Concentric and eccentric isokinetic resistance training similarly increases muscular strength, fat-free soft tissue mass, and specific bone mineral measurements in young women.

UNLABELLED: Women participated in 5 months of unilateral concentric (n = 37) or eccentric (n = 33) isokinetic resistance training of the legs and arms. Limb muscular strength increased as did total body, leg, and arm fat-free soft tissue mass, total body BMC, hip BMD, and forearm BMC and BMD. Isokinetic training benefits bone mineral acquisition. INTRODUCTION AND HYPOTHESIS: Isokinetic resistance training (IRT) is osteogenic; however, it is not known if concentric or eccentric modalities of IRT produce differential effects on bone. We tested our hypothesis that high-load eccentric versus concentric mode of IRT would produce greater increases in muscular strength, fat-free soft tissue mass (FFSTM), bone mineral density (BMD) and content (BMC) in trained legs and arms. METHODS: Participants were randomized to 5 months of concentric (n = 37) or eccentric (n = 33) training. The non-dominant leg and arm were used during training; dominant limbs served as controls. Muscular strength was measured with an isokinetic dynamometer; body composition was measured by dual-energy X-ray absorptiometry. RESULTS: Muscular strength of the concentrically and eccentrically trained leg (18.6%; 28.9%) and arm (12.5%; 24.6%) significantly increased with training. Gains in total body (TB) BMC (p < 0.05) and, in the trained limbs, total proximal femur BMD (p < 0.05) and total forearm BMD (p < 0.05) and BMC (p < 0.05) occurred in both groups. FFSTM increased for the TB and trained leg and arm (all p < 0.001) in both modes. CONCLUSION: Regardless of the mode, high-intensity, slow-velocity IRT increases muscular strength and FFSTM of trained limbs and imparts benefits to TB BMC and site-specific BMD and BMC in young women.

Relationships among bone mineral density, body composition, and isokinetic strength in young women.

The purpose of this study was to examine the relationships among bone mineral density (BMD), body composition, and isokinetic strength in young women. Subjects were 76 women (age: 20 +/- 2 yr, height: 164 +/- 6 cm, weight: 57 +/- 6 kg, body fat: 27 +/- 4%) with a normal body mass index (18-25 kg/m(2)). Total body, nondominant proximal femur, and nondominant distal forearm BMD were measured with dual-energy x-ray absorptiometry. Isokinetic concentric (CON) and eccentric (ECC) strength of the nondominant thigh and upper arm were measured at 60 deg/sec. Fat-free mass (FFM) correlated (P < 0.001) with BMD of the total body (r = 0.56) and femoral neck (r = 0.52), whereas fat mass (FM) did not relate to BMD at any site. Leg FFM, but not FM, correlated with BMD in all regions of interest at the proximal femur. Weak associations were observed between arm FFM and forearm BMD. Isokinetic strength did not relate to BMD at any site after correcting for regional FFM. In conclusion, strong, independent associations exist between BMD and FFM, but not FM or isokinetic strength, in young women.

The influence of patient strength, aerobic capacity and body composition upon outcomes after coronary artery bypass grafting.

BACKGROUND: Physical activity, physical fitness and body habitus of patients may be important predictors of outcomes after cardiac surgery. This study sought to quantify physical fitness and determine whether components of fitness enhance the prediction of outcomes in a group of patients undergoing coronary artery bypass grafting. METHODS: A group of 200 patients were evaluated prior to coronary artery bypass surgery. A Veterans Specific Activity Questionnaire (VSAQ) measured aerobic capacity. A grip dynamometer assessed strength. Skin-fold thickness was used to calculate percent body fat and lean body mass index. Patients were divided into low risk (0-2.5%) and high risk (>2.5%) groups based on the STS National Cardiac Surgery Database prediction of operative mortality. RESULTS: Patients with both a high percent body fat and a low VSAQ were at higher risk for at least one serious complication (p<0.05) and a longer postoperative length of stay (p<0.05). CONCLUSION: This study suggests: 1) An index of physical fitness can be obtained preoperatively in cardiac surgical patients; 2) This information aids in the prediction of operative risk.

Effects of combined aerobic and resistance training versus aerobic training alone in cardiac rehabilitation.

PURPOSE: This study examined the effects of performing combined resistance and aerobic training, versus aerobic training alone, in patients with coronary artery disease. METHODS: Thirty-six patients with coronary artery disease were randomized to either an aerobic-only training group (AE) or a combined aerobic and resistance training group (AE + R). Both groups performed 30 minutes of aerobic exercise 3 days/week for 6 months. In addition, AE + R group performed two sets of resistance exercise on seven different Nautilus machines after completion of aerobic training each day. Twenty patients (AE: n = 10; AE + R: n = 10) completed the training protocol with > 70% attendance. RESULTS: Strength gains for AE + R group were greater than for AE group on six of seven resistance machines (P < 0.05). VO2peak increased after training for both AE and AE + R (P < 0.01) with no difference in improvement between the groups. Resting and submaximal exercise heart rates and rate-pressure product were lower after training in the AE + R group (P < 0.01), but not in the AE group. AE + R increased lean mass in arm, trunk, and total body regions (P < 0.01), while AE increased lean mass in trunk region only (P < 0.01). Percent body fat was reduced for AE + R after training (P < 0.05) with a between group trend toward reduced body fat (P = 0.09). Lean mass gain significantly correlated with strength increase in five of seven resistance exercises for AE + R. CONCLUSIONS: Resistance training adds to the effects of aerobic training in cardiac rehabilitation patients by improving muscular strength, increasing lean body mass, and reducing body fat.

In vivo internalization of Staphylococcus aureus by embryonic chick osteoblasts.

Staphlylococcus aureus is the primary pathogen associated with osteomyelitis, an acute and recurrent bone disease. Internalization of S. aureus by cultured embryonic chick calvarial osteoblasts has been observed. The purpose of this study was to demonstrate that internalization of bacteria by embryonic chick calvarial and tibial osteoblasts occurs in vivo. In initial experiments, 10(8) colony forming units (cfu) of S. aureus, strain UAMS-1 or Cowan 1, were injected subcutaneously under the scalp skin of 17 day chick embryos. After 45 min, calvariae were harvested and processed for transmission electron microscopy (TEM). In subsequent experiments, 10(9) cfu of UAMS-1 were injected into the allantoic sac of 17 day chick embryos via a small opening in the egg shell. After 48 h, calvariae and tibiae were harvested for TEM. S. aureus cells were found in approximately 14% of the calvarial osteoblasts after subcutaneous injection and in 11% of calvarial and tibial osteoblasts following intraallantoic injection. Endosomes were observed in some cells, but most bacteria internalized appeared to be free in the cytoplasm. Osteoblasts with as few as five bacteria had a greater loss of cytoplasmic integrity and a more heterochromatic nucleus than osteoblasts with fewer bacteria or than uninfected osteoblasts. S. aureus cells in calvariae and tibiae were also observed in the cytoplasm of approximately 4% of the osteocytes in mineralized bone matrix. Thus, internalization of S. aureus by osteoblasts in vivo augments the previous observation in vitro. This study has also shown that osteoblasts with few bacteria continue differentiating into osteocytes. Results of these experiments support the hypothesis that internalization of S. aureus by osteoblasts may play a role in the etiology of osteomyelitis.

Staphylococcus aureus infection of mouse or human osteoblasts induces high levels of interleukin-6 and interleukin-12 production.

Staphylococcus aureus is the principal causative agent of the inflammatory bone disease osteomyelitis. Unfortunately, the pathogenesis of this often chronic infection is poorly understood and is complicated by the recent observation that bone-forming osteoblasts can harbor S. aureus. Such an infection presents a significant challenge for the host immune response, because osteoblasts are not known to initiate protective cell-mediated immune responses. Cultured mouse and human osteoblasts infected with S. aureus were found to express high levels of interleukin (IL)-6 and IL-12p75, on the basis of complementary investigations demonstrating both S. aureus-induced up-regulation of expression of IL-6 and IL-12p40 mRNA and secretion of IL-6 and IL-12p75 by these cells. Additionally, a quantitative bioassay demonstrated that IL-12p75 secreted after infection was biologically active. These studies are the first to demonstrate induced IL-12p75 expression by osteoblasts and suggest a previously unrecognized role for osteoblasts in initiating immune responses after S. aureus infection.

Mechanisms of Staphylococcus aureus invasion of cultured osteoblasts.

Staphylococcus aureus is a bacterial pathogen causing approximately 80% of all cases of human osteomyelitis. This bacterium can adhere to and become internalized by osteoblasts and previous studies indicate that osteoblasts are active in the internalization process. In the current study, we examined the roles of microfilaments, microtubules and clathrin-dependent receptor-mediated endocytosis in the internalization of S. aureus by MC3T3-E1 mouse osteoblast cells. Microfilament and microtubule polymerization was inhibited with cytochalasin D and colchicine. Clathrin-coated pit formation was examined by using the transaminase inhibitor, monodanslycadaverine. The results of this study indicate that mouse osteoblasts utilize actin microfilaments, microtubules and clathrin-coated pits in the internalization of S. aureus; however, microfilaments seem to play the most significant role in the invasion process.

Staphylococcus aureus adhesion to bone matrix and bone-associated biomaterials.

Staphylococcus aureus is a frequent cause of orthopedic infections in humans. The bacterium expresses several adhesins that facilitate bacterial binding to the bone matrix and to bone implant biomaterials coated with host plasma constituents. The relevant S. aureus adhesins are termed microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) and specific MSCRAMMs are involved in bone and joint infections.

Comparison of growth and metabolism of avian osteoblasts on polished disks versus thin films of titanium alloy.

The purpose of this study was to evaluate the efficacy of using high vacuum, thermal evaporation to deposit thin films of Ti-6Al-4V onto plates for subsequent cell culture investigations. Osteoblastic response to thin-film coated plates was compared to that of cells grown on Ti alloy disk inserts and uncoated culture plates. The Ti alloy disks were polished, cleaned, and passivated following a commercial protocol for orthopedic implants. Mean surface roughness was 262 nm for the Ti alloy disks and 4.756 nm for the coated culture plates. Osteoblasts isolated from 16-day chick embryo calvariae were cultured on polystyrene, thin films, and disks. At confluence, the cells were cultured an additional 48 h and were evaluated for cell number (DNA content), rate of glycolysis (lactate production), alkaline phosphatase activity (ALPase), and collagenous (3H-proline hydroxylation) and noncollagenous protein synthesis. Cell morphology was similar for the controls, disks, and thin-film groups. DNA, lactate, cell layer ALPase, 3H-hydroxyproline, and noncollagenous protein were not different (p > 0.05) among the control, thin-film, and disk groups. Medium ALPase was lower (p < 0.05) in the thin-film group compared to the control group. Although aluminum and vanadium percentages varied from nominal in the thin-film groups (11Al-2V as opposed to 6Al-4V), avian osteoblasts responded similarly to the Ti alloy thin films, disks, and uncoated culture plates for the smooth surfaces tested. The thin-film cell culture system used for elemental material studies appears to offer a promising method for the investigation of cellular response to alloyed biomaterials as well. Proper adjustments in alloy percentages before deposition, however, need to be made if thermal evaporation is utilized.

Hydrogen peroxide inhibits giant cell tumor and osteoblast metabolism in vitro.

This study investigates the efficacy of using hydrogen peroxide as adjuvant therapy after extended local curettage for benign giant cell tumors of bone. Hydrogen peroxide is used clinically as a chemical adjuvant for removal of residual tumor cells, presumably by effervescent cleansing with minimal damage to surrounding soft tissue and bone cells. This investigation examined the effects of hydrogen peroxide on giant cell tumor cells and osteoblasts grown in culture. Fresh fragments of histologically confirmed giant cell tumor tissue (six patients) and trabecular bone (one patient) were excised. Cells obtained from the fragments were grown in culture. Confluent cell cultures were exposed to saline (control) or hydrogen peroxide (0.1-1000 mm) for 2 minutes, and incubation was continued for 12, 24, or 48 hours without hydrogen peroxide. Protein content, deoxyribonucleic acid content, tartrate resistant acid phosphatase activity, and alkaline phosphatase activity were measured in the cell layers. The medium from the final 12 hours of each incubation period was used to evaluate lactate production. Cell lysis or death occurred after exposing giant cell tumor cells and osteoblasts to 100 mm and 30 mm hydrogen peroxide, respectively, concentrations substantially lower than the 3% (880 mm) hydrogen peroxide commonly used clinically. These results support the theory of using a minimal concentration of hydrogen peroxide as a chemical adjuvant in the surgical treatment of giant cell tumors of bone.

Extracellular pH modulates the activity of cultured human osteoblasts.

The effect of medium pH on the activity of cultured human osteoblasts was investigated in this study. Osteoblasts derived from explants of human trabecular bone were grown to confluence and subcultured. The first-pass cells were incubated in Hepes-buffered media at initial pHs adjusted from 7.0 to 7.8. Osteoblast function was evaluated by measuring lactate production, alkaline phosphatase activity, proline hydroxylation, DNA content, and thymidine incorporation. Changes in medium pH were determined from media pHs recorded at the beginning and end of the final 48 h incubation period. As medium pH increased through pH 7.6, collagen synthesis, alkaline phosphatase activity, and thymidine incorporation increased. DNA content increased from pH 7.0 to 7.2, plateaued from pH 7.2 to 7.6, and increased again from pH 7.6 to 7.8. The changes in the medium pH were greatest at pHs 7.0 and 7.8, modest at pHs 7.4 and 7.6, and did not change at 7.2, suggesting that the pHs are migrating towards pH 7.2. Lactate production increased at pH 7.0 but remained constant from 7.2 to 7.8. These results suggest that in the pH range from 7.0-7.6 the activity of human osteoblasts increases with increasing pH, that this increase in activity does not require an increase in glycolytic activity, and that pH 7.2 may be the optimal pH for these cells.

Calcium hydroxide ameliorates tobramycin toxicity in cultured chick tibiae.

Results from this laboratory have shown that bone metabolism is directly related to extracellular pH and that high concentrations of tobramycin released from impregnated polymethylmethacryrate (PMMA) beads has pH-dependent toxic effects on bone. In the present study, beneficial effects of calcium hydroxide-impregnated PMMA were investigated regarding tobramycin toxicity and bone metabolism in chick embryo tibiae in vitro. Also using Ca(OH)2 as a pH regulator, the antibiotic efficacy of tobramycin-impregnated PMMA was evaluated with respect to inhibition of Staphylococcus aureus growth. When Ca(OH)2 was added to PMMA beads containing tobramycin, the beads released hydroxyl and calcium ions into the culture medium and released more antibiotic than beads containing only tobramycin. Bone metabolism (glycolysis, total protein synthesis, and collagen synthesis) was enhanced by Ca(OH)2-impregnated beads with or without tobramycin. Additionally, bacterial growth was inhibited more strongly when S. aureus was incubated with tobramycin- and Ca(OH)2-impregnated PMMA disks than with disks containing only tobramycin. This study demonstrates the feasibility of adding Ca(OH)2 to tobramycin-impregnated PMMA beads as a regulator of local pH and a promoter of bone metabolism for protection of bone when high concentrations of tobramycin are used to treat osteomyelitis. It also suggests that lower concentrations of antibiotic may be effective if Ca(OH)2 and tobramycin are administered simultaneously.

Comparison of human osteoblasts and osteogenic cells from heterotopic bone.

Heterotopic ossification is a common complication in which bone develops in soft tissues. Although frequently benign, in some patients the condition becomes painful, restricts motion, and requires surgical intervention. This condition and the cells responsible for it are poorly characterized. Using cell culture techniques, this study compares the performance of osteogenic cells obtained from heterotopic ossification with trabecular osteoblasts isolated from the same patient. Cells isolated from trabecular bone samples and heterotopic ossification sites from six patients were evaluated for osteocalcin production after exposure to 1,25-dihydroxycholecalciferol, alkaline phosphatase activity, typing and synthesis of collagen, cell proliferation, and total protein content. Samples of heterotopic ossification and trabecular bone from three of the patients were examined histologically. Heterotopic ossification derived cells were shown to produce osteocalcin, Type 1 collagen, and alkaline phosphatase activity. They also had increased rates of collagen synthesis, alkaline phosphatase activity, and cell proliferation compared with the normal osteoblasts. Initial tissue from the heterotopic ossification sites showed increased numbers of osteocytes/mm2 compared with normal trabecular bone. Although heterotopic ossification derived cells functioned qualitatively like osteoblasts, they exhibited elevated levels of activities traditionally ascribed to osteoblasts, such as collagen synthesis and alkaline phosphatase activity.

Interaction of tobramycin and pH in cultured chick tibiae.

The toxicity of tobramycin at concentrations released from antibiotic-impregnated polymethylmethacrylate beads was determined using cultured embryonic chick tibiae. Because previous results from this laboratory have shown that osteoblast metabolism is inhibited at low pH and because of the potential for a low local pH in infected bone, the antibiotic was tested in medium with pH values from 6.8 to 7.4. Bone metabolism was evaluated by measuring the rates of glycolysis (lactate production), protein synthesis ([3H]proline uptake), and collagen synthesis ([3H]proline hydroxylation). Tobramycin at the concentrations released from the beads (1.0-1.5 mg/ml) contributed to lowering the pH of the medium. At pH 7.4, the antibiotic produced as much as a 30, 39, and 48% decrease in glycolysis, protein synthesis, and collagen synthesis, respectively. Tibiae exposed to tobramycin for 3 days, followed by 2 days without the antibiotic, showed only a partial recovery from its toxic effects. Although tobramycin was relatively less toxic in an acidic environment, the overall metabolic activity of the bones was poorest when the tobramycin concentration was high (2.0 mg/ml) and pH was low (6.8). The results of this study support the following conclusions: (a) tobramycin at high concentrations is toxic to bone, (b) a combination of high tobramycin concentration and low pH in the bone microenvironment may greatly inhibit bone metabolism, and (c) treatment and prevention of osteomyelitis by means of tobramycin-impregnated beads may be augmented by preventing the pH from decreasing in traumatized areas.

Cigarette smoking and its orthopedic consequences.

Cigarette smoking and its ramifications are coming under increasing scrutiny in the field of orthopedic surgery. Smoking has been implicated in impeding bone metabolism and fracture repair, and increasing the rate of postoperative infection and the incidence of nonunion. This article reviews the current body of knowledge on these topics, as well as the potential adverse effects of smoking on wound healing and microsurgical procedures. An in-depth discussion on the pathophysiologic mechanisms of nicotine is also included.

Internalization of Staphylococcus aureus by cultured osteoblasts.

Staphylococcus aureus is the most common cause of both acute and chronic osteomyelitis; however, the pathogenesis of osteomyelitis is poorly understood. We investigated the ability of S. aureus to associate with chick osteoblasts in culture and have demonstrated internalization of bacteria by the osteoblasts. Two strains of S. aureus were examined that were ingested by osteoblasts to different extents, suggesting strain differences in uptake. Initial association of S. aureus strains with osteoblasts was independent of the presence of matrix collagen produced by the osteoblasts. Internalization of bacteria required live osteoblasts, but not live S. aureus, indicating osteoblasts are active in ingesting the organisms. The bacteria were not killed by the osteoblasts, since viable bacteria were cultured several hours after ingestion.

Toxic effects of wound irrigation solutions on cultured tibiae and osteoblasts.

Irrigating wounds with solutions of antiseptic or antibiotic agents is routinely performed in orthopaedic surgery to reduce the incidence of microbial infection. The effects of these agents on healthy bone tissue is unknown. Three commonly employed antiseptic agents (hydrogen peroxide, Betadine solution, Betadine scrub) and one antibiotic solution (bacitracin) were tested on tibiae and osteoblasts isolated from embryonic chicks. Osteoblast function was evaluated by glycolytic energy metabolism (lactate production), cell number (DNA content), and collagen synthesis ([3H]proline hydroxylation). Two series of experiments were performed. To study concentration-related effects, tibiae or cells were exposed to a range of concentrations of the agents (diluted in saline, 0.9% NaCl) for 2 min, rinsed with saline, and incubated for 24 h in medium containing [3H]proline. For the recovery study, the cells were exposed to an effective, but nonlethal, concentration of the antiseptic agents for 2 min, rinsed with saline, and the incubation was continued in complete culture medium for 6, 12, 24, 48, or 72 h with [3H]proline added for the final 6 h. Solutions containing the antiseptic agents were cytotoxic to both bones and cells at concentrations well below those used clinically in irrigation solutions. In contrast, bacitracin at the concentrations tested was safe for osteoblasts and tibiae. These results suggest that the use of irrigation solutions containing H2O2, Betadine solution, or Betadine scrub on exposed bone tissue should be considered with caution.

Lumbar intervertebral disc transfer. A canine study.

STUDY DESIGN: Degenerative lumbar disc disease has been implicated as a cause of low back pain. Current treatment options for low back pain involve nonphysiologic fusion of the involved segments and have variable success rates. This is an experimental study of lumbar intervertebral disc transplantation using a canine surgical model. OBJECTIVES: This study evaluated the feasibility of lumbar disc transplantation and its effects on disc metabolism and morphology. METHOD: Eight mature mongrel dogs underwent disc transfer surgeries, in which the L2-L3 and L4-L5 intervertebral discs, with a small segment of adjacent superior and inferior vertebral body, were removed and transposed. The transplanted disc were stabilized by plates or by a flexible cable wire construct using Songer cables (DANEK, Inc., Memphis, TN). Unrestricted activity was allowed postoperatively. At 4 months, the spines were harvested, and the transplanted discs were evaluated biochemically and histologically. Intervening nontransplanted discs served as viable controls and thrice-frozen discs served as nonviable controls. Cell viability was assessed by measuring proteoglycan synthesis and DNA content. RESULTS: Proteoglycan synthesis (35S uptake normalized to DNA content) was maintained in transplanted anulus fibrosus tissue, but was decreased in nucleus pulposus samples (P < 0.05). DNA content was not altered significantly in the transplanted discs. Histologic analysis of the transplanted discs showed revascularization and remodeling of the bone adjacent to the disc and preservation of the lamellar architecture of the anulus fibrosus. The transplanted nucleus pulposus samples had chondrocyte-like cells present, but the staining characteristics of the nucleus material was variable. The contour of the transplanted disc endplates was irregular in all specimens. CONCLUSIONS: The structure and function of autograft intervertebral discs were maintained after disc transfer surgery; the transplant discs, however, were not completely normal in either their morphology or their metabolic functioning.

Medium pH modulates matrix, mineral, and energy metabolism in cultured chick bones and osteoblast-like cells.

The effects of medium pH were tested on calvariae, tibiae, and osteoblast-like cells from chick embryos. Bones and isolated cells were incubated for 5 h or 2 days in Hepes-buffered medium at pH values ranging from 6.8 to 8.2. Osteoblast function was evaluated by lactate production, oxygen consumption, alkaline phosphatase activity (AlPase), Ca and inorganic phosphate (Pi) flux, proline hydroxylation, DNA content, and thymidine incorporation. As medium pH was increased, glycolysis, collagen synthesis, and AlPase increased, while Ca efflux decreased. No effect of pH was seen on mitochondrial activity, Pi efflux, or cell number or proliferation. The importance of glycolysis as an endogenous pH regulator was demonstrated by inhibition with iodoacetic acid or glucose restriction and by adding lactate to the medium. The results suggest that the pH of bone interstitial fluid may be regulated by glycolysis and that changes in pH of this compartment may have marked effects on osteoblast function.

Smokeless tobacco contains an inhibitor of prolyl hydroxylase activity.

The effects of smokeless tobacco extract (STE) and various constituents of STE on prolyl hydroxylase activity were determined using enzyme extracted from chick embryos. STE inhibited prolyl hydroxylase activity in a concentration-dependent manner, but nicotine and anabasine had essentially no effect. Enzymatic activity was inhibited by zinc, but not by the other inorganic elements in STE; however, the zinc concentration in STE was not high enough to produce the observed inhibition. The inhibition by STE was diminished by increasing concentrations of 2-oxoglutarate, but not by increasing concentrations of other cofactors. Thus, STE contains an inhibitor of prolyl hydroxylase which may be competitive with 2-oxoglutarate.