Yang R, Davies C M, Archer C W, Richards R G
AO Research Institute, Davos, Switzerland.
Eur Cell Mater. 2003 Dec 31;6:57-71; discussion 71. doi: 10.22203/ecm.v006a06.
Trabecular bone is routinely analysed by histomorphological-histometrical and immunohistochemical techniques as means of assessing the differentiation status of bone deposition and growth. Currently few embedding resins exist for which both morphological and immunohistochemical analyses can be performed on mineralised tissue. Paraffin, the standard embedding medium for bone enzyme and immunohistochemistry, can only be used on demineralised tissue, but then trabecular structure may be badly preserved. Methyl methacrylate (MMA), the resin of choice for undecalcified bone histology can only be used for bone immunohistochemistry if the usual, highly exothermic polymerisation procedure is avoided which destroys tissue antigenicity. Consequently, most current practices involve cutting samples in half to be processed in separate resins when more than one type of analysis is required. Technovit 9100 New is a low temperature MMA embedding system that is purported to significantly improve tissue antigenicity preservation allowing polymerisation at -20 degrees C. In this study, Technovit 9100 New-embedded undecalcified trabecular bone samples (adult human, young bovine and ovine) yielded immunolabelling with several bone matrix markers and preserved morphological features in 7 microm sections when stained with Masson-Goldner, von Kossa, or toluidine blue. Bone samples from all resins used were immunolabelled with antibodies against osteocalcin, alkaline phosphatase, osteopontin, osteonectin, bone sialoprotein and procollagen type I amino-terminal propeptide. Technovit-embedded bone yielded more reliable immunolabelling of the matrix proteins when compared with heat or cold-cured LR White or standard embedded MMA samples. Technovit 9100 New provided better routine histology than LR White, and was comparable to MMA. Results demonstrated that Technovit 9100 New can be used as a low-temperature acrylic resin embedding method for routine undecalcified bone histology, as well as for immunohistochemistry.
小梁骨通常通过组织形态学 - 组织计量学和免疫组织化学技术进行分析，以此作为评估骨沉积和生长分化状态的手段。目前，可供对矿化组织同时进行形态学和免疫组织化学分析的包埋树脂很少。石蜡是骨酶和免疫组织化学的标准包埋介质，仅可用于脱矿组织，但此时小梁结构可能保存不佳。甲基丙烯酸甲酯（MMA）是未脱钙骨组织学的首选树脂，只有避免通常会破坏组织抗原性的高度放热聚合过程，才能用于骨免疫组织化学。因此，当需要进行多种类型分析时，当前大多数做法是将样本切成两半，分别用不同的树脂处理。Technovit 9100 New是一种低温MMA包埋系统，据称能显著改善组织抗原性的保存，可在-20摄氏度下聚合。在本研究中，用Technovit 9100 New包埋的未脱钙小梁骨样本（成人、幼年牛和羊），用几种骨基质标记物进行免疫标记时，用马森 - 戈德纳染色法、冯·科萨染色法或甲苯胺蓝染色时，在7微米切片中保留了形态特征。使用的所有树脂的骨样本均用抗骨钙素、碱性磷酸酶、骨桥蛋白、骨连接蛋白、骨唾液蛋白和I型前胶原氨基端前肽的抗体进行免疫标记。与热固化或冷固化的LR White或标准包埋的MMA样本相比，Technovit包埋的骨对基质蛋白产生了更可靠的免疫标记。Technovit 9100 New提供了比LR White更好的常规组织学效果，与MMA相当。结果表明，Technovit 9100 New可作为一种低温丙烯酸树脂包埋方法，用于常规未脱钙骨组织学以及免疫组织化学。
