肌腱-骨界面（TBI）整合不良是导致前交叉韧带（ACL）重建后患者愈合质量不佳的主要原因之一。最近发现H型血管通过调节骨-血管生成串扰密切调节骨形成。因此，有利于H型血管形成的策略可能是改善移植物骨整合的有希望的治疗方法。在这项研究中,我们首次报道了狭缝引导配体3（slit3）的治疗结果，一种有利于H型血管形成的新型促血管生成因子，在ACL重建小鼠的TBI愈合中。将小鼠（n=87）分为三组进行各种治疗：水凝胶微粒（HMP，对照组)，slit3@HMP,和slit3中和抗体@HMP(slit3-AB@HMP)。组织学分析,步态表现,射线照相测量，并进行生物力学测试以评估TBI愈合质量。与HMP组相比，在Slit3@HMP组中，在TBI处形成了骨向内生长增加和纤维瘢痕组织减少。同时,相对于HMP组，slit3-AB@HMP抑制骨向内生长并增加纤维瘢痕组织的形成。与HMP组相比，slit3@HMP有利于TBI的H型血管形成，而slit3-AB@HMP阻碍了它。根据Micro-CT评估,与HMP组相比，slit3@HMP显着增加了隧道周围的骨量，而slit3-AB@HMP显着减少了隧道周围的骨量。Slit3@HMP组中的小鼠在站立时间方面表现出最佳的步态表现，步幅长度,爪印区域,和立场压力。动态松弛度测量和拉伸测试显示，相对于其他两组，slit3@HMP组表现出显着降低的松弛位移和改善的破坏载荷和刚度。总的来说，注射Slit3可用于增强腱-骨整合，这可能归因于H型血管耦合的血管生成-成骨串扰的调节。
Poor tendon-bone interface (TBI) integration is one of the major causes contributing to unsatisfactory healing quality in patients after anterior cruciate ligament (ACL) reconstruction. Type H vessels have been recently found to closely modulate bone formation via regulation of the osteo-angiogenic crosstalk, so the strategies favoring type H vessel formation may be promising therapeutic approaches for improved graft osteointegration. In this
study, we reported for the first time the treatment outcome of slit guidance ligand 3 (slit3), a novel proangiogenic factor favoring type H vessel formation, in TBI healing in mice with ACL reconstruction. The mice (n = 87) were divided into three groups for various treatments: hydrogel microparticles (HMP, control group), slit3@
HMP, and slit3 neutralizing antibody@
HMP (slit3-AB@
HMP). Histological analysis, gait performance, radiographic measurement, and biomechanical testing were performed to assess the TBI healing quality. Increased bony ingrowth and reduced fibrous scar tissue was formed at the TBI in the slit3@HMP group when compared to the
HMP group. Meanwhile, the slit3-AB@HMP inhibited the osseous ingrowth and increased fibrous scar tissue formation relative to the
HMP group. Compared to the HMP group, the slit3@HMP favored type H vessel formation at the TBI while the slit3-AB@
HMP impeded it. According to micro-CT assessment, compared to the HMP group, the slit3@HMP significantly increased the peri-tunnel bone mass while the slit3-AB@HMP significantly reduced the peri-tunnel bone mass. The mice in the slit3@HMP group showed the best gait performance in terms of stance time, stride length, paw print area, and stance pressure. Dynamic laxity measurement and tensile testing showed the slit3@HMP group exhibited significantly reduced laxity displacement and improved failure load and stiffness relative to the other two groups. Collectively, the injection of slit3 could be used to enhance tendon-bone integration, which may be ascribed to modulation of angiogenesis-osteogenesis crosstalk coupled by type H vessels.
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