Urine matrix metalloproteinases (MMPs) as biomarkers for the progression of fracture healing.

Whilst the majority of fractures heal normally, it is estimated that ∼10% of fractures exhibit some level of delayed or impaired healing. Although radiography is the primary diagnostic tool to assess the progression of fracture healing, radiographic features only qualitatively correlate with tissue level increases in mineral content and do not quantitatively measure underlying biological processes that are associated with the progression of healing. Specific metaloproteinases have been shown to be essential to processes of both angiogenesis and mineralised cartilage resorption and bone remodelling at different phases of fracture healing. The aim of this study was to determine the potential of using a simple urine based assay of the activity of two MMPs as a means of assessing the biological progression of fracture healing through the endochondral phase of healing. Using a standard mid-diaphyseal murine model of femoral fracture, MMP9 and MMP13 proteins and enzymatic activity levels were quantified in the urine of mice across the time-course of fracture healing and compared to the mRNA and protein expression profiles in the calluses. Both urinary MMP9 and MMP13 protein and enzymatic activity levels, assessed by Western blot, zymogram and specific MMP fluorometric substrate assays, corresponded to mRNA expression and immunohistologic assays of the proteins within callus tissues. These studies suggest that urinary levels of MMP9 and MMP13 may have potential as metabolic markers to monitor the progression of fracture healing.

Biochemical bone markers are useful to monitor fracture repair.

UNLABELLED: Biochemical bone markers reflect bone metabolism but little is known regarding their usefulness during fracture repair. Reduced bone mineral density may influence fracture healing. We hypothesized that low bone mineral density results in decreased levels of bone markers during the acute phase of fracture healing, especially in women who are postmenopausal. We also addressed the question of different fracture types and locations resulting in different levels of bone markers. Urinary levels of N-terminal cross-linked telopeptide, deoxypyridinoline, and pyridinoline were measured preoperatively and postoperatively in patients with hip fractures, distal forearm fractures, and in 25 control subjects. Bone mineral density was determined using quantitative computed tomography of the spine. Patients with low bone mineral density, especially women who were postmenopausal, had greater concentrations of N-terminal cross-linked telopeptide when compared with patients with normal bone mineral density or men. Patients with pertrochanteric fractures had greater concentrations than patients with femoral neck fractures, as did patients with hip fractures compared with patients with fractures of the distal forearm. These results suggest that levels of bone markers increase during fracture healing despite low bone mineral density and that different fracture types and locations result in different levels of bone markers. LEVEL OF EVIDENCE: Prognostic study, Level I (high quality prospective study-all patients were enrolled at the same time with > or = 80% of followup of enrolled patients). See the Guidelines for Authors for a complete description of levels of evidence.

Metabolic rate and nitrogen balance after skeletal trauma in female and male rats.

Unequal metabolic responses to trauma by women and men have been suggested, but an explicit investigation demonstrating this conjecture has not been made. The responses of resting energy expenditure (REE) and nitrogen balance for 3 days before and 7 days after skeletal trauma were determined for female and male rats. Food intake and body weight were recorded daily, and 24-h urine samples were collected. Baseline REE and nitrogen balance were obtained for 3 consecutive days before induction of trauma. Then rats were divided into female trauma (n = 8), male trauma (n = 7), female control (n = 8), and male control (n = 7) groups. Trauma was produced by bilateral femoral fracture to anesthetized rats. Control rats were anesthetized without skeletal trauma. Traumatized rats were fed ad libitum for 7 days, and control rats were pair fed with the traumatized rats. The results showed that REE increased and nitrogen balance decreased in traumatized male rats relative to their controls. Traumatized female rats had increased REE and unchanged nitrogen balance compared with their controls. Traumatized female rats had a larger percentage increase in REE on days 5 through 7 than did traumatized male rats. These findings demonstrate a difference between female and male rats in response to trauma. Female rats use more energy and lose less nitrogen after trauma than do male rats. The results suggest that recommendations for increased energy and protein needs after trauma should consider the sex of the subject intended to be fed.

Increased rates of cortisol production and urinary free cortisol excretion in elderly women 2 weeks after proximal femur fracture.

Cortisol production rate and urinary free cortisol excretion have been measured in healthy elderly women and elderly women about two weeks after upper femur fracture. Plasma cortisol was determined mid-morning, at the start of urine collection. All three variables were higher in the injured patients than in the control subjects. Urinary free cortisol excretion showed the greatest rise and was correlated with cortisol production rate in the patients. In the control subjects there was no correlation and nearly all the points fell below the regression line for the injured patients, indicating that urinary free cortisol excretion rose in relation to cortisol production rate after injury. Measurement of creatinine clearance showed that this was not due to an increased glomerular filtration rate, and a possible explanation is decreased metabolic clearance of cortisol. Plasma cortisol was not significantly correlated with either cortisol production rate or urinary free cortisol excretion.

Immobilisation hypercalciuria after limb fractures in children.

Immobilisation hypercalciuria and hypercalcaemia following limb fractures or paralysis is a frequent occurrence in children. Assessment of calcium metabolism should be performed in such patients, since the formation of kidney stones is possible. The beneficial effect of hydrochlorothiazide (HCT) in the prevention of renal stones is most likely due to a reduction of calcium concentration in urine and a significant decrease of crystalluria. Thus, its administration is recommended for children with hypercalciuria following prolonged immobilisation especially due to fracture treatment or paralysis.

Effects of enviromental temperature and femoral fracture on wound healing in rats.

Femoral fracture, unilateral and bilateral, impaired the healing of dorsal skin incisions and formation of reparative granulation tissue in subcutaneously implanted polyvinyl alcohol sponges judged histologically and by breaking strengths and hydroxyproline contents, respectively, 1 week after injury in pair-fed rats kept at 22 degrees C. When rats were transferred to a room at 30 degrees C immediately after skin incision and sponge implants, with or without unilateral fracture, no differences in healing were observed between the two groups. Rats with skin incision, sponge implants, and either femoral fracture or sham-fracture excreted more urinary nitrogen than preoperatively when kept at 22 degrees. Counterpart groups transferred to a 30 degrees room right after operation excreted less urinary nitrogen than preoperatively, but because of lower food intakes postoperatively, the ratio of urinary nitrogen to food intake nitrogen was increased. With equivalent food intakes, pair-fed rats with fracture kept at 22 degrees postoperatively lost more weight and excreted more nitrogen than corresponding rats transfered to a 30 degrees room.