Oral Chronic Mucositis in a Known Lichen Planus Pigmentosus Patient.

Oral manifestations of mucocutaneous inflammatory diseases such as lichenoid dermatoses often affect the patient's quality of life through the symptoms of pain and burning sensation. In this case report, a patient with known lichen planus pigmentosus (LPP), a variant of lichenoid dermatoses that rarely affect oral mucosa, presented with chronic pain in the oral cavity for 2 months. Her intraoral examination revealed multiple pseudomembranous ulcers with erythematous areas and white striae similar to erosive lichen planus. The histological assessment revealed chronic mucositis, while the direct immunofluorescent examination showed negative results, contradicting the diagnosis of both lichen planus and LPP. Thus, the diagnosis was given as chronic mucositis, and the topical steroid was given. After 6 weeks of steroid application, the lesion resolved. To the best of our knowledge, this is the first report of chronic mucositis in LPP patients. This report could raise the awareness of clinicians to carefully take history as the patient with LPP may suffer from chronic mucositis in the oral cavity as well.

3D-printed TCP-HA scaffolds delivering MicroRNA-302a-3p improve bone regeneration in a mouse calvarial model.

OBJECTIVE: To demonstrate hydroxyapatite nanoparticles modified with cationic functional molecules. 3-aminopropyltriethoxysilane (HA-NPs-APTES) carrying microRNA-302a-3p (miR) in the 3D-printed tricalcium phosphate/Hydroxyapatite (TCP/HA) scaffold can increase healing of the critical-sized bone defect. MATERIALS AND METHODS: 3D-printed TCP/HA were modified with HA-NPs-APTES by two methods (M1, M2). The dispersion of particles was visualized by fluorescent microscopy. Biocompatibility of the scaffolds was tested by alizarin assay. Delivery of miR to the cells and osteogenic gene expression were evaluated by qPCR. After selecting best method (M2), scaffolds, scaffolds+HA-NPs-APTES with or without miR were implanted in 4 mm mouse calvarium defect (n = 4 per group). After 2,4 and 6 weeks, bone regeneration were evaluated by microCT and histology sections. RESULTS: Both M1 and M2 scaffolds were biocompatible with cell adhesion on its surface. M2 scaffold showed significant increase of miR, suggesting successful delivery, resulted in downregulation of its target mRNA COUP-TFII, and upregulation of RUNX2 mRNA. Calvarium defect with M2 scaffold also showed significantly higher BV/TV and higher number of filled spaces at all time points. Histomorphometry demonstrated new bone formed at the center of the HA-NPs-APTES-miR scaffold earlier than controls. CONCLUSION: TCP/HA scaffold modified with HA-NPs-APTES facilitated delivery of miR and enhanced bone regeneration.

Delivery of microRNA-302a-3p by APTES modified hydroxyapatite nanoparticles to promote osteogenic differentiation in vitro.

OBJECTIVE: To demonstrate the miRNA delivery by hydroxyapatite nanoparticles modified with APTES (HA-NPs-APTES) and promote osteogenic gene expression. MATERIALS AND METHODS: Osteosarcoma cells (HOS, MG-63) and primary human mandibular osteoblasts (HmOBs) were co-cultured with HA-NPs-APTES conjugated with miRNA-302a-3p. Resazurin reduction assay was performed to evaluate HA-NPs-APTES biocompatibility. Intracellular uptake was demonstrated by confocal fluorescent and scanning electron microscopy. The miRNA-302a-3p and its mRNA targets expression levels including COUP-TFII and other osteogenic genes were assessed by qPCR on day1 or day5 post-delivery. Calcium deposition induced by the osteogenic gene upregulation was shown by alizarin red staining on day7 and 14 post-delivery. RESULTS: Proliferation of HOS cells treated with HA-NPs-APTES was similar to that of untreated cells. HA-NPs-APTES was visualized in cell cytoplasm within 24 hours. MiRNA-302a-3p level was upregulated in HOS, MG-63 and HmOBs as compared to untreated cells. As a result, COUP-TFII mRNA expression was reduced, followed by an increase of RUNX2 and other osteogenic genes mRNA expression. Calcium deposition induced by HA-NPs-APTES-miR-302a-3p in HmOBs was significantly higher than in untreated cells. CONCLUSION: HA-NPs-APTES may support the delivery of miRNA-302a-3p into bone cells, as assessed by osteogenic gene expression and differentiation improvement once this combination is used on osteoblast cultures.

miR-302a-3p regulates RANKL expression in human mandibular osteoblast-like cells.

Receptor activator of nuclear factor kappa-B ligand (RANKL) is important substance during osteoclastogenesis that resulted in alveolar bone loss of periodontitis. MicroRNAs (miRNAs) regulate gene expression in several biological processes including osteoclastogenesis. We investigated the function of microRNA-302a-3p (miR-302a-3p) to regulate receptor activator of nuclear factor kappa-B ligand (RANKL) expression in human mandibular osteoblast-like cells (HMOBs). HMOBs were incubated with prostaglandin E2 (PGE2 ) to mimic inflammation, or with PGE2 and interferon gamma (IFNγ) to mimic homeostasis. MicroRNA (miRNA) profiles related to RANKL expression were demonstrated by PCR array, and miR-302a-3p was identified. Using TargetScanHuman 7.0, a target of miR-302a-3p was predicted. To confirm its function, miR-302a-3p was overexpressed, or silenced, by transfection with miR-302a-3p mimic, or inhibitor, respectively. Level of miR-302a-3p and RANKL mRNA was assessed by qRT-PCR. Soluble RANKL (sRANKL), and membrane-bound RANKL (mRANKL) were measured by ELISA and by Western blot, respectively. When PGE2 stimulated RANKL in HMOBs, miR-302a-3p was lower than baseline level. However, upregulation of miR-302a-3p is observed when IFNγ suppressed RANKL expression in PGE2 -stimulated HMOBs. miR-302a-3p was predicted to target PRKACB mRNA encoding the catalytic subunit in cAMP/PKA pathway. Overexpression of miR-302a-3p could decrease RANKL expression during PGE2 stimulation. In contrast, silencing of miR-302a-3p by its inhibitor increased RANKL expression in PGE2 -IFNγ conditioned HMOBs. miR-302a-3p regulates RANKL expression in HMOBs within PGE2 -IFNγ regulatory network.