Search

Background: Epigenetics is the study of changes in gene expression or cellular phenotype caused by mechanisms other than changes in the underlying DNA sequence. It is widely accepted that cancer has genetic and epigenetic origins. The idea of epigenetic reprogramming of cancer cells by an embryonic microenvironment possesses potential interest from the prospect of both basic science and potential therapeutic strategies. Chick embryo extract (CEE) has been used for the successful expansion of many specific stem cells and has demonstrated the ability to facilitate DNA demethylation. Questions/purposes: The current study was conducted to compare the status of DNA methylation in highly metastatic and less metastatic osteosarcoma cells and to investigate whether CEE may affect the epigenetic regulation of tumor suppressor genes and thus change the metastatic phenotypes of highly metastatic osteosarcoma cells. Methods: K7M2 murine OS cells were treated with CEE to determine its potential effect on DNA methylation, cell apoptosis, and invasion capacity. Results: Our current results suggest that the methylation status of tumor suppressor genes (p16, p53, and E-cadherin) is significantly greater in highly metastatic mouse ostoesarcoma K7M2 cells in comparison with less metastatic mouse osteosarcoma K12 cells. CEE treatment of K7M2 cells caused demethylation of p16, p53, and E-cadherin genes, upregulated their expression, and resulted in the reversion of metastatic phenotypes in highly metastatic osteosarcoma cells. Conclusions: CEE may promote the reversion of metastatic phenotypes of osteosarcoma cells and can be a helpful tool to study osteosarcoma tumor reversion by epigenetic reprogramming. Clinical Relevance: Demethylation of tumor suppressor genes in osteosarcoma may represent a novel strategy to diminish the metastatic potential of this neoplasm. Further studies, both in vitro and in vivo, are warranted to evaluate the clinical feasibility of this approach as an adjuvant to current therapy. [ABSTRACT FROM AUTHOR]

Duchenne muscular dystrophy (DMD) is a degenerative muscle disorder characterized by the lack of dystrophin expression at the sarcolemma of muscle fibers. In addition, DMD patients acquire osteopenia, fragility fractures, and scoliosis indicating that a deficiency in skeletal homeostasis coexists but little is known about the effects of DMD on bone and other connective tissues within the musculoskeletal system. Recent evidence has emerged implicating adult stem cell dysfunction in DMD myopathogenesis. Given the common mesenchymal origin of muscle and bone, we sought to investigate bone and other musculoskeletal tissues in a DMD mouse model. Here, we report that dystrophin-utrophin double knockout ( dko) mice exhibit a spectrum of degenerative changes, outside skeletal muscle, in bone, articular cartilage, and intervertebral discs, in addition to reduced lifespan, muscle degeneration, spinal deformity, and cardiomyopathy previously reported. We also report these mice to have a reduced capacity for bone healing and exhibit spontaneous heterotopic ossification in the hind limb muscles. Therefore, we propose the dko mouse as a model for premature musculoskeletal aging and posit that a similar phenomenon may occur in patients with DMD. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 343-349, 2013 [ABSTRACT FROM AUTHOR]

Background: G-CSF-mobilized PBPCs are routinely cryopreserved within 24 hours of collection. The ability to hold PBPCs for extended time would offer increased flexibility for patients and hospitals. Retention of PBPC properties following overnight shipping, extended liquid storage at 1 to 6 degrees C, and cryopreservation was evaluated.Study Design and Methods: PBPCs were stored in liquid at 1 to 6 degrees C up to 3 days, with and without shipping, and then cryopreserved in HES (6%), DMSO (5%), and HSA (4%). Thawed samples were assayed after two procedures, on dilution and after dilution and washing. Nucleated cells, viability, CD34+ cell number, committed progenitor colonies, and long-term culture-initiating cells were measured.Results: CD34+ cell number, committed colony-forming cells, and long-term culture-initiating cells were essentially maintained when samples were stored in liquid for 1, 2, or 3 days before cryopreservation or after thawing and dilution. Nevertheless, significant (p < 0.05, paired t test) losses in total nucleated cell numbers were observed if thawed PBPC samples were washed before assay.Conclusion: PBPCs can be maintained at 1 to 6 degrees C for up to 3 days and can be cryopreserved after extended storage with properties minimally altered. Dilution alone, without centrifugation and washing, of thawed PBPC samples is a satisfactory procedure for preparing samples for in vitro assays. [ABSTRACT FROM AUTHOR]

Introduction: Skeletal muscle relaxants have previously not been examined in multimodal anesthesia regimens following joint arthroplasty. We sought to evaluate cyclobenzaprine's effect on postoperative opioid consumption as well as surgical recovery following joint arthroplasty., Materials and Methods: In this retrospective cohort study, 471 patients undergoing 554 joint arthroplasty procedures were evaluated. Patients were divided into cohorts who did and did not receive cyclobenzaprine postoperatively, and postoperative opioid consumption and functional recovery measures were recorded in each cohort., Results: In the unadjusted model, the cyclobenzaprine cohort experienced a 1.11 increase in pain scores on postoperative day zero (95% CI (0.60, 1.62), p < 0.0001). After adjusting for age, sex, BMI, CCI, perioperative MME, laterality, procedure, anesthesia, pre-op opioid use, pre-operative other controlled substance uses and pre-op benzodiazepine use, the cyclobenzaprine cohort's pain scores were 1.08 units higher at rest (95% CI (0.59, 1.56), p < 0.0001) and 1.25 units higher with activity on postoperative-day-zero (95% CI (0.78, 1.72), p < 0.0001). Both cohorts experienced statistically insignificantly different changes in mobility scores between postoperative day zero and postoperative day one, range of motion at 6 and 12 weeks, and readmission in <90 days. Postoperative morphine milliequivalents were insignificantly different between cohorts after controlling for pain in all models (base model mean ratio: 1.06, 95% CI (0.87,1.29), p = 0.5599) (Full model mean ratio: 1.09, 95% CI (0.91,1.30), p = 0.3608)., Conclusions: Cyclobenzaprine shows utility in a multimodal anesthetic approach after joint arthroplasty in patients with higher baseline pain., Competing Interests: The authors received no funding for this study and declare no conflicts of interest., (© 2024 Professor P K Surendran Memorial Education Foundation. Published by Elsevier B.V. All rights reserved.)

»: In anterior cruciate ligament (ACL)-deficient knees, treatment of medial compartment osteoarthritis (OA) that is refractory to nonoperative modalities is a controversial subject., »: Currently available treatment options include unicompartmental knee arthroplasty (UKA) with or without ACL reconstruction (ACLR), high tibial osteotomy (HTO) with or without ACLR, and total knee arthroplasty (TKA)., »: Each treatment option has its own risks and benefits, and the evidence that is reviewed in this article suggests that patient characteristics guide treatment selection., »: Future high-quality prospective studies that directly compare all 3 of the modalities are necessary to determine the best treatment option for different patient populations., Competing Interests: Disclosures: The authors indicated that no external funding was received for any aspect of this work. The Disclosure of Potential Conflicts of Interest forms are provided with the online version of the article (http://links.lww.com/JBJSREV/A673)., (Copyright © 2021 by The Journal of Bone and Joint Surgery, Incorporated.)

Background: Total knee arthroplasty (TKA) is one of the most effective ways to treat end-stage painful conditions of the knee. However, non-standardized reporting patterns can make quantitative analysis of patient outcomes difficult., Methods: A systematic review of the literature was performed using keywords "total knee arthroplasty" and "total knee replacement." Randomized controlled trials (RCTs) meeting the inclusion criteria were sorted and reviewed. Type of study, outcome measures used to report their results, and the actual results were recorded. Quantitative analysis was performed., Results: A total of 233 RCTs were included. There was significant variability in the reporting of short term and long term outcomes in total knee arthroplasty. The most common treatment domains in order of decreasing frequency were objective knee function, subjective knee function, perioperative complications, and pain. Range of motion was the most common outcome metric reported in all the RCTs and also was the most common metric used to assess objective knee function. The most common patient reported outcome measure used to assess postoperative function was the Knee Society Score followed by Knee Injury and Osteoarthritis Outcome Score. The Visual Analog Scale was the most common measurement tool used to assess postoperative pain. Most studies assessed patient outcomes in three treatment domains. None reported outcomes in all seven domains., Conclusion: There is significant variability in outcome reporting patterns in TKA literature. Most studies do not track outcomes comprehensively, with a significant minority of the RCTs tracking outcomes in only one treatment domain., Competing Interests: None., (© 2020 Delhi Orthopedic Association. All rights reserved.)

Background: There has been a shift toward using patient-reported outcome measures (PROMs) to capture functional improvement and patient satisfaction after total hip arthroplasty (THA). Because there is no standard measure or set of measures, variability in reporting patterns makes comparison across studies difficult., Methods: We performed a review of the literature using the keywords "total hip arthroplasty" and "total hip replacement" to electronically search PubMed, using the date range August 1, 2014, to August 1, 2019. Randomized clinical trials (RCTs) that were published in 12 high-impact journals were analyzed., Results: One hundred and fifty-nine RCTs were included. The most common topic of investigation was hip implant design and materials, followed by the effect of different hip approaches on patient outcomes. The follow-up period was classified as short-term (<2 years), mid-term (2 to 10 years), or long-term (>10 years). Only 6% of the RCTs reported long-term outcomes. The comprehensiveness of studies was determined on the basis of how many of the 7 following outcome domains were assessed: subjective hip function (PROMs), objective outcome measures (examination findings, laboratory values, etc.), imaging analysis, survivorship, patient satisfaction, pain assessment, and postoperative complications. Subjective hip function and imaging findings were the most commonly reported outcome domains, while implant survivorship and patient satisfaction were the least frequently reported. There was substantial variation in outcome reporting, with 35 unique PROMs utilized to assess subjective hip function. Although the Harris hip score was the most commonly used joint-specific PROM, it was only reported in 42% of the studies. None of the RCTs reported results in all 7 outcome domains, and 13.8% of studies reported results in only 1 outcome domain., Conclusions: There is substantial variability and a lack of comprehensiveness in outcome measures used to report results in THA clinical trials, making it nearly impossible to perform cross-study comparisons., Clinical Relevance: There is an immediate need for the establishment of a standardized set of measures to allow comparison of outcomes across studies.

Background: Previous research has indicated that unicompartmental arthroplasty may be an effective treatment for focal osteonecrosis in the knee; however, these reports have been composed of small patient cohorts and without characterization of the osteonecrotic lesions. Therefore, the purpose of this study was to investigate the effectiveness of unicompartmental arthroplasty for the treatment of focal osteonecrosis within the medial femoral condyle including an assessment of lesion size., Methods: A consecutive series of >5,000 unicompartmental knee arthroplasties performed at a single institution was retrospectively reviewed to identify cases of medial femoral condyle osteonecrosis with a minimum 2-year follow-up. Lesion size was classified according to the ratio of lesion width to condylar width, as well as lesion depth relative to condylar depth. Patient-reported outcome measures and need for a revision procedure were studied., Results: Sixty-four patients (32 males, 32 females; 65 knees) with a mean age of 64 years were included. The mean patient follow-up was 5.3 years (range, 2 to 12 years). The mean ratio of lesion width to condylar width was 64%, the mean lesion depth was 1.11 cm, and 82% of cases demonstrated subchondral collapse. At the time of the latest follow-up, patients demonstrated substantial improvements in the pain, function, and clinical components of the Knee Society Score, by 36, 25, and 51, respectively. Four patients (6%) required a revision, of which only 1 was for aseptic loosening of the femoral component., Conclusions: Unicompartmental arthroplasty is an effective treatment for advanced-stage focal osteonecrosis of the medial femoral condyle. Loss of component fixation to the femoral condyle did not appear to be a substantial concern because there was only 1 femoral failure as a result of aseptic loosening, despite lesions affecting a significant portion of the femoral condyle., Level of Evidence: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Background: Thigh pain is a variably reported symptom in the postoperative period following primary total hip arthroplasty (THA) with a well-fixed cementless femoral implant. While research has identified stem size, stem alignment, and differences in modulus of elasticity between implant and host bone as potential sources of thigh pain, only one study has specifically examined the impact of variation in implant design within a single femoral stem design. The purpose of this work was to determine whether there were differences in the pain experienced by patients treated with two design variants of a femoral stem during direct anterior THA., Materials and Methods: Patients undergoing primary direct anterior THA at a single center between 2011-2015 were included in the study. Those patients suffering extensive comorbidities and postoperative complications were excluded from analysis. Study subjects completed a pain drawing and scale questionnaire for the operative hip at least one year following surgery. A cementless, single-taper wedge, titanium femoral component design available in short- and standard-length variations was used in all cases. Pain outcomes were compared between these two femoral stem options., Results: A total of 1347 patients (1536 THA) met inclusion criteria for the study and surveys were returned for 820 of these THAs. Demographic data and UCLA activity scores were similar between cohorts of patients receiving the short- and standard-length components. The most common locations of pain reported were in the lower back and trochanteric region, 28% and 24% respectively. Patients in the short-length cohort reported a significantly lower incidence of pain in the anterior thigh as compared to the standard-length cohort, 12% versus 19% respectively [p=0.007]. There was no difference in the number of patients experiencing moderate to severe intensity of anterior thigh pain between these two groups, 3% versus 5% respectively [p=0.36]. No other statistically significant differences were found in the incidence of pain in the lower back, buttock, groin, trochanter, lateral thigh, or posterior thigh regions between the two cohorts., Conclusion: While the lower back and trochanteric region may be the most frequent areas of pain experienced in patients at one-year or more postoperative from direct anterior THA, a significantly higher incidence of anterior thigh pain is found in those patients treated with a standard-length stem design as compared to the short design. This finding may be due to contact between the tip of the distal stem with the femoral diaphysis as has been theorized in previous research, which is circumvented with the short design variant.

Background: Treatment of isolated lateral compartment arthritic disease with partial knee arthroplasty remains underutilized in comparison to medial unicompartmental arthroplasty. This study examines the survival and outcome of lateral unicompartmental arthroplasty utilizing the first implant specifically developed for the lateral compartment., Materials and Methods: A retrospective review was performed to detect lateral unicompartmental arthroplasty procedures performed in our practice between January 2013 and May 2016. Patients indicated for surgery met specific preoperative clinical and radiographic criteria confirming lateral compartment arthritic disease with a correctable deformity, intact full-thickness medial cartilage, competent anterior cruciate ligament, and minimal disease in the patellofemoral compartment. A single implant design was used in all cases which consisted of a fixed-bearing tibial component specifically adapted to lateral compartment anatomy. Unicompartmental arthroplasty surgical technique was adjusted to attempt to recreate lateral compartment kinematics., Results: Fifty-two consecutive patients (56 knees) with lateral unicompartmental arthroplasty procedures meeting minimum two-year follow up were included in the study. Thirty-nine patients were female, and 93% of cases were performed for treatment of osteoarthritis. At a mean follow up of nearly three years, Knee Society clinical and functional scores improved postoperatively by a mean difference of 41 and 21, respectively. There were two reoperations, one medial unicompartmental arthroplasty for osteoarthritis progression and a superficial debridement for a non-healing wound. Thus, failure of lateral unicondylar knee arthroplasty (UKA) was less than 2% in this study. There were no other component revisions, radiographic evidence of loosening, or clinical failures., Conclusions: At early follow up, lateral unicompartmental arthroplasty using a modified surgical technique and an implant specifically designed for the lateral compartment is a reliable treatment for isolated lateral femorotibial arthritis when meeting defined indications.

Background: Despite growing interest in direct anterior approach total hip arthroplasty, perioperative femoral fracture and early aseptic loosening are increasingly recognized complications. Previous research has documented the role of surgeon experience in association with these femoral complications. The purpose of this study was to explore the relationship between femoral component design and early periprosthetic femoral complications., Methods: This was an extension of previous work with an updated patient cohort of 5090 consecutive direct anterior primary total hip arthroplasties at a single institution with a single-taper, wedge femoral stem comprising 4 variants involving length and geometry: group 1, full-length, standard profile; group 2, full-length, reduced distal profile; group 3, short-length, standard profile; and group 4, short-length, reduced distal profile. Records were reviewed retrospectively for the incidence of early postoperative periprosthetic fracture or aseptic loosening and analyzed with regard to patient demographics and femoral stem type., Results: There were 42 (0.83%) periprosthetic femur complications observed in the early postoperative period. Increased age (P < .001) and female gender (P = .023) were significantly associated with incidence of femoral complications in univariate analysis, while age maintained this significant relationship in multivariate analysis (P < .001). There was a trend toward increased complication rate in patients receiving a short stem with full profile taper (1.27%, P = .0539)., Conclusion: Despite an overall low rate of femoral complications after direct anterior total hip arthroplasty, the risk is increased in elderly patients and females. Furthermore, femoral stem design may portend an elevated risk of these complications., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Despite improvements in polyethylene bearing surface properties, only 1 previous study has examined the results of larger thickness bearings. The purpose of this study was to determine whether polyethylene thickness influenced patient outcomes and implant survival following modular total knee arthroplasty., Methods: A retrospective review was performed of patients undergoing primary total knee arthroplasty from 2003 to 2014 in a single practice database. Patients were separated into "thin" and "thick" polyethylene groups based on manufacturer polyethylene bearing sizes of 14 mm or less compared to those greater than 14 mm, respectively. Patient clinical outcomes, need for revision surgery, and overall implant survival rates were evaluated., Results: A total of 6698 primary knee arthroplasties were included, and a thin bearing was used in 96.5% of these cases. Preoperatively, patients with a thick bearing had significantly lower Knee Society clinical scores (P < .01), a trend toward lower functional scores (P = .06), and more significant coronal plane deformity. Postoperatively, patients with thick bearings exhibited better Knee Society clinical and pain scores as well as similar functional scores and University of California at Los Angeles activity scores. The overall reoperation rate and 10-year survivorship free of revision were similar between thick and thin bearings (1.7% vs 2.3%; 98.2% vs 96.1%). Patients with thin bearings were twice as likely to require a manipulation under anesthesia postoperatively (P = .02), while there were no failures in the thick bearing group due to aseptic loosening or instability., Conclusion: Patients with thick polyethylene bearings performed similarly or better in multiple clinical outcomes and survivorship compared to those with thin bearings., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Contemporary research has shown medial mobile-bearing unicompartmental knee arthroplasty to be an effective treatment in patients younger than 60 years; however, only one other study has specifically investigated unicompartmental arthroplasty outcomes in patients 50 years or younger. The purpose of this study was to determine the clinical outcomes and survivorship of medial mobile-bearing unicompartmental arthroplasty in this younger patient population., Methods: A retrospective review of patients undergoing primary unicompartmental knee arthroplasty using the Oxford partial knee from 2003 to 2014 in a single practice database was performed. Patients were included in the study if they were 50 years of age or younger with a primary diagnosis of anteromedial osteoarthritis and minimum clinical follow-up of 2 years. Patient clinical outcomes, function, and need for revision surgery were assessed., Results: The study included 340 knees. Average patient age was 46.5 years, and the mean follow-up was 6.1 years. Patients demonstrated significant improvements (P < .05) in range of motion (114.5 v 116.9), University of California Los Angeles activity score (4.4 vs 5.6), Knee Society clinical (37.3 vs 86.5) and functional scores (58.8 v 79.8). Overall, 20 patients required reoperation, and the predicted survival rate was 96% at 6 years and 86% at 10 years. Aseptic loosening occurred in 7 patients at an average of 5.6 years postoperatively, while 4 patients required conversion to total knee arthroplasty because of arthritic progression at a mean time of 6.6 years. There were no revision procedures required due to polyethylene liner wear or breakage., Conclusion: Medial mobile-bearing unicompartmental arthroplasty should be considered as a treatment option in patients younger than 50 years of age suffering from anteromedial osteoarthritis of the knee., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The case of an atraumatic dislocation of a fixed bearing in a medial unicompartmental arthroplasty with an oxinium femoral component is presented. A review of the literature pertaining to knee arthroplasty locking mechanisms is discussed. Potential modes of locking mechanisms failure are reviewed including the recognition of such failures in the clinical setting. This is the first report of a dislocated fixed-bearing medial oxinium unicompartmental arthroplasty and consequent metal arthrogram., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Arginine vasopressin 1B receptor (V 1B ) antagonists may have utility for the treatment of major depressive disorder (MDD)., Methods: The V 1B antagonist ABT-436 ( N  = 31) or matching placebo ( N  = 20) was administered to MDD subjects for 7 days. The main study objectives were to assess the safety and hypothalamic-pituitary-adrenal axis (HPA) effects of ABT-436 in MDD subjects. MDD symptoms were assessed using the 17-item Hamilton Depression Rating Scale (HAM-D-17) and the subject-rated Mood and Anxiety Symptom Questionnaire (MASQ)., Results: The most prevalent safety finding associated with ABT-436 800 mg QD was increased mild-moderate diarrhea (68% v 5%, p  < 0.001). Increased nausea (26% v 5%, p  < 0.10), decreased systolic blood pressure (3.15-3.44 mmHg, p  < 0.10) and increased heart rate (3.42-4.01 bpm, p  < 0.05) were also associated with ABT-436 800 mg QD. Basal HPA activity measured by 24-hr urine total glucocorticoids was 25% lower with ABT-436 than placebo ( p  < 0.001). The reduction was, on average, larger in subjects with higher baseline urine total glucocorticoids. Results on plasma adrenocorticotrophic hormone (ACTH), urine, serum and saliva cortisol, and saliva cortisone also showed basal HPA attenuation with ABT-436. Dynamic HPA activity measured by plasma ACTH and serum cortisol responses to corticotrophin releasing hormone (CRH) were 30-46% lower in ABT-436 subjects (all p  < 0.001). Each ABT-436 subject showed response to CRH in or near the baseline range of responses. ABT-436 was associated with more favorable symptom changes on two of five MASQ subscales (estimated effect size 1.47-1.86, p  < 0.01) but not on HAM-D-17., Conclusions: The results support further clinical study of the antidepressant potential of ABT-436.

Background: Osteosarcoma (OS) is the most common primary bone tumor, with a propensity to metastasize to the lungs. Five-year survival for metastatic OS is below 30%, and has not improved for several decades despite the introduction of multi-agent chemotherapy. Understanding OS cell migration to the lungs requires an evaluation of the lung microenvironment. Here we utilized an in vitro lung cell and OS cell co-culture model to explore the interactions between OS and lung cells, hypothesizing that lung cells would promote OS cell migration and survival. The impact of a novel anti-OS chemotherapy on OS migration and survival in the lung microenvironment was also examined., Methods: Three human OS cell lines (SJSA-1, Saos-2, U-2) and two human lung cell lines (HULEC-5a, MRC-5) were cultured according to American Type Culture Collection recommendations. Human lung cell lines were cultured in growth medium for 72 h to create conditioned media. OS proliferation was evaluated in lung co-culture and conditioned media microenvironment, with a murine fibroblast cell line (NIH-3 T3) in fresh growth medium as controls. Migration and invasion were measured using a real-time cell analysis system. Real-time PCR was utilized to probe for Aldehyde Dehydrogenase (ALDH1) expression. Osteosarcoma cells were also transduced with a lentivirus encoding for GFP to permit morphologic analysis with fluorescence microscopy. The anti-OS efficacy of Disulfiram, an ALDH-inhibitor previously shown to inhibit OS cell proliferation and metastasis in vitro, was evaluated in each microenvironment., Results: Lung-cell conditioned medium promoted osteosarcoma cell migration, with a significantly higher attractive effect on all three osteosarcoma cell lines compared to basic growth medium, 10% serum containing medium, and NIH-3 T3 conditioned medium (p <0.05). Lung cell conditioned medium induced cell morphologic changes, as demonstrated with GFP-labeled cells. OS cells cultured in lung cell conditioned medium had increased alkaline phosphatase staining., Conclusions: Lung endothelial HULEC-5a cells are attractants for OS cell migration, proliferation, and survival. The SJSA-1 osteosarcoma cell line demonstrated greater metastatic potential than Saos-2 and U-2 cells. ALDH appears to be involved in the interaction between lung and OS cells, and ALP may be a valuable biomarker for monitoring functional OS changes during metastasis.

Open tibial shaft fracture occurring below an ipsilateral total knee arthroplasty (TKA) is a unique injury pattern that presents an additional degree of complexity to an already challenging treatment algorithm. Tibial shaft fracture is a surgical emergency requiring respect for the soft-tissue envelope and consideration of the biomechanical and biologic factors involved in healing. Treatment with an intramedullary nail relative to other types of internal or external fixation methods optimizes these factors and minimizes the risks of nonunion, malunion, infection, soft-tissue compromise, and reoperation, which are prevalent after this fracture. However, tibial shaft fracture associated with an ipsilateral TKA complicates standard treatment principles and increases the risks after surgery. In many instances, this type of injury pattern in a patient with medical comorbidities that would impede fracture and wound healing would make a limb amputation the preferred method of treatment. However, in this case report, we examine treatment options for an open tibial shaft fracture in the setting of an ipsilateral TKA and propose a method of limb salvage in a patient with medical comorbidities sustaining this injury pattern.

CD133+ cells purified from hematopoietic tissues are enriched mostly for hematopoietic stem/progenitor cells, but also contain some endothelial progenitor cells and very small embryonic-like stem cells. CD133+ cells, which are akin to CD34+ cells, are a potential source of stem cells in regenerative medicine. However, the lack of convincing donor-derived chimerism in the damaged organs of patients treated with these cells suggests that the improvement in function involves mechanisms other than a direct contribution to the damaged tissues. We hypothesized that CD133+ cells secrete several paracrine factors that play a major role in the positive effects observed after treatment and tested supernatants derived from these cells for the presence of such factors. We observed that CD133+ cells and CD133+ cell-derived microvesicles (MVs) express mRNAs for several antiapoptotic and proangiopoietic factors, including kit ligand, insulin growth factor-1, vascular endothelial growth factor, basic fibroblast growth factor, and interleukin-8. These factors were also detected in a CD133+ cell-derived conditioned medium (CM). More important, the CD133+ cell-derived CM and MVs chemoattracted endothelial cells and display proangiopoietic activity both in vitro and in vivo assays. This observation should be taken into consideration when evaluating clinical outcomes from purified CD133+ cell therapies in regenerative medicine.

Umbilical cord blood-derived very small embryonic-like stem cells (UCB-VSELs) are the most primitive stem cells circulating in fetal peripheral blood. These very rare cells slightly smaller than red blood cells i) become mobilized during delivery, ii) are enriched in fraction of CD133+ Lin-CD45- cells iii) express markers of pluripotent stem cells (e.g., Oct4, Nanog, and SSEA-4) and iv) display a distinct morphology characterized by a high nuclear/ cytoplasmic ratio and undifferentiated chromatin. We envision that VSELs are released into neonatal peripheral blood as a migrating population of stem cells involved in regeneration of tissues that become damaged in the process of delivery. They may also be responsible for the occurrence of fetal-maternal chimerism. Our most recent data suggest that UCB-VSELs exhibit some characteristics of long-term repopulating hematopoietic stem cells (LT-HSCs). We propose that UCB-VSELs may eventually be employed as a source of pluripotent stem cells in regenerative medicine.

Many psychopathology research assessment tools can be used easily and productively in clinical practice. We conducted a workshop in 2009 and 2010 at the American Psychiatric Association annual meeting designed to bring clinicians some commonly used adult research measures with broad applicability to a variety of conditions. This article reviews what was most helpful to the practicing clinicians at the workshop.

The Phase I clinical study was designed to assess the safety and feasibility of a dose escalating intracoronary infusion of autologous bone marrow (BM)-derived CD133+ stem cell therapy to the patients with chronic total occlusion (CTO) and ischemia. Nine patients were received CD133+ cells into epicardial vessels supplying collateral flow to areas of viable ischemic myocardium in the distribution of the CTO. There were no major adverse cardiac events (MACE), revascularization, re-admission to the hospital secondary to angina, or acute myocardial infarction (AMI) for the 24-month period following cellular infusion. In addition, there were no periprocedural infusion-related complications including malignant arrhythmias, loss of normal coronary blood flow or acute neurologic events. Cardiac enzymes were negative in all patients. There was an improvement in the degree of ischemic myocardium, which was accompanied by a trend towards reduction in anginal symptoms. Intracoronary infusion of autologous CD133+ marrow-derived cells is safe and feasible. Cellular therapy with CD133+ cells to reduce anginal symptoms and to improve ischemia in patients with CTO awaits clinical investigation in Phase II/III trials.

Background: The International Knee Documentation Committee Subjective Knee Form (IKDC SKF) is a patient-reported knee-specific outcome measure that has been shown to be a reliable, valid, and responsive measure for patients with a variety of knee conditions. Further testing is required to compare the reliability and responsiveness of the IKDC SKF to other commonly used patient-reported outcome measures for patients with articular cartilage lesions., Hypothesis: The IKDC SKF has equal or better levels of reliability and responsiveness than the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), modified Cincinnati Knee Rating System (CKRS), and the Short Form 36 in patients with articular cartilage lesions., Study Design: Cohort study (diagnosis); Level of evidence, 2., Methods: Reliability was assessed by administering the 4 patient-reported outcome measures to 17 individuals who had undergone articular cartilage surgery 5 years before participation in this study. Responsiveness was determined by administering the 4 patient-reported outcome measures to 51 individuals with diagnosed focal articular cartilage defects who were scheduled to undergo surgical treatment. In both groups, the outcome measures were administered at baseline and at 6 and 12 months' follow-up. Participants also provided a global rating of change in comparison to baseline at the 6- and 12-month follow-ups., Results: Test-retest reliability coefficients were 0.91 and 0.93 for the IKDC SKF at the 6- and 12-month follow-ups, respectively. The effect sizes and standardized response means were large (>0.80) at 6 months after surgery for the WOMAC pain, physical function, and total scores and 12 months after surgery for the IKDC SKF; WOMAC pain, physical function, and total; and CKRS scores. Six months after surgery, significant differences between those who were improved compared with those who were unchanged or worse were found only for the IKDC SKF. Twelve months after surgery, significant differences between the improved and unchanged groups were found for all of the knee-specific patient-reported outcome measures. Finally, the IKDC SKF, WOMAC, and CKRS scores were able to differentiate between individuals who perceived themselves to be improved versus not improved and the minimum clinically important difference for the IKDC SKF was 6.3 at 6 months and 16.7 at 12 months., Conclusion: The reliability and responsiveness of the IKDC SKF is comparable with other commonly used patient-reported outcome measures for patients with articular cartilage lesions. The IKDC SKF is a suitable alternative to other commonly used knee-specific instruments for measuring symptoms, daily function, and level of symptom-free sports activity in patients undergoing articular cartilage surgery.

Background Aims: Current clinical trials utilize non-selected bone marrow (BM) mononuclear cells (MNC) to augment vasculo genesis within ischemic vascular beds. Recent reports have identified a diminished number and function of hemat-opoietic stem cells (HSC) from aged and diseased patients. Umbilical cord blood (UCB) provides a potential robust allo-geneic source of HSC for therapeutic vasculogenesis., Methods: MNC and magnetically isolated CD133(+) cells were assessed for viability (trypan blue) and surface phenotype (flow cytometry). To test in vivo functionality of the cells, NOD/SCID mice underwent ligation of the right femoral artery followed immediately by cell injection. Blood flow recovery, necrosis, BM engraftment of human cells and histologic capillary density were determined. Cells were tested for potential mechanisms mediating the in vivo effects, including migration, cytokine secretion and angiogenic augmentation (Matrigel assays)., Results: Surface expression analysis showed CD31 (PECAM) expression was greatly increased in UCB CD133(+) cells compared with BM MNC. At 28 days, perfusion ratios were highest in animals receiving UCB CD133(+) cells, while animals receiving BM CD133(+) cells and BM MNC demonstrated perfusion ratios statistically higher than in animals treated with cytokine media alone. Animals receiving CD133(+) cells showed a statistically higher capillary density, reduced severe digit necrosis and increased engraftment in the BM than animals treated with unselected BM MNC. In vitro studies showed equivalent migration to stromal-derived factor-1 (SDF-1), increased production of tumor necrosis factor alpha (TNF-alpha) and increased branch points with the co-incubation of CD133(+) cells with human umbilical vein endothelial cells (HUVEC) in the Matrigel angiogenesis assay., Conclusions: Taken together, UCB CD133(+) cells exhibit robust vasculogenic functionality compared with BM MNC in response to ischemia.

Cardiovascular disease remains a major cause of morbidity and mortality with substantial economic cost. There remains a need for therapeutic improvement for patients refractory to revascularization and those who redevelop occlusions following revascularization. Early evidence linked age-associated reductions in the levels of circulating marrow-derived hematopoietic stem cells (HSC), characterized by expression of early HSC markers CD133 and CD34, with the occurrence of cardiovascular events and associated death. Heart tissue has the endogenous ability to regenerate through the activation of resident cardiac stem cells or through recruitment of a stem cell population from other tissues, such as bone marrow. A number of clinical trials have utilized patient-derived autologous bone marrow-derived cells or whole BM uncultured mononuclear cells (MNC) infused or injected locally to augment angiogenesis. In most cases of treating animal models with human cells, the frequency of stem cell engraftment, the subsequent number of newly generated cardiomyocytes and vascular cells, and the augmentation of endogenous microvascular collateralization, either by deposition, transdifferentiation, and/or by cell fusion, appear to be too low to explain the significant cardiac improvement. Initially, it was hypothesized that cell therapy may work by cell replacement mechanisms, but recent evidence suggests alternatively that cell therapy works by providing trophic support to the injured tissues. An alternative hypothesis is that the transplanted stem cells release soluble cytokines and growth factors (i.e., paracrine factors) that function in a paracrine fashion, contributing to cardiac repair and regeneration by inducing cytoprotection and neovascularization. Another hypothesis which may also be operative is that cell therapy may mediate endogenous regeneration by the activation of resident cardiac stem cell. Well-established clinical trials have used cord blood for the treatment of hematological malignances (e.g., leukemia, lymphoma, myeloma) and nonmalignancies (e.g., in born errors of metabolism, sickle cells anemia, autoimmune diseases), but further advances in other areas of regenerative medicine (e.g., cardiac repair) will directly benefit with the use of cord blood. These clinical outcomes demonstrate that effector cells may be delivered by an allogeneic approach, where strict tissue matching may not be necessary and treatment may be achieved by making use of the trophic support capability of cell therapy and not by a cell replacement mechanism.

The reduced expression of nuclear factor of activated T cells-1 (NFAT1) protein in umbilical cord blood (UCB)-derived CD4+ T cells and the corresponding reduction in inflammatory cytokine secretion after stimulation in part underlies their phenotypic differences from adult blood (AB) CD4+ T cells. This muted response may contribute to the lower incidence and severity of high-grade acute graft-versus-host disease (aGVHD) exhibited by UCB grafts. Here we provide evidence that a specific microRNA, miR-184, inhibits NFAT1 protein expression elicited by UCB CD4+ T cells. Endogenous expression of miR-184 in UCB is 58.4-fold higher compared with AB CD4+ T cells, and miR-184 blocks production of NFAT1 protein through its complementary target sequence on the NFATc2 mRNA without transcript degradation. Furthermore, its negative effects on NFAT1 protein and downstream interleukin-2 (IL-2) transcription are reversed through antisense blocking in UCB and can be replicated via exogenous transfection of precursor miR-184 into AB CD4+ T cells. Our findings reveal a previously uncharacterized role for miR-184 in UCB CD4+ T cells and a novel function for microRNA in the early adaptive immune response.

The synthesis and photophysical evaluation of modified nucleoside analogues in which a five-membered heterocycle (furan, thiophene, oxazole and thiazole) is attached to the 5 position of 2'-deoxyuridine are reported. The furan containing derivative is identified as the most promising responsive nucleoside of this family due to its emission quantum efficiency and degree of sensitivity to its microenvironment. The furan moiety was then attached to the 5 position of 2'-deoxycytidine as well as the 8 position of adenosine and guanosine. Photophysical evaluation of these four furan containing nucleoside analogues reveal distinct differences in the absorption, emission and quantum efficiency depending upon the class of nucleoside (pyrimidine or purine). Comparing the photophysical properties of all furan containing nucleosides, identifies the furan thymidine analogue, 5-(fur-2-yl)-2'-deoxyuridine, as the best candidate for use as a responsive fluorescent probe in nucleic acids. 5-(fur-2-yl)-2'-deoxyuridine was then converted to the corresponding phosphoramidite and site specifically incorporated into DNA oligonucleotides with greater than 88% coupling efficiency. Such furan-modified oligonucleotides form stable duplexes upon hybridization to their complementary DNA strands and display favorable fluorescent features.

Background: Intracoronary mononuclear cell therapy may produce angiogenesis in chronic myocardial ischemia. Potential complications include periprocedural infarction secondary to: reduced coronary blood flow; hyperviscosity from the cell preparation; or microvascular dysfunction. To date, no studies to evaluate these potential complications have been reported. The objective of this report was to study the safety and feasibility of intracoronary injections of autologous bone marrow mononuclear cells in a porcine chronic myocardial ischemia model., Methods: Domestic pigs (n = 5) underwent ameroid cuff placement of the left circumflex artery. Bone marrow-derived mononuclear cells [15 x 10(6) cells] labeled with CM dioctadecyl tetramethylindocarbocyanine were given by intracoronary injection. Animals were sacrificed, and hearts and vital organs were inspected grossly and by histopathology, and bone marrow underwent immunofluorescence microscopy., Results: Troponin I levels, gross inspection and histopathology did not reveal evidence of myocardial infarction. Labeled cells were observed in perivascular structures in myocardium at the injection site in all animals and in the spleen from one animal. Bone marrow aspirates indicated labeled cells., Conclusions: Intracoronary injection of autologous mononuclear cells in a porcine chronic myocardial ischemia model appears safe. Intracoronary injection resulted in cell localization in the perivascular areas of myocardium supplied by the injected vessel. Cell localization was observed only in the spleen in just one animal. Labeled cells were identified in bone marrow aspirates from three animals following injection, suggesting a role for bone marrow engraftment and repopulation as a possible mechanism for progenitor cell localization in myocardium.

Endothelial precursor cells (EPCs) cultured from adult bone marrow (BM) have been shown to mediate neovasculogenesis in murine models of vascular injury. We sought to directly compare umbilical cord blood (UCB)- and BM-derived EPC surface phenotypes and in vivo functional capacity. UCB and BM EPCs derived from mononuclear cells (MNC) were phenotyped by surface staining for expression of stromal (Stro-1, CXCR4, CD105, and CD73), endothelial (CD31, CD146, and vascular endothelial [VE]-cadherin), stem cell (CD34 and CD133), and monocyte (CD14) surface markers and analyzed by flow cytometry. The nonobese diabetic/severe combined immunodeficiency murine model of hind-limb ischemia was used to analyze the potential of MNCs and culture-derived EPCs from UCB and BM to mediate neovasculogenesis. Histologic evaluation of the in vivo studies included capillary density as a measure of neovascularization. Surface CXCR4 expression was notably higher on UCB-derived EPCs (64.29%+/-7.41%) compared with BM (19.69%+/-5.49%; P=.021). Although the 2 sources of EPCs were comparable in expression of endothelial and monocyte markers, BM-derived EPCs contained higher proportions of cells expressing stromal cell markers (CD105 and CD73). Injection of UCB- or BM-derived EPCs resulted in significantly improved perfusion as measured by laser Doppler imaging at days 7 and 14 after femoral artery ligation in nonobese diabetic/severe combined immunodeficiency mice compared with controls (P<.05). Injection of uncultured MNCs from BM or UCB showed no significant difference from control mice (P=.119; P=.177). Tissue samples harvested from the lower calf muscle at day 28 demonstrated increased capillary densities in mice receiving BM- or UCB-derived EPCs. In conclusion, we found that UCB and BM-derived EPCs differ in CXCR4 expression and stromal surface markers but mediate equivalent neovasculogenesis in vivo as measured by Doppler flow and histologic analyses.

A family of simple pyrimidine analogues has been synthesized, and their photophysical properties have been investigated. The most responsive of the family was incorporated in DNA. This isosteric fluorescent DNA analogue monitors denaturation of a DNA duplex via fluorescence and positively detects the presence of abasic sites in DNA duplexes.

Background: G-CSF-mobilized PBPCs are utilized in allogeneic and autologous PBPC transplants. Homing, adhesion, and transmigration of hematopoietic CD34+ cells are required for successful engraftment. Hematopoietic CD34+ cells undergo directional migration toward the CXCR4 receptor ligand stromal-derived factor-1 (SDF-1). Limited data are available on the effects of liquid storage and cryopreservation on PBPC CD34+ cells., Study Design and Methods: Magnetic-assisted cell sorting (MACS)-selected CD34+ cells were assayed for retention of in vitro transmigration and phenotypic changes of unit-matched liquid-stored and cryopreserved PBPC samples from healthy donors. Studies evaluated whether transmigration of CD34+ cells in Iscove's modified Dulbecco's medium plus 1 percent HSA alone or in medium supplemented with SCF or allogeneic plasma was affected by overnight incubation at 37 degrees C, relative to nonincubated conditions., Results: Transmigration was maintained during liquid storage at 1 to 6 degrees C during a 2-day period and in unit-matched cryopreserved-thawed samples that had been initially liquid stored. Overnight incubation at 37 degrees C of MACS-selected unit-matched liquid-stored or cryopreserved-thawed CD34+ cells resulted in substantially increased transmigration, in particular with noncoated filters chemoattracted with the chemokine SDF-1., Conclusion: CD34+ cell transmigration was comparable between liquid-stored and cryopreserved samples, and both demonstrated similar increases after overnight incubation at 37 degrees C.

Unlike granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood, which show a single homogeneous population of CD34(+) cells, umbilical cord blood (CB) CD34(+) cells are present as multiple populations, CD34(regular) and CD34(bright) (the latter comprising 7.0-58.2% of the total CD34(+) cells), using the ProCOUNT trade mark procedure or with anti-CD34 labeling of immunoselected cells. The CD34(regular) population contains cells with high forward scatter (CD34(regular)FSC(high)) and with low forward scatter (CD34(regular) FSC(low)). Immunomagnetically selected CD34(+) cells, sorted into CD34(regular), CD34(regular) FSC(high), CD34(regular)FSC(low), and CD34(bright) cell populations, were used in in vitro assays: only the CD34(regular)FSC(high) population transmigrated and showed growth of colony-forming unit (CFU) and long-term culture initiating cells (LTC-IC) colonies. The absolute number of CD34(+) cells in CB samples was determined by ProCOUNT trade mark and Stem Kit trade mark enumeration protocols. In liquid stored CB units, ProCOUNT trade mark and Stem Kit trade mark count differences are accounted for by the enumeration of CD34(bright) cells. Differences between ProCOUNT trade mark and Stem Kit trade mark counts using cryopreserved/thawed samples are accounted for by increased CD34(regular) FSC(low) cell numbers (2.0 +/- 1.4% in liquid stored and 27.8 +/- 14.6% in cryopreserved/thawed samples). The ProCOUNT trade mark assay includes the nonfunctional CD34(bright) and CD34(regular)FSC(low) cells as part of the CD34(+) cell count, thereby elevating the absolute number of CD34(+) cells. Using the Stem Kit trade mark assay method of gating, CD34(bright) and CD34(regular)FSC(low) cells are not counted. Our data indicate that the CD34(regular)FSC(high) cell population has functional characteristics based on the in vitro assays and a more accurate count of these cells can be achieved using the Stem Kit trade mark assay.