OBJECTIVE: To explore whether microneedle pretreatment can significantly improve the efficacy and safety of 5-aminolevulinic acid (ALA)-photodynamic therapy (PDT) in the treatment of oral leukoplakia. METHODS: A non-randomized controlled clinical trial was conducted. Patients with clinical and pathological diagnosis of oral leukoplakia in the Department of Oral Mucosa, Peking University School and Hospital of Stomatology were divided into experimental group and control group. The control group was treated with conventional ALA-PDT, and the experimental group was pretreated with micro- needle buckling under superficial anesthesia with lidocaine before conventional ALA-PDT. The clinical manifestations of the two groups were recorded, the lesion area was measured, the clinical efficacy was evaluated, the number of treatment sessions and treatment unit duration were analyzed, and the pain after treatment was evaluated by visual analogue scale. The above data of the two groups were statistically analyzed. RESULTS: A total of 11 patients were included in the experimental group and 19 patients were included in the control group. The complete remission rate of the experimental group and the control group was 45.5% and 36.8%, the partial remission rate was 54.5% and 57.9%, and the no remission rate was 0% and 5%, respectively. There was no significant difference in the treatment effect between the two groups. Meanwhile, the treatment unit duration of the experimental group and the control group were (9.05±5.74) min/cm² and (21.38±15.44) min/cm², respectively, and the number of treatment sessions were (2.36±0.67) times and (3.58±1.57) times, respectively. These differences between the two groups were statistically significant (t=-3.125, P < 0.05; t=-2.932, P < 0.05). Similarly, multiple linear regression analysis with 7 factors including age, dysplastic pathology, lesion classification, etc., also confirmed that pretreatment could significantly shorten the treatment unit duration (P < 0.05). In addition, there was no significant difference in pain score (visual analogue scale) between the two groups after treatment, and the microneedle puncture pretreatment did not increase the adverse reactions of ALA-PDT treatment. CONCLUSION Microneedle pretreatment followed by conventional ALA-PDT shows a good clinical effect on oral leukoplakia, which can significantly shorten the clinical treatment time, reduce the number of visits, and save medical costs.
Humans ; Photochemotherapy/instrumentation* ; Leukoplakia, Oral/drug therapy* ; Aminolevulinic Acid/therapeutic use* ; Male ; Female ; Middle Aged ; Adult ; Needles ; Photosensitizing Agents/therapeutic use* ; Aged ; Combined Modality Therapy

As the field of oral pathology has evolved, the nomenclature and classification of oral mucosal diseases with a remarkable risk of malignant transformation have undergone several modifications. In 2005, the World Health Organization (WHO) introduced the concept of oral potentially malignant disorders (OPMDs) as an alternative to the terms for oral precancerous lesions and precancerous conditions. In the consensus report by the WHO Collaborating Center for Oral Cancer of 2021, OPMD is defined as "any oral mucosal abnormality that is associated with a statistically increased risk of developing oral cancer."This definition encompasses a range of conditions, in-cluding oral leukoplakia, oral submucous fibrosis, proliferative verrucous leukoplakia, oral lichen planus, and other lesions. In light of the complex etiology, unclear pathogenesis, and carcinogenesis of OPMDs, early and precise diagnosis and treatment can contribute to the secondary prevention of oral cancer. For this reason, this review, which aims to provide a basis for the precise clinical diagnosis of OPMDs, was performed. Its aim was achieved by reviewing the historical evolution and research progress of the nomenclature, classification, and histopathological diagnostic criteria of OPMDs.
Humans ; Mouth Neoplasms/diagnosis* ; Precancerous Conditions/diagnosis* ; Leukoplakia, Oral/diagnosis* ; Lichen Planus, Oral/pathology* ; Oral Submucous Fibrosis/pathology* ; Mouth Mucosa/pathology* ; World Health Organization

Humans ; Leukoplakia, Oral/therapy*

Objective: To evaluate the inter-observer agreement of the severity of oral epithelial dysplasia in oral leukoplakia, providing a theoretical basis for the development of a more objective grading system. Methods: This study included 60 digital pathological slides of oral leukoplakia from Oral Medicine Department of West China Hospital of Stomatology, Sichuan University, and 239 tissue microarray images of oral leukoplakia from State Key Laboratory of Oral Diseases, Sichuan University, to evaluate the agreement of grading. Besides, 1 000 patches were generated from the 60 digital pathological slides and were divided into 500 small-sized patches (224 pixel×224 pixel) and 500 large-sized patches (1 024 pixel×1 024 pixel), to evaluate the agreement of feature detection. Gradings and feature detections were completed by three pathological experts from the oral pathology departments of two Grade 3, Class A stomatological hospitals in China. Kappa coefficient was used to quantify the inter-observer agreement among pathologists. Results: Minimal agreement was found in the grading of oral epithelial dysplasia among pathologists (Kappa=0.30 in the pathological slide group, Kappa=0.30 in the tissue microarray group). None agreement was found in feature detection within the small-sized patches group (median Kappa=0.14 for architectural features, median Kappa=0.18 for cytological features), and minimal agreement was found in feature detection within the large-sized patches group (median Kappa=0.25 for architectural features, median Kappa=0.25 for cytological features). Conclusions: Generally, the agreement of grading and feature detection of oral epithelial dysplasia in oral leukoplakia is poor. Development of a more objective grading system of oral epithelial dysplasia based on artificial intelligence may be helpful to improve the agreement.
Artificial Intelligence ; China ; Humans ; Leukoplakia, Oral ; Observer Variation ; Precancerous Conditions

OBJECTIVES: To study the hypoxia response gene and microRNA (miRNA) expression profiles in the pathogenesis and progression of oral leukoplakia (OLK). METHODS: Affymetrix GeneChip human transcriptome array 2.0 was used to detect the transcriptome of normal mucosa, low-risk OLK, high-risk OLK, and early squamous cell carcinoma (SCC). Gene ontology function analysis was used to screen genes and key miRNAs whose biological role is hypoxia response. Quantitative reverse transcription polymerase ch-ain reaction (qRT-PCR) was used to verify the expression of hypoxia response genes and miRNAs. RESULTS: A total of 7 different genes of hypoxia response between normal mucosa and low-risk OLK, 10 genes between low-risk and high-risk OLK, and 21 genes between high-risk OLK and SCC were identified. The results of qRT-PCR showed that the expression of hypoxia-inducible factor 1α, chemokine cc-motif ligand 2, and matrix metalloproteinase 3 mRNA and miR-21 in normal mucosa, OLK, and SCC increased in a stepwise manner. The expression difference between OLK and SCC was statistically significant and consistent with the results of transcriptome array. CONCLUSIONS The hypoxia response gene and related miRNA play roles in the development and progression of OLK.
Carcinogenesis ; Humans ; Hypoxia ; Leukoplakia, Oral ; MicroRNAs ; Mouth Neoplasms ; Transcriptome

OBJECTIVES: To investigate the expression of Ki-67, Cyclin D1, P53, and P16 in patients with oral leukoplakia (OLK) and OLK cancerization who have aspicy diet in Chengdu. METHODS: Thirtypatients with OLK andspicy diet and 15 patients with OLK without spicy diet in Chengdu were divided into three groups: hyperplastic OLK (OLK-), OLK with mild to moderate dysplasia (OLK+), and severe dysplastic  OLK or oral squamous cell carcinoma (OSCC) transforming from OLK (OLK++/OSCC). The expression of Ki-67, Cyclin D1, P53, and P16 were detected by immunohistochemistry and statistically analyzed. RESULTS: The expression of Ki-67 and P53 in patients with or without spicy diet in the OLK+and OLK++/OSCC groups were stronger than that of the OLK- group ( CONCLUSIONS Spicy diet did not have an influence on the expression of Ki-67, Cyclin D1, P53, and P16 in patients with OLK and OSCC. The expression of Ki-67, Cyclin D1, and P53 increased with the development of OLK, whereas P16 showed opposite expression trend.
Carcinoma, Squamous Cell ; Cyclin D1 ; Diet ; Head and Neck Neoplasms ; Humans ; Ki-67 Antigen ; Leukoplakia, Oral ; Mouth Neoplasms ; Tumor Suppressor Protein p53

Oral immunosuppression caused by smoking creates a microenvironment to promote the occurrence and development of oral mucosa precancerous lesions. This study aimed to investigate the role of metabolism and macrophage polarization in cigarette-promoting oral leukoplakia. The effects of cigarette smoke extract (CSE) on macrophage polarization and metabolism were studied in vivo and in vitro. The polarity of macrophages was detected by flow cytometric analysis and qPCR. Liquid chromatography-mass spectrometry (LC-MS) was used to perform a metabolomic analysis of Raw cells stimulated with CSE. Immunofluorescence and flow cytometry were used to detect the polarity of macrophages in the condition of glutamine abundance and deficiency. Cell Counting Kit-8 (CCK-8), wound-healing assay, and Annexin V-FITC (fluorescein isothiocyanate)/PI (propidium iodide) double-staining flow cytometry were applied to detect the growth and transferability and apoptosis of Leuk-1 cells in the supernatant of Raw cells which were stimulated with CSE, glutamine abundance and deficiency. Hyperkeratosis and dysplasia of the epithelium were evident in smoking mice. M2 macrophages increased under CSE stimulation in vivo and in vitro. In total, 162 types of metabolites were detected in the CSE group. The metabolites of nicotine, glutamate, arachidic acid, and arginine changed significantly. The significant enrichment pathways were also selected, including nicotine addiction, glutamine and glutamate metabolism, and arginine biosynthesis. The results also showed that the supernatant of Raw cells stimulated by CSE could induce excessive proliferation of Leuk-1 and inhibit apoptosis. Glutamine abundance can facilitate this process. Cigarette smoke promotes oral leukoplakia via regulating glutamine metabolism and macrophage M2 polarization.
Animals ; Glutamine ; Leukoplakia, Oral ; Macrophages ; Mice ; Smoking ; Tumor Microenvironment

OBJECTIVE: To evaluate the diagnostic efficiency of oral mucosa disease, especially oral squamous cell carcinoma (OSCC) and oral potential malignant disorders (OPMDs) by DNA cytometry compared with histopathological diagnosis, so as to find a convenient, simple and low-invasive method for screening and follow-up. METHODS: 203 subjects with OSCC, OPMDs and other oral mucosa disease without dysplasia according to the inclusion criteria and exclusion criteria were recruited from Peking University School and Hospital of Stomatology. The mean age was (52.44±13.55) years, 98 males and 105 females. Brush biopsy was taken before scalpel biopsy at the same site. The brush biopsy sample was screened by moticytometer system for DNA cytometry after Feulgen stain, and histopathological examination were taken for the scalpel tissue. Data from DNA cytometry were used to calculate the parameters, such as sensitivity, specificity, positive and negative predictive values, odds ratios, Youden index (YI), positive and negative likelihood ratios, compared with the golden standard, histopathological diagnosis. DNA cytometry and histopathological diagnosis were performed back to back. RESULTS: Totally, 42 OSCC and 4 tumor in situ (TIS), 39 oral leukoplakia (OLK) with dysplasia (17 mild dysplasia, 13 medium dysplasia and 9 severe dysplasia), 29 OLK with hyperplasia, 1 verrucous OLK, 83 oral lichen planus (OLP) and 5 inflammation were included in our research. We grouped the OSCC, TIS and dysplasia as the positive group and others without dysplasia as the negative group, the sensitivity of DNA cytometry was 79.07%, the specificity was 81.20%, and the diagnostic accuracy was 80.30%,We grouped the OSCC and TIS as the tumor group, OLP, OLK with hyperplasia and inflammation as the non-tumor group, The sensitivity of DNA cytometry in diagnosing OSCC and TIS was 95.65%, and the specificity was 81.2%, The diagnostic accuracy was 85.28%. positive predictive values 66.67%, negative predictive values 97.94%, ratio odds 95, positive likelihood ratio 5.09, negative likelihood ratio 0.05, and Youden index 0.77. For the dysplasia, we grouped the different dysplasia together as the dyaplasia group, OLP, OLK with hyperplasia and inflammation as the non-tumor group, the sensitivity of DNA cytometry in diagnosing dyaplasia is 60%, the specificity is 81.2%. The diagnostic accuracy is 75.8%, positive predictive values 52.17%, negative predictive values 85.59%, ratio odds 6.48, positive likelihood ratio 3.19, negative likelihood ratio 0.49, and Youden index 0.41. CONCLUSION DNA cytometry is convenient and low-invasive, which can be used as an adjuvant method for screening the early OSCC and OPMDs, monitoring the prognosis of OSCC after surgery. Further large-scale and long period prospective studies are necessary to validate the better value of DNA cytometry.
Adult ; Aged ; Carcinoma, Squamous Cell ; DNA ; Female ; Humans ; Leukoplakia, Oral ; Lichen Planus, Oral ; Male ; Middle Aged ; Mouth Mucosa ; Mouth Neoplasms ; Prospective Studies

BACKGROUND AND OBJECTIVES: Cancer of the oral cavity is a disease of the head and neck that is difficult to treat. Periodic observation and biopsy are important for its early diagnosis once a premalignant lesion in the oral cavity is confirmed. The purpose of this study was to determine the importance of early excisional biopsy by investigating the histological features of oral leukoplakia and the rate of malignant change in the oral cavity. SUBJECTS AND METHOD: A total of 327 patients who underwent punch biopsy of oral cavity from January 2011 to December 2017 were reviewed retrospectively for the presence of initial gross lesions and for their biopsy results. The histological findings of 6 initial gross lesion groups were compared. Additional excisional biopsies were performed in the seven oral cavity subsites. RESULTS: There were 33 cases of oral leukoplakia. The punch biopsies of 3 of these cases (9.1%) showed malignancy. Additional excisional biopsies were performed in 6 cases, 4 of which were malignant (66.7%). Additional excisional biopsies of the tongue were performed in 14 cases (9.0%), 5 of which (35.7%) were malignant. The rate of atypia in leukoplakia (9.1%) was higher than in other atypia groups. Additional excisional biopsies were performed in 3 cases (100%) of atypia of leukoplakia, all of which were assessed to be malignant. CONCLUSION: For tongue leukoplakia, performing an early excisional biopsy rather than an incisional biopsy is recommendable. Moreover, additional excisional biopsies are needed when the initial biopsy is suggestive of hyperkeratosis, parakeratosis, or atypia.
Biopsy ; Early Diagnosis ; Head ; Humans ; Leukoplakia ; Leukoplakia, Oral ; Methods ; Mouth ; Neck ; Parakeratosis ; Retrospective Studies ; Tongue

With recent developments in photosensitizers and light delivery systems, topical 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) has become the fourth alternative therapeutic approach in the management of oral leucoplakia (OLK) due to its minimally invasive nature, efficacy, and low risk of systemic side effects and disfigurement. This report presents step-by-step guidelines for applying topical ALA-PDT in the management of OLK based on both the clinical experience of the authors and a systematic review of the current literature. Studies using protocols with standardized parameters and randomized clinical trials at multiple centres with adequate sample sizes and both interim and long-term follow-ups are needed before universally applicable guidelines can be produced in this field.
Aminolevulinic Acid ; administration & dosage ; therapeutic use ; Humans ; Leukoplakia, Oral ; therapy ; Photochemotherapy ; methods ; Photosensitizing Agents ; administration & dosage ; therapeutic use ; Practice Guidelines as Topic