THE STRUCTURAL RESPONSE OF MUSCLE TO LIMB LENGTHENING

Abstract

Previous studies in animal models of limb lengthening have shown a wide spectrum of histopathological changes during distraction phase. Much less is known about the structural response of muscle during the consolidation phase. This study aimed to observe and score changes in morphology, weight, length and maximal perimeter of gastrocnemius during the distraction and consolidation phases.

Thirty two immature New Zealand white rabbits were divided into two equal groups: lengthening and sham. In each group, half of the rabbits were killed at the end of lengthening and half 5 weeks later. A bilateral external fixator was applied to tibia and a mid-diaphysis osteotomy performed. The lengthening rate was 0.4 mm twice daily with an initial delay of 7 days. 30% lengthening was achieved in 4 to 5 weeks. After sacrifice, the whole gastrocnemius was taken from its attachments. Its weight, length and maximal perimeter were measured. At the middle of belly, a specimen 0.5cm in length was taken from the medial gastrocnemius for H& E and Masson trichrome staining. A scoring system was used to achieve a semi-quantitative analysis of the histopathological changes in gastrocnemius.

No abnormal changes were observed in the sham side. Degeneration, atrophy and endomysial fibrosis were all found in the lengthened side. The scores of histopathological changes between the end of lengthening and 5 weeks later showed a decreasing trend, but no significant difference. The weight and perimeter decreased and length increased in the lengthening side. The weight, perimeter and length of gastrocnemius in both lengthening and control sides increased at 5 weeks after the end of lengthening.

Muscular atrophy, as shown by a decrease in weight, perimeter and muscle fibre size, occurred and might be due to the combined effect of continuous muscle stretching and inactivity. Continuous stretching of muscles beyond a certain point produced damage. Some studies reported that damage to muscle fibres, which has been shown as degeneration and fibrosis in this study, can release and activate satellite cells. As myoblast precursors, satellite cells become myoblasts, which proliferate and fuse into the microlesioned areas, regenerating and repairing myofibrils. Also, the immature muscles have more active abilities of proliferation, regeneration, growth and healing. In this study gastrocnemius growth shown by an increase in weight, perimeter and length occurred during the consolidation phase of 5 weeks. The mean scores of histopathological changes in gastrocnemius decreased during consolidation period, indicating some recovery of damage to muscle. It is not clear whether this reflects a normal response, which would have been seen in other studies had samples been taken later or whether it is a unique response of the immature animal.