Abstract Fibroblast growth factor‐2 and parathyroid hormone are strong modulators of the maturation process of chondrocytes during endochondral ossification. To clarify whether and how these agents may exert stage‐specific effects during this process, we analyzed the responsiveness and phenotypic consequences of treatment with fibroblast growth factor‐2 or parathyroid hormone on chondrocytes at different stages of maturation. Populations of immature lower sternal, maturing upper sternal, and hypertrophic tibial growth plate chondrocytes were isolated from day 18–20 chick embryos and were allowed to resume the maturation process by growth in standard monolayer cultures. Treatment of immature lower sternal cultures with as little as 0.1 ng/ml of fibroblast growth factor‐2 or 10 −10 M parathyroid hormone prevented both the emergence of mature type‐X collagen‐synthesizing chondrocytes and the ensuing enlargement of cells that occurred in control (untreated) cultures. Similarly, the treatment of cultured early maturing upper sternal cells with these factors severely reduced the synthesis of type‐X collagen and alkaline phosphatase activity and the levels of their respective mRNAs. In sharp contrast, when the cultured upper sternal cells were allowed to grow and mature further before treatment, the responsiveness to fibroblast growth factor‐2 was markedly reduced and the responsiveness to parathyroid hormone remained strong and largely unchanged. Cultures of hypertrophic tibial growth plate cells displayed a similar reduced sensitivity to fibroblast growth factor‐2, as also indicated by the lack of mitogenic effects, and strong sensitivity to parathyroid hormone. The phenotypic changes induced by treatment with either of these factors were fully reversible when cultures that had been treated were placed in control medium. The results demonstrate that fibroblast growth factor‐2 and parathyroid hormone are equally potent in affecting the early stages of maturation but exert differential effects as the cells progress along the maturation pathway. The factors appear to be part of sequentially acting mechanisms to ensure normal progression of chondrocyte maturation during endochondral ossification.
The effects of PTH and calcitonin (CT) on the expression of mineralization-related phenotypes by chondrocytes were examined. In cultures of pelleted growth-plate chondrocytes. PTH caused 60-90% decreases in alkaline phosphatase activity, the incorporation of 45Ca into insoluble material, and the calcium content during the post-mitotic stage. These effects of PTH were dose-dependent and reversible. In contrast, CT increased alkaline phosphatase activity, 45Ca incorporation into insoluble material, and the calcium content by 1.4- to 1.8-fold. These observations suggest that PTH directly inhibits the expression of the mineralization-related phenotypes by growth-plate chondrocytes, and that CT has the opposite effects.
This study was undertaken to investigate the physicochemical aspects of the interaction between the surface of biomaterials and bacterial cell membranes in vitro, aimed at studying the mechanisms of bacterial adhesion to biomaterials. Correlations were made between the number of adherent bacterial cells (S. aureus) and each of the calculated components of surface free energy (i.e., dispersion, polarity and hydrogen bond) of biomaterials. The effect of antibodies to cell-adhesion molecules on bacterial adhesion was also studied using monoclonal antibodies to vitronectin receptor, fibronectin receptor and CD44. This study indicates the polarity component of surface free energy plays a dominant role in the process of bacterial adhesion at least in vitro. The number of cells adherent to materials decreased to 44-73% of the control value in the presence of antibodies tested, showing that cell adhesion molecules affect adherence to biomaterials. Moreover, the results suggested that bacterial adhesion was prevented by specific blockade of cell adhesion molecule receptors.
Syndecans are transmembrane heparan sulfate proteoglycans. They are known to interact with basic fibroblast growth factor (bFGF), and it has been suggested that they play important roles in the growth, morphology, and migration of a variety of cell types. We examined the expression of syndecans and fibroblast growth factor receptor type 1 (FGFR1) in periodontal ligament (PDL) cells, because these membrane proteins may play roles in the control of growth and differentiation during regeneration of PDL. Reverse-transcription/polymerase chain-reaction (RT-PCR) showed that PDL cells expressed syndecan-2 and -4 mRNAs. This was confirmed by sequence analysis of the PCR products. When PDL cells were maintained for 25 days, alkaline phosphatase (ALPase) activity gradually increased and reached a maximal level on day 20. Northern blotting analysis showed that PDL cells expressed 2.3-kb syndecan-2, 2.6-kb syndecan-4, and 2.8-kb FGFR1 mRNAs throughout the entire culture period, whereas no syndecan-1 mRNA was detectable by this method. Maximal levels of syndecan-2, -4, and FGFR1 mRNAs were observed on day 5. However, their levels were markedly decreased on days 20 and 25. Accordingly, the inhibitory effect of bFGF on ALPase activity was less on day 20 than on day 5. When PDL cells were pre-treated with heparitinase, a mitogenic response of PDL cells to bFGF was decreased. These observations indicate that PDL cells express syndecan-2, -4, and FGFR1 mRNAs, and that those levels are changed with the increase in ALPase activity in culture. The reductions in syndecan-2, -4, and FGFR1 levels may be involved in the control of growth and differentiation of PDL cells during development and regeneration.
The purpose of this study was to investigate the relationship between changes in parathyroid hormone (PTH) receptor levels and chondrocyte maturation during endochondral ossification. Chondrocytes were isolated from the growth plate of rabbit ribs and maintained in the presence of 10% serum in mass cultures. Treatment with PTH-(1-84) and a PTH-(1-34) fragment suppressed the increases in alkaline phosphatase activity and in type X collagen and 1 alpha,25-dihydroxyvitamin D3 receptor levels and abolished 45Ca incorporation into mineral, all of which occurred in parallel untreated cultures in the hypertrophic (terminal) stage. These effects of PTH were observed at low concentrations (10(-10) to 10(-9) M) and within 24-48 h of treatment. PTH-(1-84) and PTH-(1-34) also increased [35S]sulfate incorporation into newly synthesized proteoglycans. In contrast, the middle and carboxyl-terminal fragments of PTH tested had little effect on proteoglycan synthesis or terminal differentiation. The binding of 125I-PTH-(1-34) to cells in the growth plate was greater than that to cells in liver, skin, muscle, brain, or kidney. When the correlation between binding levels and stage of maturation was examined, we found that 125I-PTH-(1-34) binding to its 72-kDa receptor was low in resting and proliferating chondrocytes, increased 10-fold in matrix-forming chondrocytes, and thereafter decreased in hypertrophic chondrocytes both in vitro and in situ. Scatchard analysis revealed that the changes in PTH binding were due to changes in the number, and not in the affinity, of the receptor. The changes in PTH-(1-34) binding paralleled those in [35S]sulfate incorporation into proteoglycans. These findings suggest that stage-dependent increases in PTH/PTH-related peptide receptor levels localize the hormone stimulation of proteoglycan synthesis and inhibition of precocious hypertrophy in the matrix-forming zone of growth plates.
Compounded by the needs of an aging society, interactions between oral condition and systemic diseases may require that dentists pursue additional training in oral medicine beyond that received in dental school. The purpose of this study was to investigate whether pursuing oral medicine professional education is recognised by dental practitioners as an important factor regarding job satisfaction.A questionnaire was mailed to 1,379 dental practitioners in Japan, along with a follow-up survey to assess repeatability, in 2017. The questionnaire consisted of 19 items/questions related to the respondents' attributes and job satisfaction (5 items), willingness to learn oral medicine (4 items), willingness to learn more about dentistry (4 items), and willingness to contribute to society (6 items). Representative questions were extracted via binomial logistic regression analysis. Multivariable logistic regression analysis was performed to assess the relationships between job satisfaction and the explanatory variables.Amongst 337 respondents, multivariable logistic regression analysis showed an association between strong job satisfaction (n = 126, 37%) and willingness to learn more about oral medicine and dentistry and contribute to society, with odds ratios (95% confidence interval) of 4.22 (1.84-9.68), 3.16 (1.16-8.62), and 7.32 (3.14-17.06) and κ values of 0.38, 0.58, and 0.51, respectively.Our results from dental practitioners suggest additional benefits of oral medicine professional education for future job satisfaction.
