Comprehensive histological evaluation of bone implants

Abstract

To investigate and assess bone regeneration in sheep in combination with new implant materials classical histological staining methods as well as immunohistochemistry may provide additional information to standard radiographs or computer tomography. Available published data of bone defect regenerations in sheep often present none or sparely labeled histological images. Repeatedly, the exact location of the sample remains unclear, detail enlargements are missing and the labeling of different tissues or cells is absent. The aim of this article is to present an overview of sample preparation, staining methods and their benefits as well as a detailed histological description of bone regeneration in the sheep tibia. General histological staining methods like hematoxylin and eosin, Masson-Goldner trichrome, Movat's pentachrome and alcian blue were used to define new bone formation within a sheep tibia critical size defect containing a polycaprolactone-co-lactide (PCL) scaffold implanted for 3 months (n = 4). Special attention was drawn to describe the bone healing patterns down to cell level. Additionally one histological quantification method and immunohistochemical staining methods are described.

Osteophytes and fracture calluses share developmental milestones and are diminished by unloading

“…Ossification at each time point was evaluated by quantifying ossified tissue areas, including osteoid and mineralized tissue, within chondro/osteophytes and fracture calluses. Ossified tissue was identified as a compact structure with high Eosin content . Total ossified tissue area was normalized by total chondro/osteophyte or fracture callus areas to calculate ossification percentage.…”

“…Ossified tissue was identified as a compact structure with high Eosin content. 34 Total ossified tissue area was normalized by total chondro/osteophyte or fracture callus areas to calculate ossification percentage.…”

Image matters: Development of a novel staining method, the RGB-trichrome, as a reliable tool for the study of the musculoskeletal system

“…However, these microscopy technologies remain expensive, require highly-specialized management, and are not available in many laboratories. In the field of light microscopy, numerous staining methods have been developed, and usually several staining procedures are needed to cope with the differences in tissue components and functional status of the skeletal system 9, 15 . In this context, a general staining method should fulfill a number of requirements to provide a general overview of the microstructure and functional characteristics of the musculo-skeletal organs, at cell and tissue levels.…”

“…A large number of staining methods are used for the recognition of different components and/or functional status of the musculoskeletal system. In order to highlight these different components and to allow histomorphometry quantification several staining methods are usually applied, from general staining, such as hematoxylin and eosin, Movat pentachrome, toluidine blue, alcian blue or safranin O/fas green 9, 15 , to more specific silver impregnation 16, 17 or fluorescence-based 18, 19 methods for visualization of the osteocyte network. In addition, some specific staining methods, such as von Kossa or Masson-Goldner trichrome, are applied to undecalcified, resin-embedded, bone sections to distinguish the mineralized bone matrix from osteoid 9 .…”

The Effect of Local Application of Bisphosphonates in Osseointegration of Dental Implant: An Experimental Study

“…The ready slide(longitudinal section) was placed on a fixed temperature drying table which was regulated to about 37-42°C, so that the sections adhere to the slide. (16)…”