Earl Fu

Chitosan enhances platelet adhesion and aggregation.

In this study, chitosan (MW=50,000) was tested for its enhancing platelet activity in rabbit platelet suspensions and the possible mechanisms involved were further investigated. Our results showed that after initial (5 min) and long-term (30 min) contact of platelets with chitosan, the platelet adhesion to chitosan-coated microtiter plates was dose-dependently increased compared to that of solvent control. Similarly, chitosan also dose-dependently increased the platelet aggregation and the intracellular free Ca(2+) rise of Fura-2-AM loaded platelets. Additionally, in the presence of FITC-labeled anti-CD41/CD61, chitosan significantly enhanced the expression of platelet glycoprotein IIb/IIIa complex assayed by a flow cytometer. It is concluded that chitosan is an effective inducer for platelet adhesion and aggregation and the mechanisms of action of chitosan may be associated, at least partly, with the increasing [Ca(2+)](i) mobilization and enhancing expression of GPIIb/IIIa complex on platelet membrane surfaces.

Immunosuppressive Effect of Quercetin on Dendritic Cell Activation and Function

Dendritic cells (DCs) play a crucial role in linking innate and adaptive immunity. Thus, DCs have been regarded as a major target of immunosuppressants for the control of harmful immune responses. In this study, we examined the effect of quercetin, a natural flavonoid found in many vegetables and fruits, on the activation and function of mouse DCs. Quercetin effectively inhibited LPS-induced DC activation by reducing the production of proinflammatory cytokines/chemokines and the expression levels of MHC class II and costimulatory molecules. In addition, quercetin uniquely blocked endocytosis by DCs and the LPS-induced DC migration was diminished by quercetin treatment. Furthermore, quercetin abrogated the ability of LPS-stimulated DCs to induce Ag-specific T cell activation, both in vitro and in vivo. Remarkably, coadministration of quercetin with 2,4-dinitro-1-fluorobenzene prevented 2,4-dinitro-1-fluorobenzene–induced contact hypersensitivity, indicating the potential of quercetin for treating delayed-type hypersensitive diseases. Blockage of LPS-induced ERK, JNK, Akt, and NF-κB activation contributed to the inhibitory effect of quercetin on DCs. These results strongly suggest that quercetin may be a potent immunosuppressive agent and could be used in the prevention and therapy of chronic inflammation, autoimmunity, and transplantation via the abolishment of DC activation and function.

Chitosan inhibits prostaglandin E2 formation and cyclooxygenase-2 induction in lipopolysaccharide-treated RAW 264.7 macrophages.

Chitosan, a deacetylated chitin, has been reported to accelerate the wound healing and exert anti-inflammatory effect but the possible mechanisms involved are still unclear. Enhanced production of prostaglandin E2 (PGE2) and pro-inflammatory cytokines has been shown to contribute to immunosuppression and cytotoxicity during wound healing. In this study, we examined the effect of chitosan on cyclooxygenase pathway and cytokines production in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Our results first demonstrated that chitosans (MW=50,000, 150,000 or 300,000) significantly inhibit the overproduction of PGE2 as well as cyclooxygenase-2 (COX-2) protein expression and activity accompanied by attenuation of pro-inflammatory cytokines production such as tumor necrosis factor-alpha and interleukin-1beta formation but increase of the anti-inflammatory cytokine, IL-10, formation in LPS-treated RAW 264.7 macrophages. These results suggest that the beneficial effect of chitosan on wound healing may be associated, at least partly, with the inhibition of PGE2 production by suppressing COX-2 induction and activity as well as attenuation of pro-inflammatory/anti-inflammatory cytokines ratio in activated macrophages.

Three‐dimensional analysis of the root morphology of mandibular first molars with distolingual roots

AIM To determine the prevalence of distolingual roots in mandibular first molar teeth in Taiwanese Han Chinese, and its impact on root morphology. METHODOLOGY The presence of distolingual roots in 375 subjects (521 molars) were assessed from 624 patients attending the dental clinics of medical centres around Taiwan island from August 2004 to April 2007 using computed tomography. The following observations were made: (i) numbers of roots and canals, (ii) mesial and distal root types and (iii) levels of furca in the molars presence or absence of distolingual root. RESULTS The mean age of the subject was 45; 43% were women. Among all the examined molars, 56%, 27% and 18% were two-, three- and four-rooted, respectively. Two per cent, 72% and 26% of molars had two, three and four canals, respectively. All of the four-rooted molars had four canals, but all of the molars with four canals varied in the number of roots. All molars with distolingual roots had two mesial canals. Bilateral consistency in terms of distolingual root, root canal number, root number and root type was observed in subjects with bilateral molars. In molars with distolingual roots, a higher prevalence of two mesial roots and a shorter mesial root trunk were observed than in teeth without distolingual roots. CONCLUSIONS A distolingual root was found in 22% of molars and in 24% of the subjects examined. Most subjects with a distolingual root had them bilaterally. The presence of a distolingual root was associated with variation in the root morphology, including the furcation level, the root type and the number of roots and canals.

siRNA-targeting transforming growth factor-β type I receptor reduces wound scarring and extracellular matrix deposition of scar tissue.

Hypertrophic scarring is related to persistent activation of transforming growth factor-β (TGF-β)/Smad signaling. In the TGF-β/Smad signaling cascade, the TGF-β type I receptor (TGFBRI) phosphorylates Smad proteins to induce fibroblast proliferation and extracellular matrix deposition. In this study, we inhibited TGFBRI gene expression via TGFBRI small interfering RNA (siRNA) to reduce fibroblast proliferation and extracellular matrix deposition. Our results demonstrate that downregulating TGFBRI expression in cultured human hypertrophic scar fibroblasts significantly suppressed cell proliferation and reduced type I collagen, type III collagen, fibronectin, and connective tissue growth factor (CTGF) mRNA, and type I collagen and fibronectin protein expression. In addition, we applied TGFBRI siRNA to wound granulation tissue in a rabbit model of hypertrophic scarring. Downregulating TGFBRI expression reduced wound scarring, the extracellular matrix deposition of scar tissue, and decreased CTGF and α-smooth muscle actin mRNA expression in vivo. These results suggest that TGFBRI siRNA could be applied clinically to prevent hypertrophic scarring.

Expression of p16INK4A in Pap smears containing atypical glandular cells from the uterine cervix.

OBJECTIVE To verify one of the diagnostic dilemmas concerning atypical glandular cells (AGC) by immunocytochemical detection of p16INK4A (p16) applied to routine Pap smears with correlation of follow-up biopsies for improvement of cytologic diagnoses. STUDY DESIGN The study included 36 Pap smears in AGC diagnostic categories, all of which were correlated histologically. The cytologic diagnoses of AGC were further classified according to the 2001 Bethesda System. All Pap smears were decolorized and immunostained with the primary anti-p16 antibody, clone E6H4. Immunoreactivity for p16 was correlated with histologic sections in a semiblind fashion. RESULTS Of the 36 smears containing AGC, 22 (61%) were reclassified as general AGC and 14 (39%) as AGC--favor neoplasia. Follow-up biopsies revealed that 15 (42%) cervixes had no obvious abnormalities and that 21 (58%) cases had different cervical lesions. More than half the cases (19/36, 53%) of follow-up biopsies concerning AGC-containing smears represented significant lesions. There was a much higher proportion of significant lesions (13/14, 93%) in AGC--favor neoplasia than those (6/22, 27%) in general AGC cases. Fifteen of 36 (36%) AGC-containing cases were actually squamous abnormalities on follow-up biopsies. p16 Immunocytochemical stain was reactive in 22 (61%) of 36 smears, either weakly/sporadically (2 cases, 6%) or strongly positively (20 cases, 55%). Conversely, 14 (39%) of the smears were negative for p16 and displayed predominantly reactive changes. However, there was 1 case of high grade squamous intraepithelial lesion showing negative immunostaining for p16. From the view-point of clinical significance, this analysis was highly sensitive (sensitivity, 95%) and specific (specificity, 88%) and had favorable positive (90%) and negative (94%) predictive values. CONCLUSION On the basis of both morphologic and immunostaining patterns, there was a clear association between strong p16 immunostaining of atypical cells in smears and the presence of significant lesions in the cervix except in 1 patient. Similarly, there was a clear association between lack of p16 expression and absence of cervical lesions. p16 Immunocytochemical stain can be applied successfully to conventional Pap smears and may serve as a useful biomarker in diagnoses of AGC-containing smears. This may offer a more objective parameter to help clarify this ambiguous area of gynecologic cytopathology.

Ameliorative effect of quercetin on the destruction caused by experimental periodontitis in rats.

BACKGROUND AND OBJECTIVE The purpose of this study was to evaluate the effect of quercetin, a flavonol that exhibits anti-inflammatory properties, on experimental periodontal destruction in rats. MATERIAL AND METHODS Osteoclast formation on maxillary palatal alveolus was induced with daily lipopolysaccharide (LPS) injections (0, 1 or 5 mg/mL) for 3 d. Five days later, the osteoclasts on bony surfaces were counted after histochemical staining for tartrate-resistant acid phosphatase. The effect of intragastric quercetin on the osteoclast formation was evaluated in the following three groups: quercetin (75 mg/kg/d by oral feeding); LPS (5 mg/mL); and quercetin plus LPS. Moreover, the effect of quercetin on the ligature-induced periodontitis around maxillary second and mandibular first molars was further evaluated by microcomputerized tomography (on days 0, 4, 8 and 12) and by histometry (on day 8). RESULTS A dose-dependent increase in osteoclasts occurred after LPS injections. However, quercetin (75 mg/kg) reduced the 5 mg/mL LPS-induced osteoclasts. Using microcomputerized tomography, the bone crest levels at ligation sites were found to be significantly more apical than at the control sites on days 8 and 12; however, the apically located bone crests rebounded in rats from the quercetin-plus-ligation group. Histometry demonstrated significantly more coronal alveolar crest bone levels, less inflammatory cell-infiltrated connective tissue areas and less connective tissue attachments in the ligation-plus-quercetin group compared with those in the ligation group. CONCLUSION As the quercetin could reduce the LPS-induced osteoclast formation and the ligature-enhanced periodontal inflammation and bone loss, we suggest that it may have an ameliorative effect on periodontal destruction.

Invasive pattern grading score designed as an independent prognostic indicator in oral squamous cell carcinoma

Chang Y‐C, Nieh S, Chen S‐F, Jao S‐W, Lin Y‐L & Fu E (2010) Histopathology 57, 295–303
Invasive pattern grading score designed as an independent prognostic indicator in oral squamous cell carcinoma

Tetracycline release from tripolyphosphate–chitosan cross-linked sponge: a preliminary in vitro study

BACKGROUND AND OBJECTIVE The aim of this study was to design a tripolyphosphate-chitosan cross-linked tetracycline-containing (TPP-TC) sponge that slowly releases tetracycline, for future periodontal applications. MATERIAL AND METHODS Chitosan sponge was made by freezing and drying 2.5% chitosan solution. Tripolyphosphate-chitosan cross-linked (TPP) sponge was made by immersing the chitosan sponge in tripolyphosphate solution and air drying it. Tetracycline-containing chitosan (TC) sponge was prepared by freezing and drying a mixture of chitosan and tetracycline. TPP-TC sponge was made by immersing the TC sponge in tripolyphosphate solution. The weight, thickness and diameter of the four chitosan sponges were recorded. Their surface microstructures were inspected using scanning electron microscopy. The amount of tetracycline released from the sponges was analyzed by spectrophotometry. Antimicrobial activities of the residual sponges were tested against Staphylococcus aureus and Escherichia coli. RESULTS The topography of the scaffolds was intact after the addition of tetracycline. However, increased surface irregularities were noted. In sponges with tripolyphosphate, intensified surface folding was observed. The weight of the sponges increased after tripolyphosphate and tetracycline were added, but their thicknesses and diameters decreased after cross-linking. Tetracycline was detected in the solution containing TPP-TC sponges until day 11. On day 7, the tetracycline released from TC sponges was less than that released from TPP-TC sponges. Bacterial growth was inhibited by sponges containing tetracycline. The inhibitory effect of the TPP-TC sponges was detectable until day 11. CONCLUSION Our data showed that TPP-TC sponge was suitable as a slow-release device for tetracycline and that it maintained antimicrobial effects against the bacteria tested for up to 11 d.

Expression of fascin in oral and oropharyngeal squamous cell carcinomas has prognostic significance – a tissue microarray study of 129 cases

Aims:  To elucidate the role of fascin in oral and oropharyngeal squamous cell carcinoma (OSCC) by correlation with clinical parameters.