# Influence of different preservation methods on the mechanical in vitro stability of bones: a comparative study using porcine metatarsals

**Authors:** Martin Bäumlein, Moritz Kühlein-Lehr, Alexander M. König, Volker Ziller, Steffen Ruchholtz, Jürgen R. J. Paletta, Vanessa dos Santos

PMC · DOI: 10.1007/s00402-026-06226-2 · Archives of Orthopaedic and Trauma Surgery · 2026-03-13

## TL;DR

This study compares how different preservation methods affect the mechanical stability of bones in laboratory settings, finding that chemical preservation changes bone stiffness and fracture patterns.

## Contribution

The study provides new insights into how chemical preservation methods alter bone deformation and fracture consistency compared to physical methods.

## Key findings

- Chemically fixed bones showed significantly reduced deformation and increased stiffness compared to fresh or physically preserved bones.
- Glycerol-preserved bones showed increased maximum load and breaking strength after osteosynthesis.
- Chemically fixed bones exhibited more consistent fracture patterns than fresh or physically preserved bones.

## Abstract

Large quantities of bone specimens are required for the mechanical testing of orthopedic and trauma implants. Currently, fresh-frozen or chemically preserved bones—such as formalin-fixed—are commonly used. This study aimed to evaluate the effects of different conservation methods on mechanical bone stability under standardized conditions.

Porcine metatarsals were divided into seven groups based on the preservation technique: fresh, fresh-frozen (− 18 °C), frozen with repeated freeze-thaw cycles, vacuum-sealed and refrigerated (4 °C), and chemically fixed using formalin, Thiel-fixation, or glycerol-based methods. All specimens underwent three-point bending tests to assess mechanical stability. Following induced fractures, bones were stabilized with a plate osteosynthesis (DCP® small fragment set, Synthes®, Germany) and retested. Bone mineral density (BMD) was also measured via dual-energy X-ray absorptiometry (DXA).

Results revealed no significant differences in BMD, maximum load, or breaking strength between groups. However, chemically fixed bones—especially formalin and glycerol—showed significantly reduced deformation, indicating increased stiffness. Additionally, chemically fixed specimens exhibited more consistent fracture patterns compared to fresh or physically preserved bones. After osteosynthesis, bones preserved with glycerol showed a significant increase in both maximum load and breaking strength.

While physical preservation methods had no notable effect on mechanical stability, chemically fixed bones exhibited altered deformation characteristics and should therefore be used selectively, depending on the research question.

## Linked entities

- **Chemicals:** formalin (PubChem CID 712), glycerol (PubChem CID 753)
- **Species:** Sus scrofa (taxon 9823)

## Full-text entities

- **Diseases:** trauma (MESH:D014947), shaft fractures (MESH:D000092504), bone loss (MESH:D001847), oblique fracture (MESH:C537736), Fracture (MESH:D050723)
- **Chemicals:** Phenoxitol (-), calcium (MESH:D002118), Formaldehyde (MESH:D005557), DCP (MESH:C580746), boric acid (MESH:C032688), Ethanol (MESH:D000431), ice (MESH:D007053), formic acid (MESH:C030544), glycerol (MESH:D005990), water (MESH:D014867)
- **Species:** Homo sapiens (human, species) [taxon 9606], Sus scrofa (pig, species) [taxon 9823]
- **Mutations:** C with 7, N 301261 M

## Full text

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## Figures

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## References

1 references — full list in the complete paper: https://tomesphere.com/paper/PMC12987770/full.md

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Source: https://tomesphere.com/paper/PMC12987770