TY - JOUR A1 - Weissenberger, Manuel A1 - Weissenberger, Manuela H. A1 - Wagenbrenner, Mike A1 - Heinz, Tizian A1 - Reboredo, Jenny A1 - Holzapfel, Boris M. A1 - Rudert, Maximilian A1 - Groll, Jürgen A1 - Evans, Christopher H. A1 - Steinert, Andre F. T1 - Different types of cartilage neotissue fabricated from collagen hydrogels and mesenchymal stromal cells via SOX9, TGFB1 or BMP2 gene transfer JF - PLoS One N2 - Objective As native cartilage consists of different phenotypical zones, this study aims to fabricate different types of neocartilage constructs from collagen hydrogels and human mesenchymal stromal cells (MSCs) genetically modified to express different chondrogenic factors. Design Human MSCs derived from bone-marrow of osteoarthritis (OA) hips were genetically modified using adenoviral vectors encoding sex-determining region Y-type high-mobility-group-box (SOX)9,transforming growth factor beta (TGFB) 1or bone morphogenetic protein (BMP) 2cDNA, placed in type I collagen hydrogels and maintained in serum-free chondrogenic media for three weeks. Control constructs contained unmodified MSCs or MSCs expressing GFP. The respective constructs were analyzed histologically, immunohistochemically, biochemically, and by qRT-PCR for chondrogenesis and hypertrophy. Results Chondrogenesis in MSCs was consistently and strongly induced in collagen I hydrogels by the transgenesSOX9,TGFB1andBMP2as evidenced by positive staining for proteoglycans, chondroitin-4-sulfate (CS4) and collagen (COL) type II, increased levels of glycosaminoglycan (GAG) synthesis, and expression of mRNAs associated with chondrogenesis. The control groups were entirely non-chondrogenic. The levels of hypertrophy, as judged by expression of alkaline phosphatase (ALP) and COL X on both the protein and mRNA levels revealed different stages of hypertrophy within the chondrogenic groups (BMP2>TGFB1>SOX9). Conclusions Different types of neocartilage with varying levels of hypertrophy could be generated from human MSCs in collagen hydrogels by transfer of genes encoding the chondrogenic factorsSOX9,TGFB1andBMP2. This technology may be harnessed for regeneration of specific zones of native cartilage upon damage. KW - stem cells KW - in vitro KW - chondrogenic differentiation KW - repair KW - chondrocytes KW - transplantation KW - stimulation KW - scaffolds KW - defects KW - therapy Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230494 VL - 15 IS - 8 ER - TY - JOUR A1 - Wagenbrenner, Mike A1 - Heinz, Tizian A1 - Horas, Konstantin A1 - Jakuscheit, Axel A1 - Arnholdt, Joerg A1 - Mayer-Wagner, Susanne A1 - Rudert, Maximilian A1 - Holzapfel, Boris M. A1 - Weißenberger, Manuel T1 - Impact of Tranexamic Acid on Chondrocytes and Osteogenically Differentiated Human Mesenchymal Stromal Cells (hMSCs) In Vitro JF - Journal of Clinical Medicine N2 - The topical application of tranexamic acid (TXA) helps to prevent post-operative blood loss in total joint replacements. Despite these findings, the effects on articular and periarticular tissues remain unclear. Therefore, this in vitro study examined the effects of varying exposure times and concentrations of TXA on proliferation rates, gene expression and differentiation capacity of chondrocytes and human mesenchymal stromal cells (hMSCs), which underwent osteogenic differentiation. Chondrocytes and hMSCs were isolated and multiplied in monolayer cell cultures. Osteogenic differentiation of hMSCs was induced for 21 days using a differentiation medium containing specific growth factors. Cell proliferation was analyzed using ATP assays. Effects of TXA on cell morphology were examined via light microscopy and histological staining, while expression levels of tissue-specific genes were measured using semiquantitative RT-PCR. After treatment with 50 mg/mL of TXA, a decrease in cell proliferation rates was observed. Furthermore, treatment with concentrations of 20 mg/mL of TXA for at least 48 h led to a visible detachment of chondrocytes. TXA treatment with 50 mg/mL for at least 24 h led to a decrease in the expression of specific marker genes in chondrocytes and osteogenically differentiated hMSCs. No significant effects were observed for concentrations beyond 20 mg/mL of TXA combined with exposure times of less than 24 h. This might therefore represent a safe limit for topical application in vivo. Further research regarding in vivo conditions and effects on hMSC functionality are necessary to fully determine the effects of TXA on articular and periarticular tissues. KW - tranexamic acid KW - hMSCs KW - chondrocytes KW - osteoarthritis KW - toxicity KW - differentiation capacity Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219410 SN - 2077-0383 VL - 9 IS - 12 ER - TY - THES A1 - Weißenberger, Manuel Claudius T1 - Chondrogene Differenzierung von humanen mesenchymalen Stammzellen zur Knorpelregeneration mittels adenoviralem Indian Hedgehog-Gentransfer T1 - Mesenchymal stem cell-based cartilage regeneration - Indian hedgehog gene transfer as chondrogenic inductor in an in vitro model N2 - Ziel dieser Arbeit war es zu untersuchen, ob mittels IHH-Gentransfer aus Hüftköpfen gewonnene hMSCs chondrogen im Pelletkultursystem differenziert werden können und ob zugleich durch IHH eine Modulation der hypertrophen Enddifferenzierung der hMSCs in diesem System möglich ist. IHH bestimmt in der Wachstumsfuge zusammen mit PTHrP während der endochondralen Ossifikation die Chondrozytenreifung und -differenzierung entscheidend mit und ist daher ein interessanter Kandidat zur Induktion von hyalinem oder zumindest hyalin-ähnlichem Knorpelgewebe in der stammzellbasierten Gentherapie. Nach Gewinnung und Kultivierung der hMSCs wurden diese mit Ad.GFP, Ad.IHH, Ad.IHH+TGF-β1, Ad.IHH+SOX-9 oder Ad.IHH+BMP-2 transduziert bzw. ein Teil für die Negativkontrolle nicht transduziert und im Anschluss alle Gruppen zu Pellets weiterverarbeitet. Histologische, biochemische sowie molekularbiologische Untersuchungen wurden an verschiedenen Zeitpunkten zur Evaluierung des chondrogenen Differenzierungsgrades sowie der hypertrophiespezifischen Merkmale der kultivierten Pellets durchgeführt. Es konnte durch diese Arbeit sowohl auf Proteinebene als auch auf Genexpressionsebene reproduzierbar gezeigt werden, dass primäre hMSCs im Pelletkultursystem sowohl durch den adenoviralen Gentransfer von IHH allein als auch durch die Co-Transduktionsgruppen IHH+TGF-β1, IHH+SOX-9 und IHH+BMP-2 chondrogen differenziert werden können. Dabei zeigten alle IHH-modifizierten Pellets Col II- und CS-4-positive immunhistochemische Anfärbungen, eine gesteigerte Synthese von Glykosaminoglykanen im biochemischen GAG-Assay sowie eine Hochregulation von mit der Chondrogenese assoziierten Genen. Das Auftreten hypertropher Merkmale bei den chondrogen differenzierten MSCs konnte durch IHH-Gentransfer nach 3 Wochen in vitro-Kultivierung nicht vollkommen unterdrückt werden, war jedoch besonders stark ausgeprägt, wenn BMP-2 co-exprimiert wurde und war etwas weniger evident in der IHH+SOX-9-Gruppe. Dabei zeigte die Ad.IHH+BMP-2-Gruppe sowohl in der ALP-Färbung als auch in dem ALP-Assay und der quantitativen RT-PCR die stärkste Hochregulierung des hypertrophen Markers ALP. Möglicherweise brachte die Überexpression von IHH das fein aufeinander abgestimmte Regulationssystem zwischen IHH und PTHrP aus dem Gleichgewicht und könnte als ein Grund dafür angeführt werden, warum die Hypertrophie im Pelletkultursystem nicht vollkommen supprimiert werden konnte. Es bleibt abzuwarten, ob IHH in vivo die Chondrogenese induzieren und dabei zugleich das Phänomen der chondrogenen Hypertrophie regulieren kann. In der Zukunft würde dies letztlich der stammzellbasierten Knorpelregeneration in vivo zu Gute kommen. N2 - Introduction: The issue of final end-stage chondrogenic hypertrophy has been identified in previous studies on MSC-mediated chondrogenesis using several bone morphogenetic proteins (BMPs) following adenoviral gene transfer as one hurdle in the efforts of creating stable cartilage repair tissue. Therefore, in this in vitro study we explore, whether the growth factor Indian hedgehog (IHH), alone or in combination with TGFb1, BMP-2 or SOX-9, is able to modulate the appearance of chondrogenic hypertrophy in pellet cultures in vitro, and if IHH induces chondrogenesis in human primary mesenchymal stem cells (MSCs) via its gene-delivery. Methods: First generation adenoviral vectors encoding the cDNA of the human IHH gene were created by cre-lox recombination and used alone or in combination with Ad.TGFb1, Ad.BMP-2 and Ad.SOX-9 to transduce human bone-marrow derived MSCs at 5 x 102 infectious particles/cell (50 MOI multiplicities of infection). Thereafter 3 x 105 cells were seeded into aggregates and cultured for three weeks in serum-free chondrogenic differentiation medium (ITS, Dexa, Asc) with untransduced or marker gene transduced cultures as controls. Transgene expressions were determined by ELISA, and aggregates were analyzed histologically, immunohistochemically, biochemically and by RT-PCR for chondrogenesis and hypertrophy after 10 days and 21 days of culture. Results: IHH alone or in combination with TGFb1, BMP-2 or SOX-9 were equipotent inducers of chondrogenesis in MSCs in pellet culture (strong staining for alcian blue and collagen type II, high levels of GAG synthesis, expression of mRNAs associated with chondrogenesis, controls were not chondrogenic). IHH-modified aggregates, alone as well as the Ihh co-transduced groups with TGFb1, BMP-2 or SOX-9, showed also a tendency to progress towards hypertrophy, as judged by expression of alkaline phosphatase and immunhistochemical staining for collagen type X, while the highest levels for both markers seen in the IHH+BMP-2-group after 21 days of culture. These results were confirmed by qRT-PCR analyses that showed comparable expression of cartilage specific marker genes (Col II, SOX-9) in the induced pellet cultures and a higher expression of hypertrophy associated marker genes (ALP, Col X) in the IHH+BMP2-group. Discussion: IHH gene transfer with adenoviral vectors alone or in combination with TGFb1, BMP-2 or SOX-9 efficiently induces chondrogenesis in MSCs, however, the appearance of hypertrophy could not be completely obviated, and was strongly present when BMP-2 was co-expressed. Thus, it remains to be seen in the ongoing in vivo studies, whether IHH can induce chondrogenesis while modulating chondrogenic hypertrophy in vivo. KW - Stammzelle KW - Gentherapie KW - Wachstumsfaktor KW - Knorpel KW - Knorpelregeneration KW - Mesenchymale Stammzellen KW - Gentherapie KW - Indian Hedgehog KW - Cartilage regeneration KW - mesenchymal stem cells KW - gene therapy KW - Indian hedgehog Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-78014 ER - TY - JOUR A1 - Wagenbrenner, Mike A1 - Mayer-Wagner, Susanne A1 - Rudert, Maximilian A1 - Holzapfel, Boris Michael A1 - Weissenberger, Manuel T1 - Combinations of hydrogels and mesenchymal stromal cells (MSCs) for cartilage tissue engineering — a review of the literature JF - Gels N2 - Cartilage offers limited regenerative capacity. Cell-based approaches have emerged as a promising alternative in the treatment of cartilage defects and osteoarthritis. Due to their easy accessibility, abundancy, and chondrogenic potential mesenchymal stromal cells (MSCs) offer an attractive cell source. MSCs are often combined with natural or synthetic hydrogels providing tunable biocompatibility, biodegradability, and enhanced cell functionality. In this review, we focused on the different advantages and disadvantages of various natural, synthetic, and modified hydrogels. We examined the different combinations of MSC-subpopulations and hydrogels used for cartilage engineering in preclinical and clinical studies and reviewed the effects of added growth factors or gene transfer on chondrogenesis in MSC-laden hydrogels. The aim of this review is to add to the understanding of the disadvantages and advantages of various combinations of MSC-subpopulations, growth factors, gene transfers, and hydrogels in cartilage engineering. KW - hydrogels KW - osteoarthritis KW - cartilage defects KW - MSCs KW - cartilage regeneration KW - tissue engineering Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-250177 SN - 2310-2861 VL - 7 IS - 4 ER - TY - JOUR A1 - Steinert, Andre F. A1 - Weissenberger, Manuel A1 - Kunz, Manuela A1 - Gilbert, Fabian A1 - Ghivizzani, Steven C. A1 - Goebel, Sascha A1 - Jakob, Franz A1 - Nöth, Ulrich A1 - Rudert, Maximilian T1 - Indian hedgehog gene transfer is a chondrogenic inducer of human mesenchymal stem cells N2 - Introduction: To date, no single most-appropriate factor or delivery method has been identified for the purpose of mesenchymal stem cell (MSC)-based treatment of cartilage injury. Therefore, in this study we tested whether gene delivery of the growth factor Indian hedgehog (IHH) was able to induce chondrogenesis in human primary MSCs, and whether it was possible by such an approach to modulate the appearance of chondrogenic hypertrophy in pellet cultures in vitro. Methods: First-generation adenoviral vectors encoding the cDNA of the human IHH gene were created by cre-lox recombination and used alone or in combination with adenoviral vectors, bone morphogenetic protein-2 (Ad.BMP- 2), or transforming growth factor beta-1 (Ad.TGF-b1) to transduce human bone-marrow derived MSCs at 5 × 102 infectious particles/cell. Thereafter, 3 × 105 cells were seeded into aggregates and cultured for 3 weeks in serumfree medium, with untransduced or marker gene transduced cultures as controls. Transgene expressions were determined by ELISA, and aggregates were analysed histologically, immunohistochemically, biochemically and by RT-PCR for chondrogenesis and hypertrophy. Results: IHH, TGF-b1 and BMP-2 genes were equipotent inducers of chondrogenesis in primary MSCs, as evidenced by strong staining for proteoglycans, collagen type II, increased levels of glycosaminoglycan synthesis, and expression of mRNAs associated with chondrogenesis. IHH-modified aggregates, alone or in combination, also showed a tendency to progress towards hypertrophy, as judged by the expression of alkaline phosphatase and stainings for collagen type X and Annexin 5. Conclusion: As this study provides evidence for chondrogenic induction of MSC aggregates in vitro via IHH gene delivery, this technology may be efficiently employed for generating cartilaginous repair tissues in vivo. KW - Medizin Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-75425 ER - TY - JOUR A1 - Weißenberger, Manuel A1 - Wagenbrenner, Mike A1 - Schote, Fritz A1 - Horas, Konstantin A1 - Schäfer, Thomas A1 - Rudert, Maximilian A1 - Barthel, Thomas A1 - Heinz, Tizian A1 - Reppenhagen, Stephan T1 - The 3-triangle method preserves the posterior tibial slope during high tibial valgus osteotomy: first preliminary data using a mathematical model JF - Journal of Experimental Orthopaedics N2 - Purpose Despite much improved preoperative planning techniques accurate intraoperative assessment of the high tibial valgus osteotomy (HTO) remains challenging and often results in coronal over- and under-corrections as well as unintended changes of the posterior tibial slope. Noyes et al. reported a novel method for accurate intraoperative coronal and sagittal alignment correction based on a three-dimensional mathematical model. This is the first study examining preliminary data via the proposed Noyes approach for accurate intraoperative coronal and sagittal alignment correction during HTO. Methods From 2016 to 2020 a total of 24 patients (27 knees) underwent HTO applying the proposed Noyes method (Noyes-Group). Radiographic data was analyzed retrospectively and matched to patients that underwent HTO using the conventional method, i.e., gradual medial opening using a bone spreader under fluoroscopic control (Conventional-Group). All operative procedures were performed by an experienced surgeon at a single orthopaedic university center. Results From the preoperative to the postoperative visit no statistically significant changes of the posterior tibial slope were noted in the Noyes-Group compared to a significant increase in the Conventional-Group (p = 0.01). Regarding the axial alignment no significant differences between both groups were observed pre- and postoperatively. The number of over- and under-corrections did not differ significantly between both groups. Linear regression analysis showed a significant correlation of the postoperative medial proximal tibial angle (MPTA) with the position of the weightbearing line on the tibial plateau. Conclusion The 3-triangle method by Noyes seems to be a promising approach for preservation of the posterior tibial slope during HTO. KW - knee KW - high tibial valgus osteotomy KW - axial alignment KW - posterior tibial slope KW - weight bearing line KW - cartilage KW - triangle method KW - osteoarthritis Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-300806 SN - 2197-1153 VL - 9 ER - TY - JOUR A1 - Ebert, Regina A1 - Weissenberger, Manuel A1 - Braun, Clemens A1 - Wagenbrenner, Mike A1 - Herrmann, Marietta A1 - Müller‐Deubert, Sigrid A1 - Krug, Melanie A1 - Jakob, Franz A1 - Rudert, Maximilian T1 - Impaired regenerative capacity and senescence‐associated secretory phenotype in mesenchymal stromal cells from samples of patients with aseptic joint arthroplasty loosening JF - Journal of Orthopaedic Research N2 - Aseptic loosening of total hip and knee joint replacements is the most common indication for revision surgery after primary hip and knee arthroplasty. Research suggests that exposure and uptake of wear by mesenchymal stromal cells (MSC) and macrophages results in the secretion of proinflammatory cytokines and local osteolysis, but also impaired cell viability and regenerative capacity of MSC. Therefore, this in vitro study compared the regenerative and differentiation capacity of MSC derived from patients undergoing primary total hip arthroplasty (MSCprim) to MSC derived from patients undergoing revision surgery after aseptic loosening of total hip and knee joint implants (MSCrev). Regenerative capacity was examined by measuring the cumulative population doubling (CPD) in addition to the number of passages until cells stopped proliferating. Osteogenesis and adipogenesis in monolayer cultures were assessed using histological stainings. Furthermore, RT‐PCR was performed to evaluate the relative expression of osteogenic and adipogenic marker genes as well as the expression of markers for a senescence‐associated secretory phenotype (SASP). MSCrev possessed a limited regenerative capacity in comparison to MSCprim. Interestingly, MSCrev also showed an impaired osteogenic and adipogenic differentiation capacity compared to MSCprim and displayed a SASP early after isolation. Whether this is the cause or the consequence of the aseptic loosening of total joint implants remains unclear. Future research should focus on the identification of specific cell markers on MSCprim, which may influence complication rates such as aseptic loosening of total joint arthroplasty to further individualize and optimize total joint arthroplasty. KW - aseptic loosening KW - mesenchymal stromal cells KW - regenerative capacity KW - senescence‐associated secretory phenotype Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-238963 VL - 40 IS - 2 SP - 513 EP - 523 ER - TY - JOUR A1 - Wagenbrenner, Mike A1 - Heinz, Tizian A1 - Horas, Konstantin A1 - Jakuscheit, Axel A1 - Arnholdt, Jörg A1 - Hermann, Marietta A1 - Rudert, Maximilian A1 - Holzapfel, Boris M. A1 - Steinert, Andre F. A1 - Weißenberger, Manuel T1 - The human arthritic hip joint is a source of mesenchymal stromal cells (MSCs) with extensive multipotent differentiation potential JF - BMC Musculoskeletal Disorders N2 - Background While multiple in vitro studies examined mesenchymal stromal cells (MSCs) derived from bone marrow or hyaline cartilage, there is little to no data about the presence of MSCs in the joint capsule or the ligamentum capitis femoris (LCF) of the hip joint. Therefore, this in vitro study examined the presence and differentiation potential of MSCs isolated from the bone marrow, arthritic hyaline cartilage, the LCF and full-thickness samples of the anterior joint capsule of the hip joint. Methods MSCs were isolated and multiplied in adherent monolayer cell cultures. Osteogenesis and adipogenesis were induced in monolayer cell cultures for 21 days using a differentiation medium containing specific growth factors, while chondrogenesis in the presence of TGF-ss1 was performed using pellet-culture for 27 days. Control cultures were maintained for comparison over the same duration of time. The differentiation process was analyzed using histological and immunohistochemical stainings as well as semiquantitative RT-PCR for measuring the mean expression levels of tissue-specific genes. Results This in vitro research showed that the isolated cells from all four donor tissues grew plastic-adherent and showed similar adipogenic and osteogenic differentiation capacity as proven by the histological detection of lipid droplets or deposits of extracellular calcium and collagen type I. After 27 days of chondrogenesis proteoglycans accumulated in the differentiated MSC-pellets from all donor tissues. Immunohistochemical staining revealed vast amounts of collagen type II in all differentiated MSC-pellets, except for those from the LCF. Interestingly, all differentiated MSCs still showed a clear increase in mean expression of adipogenic, osteogenic and chondrogenic marker genes. In addition, the examination of an exemplary selected donor sample revealed that cells from all four donor tissues were clearly positive for the surface markers CD44, CD73, CD90 and CD105 by flow cytometric analysis. Conclusions This study proved the presence of MSC-like cells in all four examined donor tissues of the hip joint. No significant differences were observed during osteogenic or adipogenic differentiation depending on the source of MSCs used. Further research is necessary to fully determine the tripotent differentiation potential of cells isolated from the LCF and capsule tissue of the hip joint. KW - Hip joint KW - Osteoarthritis KW - MSCs KW - Cartilage regeneration KW - Tissue engineering KW - Ligamentum capitis femoris KW - Joint capsule KW - Bone marrow Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229497 VL - 21 IS - 1 ER - TY - JOUR A1 - Prodinger, Peter Michael A1 - Lazic, Igor A1 - Horas, Konstantin A1 - Burgkart, Rainer A1 - von Eisenhart-Rothe, Rüdiger A1 - Weissenberger, Manuel A1 - Rudert, Maximilian A1 - Holzapfel, Boris Michael T1 - Revision Arthroplasty Through the Direct Anterior Approach Using an Asymmetric Acetabular Component JF - Journal of Clinical Medicine N2 - Despite increasing numbers of primary hip arthroplasties performed through the direct anterior approach (DAA), there is a lack of literature on DAA revision arthroplasty. The present study was performed in order to evaluate outcomes and revision rates after revision through the DAA using an asymmetric acetabular component with optional intra- and extramedullary fixation. In a retrospective cohort study, we analyzed prospectively collected data of 57 patients (61 hips, 43 female, 18 male) who underwent aseptic acetabular component revision through the DAA with the abovementioned implant system between January 2015 and December 2017. The mean follow-up was 40 months (12–56). Survival rates were estimated using the Kaplan–Meier method. All complications were documented and functional outcomes were assessed pre- and postoperatively. Kaplan–Meier analysis revealed an estimated five-year implant survival of 97% (confidence interval CI 87–99%). The estimated five-year survival with revision for any cause was 93% (CI 83–98%). The overall revision rate was 6.6% (n = 4). Two patients had to undergo revision due to periprosthetic infection (3.3%). In one patient, the acetabular component was revised due to aseptic loosening four months postoperatively. Another patient suffered from postoperative iliopsoas impingement and was treated successfully by arthroscopic iliopsoas tenotomy. Two (3.3%) of the revised hips dislocated postoperatively. The mean Harris Hip Score improved from 35 (2–66) preoperatively to 86 (38–100) postoperatively (p < 0.001). The hip joint’s anatomical center of rotation was restored at a high degree of accuracy. Our findings demonstrate that acetabular revision arthroplasty through the DAA using an asymmetric acetabular component with optional intra- and extramedullary fixation is safe and practicable, resulting in good radiographic and clinical midterm results. KW - anterior approach KW - revision arthroplasty KW - hip joint KW - acetabular bone defect KW - asymmetric implant KW - anatomic center of rotation Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-213184 SN - 2077-0383 VL - 9 IS - 9 ER - TY - JOUR A1 - Boelch, Sebastian Philipp A1 - Weissenberger, Manuel A1 - Spohn, Frederik A1 - Rudert, Maximilian A1 - Luedemann, Martin T1 - Insufficient sensitivity of joint aspiration during the two-stage exchange of the hip with spacers JF - Journal of Orthopedic Surgery and Research N2 - Background: Evaluation of infection persistence during the two-stage exchange of the hip is challenging. Joint aspiration before reconstruction is supposed to rule out infection persistence. Sensitivity and specificity of synovial fluid culture and synovial leucocyte count for detecting infection persistence during the two-stage exchange of the hip were evaluated. Methods: Ninety-two aspirations before planned joint reconstruction during the two-stage exchange with spacers of the hip were retrospectively analyzed. Results: The sensitivity and specificity of synovial fluid culture was 4.6 and 94.3%. The sensitivity and specificity of synovial leucocyte count at a cut-off value of 2000 cells/μl was 25.0 and 96.9%. C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) values were significantly higher before prosthesis removal and reconstruction or spacer exchange (p = 0.00; p = 0.013 and p = 0.039; p = 0.002) in the infection persistence group. Receiver operating characteristic area under the curve values before prosthesis removal and reconstruction or spacer exchange for ESR were lower (0.516 and 0.635) than for CRP (0.720 and 0.671). Conclusions: Synovial fluid culture and leucocyte count cannot rule out infection persistence during the two-stage exchange of the hip. KW - two-stage exchange KW - hip KW - periprosthetic infection KW - joint aspiration KW - spacer Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-175576 VL - 13 IS - 7 ER - TY - JOUR A1 - Jakuscheit, Axel A1 - Schaefer, Nina A1 - Roedig, Johannes A1 - Luedemann, Martin A1 - Hertzberg-Boelch, Sebastian Philipp von A1 - Weissenberger, Manuel A1 - Schmidt, Karsten A1 - Holzapfel, Boris Michael A1 - Rudert, Maximilian T1 - Modifiable individual risks of perioperative blood transfusions and acute postoperative complications in total hip and knee arthroplasty JF - Journal of Personalized Medicine N2 - Background: The primary aim of this study was to identify modifiable patient-related predictors of blood transfusions and perioperative complications in total hip and knee arthroplasty. Individual predictor-adjusted risks can be used to define preoperative treatment thresholds. Methods: We performed this retrospective monocentric study in orthopaedic patients who underwent primary total knee or hip arthroplasty. Multivariate logistic regression models were used to assess the predictive value of patient-related characteristics. Predictor-adjusted individual risks of blood transfusions and the occurrence of any perioperative adverse event were calculated for potentially modifiable risk factors. Results: 3754 patients were included in this study. The overall blood transfusion and complication rates were 4.8% and 6.4%, respectively. Haemoglobin concentration (Hb, p < 0.001), low body mass index (BMI, p < 0.001) and estimated glomerular filtration rate (eGFR, p = 0.004) were the strongest potentially modifiable predictors of a blood transfusion. EGFR (p = 0.001) was the strongest potentially modifiable predictor of a complication. Predictor-adjusted risks of blood transfusions and acute postoperative complications were calculated for Hb and eGFR. Hb = 12.5 g/dL, BMI = 17.6 kg/m\(^2\), and eGFR = 54 min/mL were associated, respectively, with a 10% risk of a blood transfusion, eGFR = 59 mL/min was associated with a 10% risk of a complication. Conclusion: The individual risks for blood transfusions and acute postoperative complications are strongly increased in patients with a low preoperative Hb, low BMI or low eGFR. We recommend aiming at a preoperative Hb ≥ 13g/dL, an eGFR ≥ 60 mL/min and to avoid a low BMI. Future studies must show if a preoperative increase of eGFR and BMI is feasible and truly beneficial. KW - patient blood management KW - total joint arthroplasty KW - haemoglobin KW - perioperative management Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-250290 SN - 2075-4426 VL - 11 IS - 11 ER - TY - JOUR A1 - Heinz, Tizian A1 - Meller, Felix A1 - Luetkens, Karsten Sebastian A1 - Horas, Konstantin A1 - Schäfer, Thomas A1 - Rudert, Maximilian A1 - Reppenhagen, Stephan A1 - Weißenberger, Manuel T1 - Can the MRI based AMADEUS score accurately assess pre-surgery chondral defect severity according to the ICRS arthroscopic classification system? JF - Journal of Experimental Orthopaedics N2 - Purpose The AMADEUS (Area Measurement And DEpth and Underlying Structures) scoring and grading system has been proposed for the MRI based evaluation of untreated focal chondral defects around the knee. The clinical practicability, its correlation with arthroscopically assessed grading systems (ICRS – International Cartilage Repair Society) and thereby its clinical value in terms of decision making and guiding prognosis was yet to determine. Methods From 2008 to 2019 a total of 89 individuals were indicated for high tibial valgus osteotomy (HTO) due to tibial varus deformity and concomitant chondral defects of the medial compartment of the knee. All patients received a preoperative MRI (1.5 Tesla or 3.0 Tesla) and pre-osteotomy diagnostic arthroscopy. Chondral defects of the medial compartment were scored and graded with the MRI based AMADEUS by three independent raters and compared to arthroscopic defect grading by the ICRS system. Interrater and intrarater reliability as well as correlation analysis with the ICRS classification system were assessed. Results Intraclass correlation coefficients for the various subscores of the AMADEUS showed an overall good to excellent interrater agreement (min: 0.26, max: 0.80). Intrarater agreement turned out to be substantially inferior (min: 0.08, max: 0.53). Spearman correlation revealed an overall moderate correlative association of the AMADEUS subscores with the ICRS classification system, apart from the defect area subscore. Sensitivity of the AMADEUS to accurately identify defect severity according to the ICRS was 0.7 (0.69 for 3.0 Tesla MRI, 0.67 for 1.5 Tesla MRI). The mean AMADEUS grade was 2.60 ± 0.81 and the mean ICRS score 2.90 ± 0.63. Conclusions Overall, the AMADEUS with all its subscores shows moderate correlation with the arthroscopic chondral grading system according to ICRS. This suggests that chondral defect grading by means of the MRI based AMADEUS is well capable of influencing and guiding treatment decisions. Interrater reliability shows overall good agreement. KW - MRI KW - knee KW - cartilage defect KW - grading system of chondral defects KW - AMADEUS KW - ICRS Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-300781 SN - 2197-1153 VL - 9 IS - 1 ER - TY - JOUR A1 - Wagenbrenner, Mike A1 - Poker, Konrad A1 - Heinz, Tizian A1 - Herrmann, Marietta A1 - Horas, Konstantin A1 - Ebert, Regina A1 - Mayer-Wagner, Susanne A1 - Holzapfel, Boris M. A1 - Rudert, Maximilian A1 - Steinert, Andre F. A1 - Weißenberger, Manuel T1 - Mesenchymal stromal cells (MSCs) isolated from various tissues of the human arthritic knee joint possess similar multipotent differentiation potential JF - Applied Sciences N2 - (1) Background: The mesenchymal stromal cells (MSCs) of different tissue origins are applied in cell-based chondrogenic regeneration. However, there is a lack of comparability determining the most suitable cell source for the tissue engineering (TE) of cartilage. The purpose of this study was to compare the in vitro chondrogenic potential of MSC-like cells from different tissue sources (bone marrow, meniscus, anterior cruciate ligament, synovial membrane, and the infrapatellar fat pad removed during total knee arthroplasty (TKA)) and define which cell source is best suited for cartilage regeneration. (2) Methods: MSC-like cells were isolated from five donors and expanded using adherent monolayer cultures. Differentiation was induced by culture media containing specific growth factors. Transforming growth factor (TGF)-ß1 was used as the growth factor for chondrogenic differentiation. Osteogenesis and adipogenesis were induced in monolayer cultures for 27 days, while pellet cell cultures were used for chondrogenesis for 21 days. Control cultures were maintained under the same conditions. After, the differentiation period samples were analyzed, using histological and immunohistochemical staining, as well as molecularbiological analysis by RT-PCR, to assess the expression of specific marker genes. (3) Results: Plastic-adherent growth and in vitro trilineage differentiation capacity of all isolated cells were proven. Flow cytometry revealed the clear co-expression of surface markers CD44, CD73, CD90, and CD105 on all isolated cells. Adipogenesis was validated through the formation of lipid droplets, while osteogenesis was proven by the formation of calcium deposits within differentiated cell cultures. The formation of proteoglycans was observed during chondrogenesis in pellet cultures, with immunohistochemical staining revealing an increased relative gene expression of collagen type II. RT-PCR proved an elevated expression of specific marker genes after successful differentiation, with no significant differences regarding different cell source of native tissue. (4) Conclusions: Irrespective of the cell source of native tissue, all MSC-like cells showed multipotent differentiation potential in vitro. The multipotent differentiation capacity did not differ significantly, and chondrogenic differentiation was proven in all pellet cultures. Therefore, cell suitability for cell-based cartilage therapies and tissue engineering is given for various tissue origins that are routinely removed during total knee arthroplasty (TKA). This study might provide essential information for the clinical tool of cell harvesting, leading to more flexibility in cell availability. KW - knee joint KW - MSCs KW - cellular origin KW - cartilage regeneration KW - tissue engineering KW - cell-based therapies KW - osteoarthritis Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-262334 SN - 2076-3417 VL - 12 IS - 4 ER - TY - JOUR A1 - Boelch, Sebastian Philipp A1 - Rüeckl, Kilian A1 - Streck, Laura Elisa A1 - Szewczykowski, Viktoria A1 - Weißenberger, Manuel A1 - Jakuscheit, Axel A1 - Rudert, Maximilian T1 - Diagnosis of chronic infection at total hip arthroplasty revision is a question of definition JF - Biomed Research International N2 - Purpose. Contradicting definitions of periprosthetic joint infection (PJI) are in use. Joint aspiration is performed before total hip arthroplasty (THA) revision. This study investigated the influence of PJI definition on PJI prevalence at THA revision. Test quality of prerevision aspiration was evaluated for the different PJI definitions. Methods. 256 THA revisions were retrospectively classified to be infected or not infected. Classification was performed according to the 4 different definitions proposed by the Musculoskeletal Infection Society (MSIS), the Infectious Diseases Society of America (IDSA), the International Consensus Meeting (ICM), and the European Bone and Joint Infection Society (EBJIS). Only chronic PJIs were included. Results. PJI prevalence at revision significantly correlated with the applied PJI definition (p=0.01, Cramer's V=0.093). PJI prevalence was 20.7% for the MSIS, 25.4% for the ICM, 28.1% for the IDSA, and 32.0% for the EBJIS definition. For synovial fluid white blood cell count, the best ROC-AUC for predicting PJI was 0.953 in combination with the MSIS definition. Conclusion. PJI definition significantly influences the rate of diagnosed PJIs at THA revision. Synovial fluid white blood cell count is a reliable means to rule out PJI. In cases with a borderline high synovial white blood cell count before THA revision as the only sign of chronic PJI, an extended diagnostic work-up should be considered. KW - periprosthetic joint infection KW - algorithm KW - consensus Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-265762 VL - 2021 ER - TY - JOUR A1 - Weissenberger, Manuel A1 - Heinz, Tizian A1 - Rueckl, Kilian A1 - Rudert, Maximilian A1 - Klug, Alexander A1 - Hoffmann, Reinhard A1 - Schmidt-Horlohé, Kay T1 - No functional differences in anatomic reconstruction with one vs. two suture anchors after non-simultaneous bilateral distal biceps brachii tendon rupture: a case report and review of the literature JF - BMC Musculoskeletal Disorders N2 - Background Surgical reattachment of the tendon is still the gold standard for ruptures of the distal biceps brachii tendon. Several fixation techniques have been described in the literature, with suture anchors being one of the most common fixation techniques. Currently, there is no data available on how many anchors are required for a safe and stable refixation. In this case report clinical data of a patient with non-simultaneous bilateral distal biceps tendon ruptures treated with a different number of suture anchors for each side (one vs. two) are demonstrated. Case presentation A 47-year-old factory worker suffered a rupture of the distal biceps tendon on both arms following two different occasions. The left side was fixed using a single suture anchor, while refixation on the right side was performed with two anchors. The patient was prospectively followed for one year. Functional outcome was assessed using the Andrews Carson Score (ACS), the Oxford Elbow Score (OES), and the Disabilities of Arm, Shoulder and Hand (DASH) Score after six, twelve, 24 and 48 weeks. Furthermore, an isokinetic strength measurement for flexion strength was performed after 24 and 48 weeks. After 48 weeks the patient presented with excellent functional outcome scores and no follow-up complications. During the follow-up period, no differences in the functional scores nor in the isokinetic flexion strength measurement could be detected. Furthermore, no radiological complications (like heterotopic ossifications) could be detected in the postoperative radiographs after one year. Conclusions Anatomic reattachment of the distal biceps tendon is a successful operative treatment option for distal biceps tendon ruptures. Suture anchor fixation remains one of the most common techniques, as it allows fast surgery and provides good results with respect to range of motion (ROM) and functional scoring according to the current literature. However, the number of anchors required for a stable fixation remains unclear. As indicated by our presented case, we hypothesize, that there are no significant differences between a one-point or a two-point fixation. In the presented case report, no intraindividual differences between the usage of one versus two suture anchors were evident in the short-term follow-up. KW - Non-simultaneous bilateral distal biceps tendon rupture KW - Distal biceps tendon repair KW - Anatomic reattachment KW - Suture anchor KW - Case report Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229266 VL - 21 ER - TY - JOUR A1 - Armbruster, Nicole A1 - Krieg, Jennifer A1 - Weißenberger, Manuel A1 - Scheller, Carsten A1 - Steinert, Andre F. T1 - Rescued Chondrogenesis of Mesenchymal Stem Cells under Interleukin 1 Challenge by Foamyviral Interleukin 1 Receptor Antagonist Gene Transfer JF - Frontiers in Pharmacology N2 - Background: Mesenchymal stem cells (MSCs) and their chondrogenic differentiation have been extensively investigated in vitro as MSCs provide an attractive source besides chondrocytes for cartilage repair therapies. Here we established prototype foamyviral vectors (FVV) that are derived from apathogenic parent viruses and are characterized by a broad host range and a favorable integration pattern into the cellular genome. As the inflammatory cytokine interleukin 1 beta (IL1β) is frequently present in diseased joints, the protective effects of FVV expressing the human interleukin 1 receptor antagonist protein (IL1RA) were studied in an established in vitro model (aggregate culture system) of chondrogenesis in the presence of IL1β. Materials and Methods: We generated different recombinant FVVs encoding enhanced green fluorescent protein (EGFP) or IL1RA and examined their transduction efficiencies and transgene expression profiles using different cell lines and human primary MSCs derived from bone marrow-aspirates. Transgene expression was evaluated by fluorescence microscopy (EGFP), flow cytometry (EGFP), and ELISA (IL1RA). For evaluation of the functionality of the IL1RA transgene to block the inhibitory effects of IL1β on chondrogenesis of primary MSCs and an immortalized MSC cell line (TERT4 cells), the cells were maintained following transduction as aggregate cultures in standard chondrogenic media in the presence or absence of IL1β. After 3 weeks of culture, pellets were harvested and analyzed by histology and immunohistochemistry for chondrogenic phenotypes. Results: The different FVV efficiently transduced cell lines as well as primary MSCs, thereby reaching high transgene expression levels in 6-well plates with levels of around 100 ng/ml IL1RA. MSC aggregate cultures which were maintained in chondrogenic media without IL1β supplementation revealed a chondrogenic phenotype by means of strong positive staining for collagen type II and matrix proteoglycan (Alcian blue). Addition of IL1β was inhibitory to chondrogenesis in untreated control pellets. In contrast, foamyviral mediated IL1RA expression rescued the chondrogenesis in pellets cultured in the presence of IL1β. Transduced MSC pellets reached thereby very high IL1RA transgene expression levels with a peak of 1087 ng/ml after day 7, followed by a decrease to 194 ng/ml after day 21, while IL1RA concentrations of controls were permanently below 200 pg/ml. Conclusion: Our results indicate that FVV are capable of efficient gene transfer to MSCs, while reaching IL1RA transgene expression levels, that were able to efficiently block the impacts of IL1β in vitro. FVV merit further investigation as a means to study the potential as a gene transfer tool for MSC based therapies for cartilage repair. KW - mesenchymal stem cell KW - chondrogenesis KW - pellet culture KW - foamy virus KW - virus vectors KW - IL1RA KW - interleukin 1 receptor antagonist KW - arthritis Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170919 VL - 8 IS - 255 ER - TY - JOUR A1 - Boelch, Sebastian P. A1 - Gurok, Anna A1 - Gilbert, Fabian A1 - Weißenberger, Manuel A1 - Rudert, Maximilian A1 - Barthel, Thomas A1 - Reppenhagen, Stephan T1 - Why compromise the patella? Five-year follow-up results of medial patellofemoral ligament reconstruction with soft tissue patellar fixation JF - International Orthopaedics N2 - Purpose This study investigates the redislocation rate and functional outcome at a minimum follow-up of five years after medial patellofemoral ligament (MPFL) reconstruction with soft tissue patellar fixation for patella instability. Methods Patients were retrospectively identified and knees were evaluated for trochlea dysplasia according to Dejour, for presence of patella alta and for presence of cartilage lesion at surgery. At a minimum follow-up of five years, information about an incident of redislocation was obtained. Kujala, Lysholm, and Tegner questionnaires as well as range of motion were used to measure functional outcome. Results Eighty-nine knees were included. Follow-up rate for redislocation was 79.8% and for functional outcome 58.4%. After a mean follow-up of 5.8 years, the redislocation rate was 5.6%. There was significant improvement of the Kujala score (68.8 to 88.2, p = 0.000) and of the Lysholm score (71.3 to 88.4, p = 0.000). Range of motion at follow-up was 149.0° (115–165). 77.5% of the knees had patella alta and 52.9% trochlear dysplasia types B, C, or D. Patellar cartilage legions were present in 54.2%. Redislocations occurred in knees with trochlear dysplasia type C in combination with patella alta. Conclusion MPFL reconstruction with soft tissue patellar fixation leads to significant improvement of knee function and low midterm redislocation rate. Patients with high-grade trochlear dysplasia should be considered for additional osseous correction. KW - MPFL KW - medial patellofemoral ligament KW - patella instability KW - patella dislocation KW - trochlear dysplasia KW - patella alta Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-235751 SN - 0341-2695 VL - 45 ER - TY - JOUR A1 - Horas, Konstantin A1 - van Herck, Ulrike A1 - Maier, Gerrit S. A1 - Maus, Uwe A1 - Harrasser, Norbert A1 - Jakob, Franz A1 - Weissenberger, Manuel A1 - Arnholdt, Jörg A1 - Holzapfel, Boris M. A1 - Rudert, Maximilian T1 - Does vitamin D deficiency predict tumour malignancy in patients with bone tumours? Data from a multi-center cohort analysis JF - Journal of Bone Oncology N2 - Vitamin D deficiency is a global health concern that is estimated to afflict over one billion people globally. The major role of vitamin D is that of a regulator of calcium and phosphate metabolism, thus, being essential for proper bone mineralisation. Concomitantly, vitamin D is known to exert numerous extra-skeletal actions. For example, it has become evident that vitamin D has direct anti-proliferative, pro-differentiation and pro-apoptotic actions on cancer cells. Hence, vitamin D deficiency has been associated with increased cancer risk and worse prognosis in several malignancies. We have recently demonstrated that vitamin D deficiency promotes secondary cancer growth in bone. These findings were partly attributable to an increase in bone remodelling but also through direct effects of vitamin D on cancer cells. To date, very little is known about vitamin D status of patients with bone tumours in general. Thus, the objective of this study was to assess vitamin D status of patients with diverse bone tumours. Moreover, the aim was to elucidate whether or not there is an association between pre-diagnostic vitamin D status and tumour malignancy in patients with bone tumours. In a multi-center analysis, 25(OH)D, PTH and calcium levels of 225 patients that presented with various bone tumours between 2017 and 2018 were assessed. Collectively, 76% of all patients had insufficient vitamin D levels with a total mean 25(OH)D level of 21.43 ng/ml (53.58 nmol/L). In particular, 52% (117/225) of patients were identified as vitamin D deficient and further 24% of patients (55/225) were vitamin D insufficient. Notably, patients diagnosed with malignant bone tumours had significantly lower 25(OH)D levels than patients diagnosed with benign bone tumours [19.3 vs. 22.75 ng/ml (48.25 vs. 56.86 nmol/L); p = 0.04). In conclusion, we found a widespread and distressing rate of vitamin D deficiency and insufficiency in patients with bone tumours. However, especially for patients with bone tumours sufficient vitamin D levels seem to be of great importance. Thus, we believe that 25(OH)D status should routinely be monitored in these patients. Collectively, there should be an increased awareness for physicians to assess and if necessary correct vitamin D status of patients with bone tumours in general or of those at great risk of developing bone tumours. KW - bone tumour KW - vitamin D KW - hypovitaminosis D KW - vitamin D deficiency KW - malignancy KW - tumour malignancy Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230314 VL - 25 ER - TY - JOUR A1 - Heinz, Tizian A1 - Eidmann, Annette A1 - Anderson, Philip A1 - Weißenberger, Manuel A1 - Jakuscheit, Axel A1 - Rudert, Maximilian A1 - Stratos, Ioannis T1 - Trends in computer-assisted surgery for total knee arthroplasty in Germany: an analysis based on the operative procedure classification system between 2010 to 2021 JF - Journal of Clinical Medicine N2 - Alignment strategies for primary total knee arthroplasty (TKA) have changed significantly over time with a shift towards a more individualized alignment goal. At the same time, computer-assisted surgery (CAS) has gained interest for intraoperative control and accuracy in implant positioning and limb alignment. Despite the often discussed benefits and drawbacks of robotics and navigation for TKA, the routine use of these new devices on a day-to-day basis remains obscure. Therefore, nationwide hospital billing data based on the Operation Procedure Classification System (OPS) were retrieved from the Federal Statistical Office of Germany for the period from 2010 to 2021. OPS codes for primary total knee arthroplasty (OPS code: 5-822*) were further analyzed regarding the usage of computer navigation (additional OPS code: 5-988) or robotic devices (additional OPS code: 5-987). Gender and age at the time of surgery were also assessed. The results show a total of 2,226,559 primary TKAs were implanted between 2010 and 2021, of which 2,044,914 were performed conventionally (91.84% of all TKAs). A total of 170,276 TKAs were performed using navigation technique (7.65% of all TKAs) and another 11,369 TKAs were performed using robotics (0.51% of all TKAs). For the period from 2018 to 2021, a substantial increase in robot-assisted TKA (R-TKA) was observed, with an average increase rate of 84.74% per year, while the number of navigated TKAs declined (−3.67% per year). Computer-assisted surgery, and particularly robotics for TKA, are seeing growing popularity and stepwise translation into routine clinical use in Germany, with a steep increase rate of more than 80% per year since 2018. Nevertheless, the majority of TKAs are still performed using manual instrumentation, rendering conventional TKA the currently unchanged gold standard. KW - robotic KW - TKA KW - knee replacement KW - computer navigation KW - Germany Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-304879 SN - 2077-0383 VL - 12 IS - 2 ER -