The results show that a chronic tear of the human rotator cuff can be imitated in the rat model with some exclusion. The rapid self-healing response in the rat and the fatty infiltration of the human muscle are the main differences. However, tendon degeneration, inflammation and muscle atrophy combined with a persisting defect at 3 weeks after detachment are comparable to the chronic tendon tears in humans. This model can serve as a basis for further research in the field of rotator cuff repair for chronic lesions.
The acellularization of tendons using detergents (sodium dodecyl sulfate, Triton-X, tri-nitro-butyl-phosphate) is a new source of scaffolds for tissue engineering in anterior cruciate ligament (ACL) repair. In vitro testing demonstrated that acellular tendon scaffolds are biocompatible and show good biomechanical properties, but in vivo confirmation of these results is not yet available. Therefore, the aim of this study was to see in vivo if an acellular allogenic construct colonized with autologous fibroblasts improves the quality of ACL reconstruction. ACL replacement was performed in 31 New Zealand White rabbits using a standardized model. Fifteen animals received autologous semitendinosus tendon, whereas 16 animals were treated with a tissue-engineered construct. This construct was made by acellularization of allogenic semitendinosus tendons using sodium dodecyl sulfate and subsequent in vitro colonization with autologous fibroblasts. Eight weeks postoperatively, macroscopic, biomechanical (ultimate load to failure, elongation, stiffness; n = 8/9), and histological (n = 5) examinations were performed. Biomechanical testing showed decreasing strength of the constructs at 8 weeks after implantation compared with the direct postsurgical strength. However, tissue-engineered constructs (F = 19.7 +/- 20.3 N) were significantly weaker than autologous tendons (F = 61.2 +/- 31.2 N). Histologically, the autologous tendons showed signs of partial necrosis and tissue remodeling. The tissue-engineered constructs exhibited an inflammatory reaction and showed both repopulated and acellular regions. In conclusion, in vivo results were much more unfavorable than in vitro results had suggested. Further studies have to be performed to test if modifications of the acellularization process yield better results in vivo.
Polychrome sequential labeling with fluorochromes is a standard technique for the investigation of bone formation and regeneration processes in vivo. However, for human application, only tetracycline and its derivates are approved as fluorochromes. Therefore, the aim of this study was to determine the fluorescence characteristics of the different tetracycline derivates to assess the feasibility of sequential in vivo bone labeling using distinguishable fluorochromes. Eight different tetracycline derivates were injected subcutaneously into growing rats as a single dose or sequentially in different combinations. After preparation of resin-embedded undecalcified bone sections, the fluorescence properties of the tetracycline derivates in bone were analyzed using conventional fluorescence microscopy, spectral image analysis and confocal laser scanning microscopy. Each tetracycline derivate exhibited a characteristic fluorescence spectrum, but the differences between them were small. Chlortetracycline could be discriminated reliably from all other derivates and could therefore be combined with any other tetracycline derivate for reliably distinguishable double labeling. Tetracycline itself exhibited the brightest fluorescence of all the investigated derivates. Interestingly, in conventional microscopy the same tetracycline derivative can appear in different colours to the human eye, even if spectral analysis confirmed identical emission peaks. In conclusion, the data suggest that fluorescence double labeling of bone is feasible using appropriate tetracycline derivates in combination with spectral imaging modalities.
COPROGs co-lyophilized with fibrinogen are a simple basis for an injectable fibrin gluebased gene-activated matrix. The preparation can be used is complete analogy to fibrin glue preparations that are used in the clinics. However, further improvements in transgene expression levels and persistence are required to yield cartilage regeneration in the osteochondral defect model chosen in this study.
ABSTRACT:The purpose was to evaluate histological changes of the supraspinatus tendon (SSP) after refixation under continuous growth factor application over 20 days in comparison to the native healing process. In a chronic rat tendon tear model (15 rats/group), a transosseous SSP refixation was performed and growth factors (control, G-CSF, b-FGF, combination) were continuously released into the subacromial space by an osmotic pump. Tendon healing was evaluated histologically by a modified MOVIN-Score, and Collagen I/ III content was determined by immunohistology at 6 weeks. A modified MOVIN sum score showed significant lower counts for G-CSF and b-FGF in comparison to the control group (p ¼ 0.050/p ¼ 0.027) and the combined group (p ¼ 0.050/p ¼ 0.043). Collagen III was significantly reduced in the combined group compared to the control group (p ¼ 0.028). Collagen I showed no significant differences. The Collagen I/III ratio was nearly doubled for b-FGF and the combined group compared to the control. At the study endpoint, 33% of pump dislocations were detected. The continuous application of both isolated growth factors (G-CSF/b-FGF) achieved improved tendonremodeling. However, the continuous application via an osmotic pump showed a relative high dislocation rate when applied in the rat model. ß
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