Beloti Marcio Mateus, Martins Walter, Xavier Samuel Porfirio, Rosa Adalberto Luiz
School of Dentistry of Ribeirao Preto, University of Sao Paulo, Sao Paulo, Brazil.
Clin Oral Implants Res. 2008 Jan;19(1):48-54. doi: 10.1111/j.1600-0501.2007.01420.x. Epub 2007 Oct 22.
This study evaluated key parameters of the in vitro osteogenesis induced by osteoblastic cells obtained from sites submitted to sinus grafting with anorganic bovine bone (ABB) in comparison with cells derived from bone sites of the same patients.
In three patients, the augmentation of maxillary sinus was carried out using ABB (Bio-Oss). After at least 6 months, during the surgical intervention for titanium implants placement, biopsies were taken from these areas using trephine burs (grafted group). Bone fragments, of the same patients, from sites that had not received graft were also obtained with trephine burs and used as a control group. Osteoblastic cells were obtained from grafted and control groups by enzymatic digestion and cultured under standard osteogenic condition until subconfluence. First passaged cells were cultured in 24-well culture plates. Cell adhesion was evaluated at 24 h. For proliferation and viability assay, cells were cultured for 1, 3, 7, and 10 days. Total protein content and alkaline phosphatase (ALP) activity were measured at 3, 7, 10, 14, 17, and 21 days. Cultures were stained with Alizarin red S at 21 days, for detection of mineralized matrix. Data were compared by Student's t-test.
Cell adhesion and viability were not affected by cell source (P>0.05). Total protein content was greater (P<0.05) for grafted group. Cell proliferation, ALP activity, and bone-like nodule formation were all greater (P<0.05) for the control group.
Taken together, these results indicate that the in vivo long-term contact of cells with ABB downregulates the expression of osteoblast phenotype and consequently the in vitro osteogenesis.
本研究评估了用无机牛骨（ABB）进行上颌窦植骨部位获取的成骨细胞诱导的体外成骨关键参数，并与同一患者骨部位来源的细胞进行比较。
在三名患者中，使用ABB（Bio-Oss）进行上颌窦增大术。至少6个月后，在植入钛种植体的手术干预期间，使用环钻从这些区域获取活检组织（植骨组）。同样从这些患者未接受植骨的部位用环钻获取骨碎片并用作对照组。通过酶消化从植骨组和对照组获得成骨细胞，并在标准成骨条件下培养至亚汇合状态。第一代传代细胞在24孔培养板中培养。在24小时时评估细胞黏附情况。为了进行增殖和活力测定，细胞培养1、3、7和10天。在3、7、10、14、17和21天测量总蛋白含量和碱性磷酸酶（ALP）活性。在21天时用茜素红S对培养物进行染色，以检测矿化基质。数据通过Student's t检验进行比较。
细胞黏附和活力不受细胞来源影响（P>0.05）。植骨组的总蛋白含量更高（P<0.05）。对照组的细胞增殖、ALP活性和骨样结节形成均更高（P<0.05）。
综上所述，这些结果表明细胞与ABB的体内长期接触会下调成骨细胞表型的表达，从而导致体外成骨作用降低。
