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Direct effects of metabolic products and sonicated extracts of Porphyromonas gingivalis 2561 on osteogenesis in vitro.

机译:牙龈卟啉单胞菌2561的代谢产物和超声提取物对体外成骨的直接影响。

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摘要

It is well documented that oral microorganisms play a significant role in the initiation and progression of periodontal disease. By using various in vitro models, it has been shown that some bacteria considered periodontal pathogens or their products can stimulate bone resorption and some other parameters of osteoblast-like cell activity. However, the effects of these organisms and their products on osteogenesis itself are not known. This study was undertaken to determine the direct effects of metabolic products and sonicated extracts of Porphyromonas gingivalis on bone formation in the chick periosteal osteogenesis model. Cultures of P. gingivalis 2561 were grown under standard anaerobic culture conditions. The spent medium was collected, and following centrifugation, sonicated bacterial extracts were prepared from the bacterial pellet. These were added in various proportions to the chick periosteal osteogenesis cultures. Sonicated extracts were further fractionated into five molecular-size ranges and similarly tested. Parameters of osteogenesis, including alkaline phosphatase activity, calcium and Pi accumulation, and collagen synthesis, were measured on 6-day-old cultures. Compared with controls devoid of bacterial products, osteogenesis was inhibited significantly in cultures treated with either conditioned medium or extracts obtained from P. gingivalis. Various amounts of inhibitory activity were observed in the different ultrafiltration molecular-size fractions, with very profound inhibitory effects observed in the < 5-kDa range. Histological observations indicated the presence of cells, some bone, and/or new fibrous connective tissue at all concentrations, indicating that toxicity was not a factor. These results suggest that periodontal pathogens such as P. gingivalis might contribute to the bone loss in periodontal diseases not only by stimulating resorption but, possibly, by inhibiting bone formation directly.
机译:众所周知,口腔微生物在牙周疾病的发生和发展中起重要作用。通过使用各种体外模型,已显示出某些被认为是牙周病原体或其产物的细菌可以刺激骨吸收以及成骨细胞样细胞活性的其他一些参数。然而,这些生物及其产物对成骨本身的影响尚不清楚。这项研究是为了确定代谢产物和牙龈卟啉单胞菌的超声提取物对雏鸡骨膜成骨模型中骨形成的直接影响。在标准厌氧培养条件下培养P. gingivalis 2561的培养物。收集用过的培养基,离心后,从细菌沉淀物中制备超声处理的细菌提取物。将它们以不同比例添加到雏鸡骨膜成骨培养物中。超声提取物进一步分为五个分子大小范围,并进行了相似的测试。在6天龄的培养物中测量成骨参数,包括碱性磷酸酶活性,钙和Pi积累以及胶原蛋白合成。与不含细菌产物的对照相比,在用条件培养基或牙龈卟啉单胞菌提取物处理的培养物中,成骨作用明显受到抑制。在不同的超滤分子大小馏分中观察到各种抑制活性,在<5-kDa范围内观察到非常深刻的抑制作用。组织学观察表明在所有浓度下都存在细胞,一些骨骼和/或新的纤维结缔组织,表明毒性不是一个因素。这些结果表明,诸如牙龈卟啉单胞菌的牙周病原体可能不仅通过刺激吸收,而且可能通过直接抑制骨形成来促进牙周疾病中的骨丢失。

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