Dentistry Department, Federal University of Uberlândia, Uberlândia, Minas Gerais, Brazil.
Dentistry Department, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil.
Microsc Res Tech. 2022 Oct;85(10):3339-3346. doi: 10.1002/jemt.24188. Epub 2022 Jun 27.
This study aimed to assess different approaches for bone healing evaluation on histological images and to introduce a new automatic evaluation method based on segmentation with distinct thresholds. We evaluated the hyperbaric oxygen therapy (HBO) effects on bone repair in type 1 diabetes mellitus rats. Twelve animals were divided into four groups (n = 3): non-diabetic, non-diabetic + HBO, diabetic, and diabetic + HBO. Diabetes was induced by intravenous administration of streptozotocin (50 mg/kg). Bone defects were created in femurs and HBO was immediately started at one session/day. After 7 days, the animals were euthanized, femurs were removed, demineralized, and embedded in paraffin. Histological sections were stained with hematoxylin and eosin (HE) and Mallory's trichrome (MT), and evaluated using three approaches: (1) conventional histomorphometric analysis (HE images) using a 144-point grid to quantify the bone matrix; (2) a semi-automatic method based on bone matrix segmentation to assess the bone matrix percentage (MT images); and (3) automatic approach, with the creation of a plug-in for ImageJ software. The time required to perform the analysis in each method was measured and subjected to Bland-Altman statistical analysis. All three methods were satisfactory for measuring bone formation and were not statistically different. The automatic approach reduced the working time compared to visual grid and semi-automated method (p < .01). Although histological evaluation of bone healing was performed successfully using all three methods, the novel automatic approach significantly shortened the time required for analysis and had high accuracy.
本研究旨在评估组织学图像中不同的骨愈合评估方法，并介绍一种基于分割的新的自动评估方法，该方法使用不同的阈值。我们评估了高压氧疗法（HBO）对 1 型糖尿病大鼠骨修复的影响。12 只动物分为四组（n=3）：非糖尿病组、非糖尿病+HBO 组、糖尿病组和糖尿病+HBO 组。糖尿病通过静脉注射链脲佐菌素（50mg/kg）诱导。在股骨中创建骨缺损，并立即开始每天进行一次 HBO。7 天后，处死动物，取出股骨，脱矿，石蜡包埋。组织学切片用苏木精和伊红（HE）和马氏三色（MT）染色，并使用三种方法进行评估：（1）使用 144 点网格的常规组织形态计量分析（HE 图像）来量化骨基质；（2）基于骨基质分割的半自动方法来评估骨基质百分比（MT 图像）；和（3）自动方法，使用 ImageJ 软件创建插件。测量每种方法进行分析所需的时间，并进行 Bland-Altman 统计分析。所有三种方法都可用于测量骨形成，且无统计学差异。与视觉网格和半自动方法相比，自动方法减少了工作时间（p<.01）。虽然使用所有三种方法都成功地进行了骨愈合的组织学评估，但新的自动方法大大缩短了分析所需的时间，且具有较高的准确性。
