The effect of fibronectin and a bone xenograft on regenerative treatment: a feasibility study.

The purpose of this study was to evaluate the ability of fibronectin to augment the regenerative effects of a bovine-derived xenograft in human periodontal defects. Using a parallel arm, randomized double-blind design, 24 patients with an intrabony defect or a Class II furcation defect were randomly assigned to either the experimental group (xenograft plus fibronectin) or the control group (xenograft without fibronectin). Probing attachment level, pocket depth, and gingival recession were measured at baseline and at 12 months after surgery. Both treatment modalities resulted in attachment gain and pocket depth reduction compared with baseline values. Changes in clinical attachment were not significantly different between the groups (gain of 1.5 mm +/- 1.1 mm in the experimental group and 1.3 mm +/- 1.4 mm in the control). Pocket depth reduction was greater in the control (2.3 mm +/- 1.2 mm) than in the experimental group (2.1 mm +/- 1.9 mm). Gingival recession also was greater in the control (0.9 mm +/- 0.6 mm) than in the experimental group (0.6 +/- 1.5 mm). Subtraction radiography revealed no significant differences between the groups when measuring changes in the distance between the cementoenamel junction and the crest of the bone or in the estimated gain in mineralized tissue mass. It was concluded that a significant difference between the regenerative treatment modalities could not be demonstrated within the limitations of the study. Fibronectin appears to have a stabilizing or proliferative effect on the gingival soft tissue by promoting less postoperative gingival recession.

Differentially expressed protein markers in human submandibular and sublingual secretions.

Proteome analysis of secretions from individual salivary glands is important for understanding the health of the oral cavity and pathogenesis of certain diseases. However, cross-contamination of submandibular (SM) and sublingual (SL) glandular secretions can occur. The close anatomic relationship of the SM and SL ductal orifices can lead to such contamination. Additionally, these glands may share common ducts. To insure the purity of SM/SL secretions for proteomic analysis, it is important to develop unique biomarkers which could be used to verify the integrity of the individual glandular saliva. In this study, a proteomics approach based on mass spectrometry and gel electrophoresis techniques was utilized to identify and verify a set of proteins (cystatin C, calgranulin B and MUC5B mucin), which are differentially expressed in SM/SL secretions. SM/SL fluids were obtained from nine healthy subjects. Cystatin C was found to be an SM-selective protein as it was found in all SM fluids but not detected in two SL fluids. MUC5B mucin and calgranulin B, on the other hand, were found to be SL-selective proteins. All SL samples contained MUC5B mucin, whereas MUC5B mucin was not detected in four SM samples. Eight of the SL samples contained calgranulin B; however, calgranulin B was absent in eight SM samples. This set of protein markers, especially calgranulin B, can be used to determine the purity of SM/SL samples, and therefore identify potential individuals who do not exhibit cross-contaminated SM/SL secretions, an important requirement for subsequent proteome analysis of pure SM and SL secretions.

Clinical repair of an osseous defect associated with a cemental tear: a case report.

Cemental tears have been described as detachment of cementum caused by trauma or aging. They often result in severe periodontal lesions that may necessitate the extraction of the affected tooth. This case report describes the clinical resolution of a periodontal lesion associated with a cemental tear. A maxillary central incisor was subjected to endodontic treatment twice with no resolution of a deep distobuccal pocket and a palatal sinus tract from its apical region. The preoperative differential diagnosis for the condition present on the tooth included a vertical fracture and a combined periodontal-endodontic lesion. Surgical exploration of the area revealed a cemental tear on the apical third of the tooth. The cementum fragments were removed, root-end resection was performed, and the osseous lesion was treated with an osseous graft and guided tissue regeneration. Clinical examination of the area 1 year after surgery revealed resolution of both the prior pocket and sinus tract. Radiographic examination of the area showed increased radiopacity in the area of the original lesion, suggesting bone fill.

Bovine-derived bone protein extract in the treatment of mandibular Class II furcations.

This study was an initial evaluation of the use of a bovine-derived bone protein (BP) extract that contains various growth factors combined with decalcified freeze-dried bone allograft (DFDBA) as regenerative treatment for class II mandibular furcations. Twenty-five patients were divided into 5 groups according to the dosage of BP present per mg of DFDBA to be grafted: (1) 0.00 microgram/mg, (2) 3.13 micrograms/mg, (3) 6.25 micrograms/mg, (4) 12.5 micrograms/mg, and (5) 25.0 micrograms/mg. Surgical exploration of the furcation defects was performed followed by grafting with BP/DFDBA. Results at 6 months showed that attachment gain in the treated furcation areas was greatest in Groups 4 and 5, suggesting that BP has the potential to increase the effects of DFDBA in gaining clinical attachment in mandibular class II furcations.

Co-localized or randomly distributed? Pair cross correlation of in vivo grown subgingival biofilm bacteria quantified by digital image analysis.

The polymicrobial nature of periodontal diseases is reflected by the diversity of phylotypes detected in subgingival plaque and the finding that consortia of suspected pathogens rather than single species are associated with disease development. A number of these microorganisms have been demonstrated in vitro to interact and enhance biofilm integration, survival or even pathogenic features. To examine the in vivo relevance of these proposed interactions, we extended the spatial arrangement analysis tool of the software daime (digital image analysis in microbial ecology). This modification enabled the quantitative analysis of microbial co-localization in images of subgingival biofilm species, where the biomass was confined to fractions of the whole-image area, a situation common for medical samples. Selected representatives of the disease-associated red and orange complexes that were previously suggested to interact with each other in vitro (Tannerella forsythia with Fusobacterium nucleatum and Porphyromonas gingivalis with Prevotella intermedia) were chosen for analysis and labeled with specific fluorescent probes via fluorescence in situ hybridization. Pair cross-correlation analysis of in vivo grown biofilms revealed tight clustering of F. nucleatum/periodonticum and T. forsythia at short distances (up to 6 µm) with a pronounced peak at 1.5 µm. While these results confirmed previous in vitro observations for F. nucleatum and T. forsythia, random spatial distribution was detected between P. gingivalis and P. intermedia in the in vivo samples. In conclusion, we successfully employed spatial arrangement analysis on the single cell level in clinically relevant medical samples and demonstrated the utility of this approach for the in vivo validation of in vitro observations by analyzing statistically relevant numbers of different patients. More importantly, the culture-independent nature of this approach enables similar quantitative analyses for "as-yet-uncultured" phylotypes which cannot be characterized in vitro.

Systemic disease-induced salivary biomarker profiles in mouse models of melanoma and non-small cell lung cancer.

BACKGROUND: Saliva (oral fluids) is an emerging biofluid poised for detection of clinical diseases. Although the rationale for oral diseases applications (e.g. oral cancer) is intuitive, the rationale and relationship between systemic diseases and saliva biomarkers are unclear. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we used mouse models of melanoma and non-small cell lung cancer and compared the transcriptome biomarker profiles of tumor-bearing mice to those of control mice. Microarray analysis showed that salivary transcriptomes were significantly altered in tumor-bearing mice vs. controls. Significant overlapping among transcriptomes of mouse tumors, serum, salivary glands and saliva suggests that salivary biomarkers have multiple origins. Furthermore, we identified that the expression of two groups of significantly altered transcription factors (TFs) Runx1, Mlxipl, Trim30 and Egr1, Tbx1, Nr1d1 in salivary gland tissue of melanoma-bearing mice can potentially be responsible for 82.6% of the up-regulated gene expression and 62.5% of the down-regulated gene expression, respectively, in the saliva of melanoma-bearing mice. We also showed that the ectopic production of nerve growth factor (NGF) in the melanoma tumor tissue as a tumor-released mediator can induce expression of the TF Egr-1 in the salivary gland. CONCLUSIONS: Taken together, our data support the conclusion that upon systemic disease development, significant changes can occur in the salivary biomarker profile. Although the origins of the disease-induced salivary biomarkers may be both systemic and local, stimulation of salivary gland by mediators released from remote tumors plays an important role in regulating the salivary surrogate biomarker profiles.

Systematic comparison of the human saliva and plasma proteomes.

The proteome of human salivary fluid has the potential to open new doors for disease biomarker discovery. A recent study to comprehensively identify and catalog the human ductal salivary proteome led to the compilation of 1166 proteins. The protein complexity of both saliva and plasma is large, suggesting that a comparison of these two proteomes will provide valuable insight into their physiological significance and an understanding of the unique and overlapping disease diagnostic potential that each fluid provides. To create a more comprehensive catalog of human salivary proteins, we have first compiled an extensive list of proteins from whole saliva (WS) identified through MS experiments. The WS list is thereafter combined with the proteins identified from the ductal parotid, and submandibular and sublingual (parotid/SMSL) salivas. In parallel, a core dataset of the human plasma proteome with 3020 protein identifications was recently released. A total of 1939 nonredundant salivary proteins were compiled from a total of 19 474 unique peptide sequences identified from whole and ductal salivas; 740 out of the total 1939 salivary proteins were identified in both whole and ductal saliva. A total of 597 of the salivary proteins have been observed in plasma. Gene ontology (GO) analysis showed similarities in the distributions of the saliva and plasma proteomes with regard to cellular localization, biological processes, and molecular function, but revealed differences which may be related to the different physiological functions of saliva and plasma. The comprehensive catalog of the salivary proteome and its comparison to the plasma proteome provides insights useful for future study, such as exploration of potential biomarkers for disease diagnostics.