Effects of immobilization on thickness of superficial zone of articular cartilage of patella in rats

Background

Articular cartilage normally functions as a load-bearing resistant material in joints. Patella is composed of hyaline cartilage and spongy bone. Chondrocytes form only 1–5% volume of the articular cartilage. They receive their nutrition by diffusion through the matrix. The alteration in articular cartilage become apparent following immobilization, from 4 to 6 weeks. Until now, focus of research has been the whole cartilage. Zonal changes have not been studied in detail. Since superficial zone bears maximum load and is the first zone to come in contact, the present study was designed to determine changes in thickness on immobilization and remobilization in superficial zone after dividing it into proximal, central, and distal segments.

Materials and Methods

Forty male rats belonging to Sprague Dawley strain were divided into two groups. Group 1 (n=20) subdivided into an experimental subgroup of 10 rats that were immobilized in plaster of Paris (POP) for 4 weeks and a control subgroup of 10 that were not immobilized. Group 2 (n=20) subdivided into an experimental subgroup of 10 rats that were immobilized for 4 weeks and remobilized for next 4 weeks and a control subgroup of 10 animals that were not immobilized. At the end of the experimental period, the knee joint was dissected and was cut in sagittal plane. The section was fixed in 10% formalin for 48 hours. Specimen was decalcified using ethylenediaminetetraacetic acid (EDTA). The paraffin blocks of 7 µm sections were cut and stained by H and E stain for routine histology and Alcian blue stain and Mallory Trichrome for fine structural microscopy. The zones were named as superficial transitional, radial, and hypertrophic according to the shape of cells present in each zone. The superficial zone was divided into superior part, central, and inferior parts. These parts were labeled as central, proximal, and distal segments. The calibrated stage micrometer was used to calibrate the ocular micrometer under objectives of different power. The ocular micrometer was placed inside the ocular lens. It was calibrated with the stage micrometer under objective lenses of different power. The number of divisions of ocular covering each zone was calculated. These divisions were converted into micrometer and the actual thickness was calculated.

Results

The significant decrease in thickness of superficial zone in proximal, central and distal segment was observed in experimental group in comparison to control group. When the experimental subgroup of group 2 was compared with experimental subgroup of group 1 (group immobilized for 4 weeks), no significant reversal was seen in superficial zone and instead significant decrease was observed in distal segment. Fibrous connective tissue was increased adjacent to superficial zone.

Conclusion

Each segment of superficial zone behaves differentially on immobilization and remobilization. Perhaps a much longer duration of remobilization is required to reverse changes of immobilization in articular cartilage and plays a significant role in knee joint movements.