Comparative study of traumatic heterotopic ossification in mice induced by Achilles tenotomy combined with skin burn injury and single Achilles tenotomy

doi:10.12307/2023.432

Abstract

Abstract: BACKGROUND: At present, Achilles tenotomy model is an animal model mainly used for traumatic heterotopic ossification. However, this method requires a long time to form ectopic bone, and the size of the formed ectopic bone is always small. In addition, this method cannot accurately reproduce the systemic inflammatory state of most traumatic heterotopic ossification cases in clinic practice.
OBJECTIVE: To verify the validity of the animal model of traumatic heterotopic ossification induced by Achilles tenotomy combined with skin burn injury, to compare this approach with single Achilles tenotomy, and to evaluate the practicability of the two methods.
METHODS: Forty male C57BL/6 mice were randomly divided into two groups: Achilles tenotomy group (control group, n=20) and Achilles tenotomy+30% skin scald on the back group (experimental group, n=20). The survival rate and healing of surgical incisions of the mice in the two groups were recorded. Survival rate and wound healing in the two groups as well as skin recovery of burn injury in the experimental group were recorded. Micro-CT examination and Masson staining of the Achilles tendon was performed 8 weeks after surgery to observe the ectopic bone at the surgical site. Formation of ectopic bone was also observed in the two groups.
RESULTS AND CONCLUSION: There was no death and wound infection in the two groups. The skin burn injury in the experimental group recovered well without ulceration. Micro-CT findings indicated that all mice in the experimental group developed traumatic heterotopic ossification, with obvious circular high-density shadow at the surgical site, and the volume of ectopic bone was (2.72±0.04) mm3. In contrast, only 17 mice developed traumatic heterotopic ossification in the control group, and the volume of ectopic bone was (0.65±0.08) mm3. There was a significant difference in the volume of ectopic bone between the two groups (P < 0.05). Masson staining showed that ectopic bone in both groups had bone trabecular and bone marrow structures, but the area of ectopic bone in the experimental group was significantly larger than that in the control group (P < 0.05). To conclude, Achilles tenotomy combined with skin burn injury can effectively induce traumatic heterotopic ossification earlier than single Achilles tenotomy in mice. This combination method has a higher successful rate and can produce larger size of ectopic bone, which can be an ideal method to establish an animal model of traumatic heterotopic ossification.

Key words: traumatic heterotopic ossification, animal model, Achilles tenotomy, burn injury, inflammation

CLC Number:

Wang Zheng, Chen Hongshu, Yi Xinzeyu, Li Zonghuan. Comparative study of traumatic heterotopic ossification in mice induced by Achilles tenotomy combined with skin burn injury and single Achilles tenotomy[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(17): 2664-2668.

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