Impact of Harvesting Method and Donor Age on the Behavior of Human Osteoblast-Like Cells.

Autogenous particulate bone grafts are being utilized in oral implantology for minor grafting procedures. This study aimed to investigate the influence of the bone-harvesting technique, donor age, and donor site on proliferation and differentiation of human primary osteoblast-like cells in the cell culture. Autogenous bone particles (20 samples) were harvested from the maxilla and mandible during surgery using two different protocols, and two types of particulate bone grafts were collected: bone chips and bone sludge. Bone samples were cultured in growth medium and, after 2 to 3 weeks, the cells that grew from bone grafts were cultured in the normal and osteogenic medium for 0, 4, 7, and 20 days. DNA, alkaline-phosphatase (ALP), calcium-content measurements, and Alizarin red/toluidine blue staining were performed. Data were analyzed by repeated-measures analysis of variance with Bonferroni test. The level of statistical significance was set at 5% (P < .05). Total DNA, ALP, and calcium content were significantly higher for the bone chip samples compared to the bone sludge samples. Total DNA and ALP content were significantly higher for the patients in age group 1 (≤ 60 years) compared to age group 2 (> 60 years) and was significantly higher for mandibular samples than maxillary samples on day 20. However, the calcium measurement showed no significant difference concerning donor age and donor site. Data analysis revealed that harvesting technique (bone chips vs bone sludge), donor age (≤ 60 years vs > 60 years), and donor site (maxilla vs mandible) influenced the osteogenic potential of the collected particulate bone graft. The bone chips were superior in terms of osteogenic efficacy and should be considered a suitable option for particulate bone graft collection.

Prostaglandins differentially affect osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells.

Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are currently used for bone tissue engineering. AT-MSCs undergoing osteogenic differentiation respond to mechanical loading with increased cyclooxygenase-2 gene expression, a key enzyme in prostaglandin (PG) synthesis. PGs are potent multifunctional regulators in bone, exhibiting stimulatory and inhibitory effects on bone formation and resorption. PGE(2), but not PGI(2) or PGF(2), recruits osteoprogenitors from the bone marrow space and influences their differentiation. We hypothesize that PGE(2), PGI(2), and PGF(2) may differentially regulate osteogenic differentiation of human AT-MSCs. PGE(2), PGI(2), and PGF(2) (0.01-10 microM) affected osteogenic differentiation, but not proliferation of AT-MSCs after 4-14 days. Only PGF(2) (0.01-10 microM) increased alkaline phosphatase (ALP) activity at day 4. PGE(2) (10 microM), PGI(2) (0.01-10 microM), and PGF(2) (10 microM) decreased ALP activity, whereas PGF(2) (0.1 microM) increased ALP activity at day 14. PGF(2) (0.01-0.1 microM) and PGI(2) (0.01 microM) upregulated osteopontin gene expression, and PGF(2) (0.01 microM) upregulated alpha1(I)procollagen gene expression at day 4. PGE(2) and PGF(2) (10 microM) at day 4 and PGF(2) (1 microM) at day 14 downregulated runt-related transcription factor-2 gene expression. We conclude that PGE(2), PGI(2), and PGF(2) differentially affect osteogenic differentiation of AT-MSCs, with PGF(2) being the most potent. Thus, locally produced PGF(2) might be most beneficial in promoting osteogenic differentiation of AT-MSCs, resulting in enhanced bone formation for bone tissue engineering.

Salivary agglutinin/DMBT1SAG expression is up-regulated in the presence of salivary gland tumors.

Salivary agglutinin (SAG) is encoded by the gene Deleted in Malignant Brain Tumors 1 (DMBT1) and represents the salivary variant of DMBT1 (DMBT1(SAG)). While SAG is a bona fide anti-caries factor, DMBT1 was proposed as a candidate tumor-suppressor for brain, digestive tract, and lung cancer. Though DMBT1(SAG) is expressed in the salivary glands, its expression in salivary gland tumors is unknown. Here we analyzed DMBT1(SAG) expression in 20 salivary gland tumors and 14 tumor-flanking tissues by immunohistochemistry. DMBT1(SAG) in salivary gland tumors resembles the changes of expression levels known from DMBT1 in tumors in other cancer types. Particularly, DMBT1(SAG) was up-regulated in 10/14 tumor-flanking tissues, and a strong staining of the luminal content in the tumor and/or the tumor-flanking tissue was observed in 14/20 cases. This suggests that, in addition to its role in caries defense, SAG may serve as a potential tumor indicator and/or tumor suppressor in salivary gland tissue.