Phosphate as an adjunct to calcium in promoting coronary vascular calcification in chronic inflammatory states.

Bone releases calcium and phosphate in response to pro-inflammatory cytokine-mediated inflammation. The body develops impaired urinary excretion of phosphate with age and chronic inflammation given the reduction of the kidney protein Klotho, which is essential to phosphate excretion. Phosphate may also play a role in the development of the resistance of the parathyroid calcium-sensing receptor (CaSR) to circulating calcium thus contributing to calcium retention in the circulation. Phosphate can contribute to vascular smooth muscle dedifferentiation with manifestation of osteoblastogenesis and ultimately endovascular calcium phosphate precipitation. Thus phosphate, along with calcium, contributes to the calcification and inflammation of atherosclerotic plaques and the origin of these elements is likely the bone, which serves as storage for the majority of the body's supply of extracellular calcium and phosphate. Early cardiac evaluation of patients with chronic inflammation and attempts at up-regulating the parathyroid CaSR with calcimimetics or introducing earlier anti-resorptive treatment with bone active pharmacologic agents may serve to delay onset or reduce the quantity of atherosclerotic plaque calcification in these patients.

Is calcium a link between inflammatory bone resorption and heart disease?

Several epidemiologic studies associate bone-resorbing chronic inflammatory conditions with increased risk of atherosclerotic heart disease. These include post-menopausal osteoporosis, spinal cord injury, rheumatoid arthritis, and osteoarthritis. Additional studies have noted that the use of anti-resorptive agents following hip fracture, during rheumatoid arthritis, and prior to intensive care management have resulted in reduced overall mortality and mortality from cardiovascular disorders. The careful study of burn patients has allowed us to detect that children and adolescents have a mechanism that protects them from the entry of calcium into the circulation following inflammatory bone resorption. That is, they respond to pro-inflammatory cytokines by up-regulating the parathyroid calcium-sensing receptor (CaSR) with consequent development of hypocalcemic hypoparathyroidism and hypercalciuria. As extracellular calcium appears to exacerbate and/or prolong the inflammatory response, this responsiveness of the CaSR to inflammatory cytokines may be the factor that reduces cardiovascular morbidity and mortality. In adults with chronic inflammatory conditions, the ability of the CaSR to respond to pro-inflammatory cytokines is lost, suggesting that the calcium that enters the circulation following inflammatory bone resorption may persist in the circulation, entering the small coronary blood vessels and favoring the formation of coronary artery calcification, inflammation, and consequent cardiovascular disease.

Transforming Growth Factor-Beta in Skeletal Muscle Wasting.

Transforming growth factor-beta (TGF-ß) is part of a family of molecules that is present in many body tissues and performs many different functions. Evidence has been obtained from mice and human cancer patients with bony metastases and non-metastatic disease, as well as pediatric burn patients, that inflammation leads to bone resorption and release of TGF-ß from the bone matrix with paracrine effects on muscle protein balance, possibly mediated by the generation of reactive oxygen species. Whether immobilization, which confounds the etiology of bone resorption in burn injury, also leads to the release of TGF-ß from bone contributing to muscle wasting in other conditions is unclear. The use of anti-resorptive therapy in both metastatic cancer patients and pediatric burn patients has been successful in the prevention of muscle wasting, thereby creating an additional therapeutic niche for this class of drugs. The liberation of TGF-ß may be one way in which bone helps to control muscle mass, but further investigation will be necessary to assess whether the rate of bone resorption is the determining factor for the release of TGF-ß. Moreover, whether different resorptive conditions, such as immobilization and hyperparathyroidism, also involve TGF-ß release in the pathogenesis of muscle wasting needs to be investigated.

Acute and severe trabecular bone loss in a rat model of critical illness myopathy.

Prolonged mechanical ventilation for critically ill patients with respiratory distress can result in severe muscle wasting with preferential loss of myosin. Systemic inflammation triggered by lung mechanical injury likely contributes to this myopathy, although the exact mechanisms are unknown. In this study, we hypothesized that muscle wasting following mechanical ventilation is accompanied by bone loss. The objective was to determine the rate, nature, and extent of bone loss in the femora of rats ventilated up to 10 days and to relate the bone changes to muscle deterioration. We have developed a rat model of ventilator-induced muscle wasting and established its feasibility and clinical validity. This model involves pharmacologic paralysis, parenteral nutrition, and continuous mechanical ventilation. We assessed the hindlimb muscle and bone of rats ventilated for 0, 2, 5, 8, and 10 days. Routine histology, microCT, and biomechanical evaluations were performed. Hindlimb muscles developed changes consistent with myopathy, whereas the femurs demonstrated a progressive decline in trabecular bone volume, mineral density, and microarchitecture beginning Day 8 of mechanical ventilation. Biomechanical testing showed a reduction in flexural strength and stiffness on Day 10. The bone changes correlated with the loss of muscle mass and myosin. These results demonstrate that mechanical ventilation leads to progressive trabecular bone loss parallel to muscle deterioration. The results of our study suggest that mechanically ventilated patients may be at risk of compromised bone integrity and muscle weakness, predisposing to post-ventilator falls and fractures, thereby warranting interventions to prevent progressive bone and muscle decline.

Pharmacologic Treatments to Preserve Bone and Muscle Mass in Osteosarcopenia.

PURPOSE OF REVIEW: We aim to recast the diagnosis of osteosarcopenia in light of its pathophysiology rather than of the age at which it is diagnosed. We will consider why we think the diagnosis of osteosarcopenia is missed in those who are not elderly and why pharmacologic treatment based on pathophysiology rather than age may provide a more comprehensive treatment for patients with the condition. RECENT FINDINGS: We will present recent findings on the pathogenesis of osteosarcopenia from two distinct groups of patients which will highlight why pathophysiology is of paramount importance in designing treatment. We will show that in patients with cancer and burns, muscle catabolic factors are released from bone on resorption, exert a paracrine effect on muscle to cause catabolism, and can be prevented with the use of anti-resorptive drugs. New uses for anti-resorptives may result from these findings.

The Role of Bone in Muscle Wasting.

This review describes the role of bone resorption in muscle atrophy as well as in muscle protein anabolism. Both catabolic and anabolic pathways involve components of the proinflammatory cytokine families and release of factors stored in bone during resorption. The juxtaposition of the catabolic and anabolic resorption-dependent pathways raises new questions about control of release of factors from bone, quantity of release in a variety of conditions, and relation of factors released from bone. The catabolic responses involve release of calcium from bone into the circulation resulting in increased inflammatory response in intensity and/or duration. The release of transforming growth factor beta (TGF-ß) from bone suppresses phosphorylation of the AKT/mTOR pathway and stimulates ubiquitin-mediated breakdown of muscle protein. In contrast, muscle IL-6 production is stimulated by undercarboxylated osteocalcin, which signals osteoblasts to produce more RANK ligand, stimulating resorptive release of undercarboxylated osteocalcin, which in turn stimulates muscle fiber nutrient uptake and an increase in muscle mass.

Burn injury and restoration of muscle function.

Burn injury in children results in a systemic inflammatory reaction as well as a stress response. Consequences of these non-specific adaptive responses include resorptive bone loss and muscle catabolism. These adverse events can result in a post-burn fracture rate of approximately 15% and long-term muscle weakness that prolongs recovery. A randomized controlled trial of a single dose of the bisphosphonate pamidronate within the first ten days of burn injury resulted in the prevention of resorptive bone loss and continuous bone accrual. Examining the muscle protein kinetics in pediatric burn patients enrolled in that randomized controlled trial revealed that those who had been given the single dose bisphosphonate experienced preservation of muscle mass and strength. An in vitro study of mouse myoblasts incubated with serum from patients who participated in the randomized controlled study demonstrated that mouse myoblasts exposed to serum from patients given the single dose bisphosphonate exhibited greater myotube diameter than those from burned children given placebo. Moreover, the serum from bisphosphonate treated patients stimulated the protein anabolic pathways and suppressed protein catabolic pathways in these cells. Inasmuch as incubation of the myotubes with an antibody to transforming growth factor beta (TGFß) rescued myotube size in the cultures with serum from patients who received the placebo to the same magnitude as cultures with serum from patients treated with single dose bisphosphonate, we postulate that post-burn bone resorption liberates muscle catabolic factors which cause muscle wasting. Future uses of bisphosphonates could include studies designed to prevent short-term acute bone resorption in conditions that may result in muscle wasting as well as in short-term interventions in chronic inflammatory conditions which may flare and cause acute bone and muscle loss.

Early reduced bone formation following burn injury in rats is not inversely related to marrow adiposity.

OBJECTIVES: The objective of the study was to determine whether postburn reduction of bone formation occurred earlier than 2-3 weeks after burn injury and whether that reduction was inversely related to marrow adiposity. METHODS: Using a rat model of burn injury with sacrifice at 3 days postburn, we measured serum osteocalcin, a biomarker of bone formation, as well as a regulator of glucose metabolism, and counted tibial marrow adipocytes. RESULTS: Serum osteocalcin was reduced as early as 3 days postburn, coinciding with a trend toward decline in marrow adipocyte number rather than demonstrating an inverse relationship with adipocyte count. CONCLUSIONS: Factors that may be responsible for the dissociation include lack of circulating sclerostin, previously reported, increased energy demands following burn injury, increased sympathetic tone and perhaps oxidative stress. The relationship between bone formation and marrow adiposity is complex and subject to a variety of influences.

Molecular Mechanisms Responsible for the Rescue Effects of Pamidronate on Muscle Atrophy in Pediatric Burn Patients.

Not only has pamidronate been shown to prevent inflammation associated bone resorption following burn injury, it also reduces protein breakdown in muscle. The aim of this study was to identify the molecular mechanisms responsible for muscle mass rescue in pamidronate treated compared to placebo/standard of care-treated burn patients. Mature myotubes, generated by differentiating murine C2C12 myoblasts, were exposed for 48 h to 1 or 5% serum obtained from 3 groups of children: normal unburned, burned receiving standard of care, and burned receiving standard of care with pamidronate. Exposure to serum from burned patients caused dose-dependent myotube atrophy compared to normal serum as expected based on previous observations of muscle atrophy induced by burn injury in humans and animals. The size of C2C12 myotubes was partially protected upon exposure to the serum from patients treated with pamidronate correlating with the rescue of muscle size previously observed in these patients. At the molecular signaling level, serum from both pamidronate and non-pamidronate-treated burn patients increased pSTAT3/STAT3 and pERK1/2/ERK1/2 compared to normal serum with no significant differences between the two groups of burn patients indicating elevated production of inflammatory cytokines. However, serum from pamidronate-treated patients restored the phosphorylation of AKT and mTOR and reduced protein ubiquitination when compared to burn serum alone, suggesting a prevention of muscle catabolism and a restoration of muscle anabolism. Myotube atrophy induced by burn serum was partially rescued after exposure to a pan anti-TGFß-1/2/3 antibody, suggesting that this signaling pathway is partially responsible for the atrophy and that bisphosphonate protection of bones from resorption during burn injury prevents the release of muscle pro-catabolic factors such as TGFß into the circulation.

The role of the musculoskeletal system in post-burn hypermetabolism.

Burn injury results in a triad of inter-related adaptive responses: a systemic inflammatory response, a stress response, and a consequent hypermetabolic state which supports the former two. Details of what precisely triggers these responses as well as the sequence of events leading up to these responses are not clear. We review the musculoskeletal effects of burn injury to determine the precise contributions of this system in the generation and sustenance of this post-burn triad as well as the possible effects of pharmacologic intervention in the musculoskeletal response to burns on the resulting hypermetabolism. Inflammation-associated bone resorption liberates calcium, which may either prolong or intensify the systemic inflammatory response. Phosphate and magnesium liberated from bone could contribute to sustaining the increased ATP turnover in skeletal muscle that accompanies burn hypermetabolism. Reduced bone formation resulting from both pro-inflammatory cytokines and elevated endogenous glucocorticoid production results in reduced bone mass and therefore reduced osteocalcin production, which may contribute to reduced glucose uptake by skeletal muscle. Moreover, bone resorption liberates muscle catabolic factors such as transforming growth factor ß, which contribute to the muscle wasting of burn hypermetabolism. Pharmacologic intervention with anti-resorptive agents early in the process preserve bone and muscle mass post-burn and future research should address the consequences for the hypermetabolic triad duration and intensity accompanying burn injury.

Aluminum toxicity to bone: A multisystem effect?

Aluminum (Al) is the third most abundant element in the earth's crust and is omnipresent in our environment, including our food. However, with normal renal function, oral and enteral ingestion of substances contaminated with Al, such as antacids and infant formulae, do not cause problems. The intestine, skin, and respiratory tract are barriers to Al entry into the blood. However, contamination of fluids given parenterally, such as parenteral nutrition solutions, or hemodialysis, peritoneal dialysis or even oral Al-containing substances to patients with impaired renal function could result in accumulation in bone, parathyroids, liver, spleen, and kidney. The toxic effects of Al to the skeleton include fractures accompanying a painful osteomalacia, hypoparathyroidism, microcytic anemia, cholestatic hepatotoxicity, and suppression of the renal enzyme 25-hydroxyvitamin D-1 alpha hydroxylase. The sources of Al include contamination of calcium and phosphate salts, albumin and heparin. Contamination occurs either from inability to remove the naturally accumulating Al or from leeching from glass columns used in compound purification processes. Awareness of this long-standing problem should allow physicians to choose pharmaceutical products with lower quantities of Al listed on the label as long as this practice is mandated by specific national drug regulatory agencies.

The Role of Calcium in Inflammation-Associated Bone Resorption.

The aim of this mini-review is to discuss the role of calcium in the process of cytokine-mediated bone resorption in an effort to understand the role circulating calcium may play in the resorption of bone. The liberation of calcium and possibly phosphorus and magnesium by bone resorption may sustain and intensify the inflammatory response. We used a burn injury setting in humans and a burn injury model in animals in order to examine the effects on the bone of the systemic inflammatory response and identified the parathyroid calcium-sensing receptor as the mediator of increasing bone resorption, hence higher interleukin (IL)-1 production, and decreasing bone resorption, hence the lowering of circulating ionized calcium concentration. Thus, extracellular calcium, by means of the parathyroid calcium-sensing receptor, is able to modulate inflammation-mediated resorption.

Bone metabolism in pediatric burned patients: A review.

Severe burns in children can lead to growth delays, bone loss, and wasting of lean body mass and muscle with subsequent long-term effects such as osteoporosis. The following review examines 11 randomized, placebo-controlled, prospective clinical trials in pediatric burns between 1995 and 2017. These studies included approximately 250 burned children, and they were conducted to evaluate the impact of severe burn on markers of bone formation and bone metabolism. Some trials also analyzed current therapy regimens such as pamidronate and vitamin D. The clinical utility of these outlined biomarkers is uncertain with regard to acute burn care, as the current literature remains unclear. This review thus serves to address the impact of severe burn on markers of bone formation and bone metabolism in pediatric patients but will not focus on the clinical utility of the markers. The aim of this review is to summarize the findings of the trials to guide the future care of burned patients to maximize bone recovery.

The Role of Bone Secreted Factors in Burn-Induced Muscle Cachexia.

PURPOSE OF REVIEW: Burn injury results in resorptive bone loss, failure to make new bone, and muscle protein breakdown resulting in cachexia. The purpose of this review is to examine the relationship between bone loss and muscle atrophy in burn injury with a view to understanding the process at work and how it may apply to other conditions that have similar features. RECENT FINDINGS: We present data suggesting that the use of bisphosphonates in the first 10 days following the burn prevents not only the resorptive bone loss but also the muscle wasting. While an extra-osseous effect of bisphosphonates remains possible, existing evidence points to a paracrine effect of bone on maintenance of muscle mass and strength. Proposed paracrine factors produced by bone include prostaglandin E2 and components of the Wnt signaling pathway. TGFß may be a bone paracrine factor that causes oxidative damage to muscle. In the light of the pattern of evidence, burn patients suffer acute resorptive bone loss and muscle wasting. This is likely due to the effects of inflammatory cytokines and endogenous glucocorticoid production in exacerbating oxidative stress. Early use of bisphosphonates can maintain bone mass leading to a paracrine effect of bone in the maintenance of muscle mass, although one cannot completely discount a direct effect of bisphosphonate on muscle. Because investigators report this relationship in a variety of conditions in addition to burns, physicians should seriously consider the early use of bisphosphonates to maintain bone and muscle mass in a variety of neuromuscular and skeletal diseases.

Interactions of Phosphate Metabolism With Serious Injury, Including Burns.

Approximately 85% of the body's phosphate pool resides within the skeleton. The remaining 15% is stored as high-energy phosphates or in its free form, where it acts as a substrate for adenosine triphosphate (ATP) production. Accordingly, phosphate plays a crucial role in energy metabolism. Trauma and critical illness result in a hypermetabolic state in which energy expenditure increases. The impact of trauma and critical illness on the body's phosphate stores and phosphate-dependent metabolic reactions is poorly understood. We had previously observed that after severe burn trauma, increased energy expenditure is temporally related to a marked reduction in serum concentrations of both parathyroid hormone and fibroblast growth factor 23, both of which have phosphaturic effects. The aim of this article is to describe as far as is known the similarities and differences in phosphate metabolism in different types of injury and to infer what these differences tell us about possible signaling pathways that may link increased phosphate utilization and phosphate retention. © 2017 The Authors. JBMR Plus is published by Wiley Periodicals, Inc. on behalf of the American Society for Bone and Mineral Research.

Calcemic response to burns differs between adults and children: A review of the literature.

OBJECTIVES: The calcemic and parathyroid hormone (PTH) responses to severe burn injury appear to differ between children and adults. In our limited studies children exhibited hypocalcemic hypoparathyroidism consistent with up-regulation of the parathyroid calcium-sensing receptor (CaSR) while adults did not, suggesting a developmental cutoff in cytokine-mediated up-regulation of the CaSR. This difference may be clinically important as published studies indicate that extracellular calcium (Ca) may stimulate the inflammatory response. The aim of this study was to examine the existing literature on burns to see if the differences between pediatric and adult calcemic and PTH responses to burn supported our findings providing stronger evidence to support this developmental difference. METHODS: We reviewed the National Library of Medicine database using the terms burns, PTH and ionized calcium and found 9 articles from 8 different medical centers; one was eliminated due to mixing of adults and children. RESULTS: There were 245 burn patients reported from the literature, 178 pediatric and 67 adults. The data are mostly consistent with our reported findings. Of the 10 pediatric patients with severe burns that we studied, mean ionized Ca concentration was below the lower limit of normal of 1.10 mM. The 67 adult burn patients reported in the literature had a mean blood ionized Ca concentration that was within the adult normal range or was lower than normal but with secondary hyperparathyroidism. Moreover, serum PTH concentrations were uniformly low in the 178 children in the burn literature but normal or mildly elevated in the 67 adults. CONCLUSIONS: These results support the hypothesis that the difference between pediatric and adult victims is consistent with an age-related CaSR response to cytokine stimulation and may be consistent with a lower level of inflammation in children. Ionized Ca and PTH might serve as possible therapeutic targets to lower the inflammatory response in burn victims.

Why so little effort to study anti-oxidant therapy in burns?

Given that oxidative stress is an inherent response to burn injury, it is puzzling as to why investigation into anti-oxidant therapy as an adjunct to burn treatment has been limited. Both the inflammatory response and the stress response to burn injury involve oxidative stress, and there has been some limited success in studies using gamma tocopherol and selenium to improve certain consequences of burns. Much remains to be done to investigate the number, doses and combinations of anti-oxidants, their efficacy, and limitations in improving defined outcomes after burn injury.

Reversal of Growth Arrest With the Combined Administration of Oxandrolone and Propranolol in Severely Burned Children.

BACKGROUND: The hypercatabolic response in severely burned pediatric patients is associated with increased production of catecholamines and corticosteroids, decreased formation of testosterone, and reduced strength alongside growth arrest for up to 2 years after injury. We have previously shown that, in the pediatric burned population, the administration of the testosterone analog oxandrolone improves lean body mass accretion and bone mineral content and that the administration of the ß1-, ß2-adrenoceptor antagonist propranolol decreases cardiac work and resting energy expenditure while increasing peripheral lean mass. Here, we determined whether the combined administration of oxandrolone and propranolol has added benefit. METHODS: In this prospective, randomized study of 612 burned children [52% ±â€Š1% of total body surface area burned, ages 0.5-14 years (boys); ages 0.5-12 years (girls)], we compared controls to the individual administration of these drugs, and the combined administration of oxandrolone and propranolol at the same doses, for 1 year after burn. Data were recorded at discharge, 6 months, and 1 and 2 years after injury. RESULTS: Combined use of oxandrolone and propranolol shortened the period of growth arrest by 84 days (P = 0.0125 vs control) and increased growth rate by 1.7 cm/yr (P = 0.0024 vs control). CONCLUSIONS: Combined administration of oxandrolone and propranolol attenuates burn-induced growth arrest in pediatric burn patients. The present study is registered at clinicaltrials.gov: NCT00675714 and NCT00239668.

Effects of whole-body vibration exercise on bone mineral content and density in thermally injured children.

BACKGROUND: Loss of bone mass, muscle mass, and strength leads to significant disability in severely burned children. We assessed the effects of exercise combined with whole-body vibration (WBV) on bone mass, lean mass (LM), and muscle strength in children recovering from burns. METHODS: Nineteen burned children (≥30% total body surface area [TBSA] burns) were randomly assigned to a 6-week exercise regimen either alone (EX; n=10) or in combination with a 6-week WBV training regimen (EX+WBV; n=9). WBV was performed concurrent to the exercise regimen for 5days/week on a vibrating platform. Dual-energy X-ray absorptiometry quantified bone mineral content (BMC), bone mineral density (BMD), and LM; knee extension strength was assessed using isokinetic dynamometry before and after training. Alpha was set at p<0.05. RESULTS: Both groups were similar in age, height, weight, TBSA burned, and length of hospitalization. Whole-body LM increased in the EX group (p=0.041) and trended toward an increase in the EX+WBV group (p=0.055). On the other hand, there were decreases in leg BMC for both groups (EX, p=0.011; EX+WBV, p=0.047), and in leg BMD for only the EX group (EX, p<0.001; EX+WBV, p=0.26). Truncal BMC decreased in only the EX group (EX, p=0.009; EX+WBV, p=0.61), while BMD decreased in both groups (EX, p<0.001; EX+WBV group, p<0.001). Leg strength increased over time in the EX group (p<0.001) and the EX+WBV group (p<0.001; between-group p=0.31). CONCLUSIONS: Exercise in combination with WBV may help attenuate regional bone loss in children recovering from burns. Studies are needed to determine the optimal magnitude, frequency, and duration of the vibration protocol, with attention to minimizing any potential interference with wound healing and graft closure.