Differences in the microstructural and mechanical qualities of semitendinosus tendon grafts between skeletally immature and mature patients in anterior cruciate ligament reconstruction.

BACKGROUND: This study aimed to investigate the microstructural and mechanical properties of semitendinosus tendon graft tissues during anterior cruciate ligament reconstruction and the clinical outcomes in skeletally immature and mature patients. METHODS: Twenty-two patients who underwent primary anterior cruciate ligament reconstruction using a hamstring tendon graft were analyzed and divided into skeletally immature (n = 7) and mature groups (n = 15) based on magnetic resonance imaging findings of the epiphyseal plate of the distal femur. Tissue samples were collected from the mid-portion of the semitendinosus tendon. The collagen fibril diameter, maximum stress, and strain at maximum stress point in the semitendinosus tendon tissues were calculated for comparison of the microstructural and mechanical properties between the two groups. Postoperative outcomes were also assessed between the two groups. RESULTS: The mean and 60th and 80th percentiles of fibril diameters in the skeletally immature group were significantly smaller than those in the mature group (65.9 ± 13.0, 73.5 ± 19.3, and 91.3 ± 27.4 nm in the skeletally immature group; and 90.3 ± 14.7, 94.0 ± 18.4, and 125.3 ± 19.9 nm in the skeletally immature group; p = 0.001, 0.024, and 0.004, respectively). Additionally, the strain at maximum stress was higher in the skeletally immature group (237.2 ± 102.4% vs. 121.5 ± 51.9%, p = 0.024). However, there was no difference in maximum stress between the skeletally immature and mature groups (19.9 ± 14.3 MPa vs. 24.5 ± 23.4 MPa, p = 0.578). Strain was negatively correlated with the mean fibril diameter and the 60th and 80th percentiles of fibril diameters, whereas stress was positively correlated with the mean fibril diameter. The skeletally immature group had a higher pivot shift test-positive rate than the mature group at the last follow-up (p = 0.023). CONCLUSION: Semitendinosus tendon graft tissues differed microstructurally and mechanically between skeletally immature and mature patients. LEVEL OF EVIDENCE: Level Ⅳ.

Investigating the histological and structural properties of tendon gel as an artificial biomaterial using the film model method in rabbits.

PURPOSE: This study aimed to evaluate the properties of tendon gel by investigating the histological and structural differences among tendon gels under different preservation periods using a rabbit model. METHODS: Forty mature female rabbits were divided into four groups, each containing ten rabbits, on the basis of in-vivo preservation periods of tendon gels (3, 5, 10, and 15 days). We created the Achilles tendon rupture models using the film model method to obtain tendon gels. Tensile stress was applied to the tendon gel to promote maturation. Histological and structural evaluations of the tendon gel were performed before and after applying the tensile force, and the results obtained from the four groups were compared. RESULTS: Although the day-3 and day-5 tendon gels before applying tensile stress were histologically more immature than the day-10 and day-15 gels, type I collagen fibers equivalent to those of normal tendons were observed in all groups after the tensile process. Based on the surface and molecular structural evaluations, the day-3 tendon gels after the tensile process were molecularly cross-linked, and thick collagen fibers similar to those present in normal tendons were observed. Structural maturation observed in the day-3 tendon gels caused by traction was hardly observed in the day-5, -10, and -15 tendon gels. CONCLUSIONS: The day-3 tendon gel had the highest regenerative potential to become a normal tendon by applying a traction force.

Appropriate Tensile Mode and Timing of Applying Tension to Promote Tendon Gel Regeneration.

"Tendon gel" secreted from a parent tendon is regenerated for tendon repair by applying tension. However, the details of the tensile stimulus have not been clarified. This study aimed to evaluate an appropriate tensile stimulus mode and the optimal timing of applying tension to promote tendon gel regeneration. Tendon gel was prepared using a film model method in mice and was preserved in vivo for 3, 5, and 10 days. Unlike tendon gel on day 3 or day 5, a fibrous structure developed in the tendon gel on day 10 when tension was applied. Infrared spectroscopy revealed that characteristic peaks appearing for the tendon gel on days 3 and 5 disappeared on day 10. Disappearance of the peaks indicated maturity of the tendon gel, and it showed the optimal timing for tension application to the tendon gel. The effect of tensile load on tendon gel preserved for 10 days was investigated using a tensile test, a creep test, or a cycle test. In the tensile test, tendon gel was elongated into a thin cord of collagen fibers with an increase in stress, and the maximum diameter of the collagen fiber was approximately 50 times larger than that in the normal Achilles tendon of mice. The results suggest that the diameter of the oriented collagen fiber is controllable by adjusting the applied load and the time in mature tendon gel.

Mechanisms of collagen fibril alignment in tendon injury: from tendon regeneration to artificial tendon.

The process by which collagen fibrils are aligned following tendon injury remains unknown. Therefore, we analyzed the process of tendon regeneration by transmission electron microscopy, using a film model method. In mice, the Achilles tendon of medial head was transected. On day 3, after only the proximal end of the transected tendon was placed on film and kept in vivo, a translucent substance containing granules, called tendon gel, was secreted. On day 5, the granules assembled in a loose (L) layer, and coalesced tightly in a dense (D) layer, forming an L-D-L layered pattern. On day 10, granules showed high electron density in H layers, which developed into D-H-D layers on day 13. The distal end was placed on film to face the proximal end. On day 10, the tendon gel showed a D-H-D layer pattern. On day 11, mechanical stress from muscular constriction changed the tendon gel to aligned collagen fibrils (6 ± 2 nm in diameter). Thereafter, the diameter of the fibrils increased. Tendon gel harvested on day 5 or day 10 was pulled manually or by hanging weights (about 0.6 MPa). Aligned collagen fibrils (32 ± 7 nm in diameter) were created by traction using tendon gel harvested on day 10.