Lack of 5-lipoxygenase in intramembranous and endochondral 129Sv mice skeleton and intramembranous healing.

OBJECTIVE: To analyze the lack of 5-lipoxygenase (5LO) on dental socket healing and post-natal phenotype of intramembranous and endochondral bones. DESIGN: Wild type (WT) 129/SvEv (n = 20) and 5LO knockout (5LOKO) (n = 20) male mice underwent tooth extraction of the upper right incisor and were euthanized after 7, 14, and 30 day time points for the evaluation of dental socket healing and histological phenotyping of intramembranous (IM) and endochondral (EC) bones. Microscopic analysis of alveolar sockets included histopathological description, histomorphometry, and immunohistochemistry for 5LO, cyclooxygenase 2 (COX2), and tartrate resistant acid phosphatase (TRAP). RESULTS: Histological phenotyping revealed thicker cortical bone in EC bones (femur and vertebra) of 5LOKO mice compared to WTs, with no differences in collagenous content. Although dental socket healing was similarly observed in both groups, WT mice revealed increased numbers of COX-2+ and 5LO+ cells during bone maturing stage, with a decrease of TRAP+ cells at day 30. On the other hand, an increased quantity of fibroblasts was observed at day 7 in 5LOKO group, as well as increased inflammatory infiltrate and significantly decreased TRAP+ cells at final stages of alveolar socket healing in comparison to WTs. CONCLUSIONS: The lack of 5LO in 5LOKO mice resulted in thicker cortical of EC, but not of IM post natal bones. Furthermore, genetic deletion of 5LO in the 5LOKO mice directly affected the inflammatory response during socket healing, influencing initial and late phases of bone repair in a model of post-tooth extraction in 129Sv WT and 5LOKO mice.

New Surgical Model for Bone-Muscle Injury Reveals Age and Gender-Related Healing Patterns in the 5 Lipoxygenase (5LO) Knockout Mouse.

Signaling lipid mediators released from 5 lipoxygenase (5LO) pathways influence both bone and muscle cells, interfering in their proliferation and differentiation capacities. A major limitation to studying inflammatory signaling pathways in bone and muscle healing is the inadequacy of available animal models. We developed a surgical injury model in the vastus lateralis (VL) muscle and femur in 129/SvEv littermates mice to study simultaneous musculoskeletal (MSK) healing in male and female, young (3 months) and aged (18 months) WT mice compared to mice lacking 5LO (5LOKO). MSK defects were surgically created using a 1-mm punch device in the VA muscle followed by a 0.5-mm round defect in the femur. After days 7 and 14 post-surgery, the specimens were removed for microtomography (microCT), histopathology, and immunohistochemistry analyses. In addition, non-injured control skeletal muscles along with femur and L5 vertebrae were analyzed. Bones were microCT phenotyped, revealing that aged female WT mice presented reduced BV/TV and trabecular parameters compared to aged males and aged female 5LOKO mice. Skeletal muscles underwent a customized targeted lipidomics investigation for profiling and quantification of lipid signaling mediators (LMs), evidencing age, and gender related-differences in aged female 5LOKO mice compared to matched WT. Histological analysis revealed a suitable bone-healing process with osteoid deposition at day 7 post-surgery, followed by woven bone at day 14 post-surgery, observed in all young mice. Aged WT females displayed increased inflammatory response at day 7 post-surgery, delayed bone matrix maturation, and increased TRAP immunolabeling at day 14 post-surgery compared to 5LOKO females. Skeletal muscles of aged animals showed higher levels of inflammation in comparison to young controls at day 14 post-surgery; however, inflammatory process was attenuated in aged 5LOKO mice compared to aged WT. In conclusion, this new model shows that MSK healing is influenced by age, gender, and the 5LO pathway, which might serve as a potential target to investigate therapeutic interventions and age-related MSK diseases. Our new model is suitable for bone-muscle crosstalk studies.