共查询到10条相似文献,搜索用时 187 毫秒
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Patrick Sadoghi Birgit Lohberger Birgit Aigner Heike Kaltenegger Jörg Friesenbichler Matthias Wolf Tarek Sununu Andreas Leithner Patrick Vavken 《Journal of orthopaedic research》2013,31(8):1249-1253
To assess the in vitro effect of platelet‐rich plasma (PRP) on biological activity of the human rotator cuff fibroblasts and to describe the optimal dose‐response to maximize cellular stimulation while reducing potential risk. Rotator cuff (RC) fibroblasts of n = 6 patients (mean age of 65.2 years) undergoing arthroscopic cuff tear reconstruction were cultured in vitro for 21 days and stimulated with PRP in three different concentrations (1‐, 5‐, and 10‐fold). Samples were obtained for DNA and GAG measurement at 1, 7, 14, and 21 days. The biological outcomes were regressed on the PRP concentration. The application of PRP significantly influenced the fibroblast proliferation and activity of the human rotator cuff with elevated glycosaminoglycan (GAG) and DNA levels. The dosage of PRP had the significantly highest impact on this proliferation using a onefold or fivefold application. PRP has a significant effect on fibroblast proliferation of the human rotator cuff in vitro with an optimal benefit using a onefold or fivefold PRP concentration. This study justifies further in vivo investigations using PRP at the human rotator cuff. © 2013 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 31:1249–1253, 2013 相似文献
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In vitro effects of glutamate and N‐methyl‐d‐aspartate receptor (NMDAR) antagonism on human tendon derived cells 下载免费PDF全文
Benjamin John Floyd Dean Sarah J.B. Snelling Stephanie Georgina Dakin Muhammad Kassim Javaid Andrew Jonathan Carr 《Journal of orthopaedic research》2015,33(10):1515-1522
It is known that extracellular glutamate concentrations are increased in tendinopathy but the effects of glutamate upon human tendon derived cells are unknown. The primary purpose was to investigate the effect of glutamate exposure on human tendon‐derived cells in terms of viability, protein, and gene expression. The second purpose was to assess whether NMDAR antagonism would affect the response of tendon‐derived cells to glutamate exposure. Human tendon‐derived cells were obtained from supraspinatus tendon tissue obtained during rotator cuff repair (tendon tear derived cells) and from healthy hamstring tendon tissue (control cells). The in vitro impact of glutamate exposure and NMDAR antagonism (MK‐801) was measured using the Alamar blue cell viability assay, immunocytochemistry, and quantitative real‐time PCR. Glutamate reduced cell viability at 24 h in tendon tear derived cells but not in control cells at concentrations of 7.5 mM and above. Cell viability was significantly reduced after 72 h of 1.875 mM glutamate in both cell groups; this deleterious effect was attenuated by NMDAR antagonism with 10 µM MK‐801. Both 24 and 72 h of 1.875 mM glutamate exposure reduced Type 1 alpha 1 collagen (COL1A1) and Type 3 alpha 1 collagen (COL3A1) gene expression, but increased Aggrecan gene expression. We propose that these effects of glutamate on tendon derived cells including reduced cell viability and altered matrix gene expression contribute to the pathogenesis of tendinopathy. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:1515–1522, 2015. 相似文献
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《Journal of orthopaedic research》2017,35(8):1806-1815
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Hirokazu Honda Masafumi Gotoh Tomonoshin Kanazawa Hidehiro Nakamura Keisuke Ohta Kei‐ichiro Nakamura Naoto Shiba 《Journal of orthopaedic research》2016,34(9):1620-1627
We determined lidocaine's action on torn rotator cuff tendons in vitro and in vivo. For in vitro experiments, cell proliferation and viability assays were performed using tenocytes derived from human torn rotator cuff tendons. For in vivo experiments, acute rotator cuff tears were made on the supraspinatus tendons in the rats’ bilateral shoulders; before closure, lidocaine was injected into the shoulder and saline into the contralateral shoulder (control). After sacrifice, the specimens underwent biomechanical testing or histological analysis at 24 h and at 2, 4, and 8 weeks after surgery. The extent of collagen organization and apoptosis were semi‐quantitatively evaluated using collagen picrosirius red staining. Apoptosis was examined using TUNEL staining and electron microscopy. Cell proliferation decreased dose‐dependently. After exposure to 0.1% lidocaine for 24 h, cell viability decreased. Two and 4 weeks after surgery, the ultimate load to failure decreased more in the lidocaine group than in the control group, with significantly reduced stiffness in the lidocaine group 2 weeks after surgery. Collagen organization significantly decreased in the lidocaine group by 4 weeks after surgery but returned to baseline at 8 weeks. TUNEL staining detected numerous apoptotic tenocytes at the torn tendon edge exposed to lidocaine 24 h after surgery; electron microscopy confirmed the condensed cell nuclei. These changes were not observed in controls. Lidocaine caused cytotoxicity to tenocytes under both conditions, decreased biomechanical properties, and induced apoptosis and delay of collagen organization in this model. Subacromial lidocaine injections in patients with rotator cuff tears should be performed carefully. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1620–1627, 2016. 相似文献
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Platelets and plasma stimulate sheep rotator cuff tendon tenocytes when cultured in an extracellular matrix scaffold 下载免费PDF全文
Brian A. Kelly Benedikt L. Proffen Carla M. Haslauer Martha M. Murray 《Journal of orthopaedic research》2016,34(4):623-629
The addition of platelet‐rich plasma (PRP) to rotator cuff repair has not translated into improved outcomes after surgery. However, recent work stimulating ligament healing has demonstrated improved outcomes when PRP or whole blood is combined with an extracellular matrix carrier. The objective of this study was to evaluate the effect of three components of blood (plasma, platelets, and macrophages) on the in vitro activity of ovine rotator cuff cells cultured in an extracellular matrix environment. Tenocytes were obtained from six ovine infraspinatus tendons and cultured over 14 days in an extracellular matrix scaffold with the following additives: (1) plasma (PPP), (2) plasma and platelets (PAP), (3) plasma and macrophages (PPPM), (4) plasma, platelets and macrophages (PAPM), (5) phosphate buffered saline (PBS), and (6) PBS with macrophages (PBSM). Assays measuring cellular metabolism (AlamarBlue), proliferation (Quantitative DNA assay), synthesis of collagen and cytokines (SIRCOL, TNF‐α and IL‐10 ELISA, and MMP assay), and collagen gene expression (qPCR) were performed over the duration of the experiment, as well as histology at the conclusion. Plasma was found to stimulate cell attachment and spreading on the scaffold, as well as cellular proliferation. Platelets also stimulated cell proliferation, cellular metabolism, transition of cells to a myofibroblast phenotype, and contraction of the scaffolds. The addition of macrophages did not have any significant effect on the sheep rotator cuff cells in vitro. In vivo studies are needed to determine whether these changes in cellular function will translate into improved tendon healing. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:623–629, 2016. 相似文献
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《Arthroscopy》2021,37(9):2754-2755
Platelet-rich plasma (PRP) injections continue to be used at increasing rates to treat common musculoskeletal conditions. PRP has a low-risk profile and emerging in vitro evidence to support its positive effects on soft-tissue healing. PRP has been shown to be of benefit for knee osteoarthritis, but less has been published regarding the shoulder. PRP delivers a high concentration of growth factors, cytokines, and other important inflammatory modulators. Its use is appealing for treating partial-thickness rotator cuff tears, subacromial bursitis, and rotator cuff tendinopathy since rotator cuff tendons often have poor healing capacity due to intrinsic degeneration. PRP has been shown to increase cell proliferation and matrix synthesis in tenocytes, which may aid tendon regeneration and healing. Adult tendons also contain a small amount of tendon progenitor cells, which can be induced to an active state by PRP. In addition, PRP is an autologous biologic agent and easy to acquire and administer in an outpatient clinical setting. Clinical studies continue to lag and are often heterogenous in quality and in results. PRP can vary widely based on multiple intrinsic and extrinsic factors, including patient age, sex, activity level, centrifugation speed, and number of centrifugation cycles. Thus, quality research methods should include reporting using the PAW (platelets/activation/white blood cells) system. Clinicians should remain cautiously optimistic about the future role of PRP injections in the shoulder. 相似文献
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《Journal of orthopaedic research》2017,35(11):2506-2512
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Melanie Vogl Johannes Fischer Marcus Jäger Christoph Zilkens Rüdiger Krauspe Monika Herten 《Journal of orthopaedic research》2013,31(11):1786-1795
Mesenchymal progenitor cells (MSCs) are promising for cell‐based regeneration therapies. In elderly patients a reduced proliferation of MSCs has been described. Platelet‐rich plasma (PRP) contains important factors necessary for osteogenic regeneration. The aim of this study was to find out whether the age‐induced decrease in cell proliferation can be compensated by the use of supernatant of centrifuged, activated PRP (tPR). MSCs of donors of three age groups (A: young, 14–16 years, B: middle age, 36–46 years, C: older, 74–83 years) were expanded with 20% FCS alone or supplemented with thrombin‐activated platelet releasate (tPR) (1%, 2.5%, and 5%) or platelet‐poor plasma (PPP 5%). Cell proliferation and differentiation was measured on days 0, 3, and 7. Proliferation increased significantly in groups A and B with tPR, and non‐significantly in group C. The generation times of MSCs of elderly patients were significantly increased in group C compared to groups A and B. Addition of 1% or 2.5% tPR significantly reduced population doubling times of all age groups. Adding tPR stimulates the proliferation rate of MSCs independent of donor age. For juvenile and middle‐aged patients this influence was significant. Cells differentiation into osteoblasts was not influenced by addition of tPR. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31:1786–1795, 2013 相似文献
10.
Roman Dittmar Bart G. M. van Dijk Marc A. M. J. van Zandvoort Keita Ito 《Journal of orthopaedic research》2014,32(4):545-550
Regenerative medicine approaches aiming at treating degenerating intervertebral discs, a major cause of back pain, are increasingly tested in ex‐vivo disc explant models mimicking in‐vivo conditions. For assessing the efficacy of regenerative therapies, cell viability is commonly measured requiring specific labels to stain cells. Here, we demonstrate and evaluate how cellular auto‐fluorescence can be utilized to non‐invasively assess viability in disc tissue in‐situ using label‐free two‐photon microscopy. Live and dead bovine disc cells (0% and 100% cell viability) from the nucleus pulposus were seeded into collagen gels and auto‐fluorescence was characterized. Subsequently, nucleus pulposus explants were cultured for 6 days in media with different glucose supplementation (0, 0.25, 0.5, and 1 g/L) to induce different degrees of cell death. Then, samples were split and viability was assessed using label‐free two‐photon microscopy and conventional staining. Results show that live and dead nucleus pulposus cells systematically emit auto‐fluorescent light with distinct characteristics. Cell viability values obtained with label‐free microscopy did not significantly differ from those acquired with staining. In summary, monitoring auto‐fluorescence facilitates accurate cell viability assessment in nucleus tissue requiring no additional dyes. Thus, this technique may be suitable for pre‐clinical testing of regenerative therapies in nucleus pulposus cultures. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:545–550, 2014. 相似文献