Forriol Francisco, Denaro Luca, Longo Umile Giuseppe, Taira Hirofumi, Maffulli Nicola, Denaro Vincenzo
Research Unit, FREMAP Hospital, Madrid, Spain.
Strategies Trauma Limb Reconstr. 2010 Aug;5(2):71-8. doi: 10.1007/s11751-010-0083-y. Epub 2010 Mar 10.
We evaluated the morphological features of the newly formed tissue in an experimental model of tibial callotasis lengthening on 24 lambs, aged from 2 to 3 months at the time of operation. A unilateral external fixator prototype Monotube Triax(®) (Stryker Howmedica Osteonics, New Jersey) was applied to the left tibia. A percutaneous osteotomy was performed in a minimally traumatic manner using a chisel. Lengthening was started 7 days after surgery and was continued to 30 mm. The 24 animals were randomly divided into three groups of 8 animals each: in Group 1, lengthening took place at a rate of 1 mm/day for 30 days; in Group 2, at a rate of 2 mm/day for 15 days; in Group 3, at a rate of 3 mm/day for 10 days. In each group, 4 animals were killed 2 weeks after end of lengthening, and the other 4 animals at 4 weeks after end of lengthening. To assess bony formation in the distraction area, radiographs were taken every 2 weeks from the day of surgery. To study the process of vascularization, we used Spalteholz's technique. After killing, the tibia of each animal was harvested, and sections were stained with hematoxylin and eosin, Masson's trichrome, and Safranin-O. Immunohistochemistry was performed, using specific antibodies to detect collagens I and II, S100 protein, and fibronectin. A combination of intramembranous and endochondral ossification occurred together at the site of distraction. Our study provides a detailed structural characterization of the newly formed tissue in an experimental model of tibial lengthening in sheep and may be useful for further investigations on callotasis.
我们在24只2至3月龄的羔羊胫骨延长骨痂形成实验模型中，评估了新形成组织的形态学特征。手术时将单侧外固定器原型Monotube Triax(®)（史赛克豪美迪克骨科学公司，新泽西州）应用于左胫骨。使用凿子以微创方式进行经皮截骨术。术后7天开始延长，持续至30毫米。24只动物随机分为三组，每组8只：第1组，以每天1毫米的速度延长30天；第2组，以每天2毫米的速度延长15天；第3组，以每天3毫米的速度延长10天。每组中，4只动物在延长结束后2周处死，另外4只在延长结束后4周处死。为评估牵张区域的骨形成情况，从手术当天起每2周拍摄X线片。为研究血管化过程，我们采用了施帕尔托兹技术。处死动物后，采集每只动物的胫骨，切片用苏木精和伊红、马松三色染色法以及番红O染色。进行免疫组织化学检测，使用特异性抗体检测I型和II型胶原蛋白、S100蛋白和纤连蛋白。在牵张部位同时发生了膜内成骨和软骨内成骨。我们的研究提供了绵羊胫骨延长实验模型中新形成组织的详细结构特征，可能有助于对骨痂形成的进一步研究。
