Orthopaedic surgeons display a positive outlook towards artificial intelligence: A survey among members of the AGA Society for Arthroscopy and Joint Surgery.

Purpose: The purpose of this study was to evaluate the perspective of orthopaedic surgeons on the impact of artificial intelligence (AI) and to evaluate the influence of experience, workplace setting and familiarity with digital solutions on views on AI. Methods: Orthopaedic surgeons of the AGA Society for Arthroscopy and Joint Surgery were invited to participate in an online, cross-sectional survey designed to gather information on professional background, subjective AI knowledge, opinion on the future impact of AI, openness towards different applications of AI, and perceived advantages and disadvantages of AI. Subgroup analyses were performed to examine the influence of experience, workplace setting and openness towards digital solutions on perspectives towards AI. Results: Overall, 360 orthopaedic surgeons participated. The majority indicated average (43.6%) or rudimentary (38.1%) AI knowledge. Most (54.5%) expected AI to substantially influence orthopaedics within 5-10 years, predominantly as a complementary tool (91.1%). Preoperative planning (83.8%) was identified as the most likely clinical use case. A lack of consensus was observed regarding acceptable error levels. Time savings in preoperative planning (62.5%) and improved documentation (81%) were identified as notable advantages while declining skills of the next generation (64.5%) were rated as the most substantial drawback. There were significant differences in subjective AI knowledge depending on participants' experience (p = 0.021) and familiarity with digital solutions (p < 0.001), acceptable error levels depending on workplace setting (p = 0.004), and prediction of AI impact depending on familiarity with digital solutions (p < 0.001). Conclusion: The majority of orthopaedic surgeons in this survey anticipated a notable positive impact of AI on their field, primarily as an assistive technology. A lack of consensus on acceptable error levels of AI and concerns about declining skills among future surgeons were observed. Level of Evidence: Level IV, cross-sectional study.

Influence of a Concomitant Medial Meniscus Injury on Knee Joint Function and Osteoarthritis Presence after Anterior Cruciate Ligament Reconstruction.

(1) Background: The aim of this study was to investigate how a medial meniscus injury accompanying an anterior cruciate ligament rupture affects the clinical outcome 10 years after ACL reconstruction. (2) Methods: A total of 37 patients who received anterior cruciate ligament reconstruction (ACLR) were included in this retrospective study. Two groups were analyzed at a single follow-up of 10 years: (i) "isolated (ACLR)" (n = 20) and (ii) "ACLR with medial meniscal injury" (n = 17). The following clinical scores were recorded: International Knee Documentation Committee (IKDC), the Knee Injury and Osteoarthritis Outcome Score (KOOS), Lysholm Score and Tegner Activity Score. To determine the degree of osteoarthritis the Kellgren-Lawrence score was used. (3) Results: The "isolated ACLR" study group scored significantly higher (p < 0.05) on the IKDC subjective questionnaire (mean: 88.4) than the "ACLR with medial meniscus injury" group (mean: 81). The KOOS category "activities of daily living" showed significantly better results in the isolated ACLR group (p < 0.05). The "ACLR with medial meniscus injury" group had significantly higher degree of osteoarthritis (p < 0.05). No significant differences were found in all the other clinical scores. (4) Conclusions: The results of this study further indicate that patients with a concomitant medial meniscus injury have slightly more discomfort in everyday life and increased risk of developing osteoarthritis 10 years after surgery.

Culture of Hoffa fat pad mesenchymal stem/stromal cells on microcarrier suspension in vertical wheel bioreactor for extracellular vesicle production.

BACKGROUND: Mesenchymal stromal cells (MSCs) are increasingly employed in regenerative medicine approaches for their immunomodulatory and anti-inflammatory properties, which are encoded in their secretome including extracellular vesicles (EVs). The Hoffa fat pad (HFP) located infrapatellarly harbours MSCs that could assist in tissue homeostasis in osteoarthritic joints. Intraarticular injection therapies based on blood products could modulate the populations of released HFP-MSC-EVs in a quantitative manner. METHODS: To obtain amounts of HFP-MSC-derived EVs that allow pre-clinical evaluation, suitable EV production systems need to be developed. This work investigates the release of EVs from primary HFP-MSCs cultivated in a 3D environment using microcarrier suspension culture in a vertical wheel bioreactor in comparison to conventional 2D culture. To simulate an intraarticular blood product therapy, cultures were treated with citrate-anticoagulated platelet-rich plasma (CPRP) or hyperacute serum (hypACT) before EV collection. HFP-MSC-EVs are enriched via ultrafiltration and characterised via Western Blot, nanoparticle tracking analysis in scatter as well as fluorescence mode. EV potency was determined via RT-qPCR analysing the expression of type II and X collagen (COL2 and COL10), as well as inducible nitric oxide synthase (iNOS) in primary OA chondrocytes. RESULTS: Blood product supplementation elevated HFP-MSC metabolic activity as determined via XTT assay over the course of 14 days. 3D culture resulted in a roughly 100-fold EV yield compared to 2D culture and elevated number of EVs released per cell. Total protein content correlated with the EV concentration. While typical EV marker proteins such as CD9, CD63 or Alix were detected in total protein extracts, CD9 and CD73 colocalised on individual EVs highlighting their cell origin. The type of blood product treatment did not affect the size or concentration of EVs obtained from HFP-MSCs. Assessing potency of 3D culture EVs in comparison to 2D EVs revealed superior biological activity with regard to inhibition of inflammation, inhibition of chondrocyte hypertrophy and induction of cartilage-specific ECM production. CONCLUSIONS: HFP-MSCs proliferate in presence of human blood products indicating that animal serum in culture media can be avoided in the future. The culture of HFP-MSCs in the employed bioreactor was successfully used to generate quantities of EVs that could allow evaluation of HFP-MSC-EV-mediated effects in pre-clinical settings. In addition, EV potency of 3D EVs is superior to EVs obtained in conventional 2D culture flasks.

Infra-patellar fat pad-derived mesenchymal stem cells maintain their chondrogenic differentiation potential after arthroscopic harvest with blood-product supplementation.

PURPOSE: Mesenchymal stem cells/medicinal signaling cells (MSCs) possess therapeutic potential and are used in regenerative orthopaedics. The infra-patellar fat pad (IFP) is partially resected during knee arthroscopy (KASC) and contains MSCs. Heat, irrigation, and mechanical stress during KASC may decrease MSC's therapeutic potential. This study assessed MSCs' regenerative potential after arthroscopic IFP harvest and potential effects of two blood products (BP) (platelet-rich plasma (PRP), hyperacute serum (HAS)) on MSCs' viability and chondrogenic differentiation capacity. METHODS: IFP was arthroscopically harvested, isolated, and counted (n = 5). Flow cytometry was used to assess cell viability via staining with annexin V/7-AAD and stemness markers via staining for CD90, CD73, and CD105. MSCs were incubated with blood products, and metabolic activity was determined via an XTT assay. Deposition of cartilage extracellular matrix was determined in histologic sections of chondrogenically differentiated 3D pellet cultures via staining with Alcian Blue. Expression of cartilage-specific genes (SOX9, MMP3/13, ACAN, COL1/2) was analyzed via quantitative PCR. RESULTS: MSC isolation from IFP yielded 2.66*106 ± 1.49*106 viable cells from 2.7 (0.748) g of tissue. MSC markers (CD 90/105/73) were successfully detected and annexin V staining showed 81.5% viable cells. XTT showed increased metabolic activity. Within the BP groups, this increase was significant (days 0-14, p < 0.05). PCR showed expression of cartilage-specific genes in each group. COL2 (p < 0.01) as well as ACAN (p < 0.001) expression levels were significantly higher in the HAS group. Histology showed successful differentiation. CONCLUSION: Arthroscopic harvest of IFP-MSCs yields sufficient cells with maintained regenerative potential and viability. Blood products further enhance MSCs' viability.

Unicompartmental Knee Arthroplasty Provides Superior Clinical and Radiological Outcomes Compared to High Tibial Osteotomy at a Follow-Up of 5-8 Years.

BACKGROUND: Knee Osteoarthritis (OA) is a debilitating disease. Initially, the medial compartments are affected in most cases. For this pathology, joint preservation is preferable. Two surgical procedures aim to meet this goal: high-tibial osteotomy (HTO) and unicompartmental knee arthroplasty (UKA). The aim was to compare clinical and radiological outcomes of HTO versus UKA in patients with unicompartmental, medial OA. METHOD: Retrospective case series. A total of 86 (61 UKA, 25 HTO) patients that received either treatment at a single, specialized center were assessed pre-operatively and at a single follow-up examination at 77.13 months (±8.170). The Knee Society Score (KSS), range of motion (ROM), SF36 questionnaire and the Tegner score were used. The Kellgren-Lawrence score was assessed pre- and post-surgically. Survivorship with the endpoint "revision" was assessed. RESULTS: The UKA group showed significantly better improvements in KSS scores for pain (p < 0.006) and function (p < 0.001). OA progression (p < 0.02) and survivorship (p < 0.018) differed, significantly favoring UKA. ROM, SF36 and Tegner score did not differ significantly. CONCLUSIONS: The presented mid-to long-term data suggest that UKA provides superior results in selected outcomes. Nevertheless, significant differences in the demographics of treatments indicate the challenge of comparing these two treatments.

Midterm Results of High-Dose-Rate Intraoperative Brachytherapy in the Treatment of Soft Tissue Sarcomas.

INTRODUCTION: According to the literature only sparse data are available on the use of high-dose-rate intraoperative brachytherapy (IOHDR-BT) as a boost to external-beam irradiation (EBRT) in combination with a wide resection in patients with high-grade soft tissue sarcomas (STS). MATERIALS AND METHODS: Applying a retrospective study design, we investigated all patients who between 2010 and 2016 underwent marginal resection of a high-grade STS and intraoperative radiotherapy, followed by EBRT. We included only patients with a traceable follow-up time of at least two years. Of 89 patients, 35 met our inclusion criteria and showed an average follow-up of four years. RESULTS: We found an overall 2-year local control rate of 94.3%. The local recurrence rate for R0 resections was 6%, whereas recurrences occurred in 13% of R1 resections and in 100% of R2 resections. One affected patient received only intraoperative radiotherapy. The recurrence rate by tumour entity was 36% for LPS, 11% for myxofibrosarcoma and 17% for undifferentiated pleomorphic sarcoma. CONCLUSION: The treatment regimen consisting of limb-preserving surgery, IORT and pre- or postoperative radiotherapy consistently shows excellent local control rates.

Brachytherapy in the Treatment of Soft-Tissue Sarcomas of the Extremities-A Current Concept and Systematic Review of the Literature.

INTRODUCTION: Evidence on the use of brachytherapy in soft-tissue sarcoma (STS) is sparse. Therapy regimens are determined more by local interdisciplinary tumor conferences than by standardized protocols. Patient-specific factors complicate the standardized application of therapy protocols. The individuality of the treatment makes it difficult to compare results. MATERIALS AND METHODS: A comprehensive literature search was conducted, whereby the literature from a period of almost 44 years (1977-2021) was graded and included in this systematic review. For this purpose, PubMed was used as the primary database. Search string included "soft-tissue sarcoma", "brachytherapy", and "extremity." Four independent researchers reviewed the literature. Only full-text articles written in English or German were included. RESULTS: Of the 175 identified studies, 70 were eligible for analysis based on the inclusion and exclusion criteria. The key points to compare were local complications, recurrence rate and correlation with margins of resection, and the use of brachytherapy regarding tumor grading. CONCLUSION: Brachytherapy represents an important subset of radiotherapy techniques used in STSs, whose indications and applications are constantly evolving, and for which a local control rate of 50% to 96% has been reported as monotherapy, depending on risk factors. However, the best benefit is seen in the combination of further resection and brachytherapy, and most authors at many other centers agree with this treatment strategy.

Artificial-Intelligence-Aided Radiographic Diagnostic of Knee Osteoarthritis Leads to a Higher Association of Clinical Findings with Diagnostic Ratings.

BACKGROUND: Radiographic knee osteoarthritis (OA) severity and clinical severity are often dissociated. Artificial intelligence (AI) aid was shown to increase inter-rater reliability in radiographic OA diagnosis. Thus, AI-aided radiographic diagnoses were compared against AI-unaided diagnoses with regard to their correlations with clinical severity. METHODS: Seventy-one DICOMs (m/f = 27:42, mean age: 27.86 ± 6.5) (X-ray format) were used for AI analysis (KOALA software, IB Lab GmbH). Subjects were recruited from a physiotherapy trial (MLKOA). At baseline, each subject received (i) a knee X-ray and (ii) an assessment of five main scores (Tegner Scale (TAS); Knee Injury and Osteoarthritis Outcome Score (KOOS); International Physical Activity Questionnaire; Star Excursion Balance Test; Six-Minute Walk Test). Clinical assessments were repeated three times (weeks 6, 12 and 24). Three physicians analyzed the presented X-rays both with and without AI via KL grading. Analyses of the (i) inter-rater reliability (IRR) and (ii) Spearman's Correlation Test for the overall KL score for each individual rater with clinical score were performed. RESULTS: We found that AI-aided diagnostic ratings had a higher association with the overall KL score and the KOOS. The amount of improvement due to AI depended on the individual rater. CONCLUSION: AI-guided systems can improve the ratings of knee radiographs and show a stronger association with clinical severity. These results were shown to be influenced by individual readers. Thus, AI training amongst physicians might need to be increased. KL might be insufficient as a single tool for knee OA diagnosis.

EBRA Migration Analysis of a Modular, Distally Fixed Stem in Hip Revision Arthroplasty: A Clinical and Radiological Study.

Background: Massive osteolysis of the proximal femur makes stem revision a challenging procedure. EBRA-FCA provides the opportunity to determine stem migration, which is considered a predictive factor for implant survival. In this study, we aimed to analyze the migration behavior of a modular, distally fixed reconstruction prosthesis. Methods: Applying a retrospective study design, we reviewed all consecutive patients who received a cementless MP reconstruction prosthesis (Waldemar Link GmbH & Co. KG, Hamburg, Germany) at our Department between 2005 and 2019. We reviewed medical histories and performed radiological measurements using EBRA-FCA software. Results: A total of 67 stems in 62 patients (female 26; male 36) fulfilled our inclusion criteria. Mean age at surgery was 68.0 (range 38.7−88.44) years. EBRA migration analysis showed a median subsidence of 1.6 mm (range 0.0−20.6) at 24 months. The angle between stem and femur axis was 0.3° (range 0.0°−2.9°) at final follow-up. No correlation between body mass index and increased subsidence was found (p > 0.05). Overall revision-free rate amounted to 92.5% and revision-free rate for aseptic loosening to 98.5%. Furthermore, no case of material breakage was detected. Conclusions: In summary, the MP reconstruction prosthesis showed low subsidence and reduction in the migration rate over the investigated follow-up. Based on this, the modular stem can be considered as a good therapy option in challenging stem revisions offering various options to address the individual anatomical situation.

Isolation and Cultivation of Adipose-Derived Mesenchymal Stem Cells Originating from the Infrapatellar Fat Pad Differentiated with Blood Products: Method and Protocol.

Adipose-derived mesenchymal stem cells (ASCs) are a promising source for clinical application in regenerative orthopedics. ASCs derived from the infra-patellar fat pad (IFP)-a distinct adipose structure in the knee-show superior regenerative potential compared to subcutaneous-fat-derived cells. Furthermore, it has been shown that blood products enhance ASCs' viability. A major challenge for clinical translation of both ASCs and blood products is the low comparability of obtained data due to non-standardized harvesting, isolation and preparation methods. The aim of this method-paper is to provide reproducible protocols to help standardize basic research in the field to build a sound basis for clinical translation with an emphasize on practicability. The presented protocols include (i) ASC isolation from the IFP, (ii) blood product preparation and (iii) ASC incubation with blood products.

Regenerative Potential of Blood-Derived Products in 3D Osteoarthritic Chondrocyte Culture System.

Intra-articular injection of different types of blood-derived products is gaining popularity and clinical importance in the treatment of degenerative cartilage disorders such as osteoarthritis. The regenerative potential of two types of platelet-rich plasma (PRP), prepared in the presence of EDTA (EPRP) and citrate (CPRP) and an alternative blood product-hyperacute serum (hypACT) was evaluated using a 3D osteoarthritic chondrocyte pellet model by assessing the metabolic cell activity, cartilage-related gene expression and extracellular matrix deposition within the pellets. Chondrocyte viability was determined by XTT assay and it revealed no significant difference in metabolic activity of OA chondrocyte pellets after supplementation with different blood products. Nevertheless, the selection of blood products influenced the cartilage-related genes expression, ECM morphology and the tissue quality of pellets. Both PRP types had a different biological effect depending upon concentration and even though CPRP is widely used in clinics our assessment did not reveal good results in gene expression either tissue quality. HypACT supplementation resulted in superior cartilage-related genes expression together with tissue quality and seemed to be the most stable product since no remarkable changes were observed between the two different concentrations. All in all, for successful regenerative therapy, possible molecular mechanisms induced by blood-derived products should be always carefully investigated and adapted to the specific medical indications.

Effects of Extracellular Vesicles from Blood-Derived Products on Osteoarthritic Chondrocytes within an Inflammation Model.

Osteoarthritis (OA) is hallmarked by a progressive degradation of articular cartilage. One major driver of OA is inflammation, in which cytokines such as IL-6, TNF-α and IL-1ß are secreted by activated chondrocytes, as well as synovial cells-including macrophages. Intra-articular injection of blood products-such as citrate-anticoagulated plasma (CPRP), hyperacute serum (hypACT), and extracellular vesicles (EVs) isolated from blood products-is gaining increasing importance in regenerative medicine for the treatment of OA. A co-culture system of primary OA chondrocytes and activated M1 macrophages was developed to model an OA joint in order to observe the effects of EVs in modulating the inflammatory environment. Primary OA chondrocytes were obtained from patients undergoing total knee replacement. Primary monocytes obtained from voluntary healthy donors and the monocytic cell line THP-1 were differentiated and activated into proinflammatory M1 macrophages. EVs were isolated by ultracentrifugation and characterized by nanoparticle tracking analysis and Western blot. Gene expression analysis of chondrocytes by RT-qPCR revealed increased type II collagen expression, while cytokine profiling via ELISA showed lower TNF-α and IL-1ß levels associated with EV treatment. In conclusion, the inflammation model provides an accessible tool to investigate the effects of blood products and EVs in the inflammatory context of OA.

Simlukafusp alfa (FAP-IL2v) immunocytokine is a versatile combination partner for cancer immunotherapy.

Simlukafusp alfa (FAP-IL2v, RO6874281/RG7461) is an immunocytokine comprising an antibody against fibroblast activation protein α (FAP) and an IL-2 variant with a retained affinity for IL-2Rßγ > IL-2 Rßγ and abolished binding to IL-2 Rα. Here, we investigated the immunostimulatory properties of FAP-IL2v and its combination with programmed cell death protein 1 (PD-1) checkpoint inhibition, CD40 agonism, T cell bispecific and antibody-dependent cellular cytotoxicity (ADCC)-mediating antibodies. The binding and immunostimulatory properties of FAP-IL2v were investigated in vitro and compared with FAP-IL2wt. Tumor targeting was investigated in tumor-bearing mice and in a rhesus monkey. The ability of FAP-IL2v to potentiate the efficacy of different immunotherapies was investigated in different xenograft and syngeneic murine tumor models. FAP-IL2v bound IL-2 Rßγ and FAP with high affinity in vitro, inducing dose-dependent proliferation of natural killer (NK) cells and CD4+/CD8+ T cells while being significantly less potent than FAP-IL2wt in activating immunosuppressive regulatory T cells (Tregs). T cells activated by FAP-IL2v were less sensitive to Fas-mediated apoptosis than those activated by FAP-IL2wt. Imaging studies demonstrated improved tumor targeting of FAP-IL2v compared to FAP-IL2wt. Furthermore, FAP-IL2v significantly enhanced the in vitro and in vivo activity of therapeutic antibodies that mediate antibody-dependent or T cell-dependent cellular cytotoxicity (TDCC) and of programmed death-ligand 1 (PD-L1) checkpoint inhibition. The triple combination of FAP-IL2v with an anti-PD-L1 antibody and an agonistic CD40 antibody was most efficacious. These data indicate that FAP-IL2v is a potent immunocytokine that potentiates the efficacy of different T- and NK-cell-based cancer immunotherapies.

Matched retrospective analysis of three different fixation devices for chevron osteotomy.

Chevron osteotomy with consecutive fixation is a commonly performed operative treatment option for hallux valgus deformities. The present retrospective study aims to compare the clinical and radiological outcome of novel bioabsorbable magnesium screw fixation with metal screw and Kirschner wire fixation. Eighteen matched triplets were assembled according to the following criteria: female gender, age difference less than 5 years, date of operation within 4 months, difference in preoperative intermetatarsal angle less than 5°, and equal experience of the first and second surgeon. These patients, between 18 and 85 years of age and with a minimum follow-up period of 12 months, were invited to a follow-up examination, of which only 16 matched triplets of patients entirely kept the appointment. Thus, 48 feet of 44 patients were clinically evaluated using the American Orthopaedic Foot & Ankle Society scale, Foot Function Index, University of California and Los Angeles Activity Score, as well as a visual analogue scale for pain, satisfaction, cosmetic results, and functional impairment. Radiographical assessment included measuring intermetatarsal angle and first metatarsophalangeal angles. All occurring complications and revision surgeries were noted. Significant differences were observed for postoperative intermetatarsal angle between magnesium screw and pin fixation (p = 0.009). Moreover, patients receiving magnesium screw were significantly more prone to undergo the same procedure again (p = 0.03). In conclusion, if the advantages of bioabsorbable magnesium screws outweigh the drawbacks of increased costs and a higher surgical demand, this implant might serve as possible chevron osteotomy fixation method. Compression screws and Kirschner wires also show comparable satisfactory outcomes. LEVEL OF EVIDENCE: III retrospective comparative study.

[Possibilities and limits of conservative treatment for osteoarthritis : Sport advice, training therapy, orthotics and cartilage therapeutics]. / Möglichkeiten und Grenzen der konservativen Therapie der Arthrose : Sportberatung, Trainingstherapie, Orthesen und Knorpeltherapeutika.

BACKGROUND: Osteoarthritis (OA)-the progressive degeneration of synovial joints-is a worldwide problem that affects society, is associated with age and is the dominant reason for pain and immobility of the locomotor system. In the population, 70% of people over the age of 70 show some signs of OA. Overall, up to 25% of the total population is affected, due to the general aging of the population itself, and this is an increasing tendency. However, in people over the age of 40 the incidence of OA is already rising due to posttraumatic or secondary forms, like dysplasia, malalignment and other background factors. CLAIM: There is a high need for mobility and sports in all generations, so the demands on the joints are high, especially if early degenerative changes are already present. Orthopaedic physicians are challenged with the limited load capacity, pain and the progressive joint problems, on the one hand to get the patient back to full mobility, and, on the other, to prevent early joint replacement. Early diagnosis and preventive measurements, as well as conservative treatments-from medication to braces-are necessary to achieve symptom-free movement and joint preservation to avoid joint replacement.

Functional repertoire of EV-associated miRNA profiles after lipoprotein depletion via ultracentrifugation and size exclusion chromatography from autologous blood products.

Cartilage breakdown, inflammation and pain are hallmark symptoms of osteoarthritis, and autologous blood products such as citrate-anticoagulated platelet-rich plasma (CPRP) or hyperacute serum (hypACT) have been developed as a regenerative approach to rebuild cartilage, inhibit inflammation and reduce pain. However, mechanisms of action of these blood derivatives are still not fully understood, in part due to the large number of components present in these medical products. In addition, the discovery of extracellular vesicles (EVs) and their involvement in intercellular communication mediated by cargo molecules like microRNAs (miRNAs) opened up a whole new level of complexity in understanding blood products. In this study we focused on the development of an isolation protocol for EVs from CPRP and hypACT that can also deplete lipoproteins, which are often co-isolated in EV research due to shared physical properties. Several isolation methods were compared in terms of particle yield from CPRP and hypACT. To gain insights into the functional repertoire conveyed via EV-associated miRNAs, we performed functional enrichment analysis and identified NFκB signaling strongly targeted by CPRP EV miRNAs, whereas hypACT EV miRNAs affect IL6- and TGFß/SMAD signaling.

[Apophyseal injuries in sports]. / Apophysenschäden im Sport.

The number of adolescents and children in elite and high-intensity mass sports is increasing, with respect to industrial nations. High-intensity training can cause overload due to the increased traction effect, particularly on tendon and muscle insertion sites. Apophyses are the center for ossification in tendon and muscle insertions and are therefore particularly vulnerable in youths to overload-related pathologies. Core measures in the prevention are a systematic planning of training and the avoidance of mechanical overstraining in the growth period. An exact imaging enables the diagnosis of apophyseal structural damage at an early stage, which in this phase can be healed by a pause in training and conservative measures.

The Effect of Blood-Derived Products on the Chondrogenic and Osteogenic Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells Originated from Three Different Locations.

BACKGROUND: Adipose-derived mesenchymal stem cells (AD-MSCs) from fat tissue considered "surgical waste" during joint surgery may provide a potent source for regenerative medicine. Intra-articular, homologous fat tissue (Hoffa's fat pad, pouch fat) might possess a superior chondrogenic and osteogenic differentiation potential in comparison to extra-articular, nonhomologous fat. Blood products might further enhance this potential. METHODS: AD-MSCs were isolated from fat tissue of 3 donors from 3 locations each, during total knee replacement. Isolated cells were analyzed via flow cytometry. Cells were supplemented with blood products: two types of platelet-rich plasma (EPRP-PRP prepared in the presence of EDTA; CPRP-PRP prepared in the presence of citrate), hyperacute serum (hypACT), and standard fetal calf serum (FCS) as a positive control. The viability of the cells was determined by XTT assay, and the progress of differentiation was tested via histological staining and monitoring of specific gene expression. RESULTS: Blood products enhance ex vivo cell metabolism. Chondrogenesis is enhanced by EDTA-PRP and osteogenesis by citrate PRP, whereas hyperacute serum enhances both differentiations comparably. This finding was consistent in histological analysis as well as in gene expression. Lower blood product concentrations and shorter differentiation periods lead to superior histological results for chondrogenesis. Both PRP types had a different biological effect depending upon concentration, whereas hyperacute serum seemed to have a more consistent effect, independent of the used concentration. CONCLUSION: (i) Blood product preparation method, (ii) type of anticoagulant, (iii) differentiation time, and (iv) blood product concentration have a significant influence on stem cell viability and the differentiation potential, favouring no use of anticoagulation, shorter differentiation time, and lower blood product concentrations. Cell-free blood products like hyperacute serum may be considered as an alternative supplementation in regenerative medicine, especially for stem cell therapies.

Lymphedema and employability - Review and results of a survey of Austrian experts.

BACKGROUND: Literature about lymphedema and its influence on the ability to work and employability is limited. The aim of the present study was to investigate the opinion of Austrian experts on factors influencing the ability to work and employability in patients suffering from lymphedema. METHODS: A self-administered questionnaire consisting of 6 questions was sent to 12 Austrian lymphedema experts with 6 different specializations from May to August 2016. These experts were asked about suitable and unsuitable professions, the possible influence of lymphedema on the ability to work and employability as well as about existing and additional measures to improve the return to work. RESULTS: The reply rate was 100% (12 out of 12). All experts agreed that lymphedema can restrict the ability to work and employability. The leading reason for limited ability to work and employability was restricted mobility or function of the affected limb along with time-consuming therapeutic modalities, pain and psychological stress. The most suitable job named was teacher and the most unsuitable job named was cook. As easements for return to work, early rehabilitation, self-management, coping strategies, patient education, employer's goodwill and employer's cooperation were reported. Furthermore, experts stressed the need for an adjustment of the legal framework as well as low-barrier and more therapy offers. CONCLUSIONS: Adjusted work demands seem to be of greater importance to support the ability to work and employability than recommendations for specific job profiles alone. Experts suggest an adjustment of the legal framework for affected patients, claiming a right for early rehabilitation as well as for life-long therapy. Even though some clinically useful conclusions may be drawn from this article, further research in the field is warranted.

Modulation of mesenchymal stromal cell characteristics by microcarrier culture in bioreactors.

Mesenchymal stromal cells (MSCs) are promising candidates for cell therapy. Their therapeutic use requires extensive expansion to obtain a sufficiently high number of cells for clinical applications. State-of-the-art expansion systems, that is, primarily culture flask-based systems, are limited regarding scale-up, automation, and reproducibility. To overcome this bottleneck, microcarrier (MC)-based expansion processes have been developed. For the first time, MSCs from the perinatal sources umbilical cord (UC) and amniotic membrane (AM) were expanded on MCs. This study focuses on the comparison of flask- and Cytodex 1 MC-expanded MSCs by evaluating the influence of the expansion process on biological MSC characteristics. Furthermore, we tested the hypothesis to obtain more homogeneous MSC preparations by expanding cells on MCs in controlled large-scale bioreactors. MSCs were extensively characterized determining morphology, cell growth, surface marker expression, and functional properties such as differentiation capacity, secretion of paracrine factors, and gene expression. Based on their gene expression profile MSCs from different donors and sources clearly clustered in distinct groups solely depending on the expansion process-MC or flask culture. MC- and flask-expanded MSCs significantly differed from each other regarding surface markers and both paracrine factors and gene expression profiles. Furthermore, based on gene expression analysis, MC cultivation of MSCs in controlled bioreactor systems resulted in less heterogeneity between cells from different donors. In conclusion, MC-based MSC expansion in controlled bioreactors has the potential to reliably produce MSCs with altered characteristics and functions as compared to flask-expanded MSCs. These findings may be useful for the generation of MSCs with tailored properties for clinical applications.