Summary

This artical discussed whether Pletelet lysate combined with adipose-derived stem cells will improve rat tendon-bone healing and what kind of fibro cartilage develepment at the tendon bone juncture.

Abstract

Objective

To discuss whether Pletelet lysate combined with adipose-derived stem cells will improve rat tendon-bone healing. To observe fibro cartilage develepment at the tendon bone juncture and evaluate tendon-bone healing from histoloy and biomechanic.

Methods

The rat model of tendon-bone healing of Achilles tendon was established. Achilles tendon-bone interface after cleanup divided into three groups, sham operation group (G1), ADSC group (G2), ADSC +% PL group (G3), respectively, for treatment. Then use the HE staining, Masson staining and Alcian blue staining for histological observation. Tendon bone tissue were extracted when performed qPCR detection of type II collagen content at 2w, 4w, 6w. And the determination of biomechanical changes were performed at 2w, 4w, 6w.

Results

Sharpy’s fibers were formed at G1 group in HE staining, as the tide mark founded at G2 after 6w, for G3, 4w. The typical structure of four- layer conjunction was founded when 6w. Masson staining: G1 group was loose fibrous tissue disorders, collagen fiber wave -like , messy arrangement . G2 group was better aligned fibrous tissue , collagen fibers clear boundary . The G3 Group neatly arranged collagen fibers , calcified cartilage and fibrocartilage tightly . Alcian blue staining: G1 group seen tendon-bone healing after 6w, blue-stained cells around the cartilage , calcified cartilage Department has calcium deposits , G2 group seen more visible blue dye around the cartilage , chondrocytes oval, and G3 groups seen around chondrocytes blue dye darker , indicating a rich proteoglycan content . Aizen value of each image: G3 statistical analysis shows significantly higher than other groups. QPCR detection in each time period , G1 group of type II collagen mRNA expression were significantly less than the G2 and G3 , when 2w, G2 group of type II collagen mRNA expression and G3 were similar , but were significantly less than the G3 group when 4w and 6w . The maximum tensile load lowest biomechanical test: G1 group , a significant difference compared with the normal tendon bone tissue after 2w, while a significant increase in G2 and G3 than G1. But G2 and G3 were significantly decreased compared to the maximum tensile load of normal tendon bone tissue . The maximum tensile load in each group increased with time . Maximum tensile load of groups G2 and G3 is significantly greater than G1 after 6w. While when 6w, G3 maximum tensile load was significantly higher than the normal tendon bone tissue .

Conclusion

Pletelet lysate combined with adipose-derived stem cells will promote migration and chondrogenic differentiation of ADSC, result in improved rat tendon-bone healing.