Disturbed muscular architecture, fatty infiltration and muscular atrophy remain irreversible in chronic rotator cuff tears (RCT) even after repair. Poly-[ADP-ribose]-polymerase 1 (PARP-1), a nuclear factor involved in DNA damage repair, has shown to be a key element in the up-regulation of early muscle inflammation, atrophy and fat deposition. We therefore hypothesized that the absence of PARP-1 would lead to a reduction in muscular architectural damage, early inflammation, atrophy and fatty infiltration subsequent to combined tenotomy and neurectomy in a PARP-1 knock-out mouse model.

PARP-1 knock-out (KO group) and standard wild type C57BL/6 (WT group) mice were randomly allocated into three different time points (1, 6 and 12 weeks, total n=72). In all mice the supraspinatus (SSP) and infraspinatus (ISP) tendons of the left shoulder were detached and the SSP muscle was denervated according to a recently established model. Macroscopic muscle weight analysis, retraction documentation using macroscopic suture, magnetic resonance imaging, immunohistochemistry gene expression analysis using real time qPCR (RTqPCR) and histology were used to assess the differences in muscle architecture, early inflammation, fatty infiltration and atrophy between knock out and wild type mice in the supraspinatus muscle.

The SSP did retract in both groups, however; the KO muscles and tendons retracted less than the WT muscles (2.1±21mm vs 3.4±0.41mm; p=0.02). Further assessment of muscle architecture demonstrated that the pennation angle was significantly higher in the KO groups at 6 and 12 weeks (28±5 vs 36±5 and 29±4 vs 34±3; p<0.0001). Combined Tenotomy and neurectomy resulted in a significant loss of muscle mass in both groups compared to the contralateral unoperated side (KO group 62±11% and WT group 52±11%, p=0.04) at 6 weeks. But at 12 weeks postoperatively, there was a significant increase in muscle mass to near normal levels in KO group compared to the WT group (14±6% and 42±7% lower muscle mass respectively; p<0.0001) and less fatty infiltration (12.5 ± 1.82% and 19.6 ± 1.96%, p=0.027). Immunohistochemistry revealed a significant decrease in the expression of inflammatory, apoptotic, adipogenic and muscular atrophy genes at both the 1 week and 6 weeks time points, but not at 12 weeks in the KO group compared to the WT group. This was confirmed by histology.

Our study is the first to show that knocking out PARP-1 leads to decreased loss of muscle architecture, early inflammation, fatty infiltration and atrophy after combined tenotomy and neurectomy of the rotator cuff muscle. Although the macroscopic muscles reaction to injury is similar in the first 6 weeks, its ability to regenerate is much greater in the PARP-1 group leading to a near normalization of the muscle substance and muscle weight, less retraction, and less fatty infiltration after 12 weeks.

Summary

Nasal Chondrocytes are safe and feasible for tissue engineering approaches in articular cartilage repair.

Introduction: Ruptured anterior cruciate ligaments (ACL) are generally thought not to heal after treatment so that the ligament must be replaced by a graft. We developed and evaluated a surgical technique that restores knee stability using the self-healing capacity of the ruptured ACL.

Methods: The ACL was cut close to the femoral insertion in 14 sheep. The contralateral, nonoperated side served as the control. After microfracturing at the femoral foot print, the ACL was stabilized using the dynamic intraligamentary stabilization (DIS) technique. A strong intraligamentary suture was attached on the femoral side with a button and on the tibial side by means of an intraosseous spring-loaded screw with a preload of 85 N. In 7 of 14 sheep the ruptured ligament was covered with a collagen membrane. The joint was not immobilized postoperatively. Histological evaluation, magnetic resonance imaging (MRI) and biomechanical testing were performed after 3 months.

Results: Three days postoperative all animals showed no lameness and regularly used the operated leg. Macroscopically, all repaired ACLs appeared healed. Histological and MRI examinations confirmed the healing response of the ACL after DIS. The collagen membrane exhibited a more hypertrophic scare tissue reaction. Biomechanical testing showed an average increase of the operated anterior drawer 2.4 mm (range 0 mm – 4.5 mm) greater than on the control side. No lesions of the meniscus and cartilage were detected.

Conclusions: A ruptured ACL has the biological potential to heal after DIS in a sheep model. The surgical technique achieved a stable knee joint with free range of motion and full weight bearing during the healing period without signs of osteoarthritis or other intraarticular damage at follow up.

INTRODUCTION: Chronic experimental rotator cuff tears are associated with muscle retraction, atrophy, fatty infiltration, a pronounced change in the pennation angle of the muscle and consequent shortening of muscle fibres. It was the purpose of this investigation to study whether slow, continuous elongation of the musculotendinous unit can revert the pennation angle and elongate the shortened muscle fibres.

MATERIAL AND METHODS: The infraspinatus tendons of twelve sheep were released. After retraction of the tenotomised musculotendinous unit, the infra-spinatus was elongated one mm per day using a new elongation-apparatus. After restoring the approximate original length, the tendon was repaired back. Muscular architecture (retraction and pennation angle), fatty infiltration (in Hounsfield units=HU) and muscular cross sectional area (in % of the mean control side) were analyzed at start, at the time before elongation, at the time of repair and six and eighteen weeks thereafter.

RESULTS: In four sheep the elongation failed technically. In the other eight sheep, elongation could be achieved as planned. After retraction of 29mm ± 6 mm after 16 weeks (14% of original length, p=0.008), the mean traction time was 24 days ± 6 days with a mean traction distance of 19 mm ± 4 mm. At sacrifice the mean pennation angle increased in the failed sheep from 30° ± 6° up to 55° ± 14° (p=0.035). In those sheep in which traction was applied, the mean pennation angle was not different to the control side (29.8° ± 7.5° vs. 30° ± 6°, p=0.575). Compared to preoperative, there was a significant increase in fatty infiltration (36 HU, p=0.0001) and decrease of the muscular cross sectional area of 43 % (range 21% to 67%, p=0.0001) at 4 months. In the sheep in which traction could be achieved, fatty infiltration remain unchanged (36 HU ± 6 HU vs 38 HU ± 4 HU, p=0.438) and atrophy decreased by 22% (range 10% to 33%) after 6 weeks of continuous traction (p=0.008).

CONCLUSION: Continuous experimental elongation of a retracted musculotendinous unit is technically feasible and might lead to recovery of the muscle architecture, partial reversibility of atrophy and, arrest of progression of fatty infiltration.

We released the infraspinatus tendons of six sheep, allowed retraction of the musculotendinous unit over a period of 40 weeks and then performed a repair. We studied retraction of the musculotendinous unit 35 weeks later using CT, MRI and macroscopic dissection.

The tendon was retracted by a mean of 4.7 cm (3.8 to 5.1) 40 weeks after release and remained at a mean of 4.2 cm (3.3 to 4.7) 35 weeks after the repair. Retraction of the muscle was only a mean of 2.7 cm (2.0 to 3.3) and 1.7 cm (1.1 to 2.2) respectively at these two points. Thus, the musculotendinous junction had shifted distally by a mean of 2.5 cm (2.0 to 2.8) relative to the tendon. Sheep muscle showed an ability to compensate for approximately 60% of the tendon retraction in a hitherto unknown fashion. Such retraction may not be a quantitatively reliable indicator of retraction of the muscle and may overestimate the need for elongation of the musculotendinous unit during repair.