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327 result(s) for "Siebert, C. H."
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Hydrolyzed fish collagen induced chondrogenic differentiation of equine adipose tissue-derived stromal cells
Adipose-derived stromal cells (ADSCs) are multipotent cells which, in the presence of appropriate stimuli, can differentiate into various lineages such as the osteogenic, adipogenic and chondrogenic. In this study, we investigated the effect of transforming growth factor beta 1 (TGF-β1) in comparison to hydrolyzed fish collagen in terms of the chondrogenic differentiation potential of ADSCs. ADSCs were isolated from subcutaneous fat of horses by liposuction. Chondrogenesis was investigated using a pellet culture system. The differentiation medium was either supplemented with TGF-β1 (5 ng/ml) or fish collagen (0.5 mg/ml) for a 3 week period. After the 3 weeks in vitro differentiation, RT-PCR and histological staining for proteoglycan synthesis and type II collagen were performed to evaluate the degree of chondrogenic differentiation and the formation of cartilaginous extracellular matrix (ECM). The differentiation of ADSCs induced by TGF-β1 showed a high expression of glycosaminoglycan (GAG). Histological analysis of cultures stimulated by hydrolyzed fish collagen demonstrated an even higher GAG expression than cultures stimulated under standard conditions by TGF-β1. The expression of cartilage-specific type II collagen and Sox9 was about the same in both stimulated cultures. In this study, chondrogenesis was as effectively induced by hydrolyzed fish collagen as it was successfully induced by TGF-β1. These findings demonstrated that hydrolyzed fish collagen alone has the potential to induce and maintain ADSCs-derived chondrogenesis. These results support the application of ADSCs in equine veterinary tissue engineering, especially for cartilage repair.
Medicinal Chemistry Based on the Sugar Code: Fundamentals of Lectinology and Experimental Strategies with Lectins as Targets
Theoretical calculations reveal that oligosaccharides are second to no other class of biochemical oligomery in terms of coding capacity. As integral part of cellular glycoconjugates they can serve as recognitive units for receptors (lectins). Having first been detected in plants, lectins are present ubiquitously. Remarkably for this field, they serve as bacterial and viral adhesins. Following a description of these branches of lectinology to illustrate history, current status and potential for medicinal chemistry, we document that lectins are involved in a wide variety of biochemical processes including intra- and intercellular glycoconjugate trafficking, initiation of signal transduction affecting e. g. growth regulation and cell adhesion in animals. It is thus justified to compare crucial carbohydrate epitopes with the postal code ensuring correct mail routing and delivery. In view of the functional relevance of lectins the design of high-affinity reagents to occupy their carbohydrate recognition domains offers the perspective for an attractive source of new drugs. Their applications can be supposed to encompass the use as cell-type-selective determinant for targeted drug delivery and as blocking devices in anti-adhe-sion therapy during infections and inflammatory disease. To master the task of devising custom-made glycans/glycomimetics for this purpose, the individual enthalpic and entropic contributions in the molecular rendezvous between the sugar receptor under scrutiny and its ligand in the presence of solvent molecules undergoing positional rearrangements need to be understood and rationally exploited. As remunerative means to this end, cleverly orchestrated deployment of a panel of methods is essential. Concerning the carbohydrate ligand, its topological parameters and flexibility are assessed by the combination of computer-assisted molecular-mechanics and molecular-dynamics calculations and NMR-spectroscopic measurements. In the presence of the receptor, the latter technique will provide insights into conformational aspects of the bound ligand and into spatial vicinity of the ligand to distinct side chains of amino acids establishing the binding site in solution. Also in solution, the hydrogen-bonding pattern in the complex can be mapped with monodeoxy and monofluoro derivatives of the oligosaccharide. Together with X-ray crystallographic and microcalorimetric studies the limits of a feasible affinity enhancement can be systematically probed. With galactoside-binding lectins as instructive model, recent progress in this area of drug design will be documented, emphasizing the general applicability of the outlined interdisciplinary approach.
Anatomical and biomechanical investigations of the iliotibial tract
Divergent descriptions of the anatomic location and biomechanical function of the iliotibial tract (IT) can be found in the literature. This study attempted to obtain exact data regarding the anatomic course and material characteristics including the biomechanical properties of this structure. The following were its aims: (1) anatomical investigations of the IT; (2) mechanical properties of the IT; (3) femoral head centralizing force of the IT and subligamentous forces in the height of the greater trochanter in different joint positions by using a custom-made measuring prosthesis and a subligamentous positioned sensor; (4) construction of a finite element model of the proximal femur including the IT and measuring the femoral neck angle under variation. The hip joints and IT in a total of 18 unfixed corpses were evaluated. We studied the anatomic relationship to surrounding structures, as well as the material properties with the help of tensile strength testing utilizing an uniaxial apparatus. During the test, a load-displacement curve was registered, documenting the maximum load and deformation of the IT. To measure the subligamentous pressure at the height of the greater trochanter, a custom-made sensor with a power-recording instrument was constructed. Furthermore, an altered hip prosthesis with a pressure gauge at the height of the femoral neck was used to measure the forces which are directed at the acetabulum. The investigations were done in neutral-0 position and ab/adduction of the hip joint of the unfixed corpse. In addition, we varied the femoral neck angle between 115 degrees and 155 degrees in 5 degrees steps. To confirm the subligamentous forces, we did the same measurements intraoperatively at the height of the greater trochanter before and after hip joint replacement in 12 patients. We constructed a finite element model of the proximal femur and considering the IT. The acquisition of the data was done at physiological (128 degrees), varus (115 degrees), and valgus (155 degrees) femoral neck angles. The influencing forces of the IT at the height of the greater trochanter and the forces at the femoral head or the acetabulum could be measured. Our anatomical investigations revealed a splitting of the IT into a superficial and a deep portion, which covers the tensor fasciae latae. The tensor fasciae latae has an insertion on the IT. The IT continues down the femur, passing over the greater trochanter without developing an actual fixation to the bone. Part of the insertion of the gluteus maximus radiates into the IT. The IT passes over the vastus lateralis and inserts at the infracondylar tubercle of the tibia or Gerdy's tubercle, at the head of the fibula, as well as at the lateral intermuscular septum. Portions also insert on the transverse and longitudinal retinaculum of the patella. Concerning the material properties of the IT, we found a structural stiffness of 17 N/mm extension on average (D = 17 N/mm). The subligamentous measurements at the height of the greater trochanter in the unfixed corpse and intraoperatively during hip joint replacement showed an increase of the forces during adduction and a decrease during abduction of the hip joint. We found thereby a maximum increase up to 106 N with 40 degrees adduction. Concerning the femoral neck angle, we can state that valgus leads to lower subligamentous forces and varus to higher subligamentous forces. The forces directed at the acetabulum, which were measured by the prosthesis with a sensor along the femoral neck, showed a decrease with varus angles and an increase with valgus angles. The highest force of 624 N was measured with 40 degrees adduction and an angle of 155 degrees. The finite element model of the proximal femur showed a sole hip joint-centralizing force of the IT of 655 N with a femoral neck angle of 128 degrees after subtraction of the gluteal muscle force and the body weight. At 115 degrees, we found an increase up to 997 N and a decrease to 438 N at 155 degrees. Concerning the resulting forces in the acetabulum, we found opposite forces in comparison with the force of the IT at the height of the greater trochanter: at 115 degrees, a femoral head-centralizing force of 1601 N; at 128 degrees, 2360 N; and at 155 degrees, 2422 N. By our investigations, we can approximately prove the hip joint-centralizing force of the IT. By variation of the femoral neck angle and the position of the hip joint, we can predict the subligamentous force of the IT and the resulting force at the femoral head or at the acetabulum. The intraoperative measurement of the subligamentous forces of the IT is a good monitoring mechanism for the persistent hip-centralizing function of the IT in the course of hip joint replacement. The surgeon has the opportunity to check the stability of the hip joint after replacement. The finite element model gives the opportunity to check the divergent relative strength by variation of the femoral neck angle and the tension of the IT. In this way, the changes in the forces induced by a displacement osteotomy could be estimated preoperatively.
Towards Defining the Role of Glycans as Hardware in Information Storage and Transfer: Basic Principles, Experimental Approaches and Recent Progress
The term ‘code’ in biological information transfer appears to be tightly and hitherto exclusively connected with the genetic code based on nucleotides and translated into functional activities via proteins. However, the recent appreciation of the enormous coding capacity of oligosaccharide chains of natural glycoconjugates has spurred to give heed to a new concept: versatile glycan assembly by the genetically encoded glycosyltransferases endows cells with a probably not yet fully catalogued array of meaningful messages. Enciphered by sugar receptors such as endogenous lectins the information of code words established by a series of covalently linked monosaccharides as letters for example guides correct intra- and intercellular routing of glycoproteins, modulates cell proliferation or migration and mediates cell adhesion. Evidently, the elucidation of the structural frameworks and the recognition strategies within the operation of the sugar code poses a fascinating conundrum. The far-reaching impact of this recognition mode on the level of cells, tissues and organs has fueled vigorous investigations to probe the subtleties of protein-carbohydrate interactions. This review presents information on the necessarily concerted approach using X-ray crystallography, molecular modeling, nuclear magnetic resonance spectroscopy, thermodynamic analysis and engineered ligands and receptors. This part of the treatise is flanked by exemplarily chosen insights made possible by these techniques.
Use of extracorporeal shock-wave therapy (ESWT) in the treatment of non-unions: a review of the literature
Presently, extracorporeal shock-wave therapy (ESWT) is not yet a standard therapeutic technique in orthopaedics. The mechanism for the analgesic effect or the effect of shock waves on osseous defects are still unknown. With the help of a review of the literature, indications and success rates for ESWT in the treatment of non-unions are outlined, while adequate impulse and energy rates are defined according to the present state of knowledge. Non-union is defined as an absent healing process after a duration of 6 months. The aim of this study is to rate the published data. A total of 52 papers referring to ESWT of the locomotor system are reviewed, with a focus on the 635 patients from 10 publications who underwent ESWT to treat non-unions. Validation was performed for each paper dealing with this topic according to the internationally accepted system of the American Association of Spine Surgery as types A-E. Conclusions regarding possible applications in therapy were taken only from high-quality publications of types A and B. This advice can be regarded as scientifically as well as economically sound. The investigations concerning non-union hardly live up to scientific standards. No serious complications were observed. Because of the complication rate in operative treatment of non-unions, ESWT seems to be justifiable. The techniques of ESWT, energy density levels and impulse rates will be described. Atrophic non-unions seem to represent a poorer starting point in comparison with hypertrophic non-unions. Most investigations showed a consolidation of the non-union during a period of 3 months following ESWT, so that in case of treatment failure, operative treatment in the form of a re-osteosynthesis would only be delayed for this period of time. The advantages of ESWT are its non-invasiveness and low rate of complications. The primary aim of further research should be the evaluation of adequate energy density levels and impulse rates for various indications in accordance with evidence-based medicine. Long-term results need to become available before ESWT can be compared with established methods.
Biomechanical investigation of the stabilization principle of the Latarjet procedure
PurposeThe purpose of the study was to determine the biomechanical status of the different components of the Latarjet procedure. The anterior capsule reconstruction with the transferred coracoacromial ligament (CAL) and the necessity of an intact subscapularis tendon were of particular interest. We hypothesized that the anterior capsule reconstruction will have a significant effect and that the Latarjet procedure will lose its stabilizing effect if the subscapularis tendon is torn.MethodsStability testing of 12 human shoulder specimens was performed. After testing of the intact joint, a combined anterior glenoid and capsule defect was set arthroscopically. Then the Latarjet procedure was performed using an open approach and tested with and without loading of the conjoint tendons (10 N). Afterwards, the specimens were distributed into two groups and the Latarjet technique was reduced stepwise: dissection of the CAL, dissection of the conjoint tendons (group A); reduction of the coracoid segment, dissection of the subscapularis tendon (group B). Biomechanical testing was performed for each condition in two positions: 60° of glenohumeral abduction with neutral rotation and with 60° of external rotation; each with a passive humerus load of 30 N in the anterior, inferior and anteroinferior direction.ResultsThe Latarjet technique with load applied to the conjoint tendons significantly reduced translation compared with the defect condition for all tested positions in all directions. In group A, the CAL-dissection led to a significant increase of anterior translation (+5.0 mm, p = 0.003) and inferior translation (+7.3 mm, p = 0.025) in neutral rotation and of anterior translation in 60° of external rotation (+4.4 mm, p = 0.034). In group B, the reduction of the coracoid bone down to the coracoid tip resulted in a significant increase of only the anterior translation in abduction and 60° of external rotation (+4.5 mm, p = 0.05). In contrast, the detachment of the subscapularis tendon led to a significant increase of translation in all testing positions except the inferior direction in the neutral rotation.ConclusionsWe found the anterior capsule reconstruction to represent a significant contribution to the stabilizing effect of the Latarjet procedure, whereas a deficiency of the subscapularis tendon eliminates its effect.Clinical relevanceWe recommend to perform the Latarjet technique with an anterior capsule reconstruction (e.g. CAL transfer) and with a transfer of the coracoid bone block rather than a transposition of the coracoid tip. Furthermore, we were able to show that an intact subscapularis tendon is a necessary prerequisite for a reliable stabilization.
Murine Homodimeric Adhesion/Growth-Regulatory Galectins-1, -2 and -7: Comparative Profiling of Gene/ Promoter Sequences by Database Mining, of Expression by RT-PCR/Immunohistochemistry and of Contact Sites for Carbohydrate Ligands by Computational Chemistry
Following the detection of individual members of the family of galectins it is an obvious challenge to define the extent of functional overlap/divergence among these proteins. As a step to address this issue a comparative profiling has been started in the mouse as a model organism, combining sequence analysis, expression patterns and structural features in the cases of the homodimeric galectins-1, -2 and -7. Close relationship was apparent at the level of global gene organization. Scrutiny of the proximal promoter regions for putative transcription-factor-binding sites by two search algorithms uncovered qualitative and quantitative differences with potential to influence the combinatorial functionality of regulatory sequences. RT-PCR mapping with samples from an array of 17 organs revealed significant differences, separating rather ubiquitous gene expression of galectin-1 from the more restricted individual patterns of galectins-2 and -7. Using specific antisera obtained by affinity depletion including stringent controls to ascertain lack of cross-reactivity these results were corroborated at the level of galectin localization in fixed tissue sections. Nuclear presence was seen in the case of galectin-1. In addition to nonidentical expression profiles the mapping of the carbohydrate recognition domains of galectins-1 and -7 by homology modelling and docking of naturally occurring complex tetraand pentasaccharides disclosed a series of sequence deviations which may underlie disparate affinities for cell surface glycans/glycomimetic peptides. In view of applicability the presented data can serve as useful reference to delineate changes with respect to disease and in genetically engineered models. To enable more general conclusions on the galectin network it is warranted to further pursue this combined approach within this lectin family.
Late results after arthroscopic partial medial meniscectomy
In a retrospective study, 100 patients underwent a clinical and radiological follow‐up examination after a minimum of 7 years (range 7.0–9.3 years) following an arthroscopic partial medial meniscectomy. None of these patients had associated intra‐articular lesions, apart from minor chondral damage (max. grade 2 Outerbridge classification; < 1 cm 2 ) of the medial compartment at the time of the primary arthroscopy. According to the modified Marshall score, the follow‐up evaluation showed excellent clinical results in 96%. Nevertheless, the radiological outcome, as measured by comparing preoperative and postoperative X‐rays, demonstrated a development or progression of the osteoarthritis of the affected knee joint in 33%, with a statistically significant correlation between the radiological and clinical outcomes ( P < 0.05). The age of the patients at the time of operation and any angular deformity of the knee joint did not influence the radiological results. Women had a statistically significantly higher risk of developing gonarthrosis after partial medial meniscectomy than men ( P < 0.05). The arthroscopic partial medial meniscectomy led to excellent subjective and functional results but could not prevent the increase or development of late degenerative changes in the medial knee compartment.
Follow-up evaluation of open intra-articular fractures of the calcaneus
The results of a follow-up evaluation of open intra-articular calcaneal fractures are presented. A modified Merle d'Aubigné functional score and Zwipp radiographic score were used. A retrospective analysis of 35 patients with 36 open intra-articular fractures represents the basis of the study. At the time of follow-up examination (on average 44 months after the injury), 5 amputations of the affected extremity and 4 ankle arthrodeses had been carried out. The 23 patients still able to bear weight on the affected hindfoot and possessing a functional ankle joint were radiographically and functionally evaluated. No excellent results were documented. Only 6 good functional and 2 good radiographic outcomes were noted. In 17 instances, a poor functional or radiographic score was given. Devastating results were seen in the course of treatment of third-degree open joint depression or comminuted intra-articular fractures (n = 15): 9 cases of osteomyelitis, 5 amputations, 1 partial calcanectomy, 1 arthrodesis. An open reduction as part of the primary treatment (n = 6) led to local complications in all instances. The most favorable results were seen after nonoperative fracture management: complication-free course of treatment in 4 of 11 patients. All workmen's injuries led to a permanent disability, and these patients received compensation. The treatment and salvage of the soft-tissue envelope should be paramount in all therapeutic decisions. The fracture treatment must not further jeopardize these tissues. An aggressive operative treatment of local complications, including arthrodesis or amputation, is recommended.
Transient osteoporosis of the navicular bone in a runner
Transient osteoporosis or the bone marrow oedema syndrome is described as a self-limiting disorder. Transient osteoporosis usually affects only one bone, predominantly the proximal femur. Involvement of the foot is rare and often overlooked. A disorder of the navicular bone of the foot can be found twice in the literature. We report a case of transient osteoporosis of the navicular bone of the foot in a 20-year-old, female, top-level track athlete (400 m sprinter) treated with alendronate, and a review of the literature. The therapeutic options are limited, frequently consisting of non-specific, symptomatic therapy. Some authors report favourable results with core decompression, while others have reported good results with a conservative regime of symptomatic treatment and avoidance of weight-bearing until the clinical and radiological changes have resolved. In the described case, the patient had a favourable result after a short course of treatment with alendronate. She experienced almost immediate pain reduction and presented a complete resolution of the abnormal signal intensity on MRI. This rapid result makes the use of alendronate seem promising in athletes with transient osteoporosis, permitting an early return to high-level activities.