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Purpose: Longitudinal imaging studies are important in the translational process of stem cell-based therapies. Small animal imaging models are widely available and practical but insufficiently depict important morphologic detail. In contrary, large animal models are logistically challenging and costly but offer greater imaging quality. In order to combine the advantages of both, we developed an intermediate-sized rabbit animal model for cartilage imaging studies.Procedures: Rabbit mesenchymal stem cells (rMSC) were isolated as primary cultures from the bone marrow of New Zealand white rabbits. rMSC were subsequentially transduced lentivirally with eGFP and magnetically labeled with the iron oxide ferucarbotran. eGFP expression was evaluated by flow cytometry and iron uptake was analyzed by isotope dilution mass spectrometry and Prussian blue staining. Fluorescence microscopy of eGFP-transduced rMSC was performed. Viability and induction of apoptosis were assessed by XTT and caspase-3/-7 measurements. The chondrogenic potential of labeled cells was quantified by glycosaminoglycan contents in TGF-β3 induced pellet cultures. Labeled and unlabeled cells underwent magnetic resonance imaging (MRI) at 1.5 T before and after differentiation using T1-, T2-, and T2*-weighted pulse sequences. Relaxation rates were calculated. rMSCs were implanted in fibrin clots in osteochondral defects of cadaveric rabbit knees and imaged by 7 T MRI. T2* maps were calculated. Statistical analyses were performed using multiple regression models.Results: Efficiency of lentiviral transduction was greater than 90 %. Fluorescence signal was dose dependent. Cellular iron uptake was significant for all concentrations (p < 0.05) and dose dependent (3.3-56.5 pg Fe/cell). Labeled rMSC showed a strong, dose-dependent contrast on all MR pulse sequences and a significant decrease in T2 and T2* relaxation rates. Compared with non-transduced or unlabeled controls, there were no adverse effects on cell viability, rate of apoptosis, or chondrogenic differentiation. MRI of labeled rMSCs in osteochondral defects showed a significant signal of the transplant with additional high-resolution anatomical information.Conclusions: This intermediate-sized rabbit model and its bifunctional labeling technique allow for improved depiction of anatomic detail for noninvasive in vivo rMSC tracking with MRI and for immunohistological correlation by fluorescence microscopy. [ABSTRACT FROM AUTHOR]

Background: Anatomic features of the tibial plateau (ie, posterior slope and medial concavity) have been associated with an increased risk of anterior cruciate ligament (ACL) injuries. However, it remains unclear whether these findings translate to ACL injuries sustained during recreational alpine skiing. Purpose: To investigate the association in recreational alpine skiers between prominent morphological features of the tibial plateau (slope and concavity) and the risk of sustaining an ACL injury during a noncontact incident. Study Design: Case-control study; Level of evidence, 3. Methods: Magnetic resonance imaging data of 121 recreational alpine skiers (74 female, 47 male) after a noncontact knee injury were used for this study. Of these patients, 80 (71% female [n = 57]) had a complete unilateral ACL tear (rupture group), and 41 (41% female [n = 17]) had no indications of an ACL injury (intact group). Two blinded independent examiners measured the slopes of the tibial plateau in the sagittal and coronal planes along with the maximum depth of the medial tibial plateau. Measurements were compared between sexes and between groups using t tests. Logistic regression was used to assess the associations between quantified anatomic indices and the risk of ACL injuries. Results: Within 121 study patients, female skiers had greater odds of an ACL tear compared with male skiers (odds ratio, 3.5; 95% CI, 1.6-7.8; P < .001). Female skiers were more likely to have a greater lateral tibial slope (LTS) (P = .02) and medial tibial slope (MTS) (P = .02) with a shallower medial tibial depth (MTD) (P = .02) compared with male skiers. No differences between sexes were observed in the coronal tibial slope (CTS) (P = .97). Male and female skiers as a combined group showed no associations between quantified anatomic indices and the risk of sustaining an ACL tear (P > .10). Likewise, no significant differences were observed between the intact versus rupture group in any of the quantified anatomic indices (P > .10). Similar findings were observed when the analyses were repeated on male and female skiers separately. Conclusion: Despite differences between sexes in knee anatomy and the injury risk, the sagittal and coronal slopes (LTS, MTS, CTS), as well as the concavity of the medial tibial plateau (MTD), were not associated with the risk of an ACL tear during a noncontact injury among recreational alpine skiers. [ABSTRACT FROM AUTHOR]

Abstract: Bioactive implants are promising tools in regenerative medicine. Here we describe a versatile procedure for preparing a gene-activated matrix on titanium. Lyophilized copolymer-protected gene vectors (COPROGs) suspended in poly(d,l-lactide) (PDLLA) solutions in ethyl acetate were used to varnish solid surfaces. The gene-activated PDLLA surfaces were first established on polypropylene 96-well plates. Vector release from these surfaces in aqueous buffer, cell viability and gene transfer efficiency to NIH 3T3 fibroblasts was strongly dependent on the vector dose and its ratio to PDLLA film thickness. A detailed analysis of these relationships allowed establishing correlations which can be used to calculate suitable combinations of COPROGs and PDLLA yielding optimal gene transfer efficiency. This was verified with COPROG-activated PDLLA coatings on titanium foils. HEK 293 and mesenchymal stem cells expressed the BMP-2 gene comprised in the gene-activated surface in a manner that was consistent with the predicted dose–response and toxicity profiles found in NIH 3T3 cells. The systematic procedure presented here for identifying optimal coating compositions can be applied to any combination of vector type and coating material. [Copyright &y& Elsevier]

Abstract: Matrix-assisted chondrocyte transplantation (m-ACI) still lacks any standardization in its execution in terms of cell passage (P), cell yield (C) and in vitro membrane-holding time (T). It was the goal of this study to analyze the effect of shifting cell culture parameters (P, C, T) on the in vitro as well as in vivo effort of a regulated animal m-ACI. Autologous rabbit knee articular chondrocytes were seeded within bilayer collagen I/III 3-D matrices in variation of P, C and T. Each time, 2 PCT-identical by 2 PCT-identical cell-matrix-constructs (CMC)/animal were created. Simultaneously 2 (PCT-distinct) were re-implanted (CMC-e) autologous into artificial trochlear pristine chondral defects in vivo to remain for 12 weeks while the remaining 2 were harvested (CMC-i) for immediate in vitro analysis at the time of transplantation of their identical twins. mRNA of both, CMC-e regenerates and CMC-i membranes, was analyzed for Collagen-1,-2,-10, COMP, Aggrecan, Sox9 expression by use of a mixed linear model, multiple regression analysis. Generally, CMC-i values were higher than CMC-e values for differentiation targets; the opposite was true for dedifferentiation targets. Regarding individual gene expression, in vivo regenerate cell-matrix properties were significantly dependent on initial cell-matrix in vitro values as a sign of linearity. The parameter membrane-holding time (T) had strongest effects on the resulting mRNA expression with slightly less impact of the parameter passage (P), whereas cell yield (C) had clearly less effects. Noting differences between in vitro and in vivo data, in general, optimal expression patterns concerning chondrogenic differentiation were achieved by few passages, medium cellular yield, short membrane-holding time. Clinical m-ACI may benefit from optimal orchestration of the cell culture parameters passage, yield and time. [Copyright &y& Elsevier]

Summary: Integrin-mediated adhesion plays a central role in T cell trafficking and activation. Genetic inactivation of the guanine nucleotide-binding (G) protein α-subunits Gα12 and Gα13 resulted in an increased activity of integrin leukocyte-function-antigen-1 in murine CD4+ T cells. The interaction with allogeneic dendritic cells was enhanced, leading to an abnormal proliferative response in vitro. In vivo, T cell-specific inactivation of Gα12 and Gα13 caused lymphadenopathy due to increased lymph node entry and enhanced T cell proliferation, and the susceptibility toward T cell-mediated diseases was enhanced. Mechanistically, we show that in the absence of Gα12 and Gα13 the activity of the small GTPases Rac1 and Rap1 was increased, whereas signaling of the small GTPase RhoA was strongly reduced. Our data indicate that locally produced mediators signal through Gα12- and Gα13-coupled receptors to negatively regulate cell polarization and adhesiveness, thereby fine-tuning T cell trafficking, proliferation, and susceptibility toward T cell-mediated diseases. [Copyright &y& Elsevier]

Abstract: For more than 20 years, countries and their agencies which monitor radionuclide discharge sites and storage facilities have relied on the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 4355 Peruvian Soil. Its low fallout contamination makes it an ideal soil blank for measurements associated with terrestrial-pathway-to-man studies. Presently, SRM 4355 is out of stock, and a new batch of the Peruvian soil is currently under development as future NIST SRM 4355A. Both environmental radioanalytical laboratories and mass spectrometry communities will benefit from the use of this SRM. The former must assess their laboratory procedural contamination and measurement detection limits by measurement of blank sample material. The Peruvian Soil is so low in anthropogenic radionuclide content that it is a suitable virtual blank. On the other hand, mass spectrometric laboratories have high sensitivity instruments that are capable of quantitative isotopic measurements at low plutonium levels in the SRM 4355 (first Peruvian Soil SRM) that provided the mass spectrometric community with the calibration, quality control, and testing material needed for methods development and legal defensibility. The quantification of the ultra-low plutonium content in the SRM 4355A was a considerable challenge for the mass spectrometric laboratories. Careful blank control and correction, isobaric interferences, instrument stability, peak assessment, and detection assessment were necessary. Furthermore, a systematic statistical evaluation of the measurement results and considerable discussions with the mass spectroscopy metrologists were needed to derive the certified values and uncertainties. The one sided upper limit of the 95% tolerance with 95% confidence for the massic 239Pu content in SRM 4355A is estimated to be 54,000atoms/g. [Copyright &y& Elsevier]

Purpose: To evaluate the clinical outcomes of patients with a minimum 2-year follow-up following contemporary patellofemoral inlay arthroplasty (PFIA) and to identify potential risk factors for failure in a multi-center study., Methods: All patients who underwent implantation of PFIA between 09/2009 and 11/2016 at 11 specialized orthopedic referral centers were enrolled in the study and were evaluated retrospectively at a minimum 2-year follow-up. Clinical outcomes included the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, the Knee Injury and Osteoarthritis Outcome Score (KOOS), the Tegner Scale, the visual analogue scale (VAS) for pain, and subjective patient satisfaction. Pre- and perioperative risk factors were compared among failures and non-failures to determine potential risk factors., Results: A total of 263 patients (85% follow-up rate) could be enrolled. The mean age at the time of index surgery was 49 ± 12 years with a mean postoperative follow-up of 45 ± 18 months. The overall failure rate was 11% (28 patients), of which 18% (5 patients) were patients with patella resurfacing at index surgery and 82% (23 patients) were patients without initial patella resurfacing. At final follow-up, 93% of the patients who did not fail were satisfied with the procedure with a mean transformed WOMAC Score of 84.5 ± 14.5 points, a mean KOOS Score of 73.3 ± 17.1 points, a mean Tegner Score of 3.4 ± 1.4 points and a mean VAS pain of 2.4 ± 2.0 points. An increased BMI was significantly correlated with a worse postoperative outcome. Concomitant procedures addressing patellofemoral instability or malalignment, the lack of patellofemoral resurfacing at the index surgery and a high BMI were significantly correlated with failure in our patient cohort., Conclusion: Patellofemoral inlay arthroplasty shows high patient satisfaction with good functional outcomes at short-term follow-up and thus can be considered a viable treatment option in young patients suffering from isolated patellofemoral arthritis. Patellar resurfacing at index surgery is recommended to decrease the risk of failure., Level of Evidence: Retrospective case series, Level IV., (© 2021. The Author(s).)

Background: To examine the risk of injury to the popliteal neurovascular bundle (pNVB) during all-inside repair of the posterior horn of the lateral meniscus (PHLM) using Upright-MRIs., Methods: Upright-MRIs of 61 knees in extension (ext) and 90°-flexion (flex) were included. Distance D from the PHLM to the pNVB was compared between extended and 90°-flexed position, subgroups with/without joint-effusion and evaluated according to demographics. Portal safety was assessed simulating suturing of the PHLM via four arthroscopy portals. Distance d (shortest space from the simulated suturing-device trajectory lines to the pNVB) was compared among portals in increasing distances from the posterior cruciate ligament (PCL)., Results: D is longer in flex (17.3 ± 6.0 mm) than in ext (11.3 ± 4.2 mm, p < 0.0001). MRIs with joint-effusion displayed longer values of D than scans without joint-effusion (flex: 20.4 ± 7.1 mm vs. 16.1 ± 5.2 mm, p = 0.012). Shorter distances are associated with female gender, lower body weight and lower BMI. At 0 mm from the PCL, the 1 cm-lateral portal was the safest (p < 0.0001) whereas at 3 mm/6mm/9mm/12 mm the 1 cm-medial portal showed the longest d values (p < 0.0001 each)., Conclusion: All-inside suturing of the PHLM is safer in 90°-flexion, in presence of intraarticular fluid and in male patients with increasing weight/BMI. Sutures of the PHLM at 0 mm from the PCL are safer from a 1 cm-lateral portal whereas for tears located ≥ 3 mm from the PCL a 1 cm-medial portal involves a lower neurovascular risk. Upright-MRI proves excellent for preoperative planning to minimize neurovascular risks., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Purpose: To evaluate subjective and objective clinical and magnetic resonance imaging-based radiologic outcomes after short-term follow-up in patients with focal full-size cartilage lesions of the knee joint treated with all-arthroscopic hydrogel-based autologous chondrocyte transplantation., Methods: A retrospective study on patients with isolated focal cartilage defects of the knee joint who were treated with arthroscopically conducted matrix-induced autologous chondrocyte transplantation was performed. Clinical scores were assessed at baseline and final follow-up using the Tegner Score, visual analog scale, the International Knee Documentation Committee, and the 5 subscales of the Knee Injury and Osteoarthritis Outcome Score. Magnetic resonance imaging scans of the treated knee joints were evaluated with the updated MOCART (Magnetic Resonance Observation of Cartilage Repair Tissue) 2.0 scoring system at follow-up., Results: Twenty-nine consecutive patients were included in the study. Mean time to follow-up was 24.9 ± 1.1 months. Average visual analog scale decreased significantly from 6.5 ± 3.1 preoperatively to 2.3 ± 1.6 at follow-up (P < .0001). Tegner score increased from 3.1 ± 1.3 to 4.3 ± 1.2 (P < .0001) and the International Knee Documentation Committee from 43.8 ± 21.9 to 64.9 ± 18.9 (P < .0001). Also, all Knee Injury and Osteoarthritis Outcome Score subscales displayed significant improvements. Patients showed similar improvements of nearly all clinical scores independent of the defect size. Average MOCART2.0 score was 70.0 ± 13.6 and 20 patients scored ≥70 points. All 8 patients with large defects (>5 cm 2 ) scored ≥75 points., Conclusions: In this small study, injectable matrix-induced autologous chondrocyte transplantation therapy in the knee joint led to favourable clinical and radiologic short-term results with significant improvements in all clinical scores and MOCART2.0 scores, confirming morphologic integrity of the transplanted chondrocytes. Therefore, this minimally invasive procedure represents a promising operative technique for cartilage regeneration, even for large-diameter lesions., Level of Evidence: IV, therapeutic case series., (Copyright © 2021 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

Background: Commonly used cartilage repair procedures have been established for focal cartilage lesions; however, degenerative lesions with accompanying changes of other intraarticular structures are much more common in clinical practice. This stage, in which classic radiological signs of osteoarthritis are absent, is called early osteoarthritis and is characterized by impaired joint homeostasis with biomechanical and biochemical changes that can have a negative effect on regenerative cartilage therapy procedures., Indication: Cartilage repair procedures are indicated for symptomatic focal early osteoarthritis, defined as cartilage degeneration ICRS grades I or II around a focal cartilage defect ICRS grades III or IV. In more advanced osteoarthritis with significant narrowing of the joint space, cartilage repair procedures are generally contraindicated., Therapy: The most studied cartilage repair procedure for early osteoarthritis is autologous chondrocyte implantation, which has shown acceptable results in case series, although higher failure rates are to be expected compared to focal, traumatic cartilage lesions. The use of bone marrow-stimulating techniques seems to be limited in early osteoarthritis and should only be used in cases of lesion < 2 cm 2 and very little surrounding cartilage degeneration. Concomitant surgical procedures, especially unloading osteotomies, are very important.

Purpose: To evaluate whether different suture materials in meniscal repair may harm cartilage., Methods: A preloaded linear friction testing setup including porcine knees with porcine cartilage, porcine meniscus, and different suture materials (braided nonabsorbable, absorbable monofilament) was used. Five groups with different tribological pairs were tested: cartilage on meniscus (control), cartilage on cartilage (control No. 2), and cartilage on different meniscus sutures (3 groups). Cartilage integrity was analyzed macroscopically by the India ink method and histologically using Giemsa-eosin-stained undecalcified methyl methacrylate sections. Cartilage lesions were classified by using a quantitative scoring system., Results: The control groups did not show cartilage damage, either macroscopically or histologically. Loading cartilage with sutured menisci led to significant damage of the superficial radial and transitional zones with braided nonabsorbable (P = .03) and absorbable monofilament (P = .02) sutures at final examination. Menisci sutured with braided nonabsorbable material resulted in deeper damage to the cartilage. However, there were no significant differences between the suture materials. Sutures oriented perpendicular to surface motion led to a larger defect than parallel-oriented sutures., Conclusions: Braided nonabsorbable and absorbable monofilament suture materials cause significant damage to cartilage during long-term cyclic loading in vitro. The extent of damage depends on suture orientation., Clinical Relevance: This study provides data on the extent to which different suture materials in meniscus repair may harm cartilage., (Copyright © 2019 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

Anteromedial knee injury with rupture of anterior cruciate ligament (ACL) and concomitant lesion of medial collateral ligament (MCL) is common in athletes. No standardized treatment concept can be found within the literature. This study presents results of a new treatment concept for concomitant MCL lesions in patients with ACL rupture. In this study, 67 recreational athletes with ACL injury and concomitant MCL lesion were treated according to a distinct treatment concept. Patients were classified in six different types of concomitant MCL lesion depending on grade of MCL lesion and presence of anteromedial rotatory instability (AMRI). Final classification and surgical indication were determined 6 weeks posttraumatic. All patients received ACL reconstruction. MCL was treated by surgical or conservative regime due to type of concomitant MCL lesion. International Knee Documentation Committee (IKDC), AMRI, and Lysholm scores were evaluated both preoperatively and after 6 weeks, 16 weeks, 12 months, and 18 months postoperatively. All patients could be uniquely classified and treated according to the introduced treatment concept. AMRI was verifiable in patients with grade II and III MCL lesions. All patients showed good to excellent clinical results at the follow-up examinations. In all 67 patients (100%), the findings were graded as normal or nearly normal according to the IKDC knee examination form. Lysholm score averaged 93.9 at final follow-up. The introduced treatment concept showed good results on short-term outcome and provides a sufficient treatment strategy for concomitant MCL lesions in athletes with ACL rupture., Competing Interests: Disclosure The authors report no conflicts of interest in this work., (Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.)

The distribution of intramyocardially injected rabbit MSCs, labeled with the near-infrared dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbo-cyanine-iodide (DiR) using hybrid Fluorescence Molecular Tomography-X-ray Computed Tomography (FMT-XCT) and Multispectral Optoacoustic Tomography (MSOT) imaging technologies, was investigated. Viability and induction of apoptosis of DiR labeled MSCs were assessed by XTT- and Caspase-3/-7-testing in vitro . 2 × 10 6 , 2 × 10 5 and 2 × 10 4 MSCs labeled with 5 and 10 μg DiR/ml were injected into fresh frozen rabbit hearts. FMT-XCT, MSOT and fluorescence cryosection imaging were performed. Concentrations up to 10 μg DiR/ml did not cause apoptosis in vitro (p > 0.05). FMT and MSOT imaging of labeled MSCs led to a strong signal. The imaging modalities highlighted a difference in cell distribution and concentration correlated to the number of injected cells. Ex-vivo cryosectioning confirmed the molecular fluorescence signal. FMT and MSOT are sensitive imaging techniques offering high-anatomic resolution in terms of detection and distribution of intramyocardially injected stem cells in a rabbit model.

Purpose: The purpose of this study was to evaluate the outcome, return to sporting activity, and postoperative articular cartilage and bony morphology of shoulders that underwent arthroscopic suture anchor repair of bony Bankart lesions., Methods: The inclusion criteria for this retrospective study were anterior glenoid rim fractures after traumatic shoulder instability that were treated with arthroscopic suture anchor repair. Patients were surveyed by a questionnaire including sport-specific outcome, Rowe score, Western Ontario Shoulder Instability Index, and Oxford Instability Score. Three-tesla magnetic resonance imaging could be performed in 30 patients to assess osseous integration, glenoid reconstruction, and signs of osteoarthritis., Results: From November 1999 to April 2010, 81 patients underwent an anterior bony Bankart repair in our department (50 arthroscopic suture anchor repairs, 5 arthroscopic screw fixations, and 26 open repairs). The 55 arthroscopic repairs comprised a consecutive cohort of patients treated by a single surgeon. Of the 50 patients in the suture anchor group, 45 (90%) were available for evaluation. At 82 ± 31 months postoperatively, the mean Rowe score was 85.9 ± 20.5 points, the mean Western Ontario Shoulder Instability Index score was 89.4% ± 14.7%, and the mean Oxford Instability Score was 13.6 ± 5.4 points. Compared with the contralateral shoulder, all scores showed a significantly reduced outcome (P < .001, P < .001, and P < .001, respectively). A redislocation occurred in 3 patients (6.6%). Regarding satisfaction, 35 patients (78%) were very satisfied, 9 (20%) were satisfied, and 1 was partly satisfied. Overall, 95% of patients returned to any sporting activity after surgery. The number of sports disciplines (P < .001), duration (P = .005), level (P = .02), and risk category (P = .013) showed a significant reduction compared with the pretrauma condition. However, only 19% of patients reported that shoulder complaints were the reason for the reduction in activity. Nonunion occurred in 16.6%, with a higher frequency in patients with chronic lesions (P = .031). Anatomic reduction was achieved in 72%, the medial step-off in patients with nonanatomic reduction averaged 1.8 ± 0.9 mm, and the remaining glenoid defect size averaged 6.8% ± 7.3%. Full-thickness cartilage defects of the anterior glenoid were detected in 70% of patients., Conclusions: Arthroscopic suture anchor repair may enable an anatomic reduction of bony Bankart lesions with no or only minimal articular steps and provides successful midterm outcomes concerning clinical scores, recurrence, and patient satisfaction. The return to activity is limited for various, mostly non-shoulder-related causes. Chronic lesions may have an inferior healing potential; therefore early surgical stabilization of acute Bankart fragments is suggested to avoid possible nonunion., Level of Evidence: Level IV, therapeutic case series., (Copyright © 2015 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

Topical thrombins are locally active hemostatic agents that can be used to minimize blood loss during any surgery. The aim of this study was to design and investigate a thrombin-containing biodegradable hemostyptic device with an optimized drug release profile to promote local blood clot formation. It is effective with ongoing systemic antithrombotic therapy and can be used in all types of bone-related surgery, for example, in dental surgery. Thrombin-loaded poly(D,L-lactide-co-glycolide) microspheres were synthesized by means of complex (w/o/w) emulsion evaporation method. The resulting enzyme activity of the serine-protease thrombin was verified by the specific chromogenic substrate S-2238. The thrombin release profile depended on four factors: (1) thrombin dosage, (2) polymer concentration in the o-phase, (3) phase quotient w1:0 in the primary emulsion, and (4) the addition of pore-introducing agents. A collagenous sponge containing thrombin-loaded microspheres by means of lyophilization was developed. The impact of several production factors of the (w1/o/w2) solvent evaporation method to optimize thrombin encapsulation, morphology of the spheres, and desired drug release profile have been investigated. The in vitro thrombin release was dependent on the polymer-to-oil phase ratio, the polymer concentration, and the type of solvent and polymer. The porosity of the spheres and release rate of the active agent were enhanced by increasing the inner aqueous w1 phase. With this study, a new biodegradable hemostyptic device could be verified and established for a potentially safe and locally controlled thrombin release to manage postsurgical hemorrhage in patients undergoing anticoagulant therapy., (Copyright © 2010 Wiley Periodicals, Inc.)

Purpose: Gene delivery from biomaterials has become an important tool in tissue engineering. The purpose of this study was to generate a gene vector-doted fibrin glue as a versatile injectable implant to be used in gene therapy supported tissue regeneration., Methods: Copolymer-protected polyethylenimine(PEI)-DNA vectors (COPROGs), naked DNA and PEI-DNA were formulated with the fibrinogen component of the fibrin glue TISSUCOL and lyophilized. Clotting parameters upon rehydration and thrombin addition were measured, vector release from fibrin clots was determined. Structural characterizations were carried out by electron microscopy. Reporter and growth factor gene delivery to primary keratinocytes and chondrocytes in vitro was examined. Finally,chondrocyte colonized clots were tested for their potency in cartilage regeneration in a osteochondral defect model., Results: The optimized glue is based on the fibrinogen component of TISSUCOL, a fibrin glue widely used in the clinics, co-lyophilized with copolymer-protected polyethylenimine(PEI)- DNA vectors (COPROGs). This material, when rehydrated, forms vector-containing clots in situ upon thrombin addition and is suitable to mediate growth factor gene delivery to primary keratinocytes and primary chondrocytes admixed before clotting. Unprotected PEI-DNA in the same setup was comparatively unsuitable for clot formation while naked DNA was ineffective in transfection. Naked DNA was released rapidly from fibrin clots (>70% within the first seven days) in contrast to COPROGs which remained tightly immobilized over extended periods of time (0.29% release per day). Electron microscopy of chondrocytecolonized COPROG-clots revealed avid endocytotic vector uptake. In situ BMP-2 gene transfection and subsequent expression in chondrocytes grown in COPROG clots resulted in the upregulation of alkaline phosphatase expression and increased extracellular matrix formation in vitro. COPROG-fibrinogen preparations with admixed autologous chondrocytes when clotted in situ in osteochondral defects in the patellar grooves of rabbit femura gave rise to luciferase reporter gene expression detectable for two weeks (n=3 animals per group). However, no significant improvement in cartilage formation in osteochondral defects filled with autologous chondrocytes in BMP-2-COPROG clots was achieved in comparison to controls (n=8 animals per group)., Conclusions: COPROGs co-lyophilized with fibrinogen are a simple basis for an injectable fibrin gluebased gene-activated matrix. The preparation can be used is complete analogy to fibrin glue preparations that are used in the clinics. However, further improvements in transgene expression levels and persistence are required to yield cartilage regeneration in the osteochondral defect model chosen in this study.

Anamnesis: Here, the case of a 48-year-old highly trained patient without classic myocardial risk factors is described who reported on frequent and recurrent angina pectoris. In a previous examination, the test for cardiac troponin T (cTnT) was slightly positive, however, cardiac examination including myocardial perfusion scintigraphy and coronary catheterization was without pathologic findings. Worth mentioning in the past medical history was a rheumatoid arthritis with persistent Raynaud's symptoms and hemoglobin as well as hematocrit levels in the upper normal range., Examinations and Course of Events: The patient reported that symptoms would occur most likely during long-term endurance exercise. Therefore, a bicycle ergometry with 180 W and open ending was performed. After 1.55 h, the patient complained of severe angina pectoris. The ECG showed massive ST segment elevations in II, III, and aVF (Figure 2). However, coronary catheterization showed no major stenosis or occlusion. Nevertheless, cTnT increased to 0.979 ng/ml. (Therefore, the reason for the symptomatic ST elevation was believed to be a myocardial tissue damage due to coronary vasospasm and the patient received an antivasospastic medication (amlodipine, atorvastatin, and acetylsalicylic acid [ASS]). After 6 months, the patient had a relapse during moderate physical activity. The ECG showed an ST segment depression in V(4) and V(5) while markers for myocardial tissue damage including cTnT were negative. A coronary CT angiography was performed that revealed a subtotal stenosis of the proximal LAD (Fig ure 3), which was successfully treated by angioplasty with subsequent stenting. In the course of further examinations, a polycythemia vera (JAK2-V617 mutation) was diagnosed as the cause for the high hemoglobin and hematocrit levels. Since then, the patient was without further events under a medication consisting of ASS, atorvastatin, candesartan, and intermittent phlebotomy., Conclusion: Although a relative weighting is difficult, it can be assumed that the combined effects of a polycythemia- associated hypercoagulability, an increased reagibility of the coronary arteries (aggravated by physical stress) as well as a hemoconcentration following prolonged exercise, could account for symptomatic recurrent minor thrombotic coronary events as well as the subtotal occlusion of the LAD. Obviously, this could not be prevented by a healthy lifestyle, regular physical activity, and the absence of classic coronary risk factors.

Objective: Arthroscopic (re)stabilization of the unstable shoulder by anatomic refixation of the detached capsulolabral complex with suture anchors or reduction of excessive capsule volume by capsule plication., Indications: Any type of shoulder instability (anterior, posterior, inferior, or multidirectional instability). Revision stabilization (even after primary open stabilization). Bone defects affecting < 25% of the glenoid surface. Lesions of the superior biceps tendon anchor complex (SLAP lesion)., Contraindications: Preexisting bone defects of the glenoid affecting > 25% of the glenoid surface. "Engaging" Hill-Sachs defects: osseous defects of the humeral head that engage with the anterior glenoid rim in extreme external rotation/abduction and consequently lead to shoulder dislocation. Bone-related etiology, e. g., clearly increased glenoid retroversion/anteversion or glenoid dysplasias (e. g., inverse pear shape). Voluntary shoulder dislocation in young patients until the end of the growth period., Surgical Technique: Diagnostic arthroscopy and additional procedures based on clinical and intraoperative findings. For anterior-inferior instability, an anterior-superior approach is made with mobilization of the labrum and decortication of the glenoid. Creation of deep anterior-inferior portal and insertion of the anchors in 5.30, 4.30 and 3.00 o'clock position. The sutures are pulled through the capsulolabral complex and tied arthroscopically. Reconstruction of the inferior glenohumeral ligament is especially important. Lesions of the superior biceps tendon anchor and/or posterior labrum detachment can be treated by the same technique. Capsule plication with PDS sutures can be performed to decrease a large rotator interval or excessive capsule volume. The range of motion at the shoulder is limited for 6 weeks postoperatively (depending on the initial direction of the instability)., Results: At the authors' hospital over 600 arthroscopic shoulder stabilizations using the deep anterior-inferior portal have been completed so far. The redislocation rate for the first 147 patients (average follow-up of 3 years) treated with the technique described here is 6.1% and is slightly higher for arthroscopic revision stabilizations (n=43; of these, redislocation n=3 and reinstability n=3). There were no instances of axillary nerve lesion.

Cardiovascular deaths during or following sport activities repeatedly raise the question about a practicable preparticipation screening for athletes to prevent such adverse events. In Germany and most European countries, well-equipped sports medicine centers evaluate the health of the Olympic athletes through regular checkups, which include a detailed medical history and thorough physical examination as well as an ECG at rest, a stress ECG, and an echocardiography. In professional sports, guidelines for this screening differ according to the federations, however, most of them intend to follow the recommendations of the Olympic sports system. For nonprofessional competitive sports, there are no guidelines for preparticipation screening, although these athletes train at the same level of intensity as professional athletes. The main issue in this international debate is the question of cost-effectiveness and how to finance preventive measures.