Quantitative assessment of neural elements in a rat model using nerve growth factor after remnant-preserving anterior cruciate ligament reconstruction: a histological and immunofluorescence pilot study.

BACKGROUND: Immunofluorescence analyses of anterior cruciate ligament (ACL) allografts following remnant-preserving ACL reconstruction using Achilles tendon allografts have provided evidence for the presence of neural elements. In this study, we aimed to examine the expression of neural elements and quantify the presence of neural cells in ACL remnants and Achilles allografts using nerve growth factor (NGF) therapy after remnant-preserving ACL reconstruction. METHODS: Experiments were conducted on 5 pairs of rats (approximately 8 weeks old and weighing 320 g at the time of surgery). Longitudinally, split Achilles tendons from the paired rats were freshly frozen and later defrosted with warm saline and allografted onto the right ACL of the other rat that was partially detached at the femoral attachment site. A sham operation was conducted on the left knee to be used as a control. NGF was injected into both knee joints every week for 6 weeks after surgery. The presence of neural cells in the ACL of the sham-operated knee, allografted Achilles tendon, and ACL remnant was examined 6 weeks post-surgery using H and E and immunofluorescent staining. RESULTS: H and E staining did not reveal neural cells in any of the three groups. However, immunofluorescence analysis showed the presence of nestin-positive neural elements in the normal ACL tissues as well as ACL remnants. Additionally, neural elements were examined in 7 of the 8 (87.5%) allograft tissues. Quantitative analysis showed no difference in the number or area of nuclei among the three groups. However, the number and area of neural cells in the Achilles allografts were significantly lower than those in the other two groups (p = 0.000 and p = 0.001, respectively). CONCLUSION: Our observations indicate that ACL remnants promote the new ingrowth and persistence of neural cells. We suggest that the ingrowth of neural elements can support the persistence and new ingrowth of mechanoreceptors, thereby enhancing the functional stability of knee joints. Moreover, the expression of neural cells in the Achilles allografts was lower than that in normal ACL tissues and ACL remnants in the quantitative evaluation, thereby confirming the essential role of ACL remnants in knee joint functionalization.

Histological assessment of mechanoreceptors in Achilles allografts after anterior cruciate ligament reconstruction.

BACKGROUND: There is a lack of histological studies investigating the presence of mechanoreceptors in anterior cruciate ligament (ACL) allografts. HYPOTHESIS: Mechanoreceptors would not grow in Achilles allografts after ACL reconstruction. STUDY DESIGN: Case series study; Level of evidence, 4. METHODS: Tissue samples were obtained from 11 patients who underwent ACL reconstruction using Achilles tendon allografts. They underwent biopsies during second-look arthroscopies. The mean period from ACL reconstruction to harvesting tissue was 26.63 months (range, 12-120 months). The control group consisted of 2 normal ACLs procured from 42- and 45-year-old men who underwent amputation above the knee due to trauma. RESULTS: Ruffini corpuscles and free nerve endings were shown to be present in the specimens of the control group by processing hematoxylin-eosin stains and immunohistochemical stains with monoclonal antibodies against S-100. In the Achilles allografts, mechanoreceptors were not observed. However, fibroblasts, collagen fibers, and vessels that were not present in fresh-frozen Achilles allografts before surgery were observed. CONCLUSION: The results demonstrate that Achilles tendon allografts appeared similar to normal ligaments except for the lack of histological evidence of mechanoreceptors. In other words, there are no newly ingrown mechanoreceptors in ACL allografts.