Markus Dietmar Schofer

Electrospun PLLA Nanofiber Scaffolds and Their Use in Combination with BMP-2 for Reconstruction of Bone Defects

Introduction Adequate migration and differentiation of mesenchymal stem cells is essential for regeneration of large bone defects. To achieve this, modern graft materials are becoming increasingly important. Among them, electrospun nanofiber scaffolds are a promising approach, because of their high physical porosity and potential to mimic the extracellular matrix (ECM). Materials and Methods The objective of the present study was to examine the impact of electrospun PLLA nanofiber scaffolds on bone formation in vivo, using a critical size rat calvarial defect model. In addition we analyzed whether direct incorporation of bone morphogenetic protein 2 (BMP-2) into nanofibers could enhance the osteoinductivity of the scaffolds. Two critical size calvarial defects (5 mm) were created in the parietal bones of adult male Sprague-Dawley rats. Defects were either (1) left unfilled, or treated with (2) bovine spongiosa, (3) PLLA scaffolds alone or (4) PLLA/BMP-2 scaffolds. Cranial CT-scans were taken at fixed intervals in vivo. Specimens obtained after euthanasia were processed for histology, histomorphometry and immunostaining (Osteocalcin, BMP-2 and Smad5). Results PLLA scaffolds were well colonized with cells after implantation, but only showed marginal ossification. PLLA/BMP-2 scaffolds showed much better bone regeneration and several ossification foci were observed throughout the defect. PLLA/BMP-2 scaffolds also stimulated significantly faster bone regeneration during the first eight weeks compared to bovine spongiosa. However, no significant differences between these two scaffolds could be observed after twelve weeks. Expression of osteogenic marker proteins in PLLA/BMP-2 scaffolds continuously increased throughout the observation period. After twelve weeks osteocalcin, BMP-2 and Smad5 were all significantly higher in the PLLA/BMP-2 group than in all other groups. Conclusion Electrospun PLLA nanofibers facilitate colonization of bone defects, while their use in combination with BMP-2 also increases bone regeneration in vivo and thus combines osteoconductivity of the scaffold with the ability to maintain an adequate osteogenic stimulus.

Improved healing response in delayed unions of the tibia with low-intensity pulsed ultrasound: results of a randomized sham-controlled trial.

BackgroundWe compared the healing response of tibial delayed unions between subjects treated with low-intensity pulsed ultrasound (LIPUS) (n = 51) and subjects treated with a sham device (n = 50). Fracture age was ≥ 4 months in all cases. Study personnel and participants were blinded to random treatment assignment throughout the study.MethodsThis multi-center randomized sham-controlled trial was undertaken at six hospitals in Germany. Adult patients who had sustained a tibial shaft fracture that subsequently showed inadequate progress toward healing (i.e., delayed union) were enrolled and randomized to receive either LIPUS (Exogen 2000/2000+, Smith & Nephew GmbH, Schenefeld, Germany) or an identical nonoperative sham device. The daily treatment duration was 20 minutes, for a period of 16 weeks. Subjects randomly assigned to active treatment had the ultrasound pressure wave signal set at the following parameters: 1.5 MHz frequency, 1 kHz repetition rate, 200 μs pulse duration, 30 mW/cm2 spatial intensity. Progress toward healing was estimated from changes in bone mineral density (BMD) and gap area as determined from computed tomography scans. Intention-to-treat analysis was conducted using a multiple imputation methodology.ResultsBased on log-transformed data, mean improvement in BMD was 1.34 (90% confidence interval (CI) 1.14 to 1.57) times greater for LIPUS-treated subjects compared to sham (p = 0.002). A mean reduction in bone gap area also favored LIPUS treatment (p = 0.014).ConclusionsThese findings demonstrate significantly greater progress toward bone healing after LIPUS treatment compared to no LIPUS treatment in subjects with established delayed unions of the tibia.

Biomechanical Comparison of Arthroscopically Performable Techniques for Suprapectoral Biceps Tenodesis

PURPOSE The aim of this study was to biomechanically compare the cyclic and ultimate failure load (UFL) of 4 widely used techniques for arthroscopically performable suprapectoral tenodesis of the long head of the biceps tendon (LHB). METHODS We used 28 fresh-frozen human cadaveric specimens (mean age, 65 years [range, 43 to 78 years; SD, 6.7 years]; 43% male specimens) to investigate 4 different techniques for LHB tenodesis. All techniques were performed in an open manner, with localization at the entrance of the bicipital groove. Two suture anchor techniques (Healix [DePuy Mitek, Raynham, MA], 5.5 mm, with modified lasso-loop stitch; BioSwiveLock [Arthrex, Naples, FL], 5.5 mm, with interlocking Krackow stitch) and two techniques using tenodesis screws (Bio-Tenodesis screw [Arthrex], 8 × 23 mm; Biceptor [Smith & Nephew, Andover, MA], 8 × 25 mm) were investigated. Under a 10-N preload, an axial cyclic load with 100 cycles, 1-Hz frequency, and 50-N maximum load was applied. UFL was evaluated with an axial traction of 0.2 mm/s. LHB displacement during testing was measured by 3-dimensional photogrammetry. RESULTS All techniques had a mean displacement of less than 3 mm after cyclic loading. The highest UFL was measured with the Bio-Tenodesis screw (mean, 218.3 N; range, 134.0 to 313.0 N; SD, 59.7 N) and the lowest with the BioSwiveLock (mean, 111.2 N; range, 60.0 to 156.8 N; SD, 32.3 N). The Healix had the second highest UFL (mean, 187.1 N; range, 144.7 to 245.0 N; SD, 35.5 N), followed by the Biceptor (mean, 173.9 N; range, 147.0 to 209.3 N; SD, 27.2 N). There was no significant difference between the Healix, Bio-Tenodesis screw, and Biceptor (P > .05), but the Healix and Bio-Tenodesis screw had a significantly higher UFL than the BioSwiveLock (P < .01). The failure mode was either suture cutout or failure at the anchor-suture-bone interface or of the tendon itself and was generally dependent on technique. CONCLUSIONS All techniques resisted cyclic testing without a higher grade of displacement, and all devices except the BioSwiveLock had a satisfactory UFL whereas different failure mechanisms were present. The modified lasso-loop stitch provides sufficient tendon fixation and is equivalent to interference screws. CLINICAL RELEVANCE The lasso-loop suture anchor technique is an appropriate alternative for suprapectoral LHB tenodesis compared with tenodesis screw techniques.

Closing-wedge high tibial osteotomy: survival and risk factor analysis at long-term follow up

BackgroundClosing-wedge high tibial osteotomy (HTO) is successful for the treatment of medial osteoarthritis with varus malalignment. Preoperative risk factors for HTO failure are still controversial. The aim of this study was to elucidate the outcome and assess the influence of risk factors on long term HTO survival.Methods199 patients were retrospectively studied with a mean follow-up period of 9.6 years after HTO. HTO failure was defined as the need for conversion to TKA. Survival was analyzed with the Kaplan-Meier method. Knee function was evaluated by the Hospital for Special Surgery (HSS) score. HTO-associated complications were also assessed. Univariate, multivariate, and logistic regression analysis were performed to evaluate the influence of age, gender, BMI, preoperative Kellgren-Lawrence osteoarthritis grade, and varus angle on HTO failure.Results39 complications were recorded. Thus far, 36 HTOs were converted to TKA. The survival of HTO was 84% after 9.6 years. Knee function was considered excellent or good in 64% of patients. A significant preoperative risk factor for HTO failure was osteoarthritis, Kellgren-Lawrence grade >2.ConclusionHTO provides good clinical results in long-term follow-up. Preoperative osteoarthritis Kellgren-Lawrence grade >2 is a significant predictive risk factor for HTO failure. Results of HTO may be improved by careful patient selection. Complications associated with HTO should not be underestimated.

Influence of Poly(L-Lactic Acid) Nanofibers and BMP-2–Containing Poly(L-Lactic Acid) Nanofibers on Growth and Osteogenic Differentiation of Human Mesenchymal Stem Cells

The aim of this study was to characterize synthetic poly-(L-lactic acid) (PLLA) nanofibers concerning their ability to promote growth and osteogenic differentiation of stem cells in vitro, as well as to test their suitability as a carrier system for growth factors. Fiber matrices composed of PLLA or BMP-2–incorporated PLLA were seeded with human mesenchymal stem cells and cultivated over a period of 22 days under growth and osteoinductive conditions, and analyzed during the course of culture, with respect to gene expression of alkaline phosphatase (ALP), osteocalcin (OC), and collagen I (COL-I). Furthermore, COL-I and OC deposition, as well as cell densities and proliferation, were analyzed using fluorescence microscopy. Although the presence of nanofibers diminished the dexamethasone-induced proliferation, there were no differences in cell densities or deposition of either COL-I or OC after 22 days of culture. The gene expression of ALP, OC, and COL-I decreased in the initial phase of cell cultivation on PLLA nanofibers as compared to cover slip control, but normalized during the course of cultivation. The initial down-regulation was not observed when BMP-2 was directly incorporated into PLLA nanofibers by electrospinning, indicating that growth factors like BMP-2 might survive the spinning process in a bioactive form.

Characterization of a PLLA-collagen I blend nanofiber scaffold with respect to growth and osteogenic differentiation of human mesenchymal stem cells.

The aim of this study was to enhance synthetic poly(L-lactic acid) (PLLA) nanofibers by blending with collagen I (COLI) in order to improve their ability to promote growth and osteogenic differentiation of stem cells in vitro. Fiber matrices composed of PLLA and COLI in different ratios were characterized with respect to their morphology, as well as their ability to promote growth of human mesenchymal stem cells (hMSC) over a period of 22 days. Furthermore, the course of differentiation was analyzed by gene expression of alkaline phosphatase (ALP), osteocalcin (OC), and COLI. The PLLA-COLI blend nanofibers presented themselves with a relatively smooth surface. They were more hydrophilic as compared to PLLA nanofibers alone and formed a gel-like structure with a stable nanofiber backbone when incubated in aqueous solutions. We examined nanofibers composed of different PLLA and COLI ratios. A composition of 4:1 ratio of PLLA:COLI showed the best results. When hMSC were cultured on the PLLA-COLI nanofiber blend, growth as well as osteoblast differentiation (determined as gene expression of ALP, OC, and COLI) was enhanced when compared to PLLA nanofibers alone. Therefore, the blending of PLLA with COLI might be a suitable tool to enhance PLLA nanofibers with respect to bone tissue engineering.

TKA following high tibial osteotomy versus primary TKA - a matched pair analysis

BackgroundHigh tibial osteotomy (HTO) is a well established technique for the treatment of medial osteoarthritis of the knee with varus malalignment. Results of total knee arthroplasty (TKA) after previous HTO are still discussed controversially. The aim of this study was to elucidate the clinical and radiological results as well as perioperative data of prior HTO on TKA.MethodsForty-one TKA after HTO were compared to 41 primary TKA at minimum of six years follow-up. Patients were matched according to age, gender, follow-up, etiology, and prosthetic design. Surgical data and complications were evaluated. Clinical outcome was assessed using a number of clinical scores and the visual analogue scale (VAS) for pain. X-rays were evaluated by the method of the American Knee Society. The patellar position was measured by the Insall-Salvati ratio.ResultsThere was no significant difference in mean operation time (p = 0.47) and complication rate (p = 0.08). The Knee Score of the KSS (p = 0.0007) and the ROM (p = 0.006 for extension and p = 0.004 for flexion, respectively) were significantly better in the control group. Mid-term results of the VAS, WOMAC, Lequesne, UCLA, Fellers Patellar Score and SF-36 showed no significant difference. Femoral and tibial component alignment were similar in both groups. One tibial component showed suspect radiolucencies in the HTO group. The Insall-Salvati ratio showed three patients with patella alta and one patient with patella baja in the HTO group. At latest follow-up all implants were still in place.ConclusionsEvaluating the clinical and radiological outcome, significant differences were only detected for range of motion and the Knee Score of the KSS. The present study suggests that the results of TKA with and without prior HTO are mainly identical. Although patients with a previous HTO had more complications, no statistically significant differences were noted with this group size.

Influence of nanofibers on growth and gene expression of human tendon derived fibroblast

BackgroundRotator cuff tears are a common and frequent lesion especially in older patients. The mechanisms of tendon repair are not fully understood. Common therapy options for tendon repair include mini-open or arthroscopic surgery. The use of growth factors in experimental studies is mentioned in the literature. Nanofiber scaffolds, which provide several criteria for the healing process, might be a suitable therapy option for operative treatment. The aim of this study was to explore the effects of nanofiber scaffolds on human tendon derived fibroblasts (TDFs), as well as the gene expression and matrix deposition of these fibroblasts.MethodsNanofibers composed of PLLA and PLLA/Col-I were seeded with human tendon derived fibroblasts and cultivated over a period of 22 days under growth-inductive conditions, and analyzed during the course of culture, with respect to gene expression of different extra cellular matrix components such as collagens, bigylcan and decorin. Furthermore, we measured cell densities and proliferation by using fluorescene microscopy.ResultsPLLA nanofibers possessed a growth inhibitory effect on TDFs. Furthermore, no meaningful influence on the gene expression of collagen I, collagen III and decorin could be observed, while the expression of collagen X increased during the course of cultivation. On the other hand, PLLA/Col-I blend nanofibers had no negative influence on the growth of TDFs. Furthermore, blending PLLA nanofibers with collagen had a positive effect on the gene expression of collagen I, III, X and decorin. Here, gene expression indicated that focal adherence kinases might be involved.ConclusionThis study indicates that the use of nanofibers influence expression of genes associated with the extra cellular matrix formation. The composition of the nanofibers plays a critical role. While PLLA/Col-I blend nanofibers enhance the collagen I and III formation, their expression on PLLA nanofibers was more comparable to controls. However, irrespective of the chemical composition of the fibres, the collagen deposition was altered, an effect which might be associated with a decreased expression of biglycanes.

Co-occurrence of outlet impingement syndrome of the shoulder and restricted range of motion in the thoracic spine - a prospective study with ultrasound-based motion analysis

BackgroundShoulder complaints, and especially the outlet-impingement syndrome, are a common condition. Among other things, poor posture has been discussed as a cause. A correlation between impingement syndrome and restricted mobility of the thoracic spine (T) has been described earlier, but there has been no motion analysis of the thoracic spine to show these correlations. In the present prospective study, we intended to find out whether there is a significant difference in the thoracic sagittal range of motion (ROM) between patients with a shoulder outlet impingement syndrome and a group of patients who had no shoulder pathology. Secondly, we wanted to clarify whether Otts sign correlates with ultrasound topometric measurements.MethodsTwo sex- and age-matched groups (2 × n = 39) underwent a clinical and an ultrasound topometric examination. The postures examined were sitting up straight, sitting in maximal flexion and sitting in maximal extension. The disabilities of the arm, shoulder and hand (DASH) score (obtained by means of a self-assessment questionnaire) and the Constant score were calculated. Lengthening and shortening of the dorsal projections of the spine in functional positions was measured by tape with Otts sign.ResultsOn examination of the thoracic kyphosis in the erect seated posture there were no significant differences between the two groups (p = 0.66). With ultrasound topometric measurement it was possible to show a significantly restricted segmental mobility of the thoracic spine in the study group compared with the control group (p = 0.01). An in-depth look at the mobility of the subsegments T1-4, T5-8 and T9-12 revealed that differences between the groups in the mobility in the lower two sections of the thoracic spine were significant (T5-8: p = 0.03; T9-12: p = 0.02). The study group had an average Constant score of 35.1 points and the control group, 85.5 (p < 0.001). On the DASH score the patient group reached 34.2 points and the control group, 1.4 (p < 0.001). The results of Otts sign differed significantly between the two collectives (p = 0.0018), but showed a weak correlation with the ultrasound topometric measurements (study group flexion/extension: r = 0.36/0.43, control group flexion/extension: r = 0.29/0.26).ConclusionThe mobility of the thoracic spine should receive more attention in the diagnosis and therapy of patients with shoulder outlet impingement syndrome.

Increased glenohumeral translation and biceps load after SLAP lesions with potential influence on glenohumeral chondral lesions: a biomechanical study on human cadavers.

PurposeThe aim of the study was to evaluate the stabilizing function of the long head of biceps tendon (LHB) and its tension, both without and with the presence of SLAP lesion to analyze a potentially occurring humeral chondral print of LHB with consecutive glenohumeral chondral lesions in SLAP lesions.MethodsTestings were performed on 21 fresh frozen human cadaver shoulders with intact shoulder girdle by a 5 axis industrial robot with a force/moment sensor and 20 N joint compression, 50 N force in anterior, posterior, anterosuperior, and anteroinferior direction, and 0°, 30°, 60° of abduction. LHB was connected over a force measuring sensor with 5 N and 25 N preload. A type IIC SLAP lesion was created arthroscopically.ResultsA significant increase in anterior and anteroinferior translation was evaluated, whereas the LHB tension increased significantly in at most anterior and anterosuperior direction. The highest increase in translation and LHB tension after SLAP lesion was measured in anterior translation in at most 60° of abduction.The glenohumeral translation was significantly higher in SLAP lesions without LHB tenotomy than after isolated LHB tenotomy.ConclusionsSLAP lesions lead to increased glenohumeral translation and concurrently LHB tension and load in at most anterior direction. The increased anterior glenohumeral instability and the increased LHB load pressing on the humeral head might cause glenohumeral chondral lesions with a typical chondral print-like lesion on the humeral head underneath the LHB.