In vitro method for the screening and monitoring of estrogen-deficiency osteoporosis by targeting peripheral circulating monocytes

Bone loss occurs insidiously and initially asymptomatically; therefore, osteoporosis is frequently diagnosed only after the first clinical fracture. The aim of this study was to test the hypothesis is that by simply observing the behavior of cultured peripheral monocytes, it might be possible to diagnose altered bone remodeling and, therefore, limit the complications associated with osteoporosis, especially fractures. Monocytes isolated as mononuclear precursors from healthy and ovariectomized rats were cultured both in basal and differentiation medium for up to 3 weeks. Viability and differentiation capability towards the osteoclastic phenotype was checked by light microscopy at early times, whereas differentiation state and synthetic activity (tartrate-resistant acid phosphatase (TRAP) staining; phalloidin, fluorescin isothiocynate (FITC) staining, cathepsin K, metalloproteinase 7 and 9, MMP-7 and MMP-9) were measured at 1, 2, and 3 weeks. Compared to their controls, monocytes isolated from ovariectomized rats proliferate and lean toward the osteoclastic phenotype in the absence of differentiating factors. In both culture conditions, osteoclasts from ovariectomized rats showed significantly higher productions of cathepsin K, MMP-7, and MMP-9 than those of cells isolated from healthy rats, steadily over time. These results obtained in an animal osteoporotic model, if confirmed by clinical studies, open up the possibility to assess the presence of an alteration in bone remodeling with a simple in vitro diagnostic test requiring a small blood sample and less than 48 h. This might allow to early select patients with a spontaneous viability and differentiation of monocytes to osteoclasts for further diagnostic techniques.     

Surface analysis and effects on interfacial bone microhardness of collagen-coated titanium implants: a rabbit model

PURPOSE: The aim of this study was to evaluate the surface chemistry and the microhardness at the implant-bone interface using a recently developed collagen-coated titanium implant in a short-term rabbit model. MATERIALS AND METHODS: Surface chemistry was evaluated by x-ray photoelectron spectroscopy (XPS), while in vivo studies involved 4-week implants mid-diaphysis in the lateral femurs of adult male rabbits. After conventional embedding and evaluation of histologic sections, the resinembedded blocks containing the implanted screws were used to measure bone hardness by means of an indentation test. RESULTS: Decomposition of the C1s peak obtained by XPS analysis confirmed that surface-immobilized collagen retained all the molecular features of the control, nonimmobilized reference. As to microhardness measurement, newly formed bone at the collagen-coated-implant/bone interface was significantly harder than bone at the interface of the uncoated control implant and bone. DISCUSSION: These results suggested that collagen coating significantly improves bone maturation and mineralization at the interface in comparison with uncoated commercially pure titanium. Surface modification of titanium implants by collagen coating has recently been discussed as a promising approach to the biochemical modification of implant surfaces. The present results support previous histologic findings and demonstrated that the biomolecular layer linked over the titanium implant can increase the bone healing rate, at least in this animal model. CONCLUSIONS: The present microhardness measurement at the bone-implant interface showed that collagen coating can significantly improve bone maturation and mineralization at the interface in comparison with uncoated commercially pure titanium, confirming and substantiating previous findings by histomorphometric measurements from the same model.     

Maxillary sinus augmentation: histologic and histomorphometric analysis

PURPOSE: Implant placement in the posterior maxilla may often be contraindicated because of insufficient bone volume and the presence of the maxillary sinus. In these situations, sinus floor lifting and grafting frequently have been proposed as the best treatment. The aim of this study was to compare histologically the use of 100% autogenous bone versus a combination of autogenous bone and corticocancellous pig bone for maxillary sinus augmentation. MATERIALS AND METHODS: Eighteen patients requiring bilateral maxillary sinus augmentation were selected for this study. Bone for grafting was harvested from the iliac crest. Each patient received 100% autogenous bone in 1 randomly selected sinus (control side) and a 1:1 mixture of autogenous bone and corticocancellous pig bone particles in the contralateral sinus (test side). Five months after the augmentation procedure, bone biopsy specimens were taken at the time of implant placement. RESULTS: No complications were observed during the surgical procedures; all patients healed uneventfully. No signs or symptoms of maxillary sinus disease were observed during the 5 months after surgery. No significant differences in bone percentages were observed in the bone biopsies from test and control sides. DISCUSSION AND CONCLUSION: It could be concluded from this study that corticocancellous pig bone particles can be successfully used in a 1:1 mixture with autogenous bone from the iliac crest for maxillary sinus augmentation in cases of severely atrophic maxilla.     

The rare condition of maxillary osteomyelitis

Osteomyelitis is an acute or chronic inflammatory process that can involve cortical and trabecular aspects of bone or bone marrow. Cranial bones are infrequently involved, but spreading of inflammation with involvement of surrounding structures represent important risk, as are cerebral abscess, encephalitis, or meningitis. We present a case of osteomyelitis of right maxillary sinus in an adult caused by a spreading of contiguous inflammation sustained by a chronic intrasinusal polyp; the complete resolution of infection was gained with a combination of surgical treatment and antibiotic therapy. The aims of this article are to illustrate diagnostic patterns and surgical treatment experienced in a case of maxillary osteomyelitis and to report radiographic and histopathologic findings.     

Histologic and histomorphometric evaluation of alveolar ridge augmentation using bone grafts and titanium micromesh in humans

BACKGROUND: Recently, the use of titanium micromesh for alveolar bone augmentation has drawn interest; however, only limited histologic data are available on the quality of the bone regenerated. Therefore, this study compared the use of 100% intraoral autogenous bone to a combination of intraoral autogenous bone (70%) and bovine porous bone mineral (BPBM) (30%) for alveolar ridge augmentation with titanium micromesh histologically and histomorphometrically. METHODS: Twelve partially edentulous patients required alveolar bone augmentation before implant insertion because of ridge resorption. The defect sites, six in the maxilla and six in the mandible, were reconstructed with particulate autologous bone (control group, N = 6) or a mixture of autologous bone and BPBM (test group, N = 6) in combination with titanium micromesh. Core biopsies were taken from the defect sites 8 to 9 months after grafting at the time of implant insertion. RESULTS: Newly formed compact bone with a well-organized lamellar pattern was identified in all specimens. In the samples taken from the test group, the BPBM particles were surrounded completely by newly formed bone with no signs of resorption. The mean total bone volume was 62.38% +/- 13.02% in the control group and 52.88% +/- 11.47% in the test group. The soft tissue volume was 37.61% +/- 13.02% and 29.96% +/- 12.58%, respectively, and the residual BPBM volume was 17.15% +/- 2.72% in the test group. No statistical difference was observed in the histologic parameters evaluated, irrespective of graft type and site (P >0.05). CONCLUSION: Within the limits of this study, BPBM (30%) in combination with autogenous bone (70%) did not yield a lower percentage of new bone formed compared to autogenous bone alone in ridge augmentation with titanium micromesh.     

Association between CD226 polymorphism and soluble levels in rheumatoid arthritis: Relationship with clinical activity

Objective: To study the relation between CD226 rs763361 gene polymorphism and CD226 serum level and to evaluate their role in susceptibility and disease activity of RA in a cohort of Egyptian individuals.

Methods: The serum level of CD226 was measured using a suitable ELISA kit and the CD226 rs763361 gene polymorphism was typed by PCR-RFLP for 112 RA patients and 100 healthy controls.

Results: Significant association with RA was found with CD226 T allele (OR (95%CI) = 1.6 (1.04–2.4), P = 0.032), and higher CD226 serum level (P = 0.001). Higher CD226 levels were associated with higher ESR values (P = 0.035), positive CRP (0.048), increased number of tender joints (P = 0.045), and higher DAS score (P = 0.035). Serum CD226 is an independent risk factor for the prediction of RA (P = 0.001). No correlations were found between the serum level of CD226 and different CD226 genotypes and also between them and RA activity grades.

Conclusion: The CD226 T allele may be susceptibility risk factors for the development of RA and the higher serum level of CD226 may be involved in the pathogenesis of RA in Egyptian patients. The serum level of CD226 and not CD226 genotypes could be considered as an independent risk factor for the prediction of RA within healthy individuals and also for RA disease activity.     

Treatment of normal individuals with granulocyte-colony-stimulating factor: donor experiences and the effects on peripheral blood CD34+ cell counts and on the collection of peripheral blood stem cells

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) has been used in patients to increase the level of circulating hematopoietic progenitors. Although G-CSF has been administered to some healthy individuals, the kinetics of mobilization of peripheral blood stem cells (PBSCs), the optimum dose schedule and the incidence and nature of adverse reactions in normal individuals are not completely defined. STUDY DESIGN AND METHODS: Normal individuals (n = 102) who received G- CSF for 5 or 10 days at doses of 2, 5, 7.5, or 10 micrograms per kg per day were studied. The subjects were observed for symptoms and physical changes, and blood samples were obtained for a variety of laboratory tests. After 5 or 10 days of G-CSF treatment, PBSCs were collected by apheresis and analyzed. RESULTS: Overall, 89 percent of the individuals completed the 5-day treatment protocol and 88 percent completed the 10- day protocol without modification of the dose of G-CSF administered. Ninety percent of donors experienced some side effect of G-CSF. The most frequent effects noted were bone pain (83%), headache (39%), body aches (23%), fatigue (14%), and nausea and/or vomiting (12%). The dose of G-CSF administered directly affected the proportion of people with bone pain (p = 0.025) or body aches (p = 0.045) or who were feeling hot or having night sweats (p = 0.02) or taking analgesics (p = 0.01). With the 5-day dose schedule, several changes in serum chemistries occurred, including increases in alkaline phosphatase (p = 0.001), alanine aminotransferase (p = 0.0013), lactate dehydrogenase (p = 0.0001), and sodium (p = 0.0001). Decreases occurred in glucose (p = 0.045), potassium (p = 0.0004), bilirubin (p = 0.001), and blood urea nitrogen (p = 0.0017). In donors who received G-CSF for 5 days, the absolute neutrophil count was increased after one G-CSF dose, and it reached a maximum on Day 6, as did the number of CD34+ cells (64.6 +/? 55.9 × 10(6) cells/L). In those same donors, the platelet count after apheresis on Day 6 was 32 +/? 13 percent lower than pretreatment values (250 +/? 42 × 10(9) cells/L). In donors receiving G-CSF for 10 days, the neutrophil count reached a maximum on Day 8, but the number of CD34+ cells peaked on Day 6 (58.3 +/? 52.1 × 10(5) cells/L) and then declined. The platelet count decreased from pretreatment values by 28 +/? 12 percent prior to apheresis on Day 11. When individuals were treated for 5 days with G-CSF, the quantity of CD34+ cells collected was directly related to the G-CSF dose. When 5 micrograms per kg per day was given, 2.80 +/? 1.81 × 10(8) cells were collected, compared with collection of 4.67 +/? 3.11 × 10(8) cells when 10 micrograms per kg per day was given (p = 0.04). More important, PBSCs collected after 10 days of G-CSF administration (5 micrograms/kg/day) had significantly fewer CD34+ cells (0.82 +/? 0.37 × 10(8) cells, p = 0.01) than did PBSCs collected after 5 days of G-CSF (5 micrograms/kg/day). CONCLUSION: Most normal donors receiving G-CSF experience side effects, but these are mild to moderate in degree. Some alterations in blood chemistries occur, but none were clinically serious. Because of the symptoms associated with G-CSF, these individuals must be monitored closely. The treatment of normal donors with G-CSF for more than 5 days significantly decreased the number of circulating CD34+ cells and the quantity collected by apheresis.     

Model for the physics and physiology of fluid administration

This article describes a model designed to provide an understanding of fluid flow in intravenous systems and human subjects. Experiments were developed which demonstrate that the model can represent common clinical situations. The model depicts physical devices as ideal resistors, pressure sources, and flow sources. The patient's venous system is depicted as a combination of ordinary and Starling resistors. For flows between 0 and 300 ml/hr, both physical devices and patients are adequately represented by a straight line representing the pressure-flow relationship (PFR): pressure = opening pressure + flow × resistance, where the slope is the resistance to fluid flow and the intercept is the opening pressure. The PFR for a normal vein is characterized by a flat slope (vein resistance =22±20 mm Hg/L/hr, mean ± SD) and a low intercept (opening pressure =15±8 mm Hg). The PFR for a partially obstructed vein has a resistance equal to that of an unobstructed vein and an opening pressure elevated approximately equal to the pressure obstructing the vein. For perivascular tissue, the PFR has a steep slope (tissue resistance =1,125±1,376 mm Hg/L/hr), while tissue opening pressure depends on the amount of fluid infused. At the onset of fluid extravasation (infiltration), tissue pressure usually is lower than venous pressure (8±8 versus 15±8 mm Hg), until fluid fills the distensible tissue compartment. In clinical practice, when infiltration or obstruction occurs, flow decreases and the clinician adjusts the roller clamp until correct flow resumes; no problem is obvious. The combined model for the intravenous tubing and venous systems explains the behavior of current clinical infusion devices.Presented in part at the Sixth Medical Monitoring Technology Conference, Vail, CO, March 1986; at the Annual Meeting of the American Society of Anesthesiologists, Las Vegas, NV, October 1986; at the Seventh Medical Monitoring Technology Conference, Vail, CO, March 1987; at a meeting on Computers in Critical Care and Pulmonary Medicine, San Diego, CA, June 1987; at the Annual Meeting of the American Society of Anesthesiologists, Atlanta, GA, October 1987; at the Regional Meeting of the Association for the Advancement of Medical Instrumentation, Cincinnati, OH, October 1987; at the Institute of Electrical and Electronics Engineers Ninth Annual Conference of the Engineering in Medicine and Biology Society, Boston, MA, November 1987; and at a meeting of the American Society of Hospital Pharmacists, Atlanta, GA, December 1987.Supported in part by grants from IVAC Corp.The author thanks the following individuals for important intellectual and/or technical assistance: Peter Basser, PhD, Avital Cnaan, PhD, Adriane Concus, MD, John Fox, MD, David Gissen, MD, David Joseph, MD, Anne Kamara, David Leith, MD, Leonard Lind, MD, Richard Morris, MB, BS, Barbara Orlowitz, MEE, Mary Anne Palleiko, RN, Beverly Philip, MD, Daniel Raemer, PhD, David Scott, MB, BS, John Stelling, MPH, and Marie vanRensberg, MB. At IVAC Corp: Walter Bochenko, BSEE, MBA, Robert Butterfield, BSEE, Douglas Christian, RPh, MBA, Alan Davison, BS, David Doan, PhD, Alan Somerville, BSEE, MS, Robin Wernick, BSEE, MS.     

Autoantibodies against bactericidal/permeability-increasing protein in patients with cystic fibrosis

Cystic fibrosis (CF), a genetic disorder, is characterized by chronic pulmonary infection/inflammation which leads to respiratory failure. The presence of anti-neutrophil cytoplasmic autoantibodies (ANCA) has previously been observed in the sera of patients with CF. In view of the known relationship of ANCA with primary vasculitis and of their putative pathogenetic role in these disorders, we studied the presence, specificity and isotype of ANCA and their clinical associations in 66 adult CF patients. None of the 66 CF samples had autoantibodies to the major ANCA antigens, proteinase 3 or myeloperoxidase. However, 60/66 (91%) CF samples contained IgG and 55/66 (83%) IgA, autoantibodies to bactericidal/permeability increasing protein (BPI), a recently characterized ANCA specificity. All the IgA anti-BPI-positive samples were also IgG anti-BPI-positive. The autoantibody specificity was confirmed by inhibition assay and immunoblotting of CF sera against a neutrophil granule preparation. Furthermore, in this cross-sectional study, anti-BPI levels were inversely correlated with the observed reductions in FEV1 and FVC (IgA anti-BPI and FEV1: r = 0.508, <it>p</it> &lt; 0.0001), and both IgG and IgA anti-BPI levels were higher in CF patients with secondary vasculitis (<it>n</it> = 6) than in those without (<it>p</it> &lt; 0.05). ANCA with specificity for BPI were present in the majority of CF sera in this study and autoimmune processes may be associated with the development of pulmonary injury in CF.      

The metabolic syndrome in women: implications for therapy

It is becoming increasingly clear that hypertension and metabolic risk factors in women are inter-related and often share underlying causes. Menopause acts explicitly as a risk factor by reducing the direct beneficial effect of ovarian hormones upon cardiovascular functions and indirectly by negatively influencing other risk factors for coronary artery disease--i.e. hyperinsulinaemia, blood cholesterol, blood pressure, coagulation etc. Adverse changes in one factor may induce adverse changes in a variety of other risk factors and it is important to consider co-ordinated changes when evaluating these patients rather than attempt to isolate independent factors. Similarly with treatment, the prevalence of metabolic syndrome in postmenopausal hypertensive women has important implications and some antihypertensive drugs may worsen the already altered metabolic profile of these patients while others may be beneficial. Centrally-acting sympatholytic agents, e.g. moxonidine, are therefore important to consider in hypertensive postmenopausal women who experience other symptoms of metabolic syndrome.