Pre-osteoblastic proliferation assessed with BrdU in undecalcified, epon-embedded adult rat trabecular bone
Pre-osteoblastic proliferation assessed with BrdU in undecalcified, epon-embedded adult rat trabecular bone
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DOI：
10.1177/002215549704500902
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发表时间：
1997-09-01
影响因子：
3.2
通讯作者：
LafageProust, MH
中科院分区：
文献类型：
--
作者：
Barou, O;Laroche, N;LafageProust, MH
We evaluated bromodeoxyuridine (BrdU) immunohistochemistry in undecalcified adult rat tibiae to study cell kinetics in various bone compartments: primary and secondary spongiosae, periosteum, and bone marrow. Several regimens of BrdU administration were tested (IF injections and osmotic minipumps). We compared LR White resin, methylmethacrylate, and Epon-araldite embedding, microwave irradiation for antigen retrieval, several concentrations of sodium ethoxide for deplastification, and various DNA denaturation procedures. Paraffin-embedded decalcified tibiae and Epon-embedded bowel were used as positive controls. The best results were obtained in rats labeled with 40 mg of BrdU for 72 hr using osmotic minipumps. The procedure using a Microprobe system in Epon-embedded bone tissue with a sodium ethoxide concentration of 50% for two intervals of 20 min provided the best staining quality and tissue preservation. Labeled pre-osteoblastic cells and bone marrow cells could be counted. Epon embedding allowed preservation of tetracycline double labeling performed 1 to 5 days before sacrifice. The number of labeled pre-osteoblastic cells was correlated with the double-labeled surface area measured histomorphometrically.
