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RNA interference (RNAi) is mediated by an ∼21-nt double-stranded small interfering RNA (siRNA) and shows great promise in delineating gene functions and in developing therapeutics for human diseases. However, effective gene silencing usually requires the delivery of multiple siRNAs for a given gene, which is often technically challenging and time-consuming. In this study, by exploiting the type IIS restriction endonuclease-based synthetic biology methodology, we developed the fast assembly of multiplex siRNAs (FAMSi) system. In our proof-of-concept experiments, we demonstrated that multiple fragments containing three, four, or five siRNA sites targeting common Smad4 and/or BMPR-specific Smad1, Smad5, and Smad8 required for BMP9 signaling could be assembled efficiently. The constructed multiplex siRNAs effectively knocked down the expression of Smad4 and/or Smad1, Smad5, and Smad8 in mesenchymal stem cells (MSCs), and they inhibited all aspects of BMP9-induced osteogenic differentiation in bone marrow MSCs (BMSCs), including decreased expression of osteogenic regulators/markers, reduced osteogenic marker alkaline phosphatase (ALP) activity, and diminished in vitro matrix mineralization and in vivo ectopic bone formation. Collectively, we demonstrate that the engineered FAMSi system provides a fast-track platform for assembling multiplexed siRNAs in a single vector, and thus it may be a valuable tool to study gene functions or to develop novel siRNA-based therapeutics., Graphical Abstract, RNA interference shows great promise in delineating gene functions and in developing therapeutics for human diseases. He et al. exploited the type IIS restriction endonuclease-based synthetic biology and developed the fast assembly of multiplex siRNAs (FAMSi) system, which streamlines RNAi-based gene function studies and the development of siRNA-based therapeutics.

Mesenchymal stem cells (MSCs) are multipotent progenitors that have the ability to differentiate into multiple lineages, including bone, cartilage, and fat. We previously demonstrated that the least known bone morphogenetic protein (BMP)9 (also known as growth differentiation factor 2) is one of the potent osteogenic factors that can induce both osteogenic and adipogenic differentiation of MSCs. Nonetheless, the molecular mechanism underlying BMP9 action remains to be fully understood. Leptin is an adipocyte-derived hormone in direct proportion to the amount of body fat, and exerts pleiotropic functions, such as regulating energy metabolism, bone mass, and mineral density. In this study, we investigate the potential effect of leptin signaling on BMP9-induced osteogenic differentiation of MSCs. We found that exogenous leptin potentiated BMP9-induced osteogenic differentiation of MSCs both in vitro and in vivo, while inhibiting BMP9-induced adipogenic differentiation. BMP9 was shown to induce the expression of leptin and leptin receptor in MSCs, while exogenous leptin upregulated BMP9 expression in less differentiated MSCs. Mechanistically, we demonstrated that a blockade of JAK signaling effectively blunted leptin-potentiated osteogenic differentiation induced by BMP9. Taken together, our results strongly suggest that leptin may potentiate BMP9-induced osteogenesis by cross-regulating BMP9 signaling through the JAK/STAT signaling pathway in MSCs. Thus, it is conceivable that a combined use of BMP9 and leptin may be explored as a novel approach to enhancing efficacious bone regeneration and fracture healing.

Introduction:Many institutions are attempting to implement patient-reported outcome (PRO) measures. Because PROs often change clinical workflows significantly for patients and providers, implementation choices can have major impact. While various implementation guides exist, a stepwise list of decision points covering the full implementation process and drawing explicitly on a sociotechnical conceptual framework does not exist.Methods:To facilitate real-world implementation of PROs in electronic health records (EHRs) for use in clinical practice, members of the EHR Access to Seamless Integration of Patient-Reported Outcomes Measurement Information System (PROMIS) Consortium developed structured PRO implementation planning tools. Each institution pilot tested the tools. Joint meetings led to the identification of critical sociotechnical success factors.Results:Three tools were developed and tested: (1) a PRO Planning Guide summarizes the empirical knowledge and guidance about PRO implementation in routine clinical care; (2) a Decision Log allows decision tracking; and (3) an Implementation Plan Template simplifies creation of a sharable implementation plan. Seven lessons learned during implementation underscore the iterative nature of planning and the importance of the clinician champion, as well as the need to understand aims, manage implementation barriers, minimize disruption, provide ample discussion time, and continuously engage key stakeholders.Conclusions:Highly structured planning tools, informed by a sociotechnical perspective, enabled the construction of clear, clinic-specific plans. By developing and testing three reusable tools (freely available for immediate use), our project addressed the need for consolidated guidance and created new materials for PRO implementation planning. We identified seven important lessons that, while common to technology implementation, are especially critical in PRO implementation.

Magnetic resonance imaging (MRI) is generally considered the most sensitive imaging for diagnosis of osteomyelitis; however, it is associated with significant cost and is at times ordered as initial screening imaging when a less resource-intensive test would suffice. The purpose of this retrospective cohort study was to examine the differences between patients with osteomyelitis of the foot and ankle, and their subsequent treatment course, who underwent MRI compared with those who did not. Financial impact of MRI as it relates to clinical decision-making was also calculated. Patients treated for a diagnosis of osteomyelitis of the foot and ankle from 2009 to 2015 were retrospectively identified. Demographics, imaging modalities, and operative procedures for each patient were collected. An “impact MRI” was defined as one that led to a subsequent operative procedure within the same admission. The impact cost of an MRI was estimated using the equation: (average MRI cost) × (total MRIs/impact MRIs). A total of 144 patients underwent 220 MRIs, and 399 patients did not have MRIs. The operative rate between the 2 groups was similar (70.8% versus 70.4%, p = .93). Multiple linear regression showed that MRI was not a significant predictor of operation (p = .50). However, we found a significant correlation between MRI use and operative intervention for patients with increased comorbidities. From 2011 to 2015, there was a significant increase in impact cost, while controlling for average MRI cost ($8172 to $15,292, p ≤ .05). Over the study period, the impact cost of an MRI significantly increased from 1.8 to 5.0 times the average cost.

Although bone morphogenetic proteins (BMPs) initially showed effective induction of ectopic bone growth in muscle, it has since been determined that these proteins, as members of the TGF-β superfamily, play a diverse and critical array of biological roles. These roles include regulating skeletal and bone formation, angiogenesis, and development and homeostasis of multiple organ systems. Disruptions of the members of the TGF-β/BMP superfamily result in severe skeletal and extra-skeletal irregularities, suggesting high therapeutic potential from understanding this family of BMP proteins. Although it was once one of the least characterized BMPs, BMP9 has revealed itself to have the highest osteogenic potential across numerous experiments both in vitro and in vivo, with recent studies suggesting that the exceptional potency of BMP9 may result from unique signaling pathways that differentiate it from other BMPs. The effectiveness of BMP9 in inducing bone formation was recently revealed in promising experiments that demonstrated efficacy in the repair of critical sized cranial defects as well as compatibility with bone-inducing bio-implants, revealing the great translational promise of BMP9. Furthermore, emerging evidence indicates that, besides its osteogenic activity, BMP9 exerts a broad range of biological functions, including stem cell differentiation, angiogenesis, neurogenesis, tumorigenesis, and metabolism. This review aims to summarize our current understanding of BMP9 across biology and the body.

Highlights • Patients, when asked if they would complete another telemedicine encounter again, 92.9% (278/299) of patients reported they would. • Physicians reported high satisfaction, and that 78.4% of the time a telemedicine encounter was successful in replacing an in-person visit. • During our study period, over 600 telemedicine encounters were performed with extremely favourable ratings from patients and physicians., Background Telemedicine provides a safe and effective means for the delivery of care by physicians amongst many subspecialties. Historically, orthopaedic practices in the United States have not widely utilized telemedicine for the delivery of orthopaedic care. As technology improves the adoption and utilization of telemedicine will likely grow, especially in light of the novel coronavirus (COVID-19) pandemic. Our study aims to assess patient and surgeon satisfaction and efficacy of telemedicine during a rapid adoption due to the global pandemic. Methods All patients who completed a telemedicine encounter (telephone or video) with an orthopaedic surgeon were contacted. Patients were individually contacted after their visit, and a standardized validated post-visit satisfaction survey was completed. Orthopaedic surgeons completed a standardized post-encounter survey after each visit. Pre-COVID-19 patient satisfaction data was used for comparison. Results Orthopaedic surgeons completed 612 telehealth encounters either via phone or video consultation between April 6, 2020 and May 22, 2020. 95% of patients rated both surgeon sensitivity to their needs and response to their concerns as ‘good’ or ‘very good.’ 93% of patients reported they would participate in a telemedicine encounter again. Surgeons reported high satisfaction with telemedicine encounters (80%, 86% phone and video respectively), and that 78.4% of the time a telemedicine encounter was successful in replacing an in-person visit. Conclusion Patients and orthopaedic surgeons documented high levels of satisfaction with telehealth encounters during the novel coronavirus (COVID-19) pandemic. Telemedicine does not appear to be a replacement for all in-person clinic encounters, however, when used in the appropriate context demonstrated favourable results. Level of Evidence Level 4 Study.

Notch is a cell–cell signaling pathway that is involved in a host of activities including development, oncogenesis, skeletal homeostasis, and much more. More specifically, recent research has demonstrated the importance of Notch signaling in osteogenic differentiation, bone healing, and in the development of the skeleton. The craniofacial skeleton is complex and understanding its development has remained an important focus in biology. In this review we briefly summarize what recent research has revealed about Notch signaling and the current understanding of how the skeleton, skull, and face develop. We then discuss the crucial role that Notch plays in both craniofacial development and the skeletal system, and what importance it may play in the future.

Background Establishing the diagnosis of peri-prosthetic shoulder infection prior to revision shoulder arthroplasty can be difficult. The objectives of the present study were (i) to describe the technique of fluoroscopic capsular needle biopsy for the diagnosis of peri-prosthetic shoulder infection and (ii) to determine the feasibility and preliminary accuracy of the test in a pilot sample of patients undergoing revision shoulder arthroplasty. Methods Eighteen patients, comprising eight females and nine males with a mean age of 61 years (range 37 years to 81 years) underwent capsular needle biopsy during the work-up of suspected chronic arthroplasty-related glenohumeral infection. Intra-operative tissue samples were taken from a minimum of three regions of the joint capsule during revision surgery. Standard serum indices were obtained. Results Of 17 patients with possible infection, five had confirmed culture positive infections based on intra-operative biopsies. Of these five patients, four (80%) had positive cultures from fluoroscopic capsular needle biopsy, with matching cultures. There were no complications. No culture-positive patients had elevated serum indices for infection. Conclusions The technique for fluoroscopic capsular needle biopsy appears to be feasible and the preliminary results for this technique appear to be promising, with a sensitivity of 80% and a specificity of 100%. Level of evidence: Level II: diagnostic test

Category: Ankle, Ankle Arthritis Introduction/Purpose: Patients living with end stage ankle arthritis suffer significant disability. They are two standard deviations below population scores for all symptom and function SF-36 sub-scales. In addition, they tend to be younger and still active in the workforce. The economic impact of disability attributable to end0stage ankle arthritis has not been quantified. In the era of cost- effectiveness, policy makers want to see tangible evidence to justify the cost of treatment. One common metric has been quality adjusted life years (QALYs). The purpose of this study is to determine the gain in QALYs attributable to surgical treatment of end- stage arthritis. Methods: This study is based on an existing longitudinal cohort of end stage ankle arthritis patients. The study cohort included 180 subjects (45% female, 55% male) with an average age of 61 years, treated with either ankle arthrodesis or arthroplasty. Patients responses to the SF-36 questionnaire were transformed to SF-6D using SF-6D preference based algorithm licensed software (University of Sheffield, UK) to calculate the health utility value at time of surgical consent and at one year postoperatively. The difference in health utility value was assumed to be the gain in health attributable to end-stage ankle arthritis surgery. Using three theoretical valuations per QALY; $20000, $50000, and $100000, based on previously published cost per QUALY for other orthopedic conditions, we calculated the valuation per QALY for treating ankle arthritis at one year post surgery. Results: The mean pre-surgical health utility value was 0.603, and post-surgically it was 0.690, an average gain in health in one year of 0.087 QALY. This is greater than what is considered the minimally important difference threshold. We then calculated the economic gain at QALY valuations of $20000, $50000, and $100000; at $20000/QALY the cost per QALY is $1740. At $50000 the gain is $4350 cost per QALY, and at $100000 it is $8700 per QALY. Conclusion: From this study cohort, surgically treating end-stage ankle arthritis has a meaningful health benefit for a range of cost per QALYs. Moreover, spending on surgical treatment of end-stage ankle arthritis is cost-effective. These results from this cohort provide valuable empirical insights into treating end-stage ankle arthritis - these findings are meaningful for policy makers faced with resource allocation demands.