Radiation-induced bone loss in mice is ameliorated by inhibition of HIF-2α in skeletal progenitor cells.

Radiotherapy remains a common treatment modality for cancer despite skeletal complications. However, there are currently no effective treatments for radiation-induced bone loss, and the consequences of radiotherapy on skeletal progenitor cell (SPC) survival and function remain unclear. After radiation, leptin receptor-expressing cells, which include a population of SPCs, become localized to hypoxic regions of the bone and stabilize the transcription factor hypoxia-inducible factor-2α (HIF-2α), thus suggesting a role for HIF-2α in the skeletal response to radiation. Here, we conditionally knocked out HIF-2α in leptin receptor-expressing cells and their descendants in mice. Radiation therapy in littermate control mice reduced bone mass; however, HIF-2α conditional knockout mice maintained bone mass comparable to nonirradiated control animals. HIF-2α negatively regulated the number of SPCs, bone formation, and bone mineralization. To test whether blocking HIF-2α pharmacologically could reduce bone loss during radiation, we administered a selective HIF-2α inhibitor called PT2399 (a structural analog of which was recently FDA-approved) to wild-type mice before radiation exposure. Pharmacological inhibition of HIF-2α was sufficient to prevent radiation-induced bone loss in a single-limb irradiation mouse model. Given that ~90% of patients who receive a HIF-2α inhibitor develop anemia because of off-target effects, we developed a bone-targeting nanocarrier formulation to deliver the HIF-2α inhibitor to mouse bone, to increase on-target efficacy and reduce off-target toxicities. Nanocarrier-loaded PT2399 prevented radiation-induced bone loss in mice while reducing drug accumulation in the kidney. Targeted inhibition of HIF-2α may represent a therapeutic approach for protecting bone during radiation therapy.

Multi-Functional Small Molecule Alleviates Fracture Pain and Promotes Bone Healing.

Bone injuries such as fractures are one major cause of morbidities worldwide. A considerable number of fractures suffer from delayed healing, and the unresolved acute pain may transition to chronic and maladaptive pain. Current management of pain involves treatment with NSAIDs and opioids with substantial adverse effects. Herein, we tested the hypothesis that the purine molecule, adenosine, can simultaneously alleviate pain and promote healing in a mouse model of tibial fracture by targeting distinctive adenosine receptor subtypes in different cell populations. To achieve this, a biomaterial-assisted delivery of adenosine is utilized to localize and prolong its therapeutic effect at the injury site. The results demonstrate that local delivery of adenosine inhibited the nociceptive activity of peripheral neurons through activation of adenosine A1 receptor (ADORA1) and mitigated pain as demonstrated by weight bearing and open field movement tests. Concurrently, local delivery of adenosine at the fracture site promoted osteogenic differentiation of mesenchymal stromal cells through adenosine A2B receptor (ADORA2B) resulting in improved bone healing as shown by histological analyses and microCT imaging. This study demonstrates the dual role of adenosine and its material-assisted local delivery as a feasible therapeutic approach to treat bone trauma and associated pain.

Extracellular adenosine signaling in bone health and disease.

Purinergic signaling is a key molecular pathway in the maintenance of bone health and regeneration. P1 receptor signaling, which is activated by extracellular adenosine, has emerged as a key metabolic pathway that regulates bone tissue formation, function, and homeostasis. Extracellular adenosine is mainly produced by ectonucleotidases, and alterations in the function of these enzymes or compromised adenosine generation can result in bone disorders, such as osteoporosis and impaired fracture healing. This mini review discusses the key role played by adenosine in bone health and how its alterations contribute to bone diseases, as well as potential therapeutic applications of exogenous adenosine to combat bone diseases like osteoporosis and injury.

Microgel-Assisted Delivery of Adenosine to Accelerate Fracture Healing.

Extracellular adenosine plays a key role in promoting bone tissue formation. Local delivery of adenosine could be an effective therapeutic strategy to harness the beneficial effect of extracellular adenosine on bone tissue formation following injury. Herein, we describe the development of an injectable in situ curing scaffold containing microgel-based adenosine delivery units. The two-component scaffold includes adenosine-loaded microgels and functionalized hyaluronic acid (HA) molecules. The microgels were generated upon copolymerization of 3-acrylamidophenylboronic acid (3-APBA)- and 2-aminoethylmethacrylamide (2-AEMA)-conjugated HA (HA-AEMA) in an emulsion suspension. The PBA functional groups were used to load the adenosine molecules. Mixing of the microgels with the HA polymers containing clickable groups, dibenzocyclooctyne (DBCO) and azide (HA-DBCO and HA-Azide), resulted in a 3D scaffold embedded with adenosine delivery units. Application of the in situ curing scaffolds containing adenosine-loaded microgels following tibial fracture injury showed improved bone tissue healing in a mouse model as demonstrated by the reduced callus size, higher bone volume, and increased tissue mineral density compared to those treated with the scaffold without adenosine. Overall, our results suggest that local delivery of adenosine could potentially be an effective strategy to promote bone tissue repair.

pH-Sensitive nanocarrier assisted delivery of adenosine to treat osteoporotic bone loss.

Bone tissue undergoes continuous remodeling via osteoclast-mediated bone resorption and osteoblast-mediated bone formation. An imbalance in this process with enhanced osteoclastic activity can lead to excessive bone resorption, resulting in bone thinning. Once activated, osteoclasts bind to the bone surface and acidify the local niche. This acidic environment could serve as a potential trigger for the delivery of therapeutic agents into the osteoporotic bone tissue. To this end, we developed a pH-responsive nanocarrier-based drug delivery system that binds to the bone tissue and delivers an osteoanabolic molecule, adenosine. Adenosine is incorporated into a hyaluronic acid (HA)-based nanocarrier through a pH-sensitive ketal group. The HA-nanocarrier is further functionalized with alendronate moieties to improve binding to the bone tissues. Systemic administration of the nanocarrier containing adenosine attenuated bone loss in ovariectomized mice and showed comparable bone qualities to that of healthy mice. Delivery of osteoanabolic small molecules that can contribute to bone formation and inhibit excessive osteoclast activity by leveraging the tissue-specific milieu could serve as viable therapeutics for osteoporosis.

Resolution of inflammation in bone regeneration: From understandings to therapeutic applications.

Impaired bone healing occurs in 5-10% of cases following injury, leading to a significant economic and clinical impact. While an inflammatory response upon injury is necessary to facilitate healing, its resolution is critical for bone tissue repair as elevated acute or chronic inflammation is associated with impaired healing in patients and animal models. This process is governed by important crosstalk between immune cells through mediators that contribute to resolution of inflammation in the local healing environment. Approaches modulating the initial inflammatory phase followed by its resolution leads to a pro-regenerative environment for bone regeneration. In this review, we discuss the role of inflammation in bone repair, the negative impact of dysregulated inflammation on bone tissue regeneration, and how timely resolution of inflammation is necessary to achieve normal healing. We will discuss applications of biomaterials to treat large bone defects with a specific focus on resolution of inflammation to modulate the immune environment following bone injury, and their observed functional benefits. We conclude the review by discussing future strategies that could lead to the realization of anti-inflammatory therapeutics for bone tissue repair.

Bone targeting nanocarrier-assisted delivery of adenosine to combat osteoporotic bone loss.

Extracellular adenosine has been shown to play a key role in maintaining bone health and could potentially be used to treat bone loss. However, systemic administration of exogenous adenosine to treat bone disorders remains a challenge due to the ubiquitous presence of adenosine receptors in different organs and the short half-life of adenosine in circulation. Towards this, we have developed a bone-targeting nanocarrier and determined its potential for systemic administration of adenosine. The nanocarrier, synthesized via emulsion suspension photopolymerization, is comprised of hyaluronic acid (HA) copolymerized with phenylboronic acid (PBA), a moiety that can form reversible bonds with adenosine. The bone binding affinity of the nanocarrier was achieved by alendronate (Aln) conjugation. Nanocarriers functionalized with the alendronate (Aln-NC) showed a 45% higher accumulation in the mice vertebrae in vivo compared to those lacking alendronate molecules (NCs). Systemic administration of adenosine via bone-targeting nanocarriers (Aln-NC) attenuated bone loss in ovariectomized (OVX) mice. Furthermore, bone tissue of mice treated with adenosine-loaded Aln-NC displayed trabecular bone characteristics comparable to healthy controls as shown by microcomputed tomography, histochemical staining, bone labeling, and mechanical strength. Overall, our results demonstrate the use of a bone-targeting nanocarrier towards systemic administration of adenosine and its application in treating bone degenerative diseases such as osteoporosis.

Obesity alters the collagen organization and mechanical properties of murine cartilage.

Osteoarthritis is a debilitating disease characterized by cartilage degradation and altered cartilage mechanical properties. Furthermore, it is well established that obesity is a primary risk factor for osteoarthritis. The purpose of this study was to investigate the influence of obesity on the mechanical properties of murine knee cartilage. Two-month old wild type mice were fed either a normal diet or a high fat diet for 16 weeks. Atomic force microscopy-based nanoindentation was used to quantify the effective indentation modulus of medial femoral condyle cartilage. Osteoarthritis progression was graded using the OARSI system. Additionally, collagen organization was evaluated with picrosirius red staining imaged using polarized light microscopy. Significant differences between diet groups were assessed using t tests with p < 0.05. Following 16 weeks of a high fat diet, no significant differences in OARSI scoring were detected. However, we detected a significant difference in the effective indentation modulus between diet groups. The reduction in cartilage stiffness is likely the result of disrupted collagen organization in the superficial zone, as indicated by altered birefringence on polarized light microscopy. Collectively, these results suggest obesity is associated with changes in knee cartilage mechanical properties, which may be an early indicator of disease progression.

Glutamine Metabolism Regulates Proliferation and Lineage Allocation in Skeletal Stem Cells.

Skeletal stem cells (SSCs) are postulated to provide a continuous supply of osteoblasts throughout life. However, under certain conditions, the SSC population can become incorrectly specified or is not maintained, resulting in reduced osteoblast formation, decreased bone mass, and in severe cases, osteoporosis. Glutamine metabolism has emerged as a critical regulator of many cellular processes in diverse pathologies. The enzyme glutaminase (GLS) deaminates glutamine to form glutamate-the rate-limiting first step in glutamine metabolism. Using genetic and metabolic approaches, we demonstrate GLS and glutamine metabolism are required in SSCs to regulate osteoblast and adipocyte specification and bone formation. Mechanistically, transaminase-dependent α-ketoglutarate production is critical for the proliferation, specification, and differentiation of SSCs. Collectively, these data suggest stimulating GLS activity may provide a therapeutic approach to expand SSCs in aged individuals and enhance osteoblast differentiation and activity to increase bone mass.

Functional outcomes after peroneal tendoscopy in the treatment of peroneal tendon disorders.

PURPOSE: The primary purpose of this study was to evaluate clinical outcomes following peroneal tendoscopy for the treatment of peroneal pathology. Correlation between pre-operative magnetic resonance imaging (MRI) and peroneal tendoscopic diagnostic findings was also assessed. METHODS: Twenty-three patients with a mean age of 34 ± 8.8 years undergoing peroneal tendoscopy were pre- and post-operatively assessed with the foot and ankle outcome score (FAOS) and the Short Form-12 (SF-12) outcome questionnaires. Follow-up was over 24 months in all patients. The sensitivity and specificity of MRI were calculated in comparison with peroneal tendoscopy, including the positive predictive value (PPV). RESULTS: Both the FAOS and the SF-12 improved significantly (p < 0.05) at a mean follow-up of 33 ± 7.3 months significantly. MRI showed an overall sensitivity of 0.90 (95% confidence interval (CI) = 0.82-0.95) and specificity of 0.72 (95% CI 0.62-0.80). The PPV for MRI diagnosis of peroneal tendon pathology was 0.76 (95% CI 0.68-0.83). CONCLUSIONS: The current study found good clinical outcomes in patients with peroneal tendon disorders, treated with peroneal tendoscopy. Although a relatively small number of patients were included, the study suggests good correlation between tendoscopic findings and pre-operative MRI findings of peroneal tendon pathology, supporting the use of MRI as a useful diagnostic modality for suspected peroneal tendon disorders. LEVEL OF EVIDENCE: Level IV, retrospective case series.

Autologous Osteochondral Transplantation for Osteochondral Lesions of the Talus.

BACKGROUND: Autologous osteochondral transplantation (AOT) is used to treat osteochondral lesions (OCLs) of the talus, typically reserved for lesions greater than 150 mm(2). Few studies exist examining the functional and magnetic resonance imaging (MRI) outcomes following this procedure. The purpose of this study was to investigate functional and MRI outcomes, including quantitative T2 mapping following AOT. METHODS: Eighty-five consecutive patients who underwent AOT were identified. Functional outcomes were assessed pre- and postoperatively using the Foot and Ankle Outcome Score (FAOS). The Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) score was used to assess cartilage incorporation. Quantitative T2 MRI relaxation time of graft tissue and adjacent normal cartilage values were recorded in a subset of 61 patients. The mean clinical follow-up was 47.2 months, with mean MRI follow-up of 24.8 months. RESULTS: Mean FAOS improved pre- to postoperatively from 50 to 81 (P < .001). The mean MOCART score was 85.8. Lesion size was negatively correlated with MOCART score (r = -0.36, P = .004). Superficial T2 values in graft tissue were higher than control tissue (42.0 vs 35.8, P < .001). Deep T2 values in graft tissue were similar to the control values (30.9 vs 30.0, P = .305). Functional outcomes were similar in patients irrespective of whether they had previous microfracture or concomitant procedures. CONCLUSION: AOT was an effective treatment for large OCLs of the talus in the current study. MOCART scoring indicated good structural integrity of the graft. Quantitative T2 mapping suggests that graft tissue may not always mirror native hyaline cartilage. The long-term implications of this are not yet known. LEVEL OF EVIDENCE: Level IV, case series.

Co-evolution of Bacterial Ribosomal Protein S15 with Diverse mRNA Regulatory Structures.

RNA-protein interactions are critical in many biological processes, yet how such interactions affect the evolution of both partners is still unknown. RNA and protein structures are impacted very differently by mechanisms of genomic change. While most protein families are identifiable at the nucleotide level across large phylogenetic distances, RNA families display far less nucleotide similarity and are often only shared by closely related bacterial species. Ribosomal protein S15 has two RNA binding functions. First, it is a ribosomal protein responsible for organizing the rRNA during ribosome assembly. Second, in many bacterial species S15 also interacts with a structured portion of its own transcript to negatively regulate gene expression. While the first interaction is conserved in most bacteria, the second is not. Four distinct mRNA structures interact with S15 to enable regulation, each of which appears to be independently derived in different groups of bacteria. With the goal of understanding how protein-binding specificity may influence the evolution of such RNA regulatory structures, we examine whether examples of these mRNA structures are able to interact with, and regulate in response to, S15 homologs from organisms containing distinct mRNA structures. We find that despite their shared RNA binding function in the rRNA, S15 homologs have distinct RNA recognition profiles. We present a model to explain the specificity patterns observed, and support this model by with further mutagenesis. After analyzing the patterns of conservation for the S15 protein coding sequences, we also identified amino acid changes that alter the binding specificity of an S15 homolog. In this work we demonstrate that homologous RNA-binding proteins have different specificity profiles, and minor changes to amino acid sequences, or to RNA structural motifs, can have large impacts on RNA-protein recognition.

Anterolateral tibial osteotomy for accessing osteochondral lesions of the talus in autologous osteochondral transplantation: functional and t2 MRI analysis.

BACKGROUND: Autologous osteochondral transplantation (AOT) is a primary treatment strategy for large or cystic osteochondral lesions of the talus (OLT) or a secondary replacement strategy after failed bone marrow stimulation. The technique requires perpendicular access to the talar dome, which is often difficult to obtain for posterior or lateral lesions. Traditional methods to access these areas have required disruption of the syndesmotic complex with concern over osteotomy reduction, malalignment, and ligament disruption. An alternate to these traditional methods of access is an anterolateral tibial osteotomy. The purpose of this study was to report functional and magnetic resonance imaging (MRI) outcomes in a series of patients that underwent AOT for treatment of an OLT via an anterolateral tibial osteotomy. METHODS: Records of patients that underwent an anterolateral tibial osteotomy for AOT were retrospectively reviewed. Pre- and postoperative Foot and Ankle Outcome Scores (FAOS) and demographic data were recorded. Magnetic resonance observation of cartilage repair tissue (MOCART) was used to assess morphologic state of tibial cartilage at the repair site of the osteotomy. Quantitative T2 mapping MRI was analyzed in the superficial and deep cartilage layers of the repair site of the osteotomy and in adjacent normal cartilage to serve as control tissue. Seventeen patients with a mean age of 36.9 (range, 17-76) years underwent anterolateral tibial osteotomy with a mean follow-up of 64 (range, 29 to 108) months. MOCART data were available in 9 of 17 patients, and quantitative T2 mapping was available in 6 patients. RESULTS: FAOS significantly improved from an average 39.2 (range, 14 to 66) out of 100 points preoperatively to 81.2 (range, 19 to 98) postoperatively (P < .01). The average MOCART score was 73.9 out of 100 points (range, 40 to 100). Quantitative T2 analysis demonstrated relaxation times that were not significantly different from the normal native cartilage in both the deep half and superficial half of interface repair tissue (P > .05). CONCLUSION: This study demonstrated that the anterolateral tibial osteotomy was a reasonable alternative for accessing centrolateral or posterolateral OLT for AOT with limited morbidity associated with the osteotomy. The evidence demonstrated adequate osteotomy and cartilaginous healing with improvement in functional outcome scores at medium-term follow-up. LEVEL OF EVIDENCE: Level IV, retrospective case series.

A single platelet-rich plasma injection for chronic midsubstance achilles tendinopathy: a retrospective preliminary analysis.

UNLABELLED: The purpose of this study was to evaluate a series of patients undergoing a single platelet-rich plasma (PRP) injection for the treatment of chronic midsubstance Achilles tendinopathy, in whom conservative treatment had failed. Thirty-two patients underwent a single PRP injection for the treatment of chronic midsubstance Achilles tendinopathy and were evaluated at a 6-month final follow-up using the Foot and Ankle Outcome Score and Short Form 12 general health questionnaire. Magnetic resonance imaging was performed on all patients prior to and 6 months after injection. Twenty-five of 32 patients (78%) reported that they were asymptomatic at the 6-month follow-up visit and were able to participate in their respective sports and daily activities. The remaining 7 patients (22%) who reported symptoms that did not improve after 6 months ultimately required surgery. Four patients went on to have an Achilles tendoscopy, while the other 3 had an open debridement via a tendon splitting approach. A retrospective evaluation of patients receiving a single PRP injection for chronic midsubstance Achilles tendinopathy revealed that 78% had experienced clinical improvement and had avoided surgical intervention at 6-month follow-up. LEVELS OF EVIDENCE: Therapeutic, Level IV: Retrospective case series.

A systematic review on the reporting of outcome data in studies on autologous osteochondral transplantation for the treatment of osteochondral lesions of the talus.

PURPOSE: The purpose of this study was to systematically review studies on autologous osteochondral transplantation (AOT) for osteochondral lesions (OCLs) of the talus and descriptively analyze the outcome data reported to determine whether it is consistent from one study to another and able to be pooled for systematic review. METHODS: A systematic electronic search was performed using the MEDLINE and EMBASE databases. Studies that were published between January 1966 and June 2011 were included in the review. Only studies that reported data specifically on AOT for treatment of OCLs of the talus and written in English were included in this review. A predefined data sheet with 36 variables was created, and it was determined whether or not each of those variables were reported or not reported. The 36 variables were then grouped into 6 categories, and the categorical means were reported. RESULTS: A total of 20 studies were included in this systematic review. The categories of general demographics and study design were generally well reported (each more than 80% of studies). Patient-reported outcomes and clinical variables were reported less in 73% and 67% of studies, respectively. The least-reported categories were patient history (45%) and imaging data (49%). CONCLUSIONS: Inconsistencies and an underreporting of data were apparent between studies, such that pooling was deemed not possible. An effort must be made by investigators to ensure that there is adequate reporting of data in studies of AOT treatment for OCLs of the talus.