Willem van der Merwe

The COL5A1 Gene Is Associated With Increased Risk of Anterior Cruciate Ligament Ruptures in Female Participants

Background Anterior cruciate ligament ruptures, especially to young female athletes, are a cause of major concern in the sports medicine fraternity. The major structural constituents of ligaments are collagens, specifically types I and V. Recently, the gene that encodes for the α1 chain of type I collagen (COL1A1) has been shown to be associated with an increased risk of cruciate ligament ruptures. The COL5A1 gene, which encodes for the α1 chain of type V collagen, has been shown to be associated with Achilles tendon injuries. Purpose The study was conducted to determine (1) if 2 sequence variants (BstUI and DpnII restriction fragment length polymorphisms [RFLPs]) within the COL5A1 gene are associated with an increased risk of anterior cruciate ligament ruptures, and (2) if there were any gender-specific positive associations between the 2 COL5A1 sequence variants and risk of anterior cruciate ligament ruptures. Study Design Case control study; Level of evidence, 3. Methods A total of 129 white participants (38 women) with surgically diagnosed anterior cruciate ligament ruptures and 216 physically active control participants (84 women) without any history of ACL injury were included in this case-control genetic association study. All participants were genotyped for the COL5A1 BstUI and DpnII RFLPs. Results There was a significant difference in the BstUI RFLP genotype frequency between the anterior cruciate ligament rupture and physically active control groups among the female participants, but not the male participants. The CC genotype in the female participants was significantly underrepresented in the anterior cruciate ligament rupture group compared with the controls (27.4% vs 5.6%; odds ratio = 6.6; 95% confidence interval, 1.5–29.7; P = .006). There were no differences in the DpnII RFLP genotype distributions between the anterior cruciate ligament rupture and physically active control groups. Conclusion The CC genotype of the COL5A1 BstUI RFLP was underrepresented in female participants with anterior cruciate ligament ruptures. Clinical Relevance This is the first study to show that there is a specific genetic risk factor associated with risk of anterior cruciate ligament ruptures in female athletes.

The association between the COL12A1 gene and anterior cruciate ligament ruptures

Background Anterior cruciate ligament (ACL) ruptures are among the most severe musculoskeletal soft tissue injuries. However, the exact mechanisms which cause these acute injuries are unknown. Recently, sequence variants within two genes, namely COL1A1 and COL5A1, which code for the α1 chains of types I and V collagen respectively, were shown to be associated with ACL ruptures. Type XII collagen, similarly to types I and V collagen, is a structural component of the ligament fibril and is encoded by a single gene, COL12A1. Objective The aim of this study was to investigate whether sequence variants within COL12A1 are associated with ACL ruptures. Methods One hundred and twenty-nine (38 female) participants with clinically and surgically diagnosed ACL ruptures, as well as 216 (83 female) physically active controls participants (CON) without any history of ACL injury were included in this case-control genetic association study. All participants were genotyped for the AluI and BsrI restriction fragment length polymorphisms (RFLPs) within COL12A1. Results The AA genotype of the COL12A1 AluI RFLP was significantly over-represented in the female (OR=2.4, 95% CI 1.0 to 5.5, p=0.048), but not male (p=0.359) ACL participants. There were no genotype differences between the ACL and CON group for the BsrI RFLP. Conclusion The COL12A1 AluI RFLP is associated with ACL ruptures among female participants in this study. The results suggest that females with an AA genotype are at increased risk of ACL ruptures. These initial genetic association studies should be explored further and, if repeated, incorporated into multifactorial models developed to identify predisposed individuals.

Knee Rotational Laxity in a Randomized Comparison of Single- Versus Double-Bundle Anterior Cruciate Ligament Reconstruction

Background: While single-bundle anterior cruciate ligament reconstruction reduces anterior-posterior laxity, studies have demonstrated residual rotational instability. Improved pivot-shift results have been shown with the double-bundle graft; however, no study has compared rotational laxity outcome of these surgical techniques in vivo under quantified, isolated torsional loading. Hypothesis: The anterior cruciate ligament–deficient knee exhibits greater rotational laxity than the contralateral uninjured knee. The double-bundle reconstruction restores rotational joint stability to a greater extent than single-bundle surgery. Study Design: Controlled laboratory study. Methods: Rotational laxity of 32 patients with unilateral anterior cruciate ligament injury was assessed in both knees at full extension and 30° of flexion using a magnetic resonance imaging–compatible torsional loading device. Patients were randomly allocated either a single- or double-bundle reconstruction and reassessed 5 months after surgery. Results: The anterior cruciate ligament–deficient knees demonstrated greater laxity to internal rotational torque in the extended position, but not in the 30° flexed position. No significant differences in rotational laxity were found between single- and double-bundle reconstructions. In extension, excessive internal rotational laxity of injured compared with contralateral knees was reduced by anterior cruciate ligament reconstruction. The single-bundle reconstruction did not affect internal rotation compared with contralateral or preoperative groups. In response to internal rotational torque in the flexed knee position, the double-bundle reconstruction reduced laxity to 10.8° from the pre-operative value of 15.3° (P = .058); postoperative rotation was also significantly less than the contralateral laxity of 16.4° (P = .022). Conclusion: The ruptured anterior cruciate ligament resulted in increased internal rotational laxity only in the extended position. The single-bundle reconstruction did not affect rotational restraint compared with contralateral or preoperative groups. The double-bundle procedure significantly reduced internal laxity in the flexed position when compared with normal. Clinical Relevance: As the anterior cruciate ligament is not the primary restraint to rotation, its contribution to joint stability is limited under isolated torsional load. While the double-bundle graft demonstrates superior rotational constraint, this may be excessive for isolated anterior cruciate ligament rupture.

Rotatory knee laxity tests and the pivot shift as tools for ACL treatment algorithm.

AbstractThe goal of anterior cruciate ligament (ACL) reconstruction surgery is to eliminate the pivot shift phenomenon. Different injury mechanisms and injury patterns may lead to specific knee laxity patterns. Computer navigation is helpful for the surgeon during examination under anesthesia. Surgical treatment may have to be altered if high-grade laxity is detected preoperatively for example by utilizing a computer navigation that is a helpful adjunct for surgeons during examination under anesthesia. A typical case for revision ACL reconstruction is presented. This article describes several techniques of laxity assessments. Based on the type and degree of pathologic laxity, a treatment algorithm has been developed. Level of evidence V.

The association of genes involved in the angiogenesis‐associated signaling pathway with risk of anterior cruciate ligament rupture

Angiogenesis‐associated signaling is a fundamental component in the remodeling of the extracellular matrix in response to loading. Genes encoding protein components within this signaling cascade are therefore suitable candidates for investigation into ACL injury susceptibility: namely, vascular endothelial growth factor A isoform (VEGFA), kinase insert‐domain receptor (KDR), nerve growth factor (NGF), and hypoxia inducible factor‐1α (HIF1A). A case‐control genetic association study was conducted on 227 asymptomatic control participants and 227 participants with surgically diagnosed ACL ruptures of which 126 participants reported a non‐contact mechanism of rupture. All participants were genotyped for seven polymorphisms within the four genes. The VEGFA rs699947 CC genotype (p = 0.010, OR: 1.92, 95% CI: 1.17–3.17) was significantly over‐represented within participants with non‐contact ACL ruptures. The VEGFA rs1570360 GA genotype was significantly over‐represented in the CON group (p = 0.007, OR: 1.70, 95% CI: 1.16–2.50). Furthermore, the KDR rs2071559 GA genotype was significantly over‐represented in the female controls (p = 0.023, OR: 2.16, 95% CI: 1.11–4.22). Inferred haplotype analyses also implicated genomic regions spanning the VEGFA and KDR genes. These novel findings suggest that regions within VEGFA and KDR may be implicated in the pathophysiology of ACL ruptures; highlighting the potential biological significance of angiogenesis‐associated signaling in the aetiology of ACL ruptures.

Genes encoding proteoglycans are associated with the risk of anterior cruciate ligament ruptures

Background Genetic variants within genes involved in fibrillogenesis have previously been implicated in anterior cruciate ligament (ACL) injury susceptibility. Proteoglycans also have important functions in fibrillogenesis and maintaining the structural integrity of ligaments. Genes encoding proteoglycans are plausible candidates to be investigated for associations with ACL injury susceptibility; polymorphisms within genes encoding the proteoglycans aggrecan (ACAN), biglycan (BGN), decorin (DCN), fibromodulin (FMOD) and lumican (LUM) were examined. Methods A case–control genetic association study was conducted. 227 participants with surgically diagnosed ACL ruptures (ACL group) and 234 controls without any history of ACL injury were genotyped for 10 polymorphisms in 5 proteoglycan genes. Inferred haplotypes were constructed for specific regions. Results The G allele of ACAN rs1516797 was significantly under-represented in the controls (p=0.024; OR=0.72; 95% CI 0.55 to 0.96) compared with the ACL group. For DCN rs516115, the GG genotype was significantly over-represented in female controls (p=0.015; OR=9.231; 95%CI 1.16 to 73.01) compared with the ACL group and the AA genotype was significantly under-represented in controls (p=0.013; OR=0.33; 95% CI 0.14 to 0.78) compared with the female non-contact ACL injury subgroup. Haplotype analyses implicated regions overlapping ACAN (rs2351491 C>T-rs1042631 T>C-rs1516797 T>G), BGN (rs1126499 C>T-rs1042103 G>A) and LUM-DCN (rs2268578 T>C-rs13312816 A>T-rs516115 A>G) in ACL injury susceptibility. Conclusions These independent associations and haplotype analyses suggest that regions within ACAN, BGN, DCN and a region spanning LUM–DCN are associated with ACL injury susceptibility. Taking into account the functions of these genes, it is reasonable to propose that genetic sequence variability within the genes encoding proteoglycans may potentially modulate the ligament fibril properties.

Interactions between collagen gene variants and risk of anterior cruciate ligament rupture

Abstract The COL5A1 and COL12A1 variants are independently associated with modulating the risk of anterior cruciate ligament (ACL) rupture in females. The objective of this study was to further investigate if COL3A1 and COL6A1 variants independently, as well as, collagen gene–gene interactions, modulate ACL rupture risk. Three hundred and thirty-three South African (SA, n = 242) and Polish (PL, n = 91) participants with diagnosed ACL ruptures and 378 controls (235 SA and 143 PL) were recruited. Participants were genotyped for COL3A1 rs1800255 G/A, COL5A1 rs12722 (T/C), COL6A1 rs35796750 (T/C) and COL12A1 rs970547 (A/G). No significant associations were identified between COL6A1 rs35796750 and COL3A1 rs1800255 genotypes and risk of ACL rupture in the SA cohort. The COL3A1 AA genotype was, however, significantly (p = 0.036) over-represented in the PL ACL group (9.9%, n = 9) when compared to the PL control (CON) group (2.8%, n = 4). Although there were genotype distribution differences between the SA and PL cohorts, the T+A-inferred pseudo-haplotype constructed from COL5A1 and COL12A1 was significantly over-represented in the female ACL group when compared to the female CON group within the SA (T+A ACL 50.5%, T+A CON 38.1%, p = 0.022), PL (T+A ACL 56.3%, T+A CON 36.3%, p = 0.029) and combined (T+A ACL 51.8%, T+A CON 37.5%, p = 0.004) cohorts. In conclusion, the novel main finding of this study was a significant interaction between the COL5A1 rs12722 T/C and COL12A1 rs970547 A/G variants and risk of ACL injury. These results highlight the importance of investigating gene–gene interactions in the aetiology of ACL ruptures in multiple independent cohorts.

Kinematic and kinetic gait deviations in males long after anterior cruciate ligament reconstruction

Background: Biomechanical deviations long (approx. 5 years) after anterior cruciate ligament reconstruction have not been quantified in males, despite their distinct risk profile as compared to females. These deviations can indicate altered joint loading during chronic, repetitive motions. Methods: Cross‐sectional study, comparing kinematic and kinetic variables between 15 male anterior cruciate ligament reconstructed patients and 15 healthy controls. During walking and running gait, measurements were taken of impact dynamics, knee and hip sagittal plane angles and moments, and knee varus angles and adduction moments. Findings: Comparing affected limbs to control limbs, significantly lower maximum (P = 0.001) and initial (P = 0.003) loading rates were found during running, but not in walking. Hip angles were lower for affected limbs of patients compared to the control group (P = 0.039) in walking, but not during running. Between‐limb comparisons showed important differences in symmetry of the affected patients. Maximum force during running was higher in the unaffected limb (P = 0.015), which was linked with a higher loading rate (P = 0.008). Knee flexion angle was reduced by 2° on average for the affected limb during running (P = 0.010), and both walking and running knee and hip moments showed differences. Knee varus angle showed a 1° difference during walking (P < 0.001), but not during running. Knee adduction moment was significantly lower (more valgus) during both walking and running. Interpretation: Male anterior cruciate ligament reconstructed patients demonstrate persistent, clinically important gait asymmetries and differences from healthy controls long after surgery in kinematics, kinetics, and impact biomechanics. HIGHLIGHTSAnterior cruciate ligament reconstructed females show persistent gait deviations.No previous research has investigated these during walking/running in male patients.Knee adduction moments and angles were lower in affected than unaffected limbs.Reduced knee flexion angles and moments were also found, but only during running.Deviations from controls were also found in impact during foot strike.

Investigation of angiogenesis genes with anterior cruciate ligament rupture risk in a South African population

ABSTRACT The angiogenesis-signalling pathway is a physiological response after mechanical loading to promote matrix remodelling and thereby maintain tissue homeostasis. Studies have shown increased expression of angiogenic molecules in response to loading and in ruptured ligaments. Recently, polymorphisms within the vascular endothelial growth factor A (VEGFA) and kinase insert-domain receptor (KDR) genes were associated with risk of anterior cruciate ligament (ACL) ruptures and Achilles tendinopathy in Caucasian study groups. A case-control genetic association study was conducted on 100 controls and 98 participants with surgically-diagnosed ACL ruptures; of which 51 participants reported non-contact mechanism of injury (NON). All participants were genotyped for five functional polymorphisms: VEGFA (rs699947, rs1570360, rs2010963) and KDR (rs2071559, rs1870377). Haplotypes were inferred. In the male participants, the KDR rs2071559 AG genotype was significantly over-represented (P = 0.048, OR: 1.90, 95% CI: 1.00–3.59) in the controls. Furthermore, the GG genotype was significantly under-represented in the male controls compared to the male ACL group (P = 0.018, OR: 2.77, 95% CI: 1.17–6.55) and the male NON subgroup (P = 0.013, OR: 3.26, 95% CI: 1.24–8.58). Haplotype analysis implicated the KDR gene in all participants and in male participants separately. Collectively, these results implicate the angiogenesis-signalling pathway as a potentially key biological pathway contributing to ACL injury susceptibility.

Modulators of the extracellular matrix and risk of anterior cruciate ligament ruptures

OBJECTIVES The extracellular matrix (ECM) of ligaments continuously undergoes remodelling in order to maintain tissue homeostasis. Several key mediators of ECM remodelling were chosen for investigation in the present study. It is thought that polymorphisms within genes encoding signalling molecules may contribute to inter-individual variation in the responses to mechanical loading, potentially altering risk of injury. DESIGN A genetic association study was conducted on 232 asymptomatic controls (CON) and 234 participants with surgically diagnosed anterior cruciate ligament (ACL) ruptures; of which 135 participants reported a non-contact mechanism of injury (NON subgroup). METHODS All participants were genotyped for ten variants in eight genes encoding ECM remodelling proteins. Haplotypes and allele combinations were also inferred. RESULTS The CASP8 rs3834129 ins allele was significantly over-represented in the male CON group compared to the male NON subgroup (p=0.047, OR: 1.46, 95% CI: 1.01-2.12). In female participants, the IL1B rs16944 TT genotype was significantly under-represented in the CON group compared to the NON subgroup (p=0.039, OR: 3.06, 95% CI: 1.09-8.64). Haplotype analysis revealed an under-representation of the CASP8 rs3834129-rs1045485 del-G haplotype in the CON group compared to both the ACL group (p=0.042; haplo.score:2.03) and the NON subgroup (p=0.037; haplo.score:2.09). Furthermore, following a pathway-based approach, genetic variants involved in the cell signalling cascade were associated with ACL injury risk. CONCLUSIONS The novel independent associations and allele combinations observed implicate the apoptosis and cell signalling cascades as potential contributors to ACL injury susceptibility. Furthermore, these genetic variants may potentially modulate ECM remodelling in response to loading and ultimately contribute to ligament capacity.