients, the use of anti - TNF therapy may be promising.

The oral mycobiome: Oral epithelial dysplasia and oral squamous cell carcinoma.

Fungi, a diverse group of eukaryotic organisms, play distinct roles in health and disease. Recent advances in the field of mycobiology have enabled the characterization of the "human mycobiome." The human mycobiome has extensively been studied in various disease models. However, to date, the role of the oral mycobiome in oral carcinogenesis has yet to be elucidated. Candida albicans, the most common oral colonizer, has been speculated to display tumorigenic effects; however, the literature lacks consistent documentation from mechanistic studies on whether oral mycobiota act as drivers, facilitators, or passive colonizers of oral premalignancy and cancer. This review article provides an overview of existing hypothesis-driven mechanistic models that outline the complex interplay between the oral mycobiome and oral epithelial dysplasia as well as their potential clinical implications.

Oral and cutaneous pemphigus vulgaris: an atypical clinical presentation.

Pemphigus vulgaris (PV) is a rare painful and blistering autoimmune mucocutaneous disorder that appears in middle-aged adults. Oral lesions typically precede cutaneous involvement and tend to be more recalcitrant to therapy. The objective of this article is to present a case of oral and cutaneous PV in an atypical patient, a 23-year-old woman. The case was distinguished by the patient's age, which was 2 to 3 decades younger than the reported mean age of onset, and its coincidence with celiac disease, an immunopathologic process rarely seen in association with PV. Intravenous administration of the monoclonal antibody rituximab provided rapid clinical improvement in the cutaneous lesions and gradual improvement in the oral lesions after 2 infusions. Dental practitioners should remain vigilant for oral manifestations of dermatologic disease and refer affected patients to appropriate healthcare providers for long-term management.

Premature Ejaculatory Dysfunction in Rheumatoid Arthritis (PED-RA Study).

Objective: To determine the frequency of premature ejaculatory dysfunction in rheumatoid arthritis patients. Methods: After approval from IRB, cross sectional study was conducted from November 1st, 2020 to August 1st, 2021 at Department of Rheumatology, Indus Medical College, Pakistan. RA patients were included, written and informed consents were taken. Demographic data was noted and detailed history and examination was carried out. Each participant BMI and Blood pressure was measured. Afterward 5-ml of blood was drawn by a trained phlebotomist for CBC, ESR, fasting blood sugar levels, HBs Ag and anti HCV Antibody test.DAS-28 Calculator was used for RA clinical activity. Premature ejaculation diagnostic tool was used for the assessment of premature ejaculation. Results: Total 168 patients with mean age 32.27 (SD=±9.49) and mean disease duration of 6.35 (SD=±3.95) years were included,. Prevalence of premature ejaculation dysfunction was (44.6%), with mean PED of score 9.17 (SD=±5.23). Hypertension, HCV, Hakeem medications, use of DMARDS had positive association while High BMI and higher DAS-28 has negative association with PED (p<0.05). Conclusion: There is high prevalence of PED in RA, it needs proper evaluation, treatment and urgent research is needed to know more about it.

Pterostilbene as a Phytochemical Compound Induces Signaling Pathways Involved in the Apoptosis and Death of Mutant P53-Breast Cancer Cell Lines.

Pterostilbene is a natural nonflavonoid polyphenolic compound. It shows a remarkable range of biological activities, including antiproliferative, antiinflammatory, and antioxidant activity. However, the mechanism of action of PT in breast cancer cells containing mutant p53 protein has not been fully elucidated. Therefore, the present study was aimed at investigating the influence of PT on signaling pathways involved in the apoptosis of mutant p53-breast cancer cell lines. Immunocytochemistry and Western Immunoblotting techniques were used in this study. The present data showed that the viabilities and the proliferations of MDA-MB-231 and T-47D decreased significantly (P < 0.001) after treatment with different concentrations of PT. In addition, the morphological characteristics of both cell lines were changed after treatment with PT. Decreased protein expression of mutant p53 (R280 K, L194F) in MDA-MB-231 and T-47D breast cancer cell lines has also been achieved. In addition, overexpression of pro-apoptotic (Bax) protein, caspase-3 activity and histone release were increased after treatment of both cell lines with different PT concentrations. Furthermore, the protein expressions of cyclin D1, mTOR, and oncogenic ß-catenin were significantly downregulated after treatment of both cell lines with PT. In conclusion, downregulations of protein expression of mutant p53, cyclin D1, mTOR, and ß-catenin were increased after both cell lines had been treated with pterostilbene. PT could point to a promising use against the development and the progression of breast cancer as a natural therapeutic agent.

Electronic nicotine delivery systems: Oral health implications and oral cancer risk.

There is a paucity of evidence surrounding the potential detrimental effects of electronic nicotine delivery systems (ENDS) for both systemic and oral health. The effects of conventional cigarettes on the development of oral cancer are well known; however, the role of ENDS in oral carcinogenesis is yet to be elucidated. Furthermore, the exponential rise of the use of ENDS by the general public means that dental healthcare providers are more likely to encounter questions on their safety in the oral cavity, and on their effectiveness as a smoking cessation aid. Herein, we review the most up to date literature on the systemic and oral health complications of ENDS. Moreover, evidence-based recommendations on the use of ENDS as a smoking cessation tool within the dental setting are discussed.

Long-term effects of one-anastomosis gastric bypass on liver histopathology in NAFLD cases: a prospective study.

PURPOSE: Weight reduction can effectively improve nonalcoholic fatty liver disease (NAFLD), which is a constant companion of severe obesity. This study aimed to determine the effect of one-anastomosis gastric bypass (OAGB) on pathological liver changes in severely obese cases with NAFLD. METHODS: The present prospective research comprised 67 subjects with morbid obesity scheduled for OAGB during the period from February 2015 to August 2018. Clinical, biological, and histologic data were evaluated pre and 15 months postoperatively. RESULTS: Fifteen months after surgery, a considerable reduction was noted in the grades of fat deposition, liver cell ballooning, and lobular inflammatory changes, in addition to the total NAS score. Fifteen months after surgery, nonalcoholic steatohepatitis (NASH) disappeared in 42% of the patients. A significant regression of fibrosis stage occurred after surgery in 79.1% of patients (p < 0.001). After surgery, patients had substantial reductions in aspartate aminotransferase, alanine aminotransferase, γ-glutamyltransferase, HbA1c, total cholesterol, and Low-density lipoprotein (p < 0.001, for all comparisons). Diabetes mellitus, hypertension, and dyslipidemia resolved in 54%, 59%, and 69% of the patients, respectively. CONCLUSION: OAGB resolved NASH from nearly 42% of patients and reduced the histological features of NAFLD 15 months after surgery. Bariatric procedures might be adopted as a therapeutic modality in severely obese cases with NAFLD after the failure of lifestyle modifications.

Impact of laparoscopic sleeve gastrectomy on fibrosis stage in patients with child-A NASH-related cirrhosis.

PURPOSE: Bariatric surgery can improve non-alcoholic fatty liver disease (NAFLD). Yet data on the effect on fibrosis are insufficient and controversial. This work endeavored to evaluate the safety of laparoscopic sleeve gastrectomy (LSG) in cases that have compensated non-alcoholic steatohepatitis (NASH)-related cirrhosis and its impact on fibrosis stage. METHODS: The current prospective work involved 132 cases with Child-A NASH-related cirrhosis suffering from morbid obesity scheduled for LSG. They were subjected to preoperative assessment, wedge biopsy, and ultrasound-guided true-cut liver biopsy after 30 months. Patients were included if proved to have F4 fibrosis initially. The liver condition was assessed based on the NALFD Activity Score (NAS). The primary outcome measure was the impact of LSG on fibrosis stage and its relation to weight loss. RESULTS: The analysis included only 71 patients who completed the 30-month follow-up period. By the end of the follow-up interval, there was a substantial weight loss with a reasonable resolution of comorbidities. The median NAS decreased significantly from 6 (1-8) to 3 (0-6) after surgery. Fibrosis score regressed to F2 in 19 patients (26.8%) and F3 in 29 (40.8%). Patients with improved scores had a significantly higher amount of weight loss (p < 0.001). Improvement was more frequent in males (p = 0.007). By 30 months after treatment, 53.8% of cases with borderline NASH and 36.8% of those with probable NASH showed complete resolution, and 44.7% of patients with NASH showed improvement. Steatosis improved in 74.6% of patients (p < 0.001). CONCLUSION: In patients with NASH-related liver cirrhosis of Child class A, LSG may be a secure approach for the management of morbid obesity. It has a long-term benefit for both obesity and liver condition with significant improvement of steatosis, steatohepatitis, and fibrosis.

Effects of obesity and diabesity on heart rhythm in the Zucker rat.

Obesity and type 2 diabetes mellitus are risk factors for hypertension, coronary heart disease, cardiac arrhythmias including atrial fibrillation, heart failure and sudden cardiac death. The effects of obesity and diabesity on heart rhythm were investigated in the Zucker diabetic fatty (ZDF) and Zucker fatty (ZF) compared to the Zucker lean (ZL) control rat. In vivo biotelemetry techniques were used to assess the electrocardiogram and other cardiac and metabolic parameters. ZDF rats were characterized by age-dependent elevations in fasting and non-fasting blood glucose, glucose intolerance and weight gain and ZF rats were characterized by smaller elevations in fasting and non-fasting blood glucose and greater weight gain compared to ZL rats. Heart rate (HR) was progressively reduced in ZDF, ZF and ZL rats. At 195 days (6.5 months) of age there were significant differences in HR between ZDF (265 ± 8 bpm, n = 10), ZF (336 ± 9 bpm, n = 10) and ZL (336 ± 10 bpm, n = 10) rats and significant differences in HRV between ZDF (22 ± 1 bpm, n = 10), ZF (27 ± 1 bpm, n = 10) and ZL (31 ± 1 bpm, n = 10) rats. Power spectral analysis revealed no significant (P > 0.05) differences in HRV at low frequencies, reduced HRV at high frequencies and increased sympathovagal balance in ZDF compared to ZF and ZL rats. HR was reduced by ageing and additionally reduced by diabesity in the absence of changes in physical activity and body temperature. Reductions in HRV associated with altered sympathovagal drive might partly underlie disturbed HR in the ZDF rat. Possible explanations for reduced HR and future mechanistic studies are discussed.

Manuka honey enhanced sensitivity of HepG2, hepatocellular carcinoma cells, for Doxorubicin and induced apoptosis through inhibition of Wnt/ß-catenin and ERK1/2.

BACKGROUND: Recently, there is increasing awareness focused on the identification of naturally occurring anticancer agents derived from natural products. Manuka honey (MH) has been recognized for its biological properties as antimicrobial, antioxidant, and anticancer properties. However, its antiproliferative mechanism in hepatocellular carcinoma is not investigated. The current study focused mainly on investigating the molecular mechanism and synergistic effect of anticancer properties of MH on Doxorubicin (DOX)-mediated apoptotic cell death, using two different p53 statuses (HepG2 and Hep3B) and one non-tumorigenic immortalized liver cell line. RESULTS: MH treatment showed a proliferative inhibitory effect on tested cells in a dose-dependent manner with IC50 concentration of (6.92 ± 0.005%) and (18.62 ± 0.07%) for HepG2 and Hep3B cells, respectively, and induced dramatic morphological changes of Hep-G2 cells, which considered as characteristics feature of apoptosis induction after 48 h of treatment. Our results showed that MH or combined treatments induced higher cytotoxicity in p53-wild type, HepG2, than in p53-null, Hep3B, cells. Cytotoxicity was not observed in normal liver cells. Furthermore, the synergistic effect of MH and Dox on apoptosis was evidenced by increased annexin-V-positive cells and Sub-G1 cells in both tested cell lines with a significant increase in the percentage of Hep-G2 cells at late apoptosis as confirmed by the flow cytometric analysis. Consistently, the proteolytic activities of caspase-3 and the degradation of poly (ADP-ribose) polymerase were also higher in the combined treatment which in turn accompanied by significant inhibitory effects of pERK1/2, mTOR, S6K, oncogenic ß-catenin, and cyclin D1 after 48 h. In contrast, the MH or combined treatment-induced apoptosis was accompanied by significantly upregulated expression of proapoptotic Bax protein and downregulated expression of anti-apoptotic Bcl-2 protein after 48 h. CONCLUSIONS: Our data showed a synergistic inhibitory effect of MH on DOX-mediated apoptotic cell death in HCC cells. To our knowledge, the present study provides the first report on the anticancer activity of MH and its combined treatment with DOX on HCC cell lines, introducing MH as a promising natural and nontoxic anticancer compound.