Open source software for semi-automated histomorphometry of bone resorption and formation parameters

Area of Science:

• Biomedical Engineering
• Skeletal Biology
• Image Analysis

Background:

• Micro-computed tomography (micro-CT) is standard for bone volume and architecture in small animals.
• Micro-CT cannot assess crucial bone turnover parameters like bone formation rate or osteoclast (OC) activity.
• Traditional histomorphometry is essential but time-consuming and challenging for OC detection in mouse bones.

Purpose of the Study:

• To develop and validate a rapid, semi-automated method for static and dynamic bone histomorphometry.
• To create user-friendly, open-source image analysis packages for bone research.
• To improve the efficiency and accuracy of bone turnover assessment in small animal models.

Main Methods:

• Adaptation of established staining protocols for bone histomorphometry.
• Development of three novel, open-source ImageJ-based software packages: TrapHisto, OsteoidHisto, and CalceinHisto.
• Semi-automated analysis of histomorphometric parameters including bone resorption, osteoid, and calcein double labeling.

Main Results:

• Successful adaptation of staining protocols and development of user-friendly image analysis software.
• TrapHisto, OsteoidHisto, and CalceinHisto enable rapid, semi-automated measurement of key histomorphometric parameters.
• The developed methods simplify the complex image analysis underlying histomorphometry.

Conclusions:

• The novel software packages offer an efficient and accessible approach to bone histomorphometry.
• These tools address the limitations of micro-CT by enabling precise assessment of bone turnover.
• The developed methods facilitate more comprehensive skeletal analysis in small animal research.