Rammelt S, Corbeil D, Manthey S, Zwipp H, Hanisch U
Department of Trauma and Reconstructive Surgery, University Hospital Carl Gustav Carus, Dresden, Germany.
J Biomed Mater Res A. 2007 Nov;83(2):313-22. doi: 10.1002/jbm.a.31243.
When investigating the tissue reaction on orthopedic implants, the cellular activity at the bone-implant interface is of special interest. Preparation of undecalcified bone sections with methylmetacrylate (MMA)-based resins allows evaluation of the host tissue reactions with the implant in situ. However, the technical workup is demanding and few reports exist on the immunohistochemical characterization of these sections. Rat (R), sheep (S), and human (H) samples were investigated. R specimens contained intramedullary rods in the rat tibia. S specimens were sheep tibiae with an external fixator. H specimens were obtained from deceased patients. Specimens were embedded in MMA-based Technovit 9100N using cold polymerization. Sections of 10-15 microm thickness were obtained and prepared for immunohistochemical staining. Good morphological detail was preserved in all specimens providing information about mineralization, recent bone formation, and bone-implant contact. The following antibodies could reproducibly be detected specifically: Osteopontin (R, S, H), Osteonectin, Cathepsin D (R, S), von Willebrand factor (R, H), Osteocalcin, ED 1 (R), CD 3, CD 68, Keratin (H). Control procedures without adding primary antibodies showed no unspecific staining. Reliable detection of immunohistochemical markers of bone resorption, bone formation, inflammation, and angiogenesis at undecalcified sections with the implant in situ appears promising in enhancing our understanding of the cellular activity and cell-matrix interactions at the bone-implant interface.
在研究骨科植入物的组织反应时，骨 - 植入物界面处的细胞活性特别令人关注。用甲基丙烯酸甲酯（MMA）基树脂制备未脱钙骨切片，可在原位评估宿主组织与植入物的反应。然而，技术操作要求较高，关于这些切片免疫组织化学特征的报道较少。对大鼠（R）、绵羊（S）和人类（H）样本进行了研究。R样本为大鼠胫骨内的髓内棒。S样本是带有外固定器的绵羊胫骨。H样本取自已故患者。样本采用冷聚合方法包埋于MMA基Technovit 9100N中。获得10 - 15微米厚的切片并准备进行免疫组织化学染色。所有样本均保留了良好的形态细节，提供了有关矿化、近期骨形成和骨 - 植入物接触的信息。以下抗体能够被特异性地重复检测到：骨桥蛋白（R、S、H）、骨连接蛋白、组织蛋白酶D（R、S）、血管性血友病因子（R、H）、骨钙素、ED1（R）、CD3、CD68、角蛋白（H）。不添加一抗的对照程序未显示非特异性染色。在未脱钙切片上原位植入物的情况下，可靠检测骨吸收、骨形成、炎症和血管生成的免疫组织化学标志物，对于增强我们对骨 - 植入物界面处细胞活性和细胞 - 基质相互作用的理解似乎很有前景。
