PubMed Health⌕ Search

Biomedical subjects

M J Silva

Publications and source records attributed to M J Silva.

At least 19 recordsLinked to original sources
PubMed2006

DNA damage in human sperm is related to urinary levels of phthalate monoester and oxidative metabolites.

BACKGROUND: The ubiquitous use of phthalate esters in plastics, personal care products and food packaging materials results in widespread general population exposure. In this report, we extend our preliminary study on the relationship between urinary concentrations of phthalate metabolites and sperm DNA damage among a larger sample of men and include measurements of mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) and mono-(2-ethyl-5-oxohexyl) phthalate (MEOHP), two oxidative metabolites of di-(2-ethylhexyl) phthalate (DEHP). METHODS: Among 379 men from an infertility clinic, urinary concentrations of phthalate metabolites were measured using isotope-dilution high-performance liquid chromatography-tandem mass spectrometry. Sperm DNA damage measurements, assessed with the neutral comet assay, included comet extent (CE), percentage of DNA in tail (Tail%) and tail distributed moment (TDM). RESULTS: Monoethyl phthalate (MEP), a metabolite of diethyl phthalate, was associated with increased DNA damage, confirming our previous findings. Mono-(2-ethylhexyl) phthalate (MEHP), a metabolite of DEHP, was associated with DNA damage after adjustment for the oxidative DEHP metabolites. After adjustment for MEHHP, for an interquartile range increase in urinary MEHP, CE increased 17.3% [95% confidence interval (CI) = 8.7-25.7%], TDM increased 14.3% (95% CI = 6.8-21.7%) and Tail% increased 17.5% (95% CI = 3.5-31.5%). CONCLUSIONS: Sperm DNA damage was associated with MEP and with MEHP after adjusting for DEHP oxidative metabolites, which may serve as phenotypic markers of DEHP metabolism to 'less toxic' metabolites. The urinary levels of phthalate metabolites among these men were similar to those reported for the US general population, suggesting that exposure to some phthalates may affect the population distribution of sperm DNA damage.

Adult↗
PubMed2006

Nicotine delays tendon-to-bone healing in a rat shoulder model.

BACKGROUND: Many studies have shown that nicotine negatively impacts fracture healing and bone fusion processes. However, very little is known about its effect on tendon and ligament healing. The goal of the present study was to evaluate the effect of nicotine on tendon-to-bone healing. METHODS: Supraspinatus tendons in both shoulders of seventy-two rats were transected and repaired to the humeral head. Osmotic pumps were implanted subcutaneously, and nicotine or saline solution was delivered for ten, twenty-eight, or fifty-six days. Cell morphology was evaluated with use of histologic sections. Cells were counted, and proliferating cell nuclear antigen (PCNA) immunohistochemistry was performed to assess cellular proliferation. In situ hybridization was performed to measure type-I collagen mRNA expression. Biomechanical and geometric properties were assessed. RESULTS: Inflammation persisted longer in the nicotine group than in the saline solution group. Cellular proliferation was higher in the saline solution group than in the nicotine group at the early time-points. Type-I collagen expression was higher in the saline solution group at twenty-eight days. Mechanical properties increased over time in both groups. Maximum stress was significantly lower in the nicotine group than in the saline solution group at ten days. Maximum force was significantly lower in the nicotine group than in the saline solution group at twenty-eight days. Maximum force was significantly higher in the nicotine group than in the saline solution group at fifty-six days. Stiffness was not different between the groups at any time-point. CONCLUSIONS: Nicotine caused a delay in tendon-to-bone healing in a rat rotator cuff animal model. Mechanical properties increased over time in both groups, but the properties in the nicotine group lagged behind those in the saline solution group. Chronic inflammation and decreased cell proliferation may partly explain the inferior biomechanical properties in the nicotine group as compared with the saline solution group. CLINICAL RELEVANCE: Failure of rotator cuff repair is a major clinical problem. The adverse effect of nicotine on rotator cuff healing noted in this clinically appropriate animal model may be an important clinical consideration.

Animals↗
PubMed2005

Delayed repair of tendon to bone injuries leads to decreased biomechanical properties and bone loss.

INTRODUCTION: Repair of the torn rotator cuff tendon is a common procedure performed in the shoulder. In the clinical setting, a significant delay between rotator cuff tear and subsequent repair often exists. The purpose of this study was to investigate the biomechanical properties and bone density of the tendon to bone repair site after acute and delayed repair. METHODS: The supraspinatus tendons in bilateral shoulders of 60 rats were transected from the bone. In the acute group, the tendons were immediately repaired with suture. In the delayed group, the tendons were allowed to retract and repaired in a second procedure after a 3-week delay. Cross sectional area and biomechanical properties were evaluated. Bone density of the humeral head was assessed using peripheral quantitative computed tomography. Histologic sections were obtained and examined. RESULTS: At 10 days the repair tissue displayed vascular and fibroblast proliferation accompanied by predominantly mononuclear infiltrate. At 28 days the inflammatory process gradually decreased. No significant histologic differences were noted between the acute and delayed repair specimens. Cross-sectional area was higher in the delayed group at the early time points (44% at 10 days and 31% at 28 days). Viscoelastic properties were greater in the acute group at the early time points and significantly less at the latest time point, compared to the delayed group. Bone density was markedly decreased (8% and 12%, 28 and 56 days respectively) in the delay group. DISCUSSION: Inferior rotator cuff healing was demonstrated when there was a delay between injury and repair. Viscoelastic properties of the acute repairs were increased compared to the delayed group at 10 days, indicating tendon stiffening during the 3-week delay before repair. Viscoelastic properties of the acute repairs were decreased compared to the delayed group at 56 days indicating deterioration of properties over time in the delayed group. The deterioration in properties in the delayed group coincide with bone density decreases in the greater tuberosity. These results indicate that bone loss may a significant factor in poor healing.

Animals↗
PubMed2005

Supplementary core sutures increase resistance to gapping for flexor digitorum profundus tendon to bone surface repair - an in vitro biomechanical analysis.

We evaluated the effects of two types of supplementary core sutures on the tensile properties and resistance to gap formation of flexor digitorum profundus (FDP) tendon-bone repairs. Forty-five human cadaver FDP tendons were sharply released from their insertion sites and repaired to bone utilizing one of three repair techniques: four-strand modified Becker core suture (Becker only), modified Becker plus a figure-of-eight supplementary core suture (Becker plus figure-of-eight), and modified Becker plus a supplementary core suture using a bone anchor (Becker plus anchor). Ultimate (maximum) force did not differ between repair groups. However, addition of a supplementary suture significantly increased repair-site stiffness and the 1, 2 and 3 mm gap forces, while decreasing the gap at 20 N compared to the Becker only suture (P<0.05). The only difference between the two supplementary suture groups was that the Becker plus anchor group had increased stiffness compared to the Becker plus figure-of-eight group. In conclusion, a supplementary figure-of-eight suture and a supplementary suture using a bone anchor provide enhanced resistance to gap formation for FDP tendon-bone repairs.

Adult↗
PubMed2005

Ligamentous versus physeal failure in murine medial collateral ligament biomechanical testing.

This study examines the age at which a femoral physeal failure ceased to occur in a mouse model of medial collateral ligament (MCL) testing. Biomechanical testing of the MCL with load to failure can result in physeal failure rather than MCL failure in skeletally immature animals. Failure mode depended significantly on age (p<0.05). Sixty percent of the knees tested at 4 months failed at the physis rather than at the ligament, whereas, only ten percent of the knees tested at 5 and 6 months failed at the physis. The mean ultimate force to failure for the specimens in which the failure occurred at the ligament was 8.1 N with a higher values for the right side versus the left (p<0.05). For the specimens in which the failure occurred at the physis, the mean ultimate force to failure was 11.2 N. We now consider that 5 month old mice are functionally skeletally mature and old enough to be tested biomechanically with few failures at the physis.

Age Factors↗
PubMed2004

Experimental and finite element analysis of the rat ulnar loading model-correlations between strain and bone formation following fatigue loading.

The rat forelimb compression model has been used widely to study bone response to mechanical loading. We used strain gages to assess load sharing between the ulna and radius in the forelimb of adult Fisher rats. We used histology and peripheral quantitative computed tomography (pQCT) to quantify ulnar bone formation 12 days after in vivo fatigue loading. Lastly, we developed a finite element model of the ulna to predict the pattern of surface strains during compression. Our findings indicate that at the mid-shaft the ulna carries 65% of the applied compressive force on the forelimb. We observed large variations in fatigue-induced bone formation over the circumference and length of the ulna. Bone formation was greatest 1-2 mm distal to the mid-shaft. At the mid-shaft, we observed woven bone formation that was greatest medially. Finite element analysis indicated a strain pattern consistent with a compression-bending loading mode, with the greatest strains occurring in compression on the medial surface and lesser tensile strains occurring laterally. A peak strain of -5190 microepsilon (for 13.3N forelimb compression) occurred 1-2 mm distal to the mid-shaft. The pattern of bone formation in the longitudinal direction was highly correlated to the predicted peak compressive axial strains at seven cross-sections (r2 = 0.89, p = 0.014). The in-plane pattern of bone formation was poorly correlated to the predicted magnitude of axial strain at 51 periosteal locations (r2 = 0.21, p < 0.001), because the least bone formation was observed where tensile strains were highest. These findings indicate that the magnitude of bone formation after fatigue loading is greatest in regions of high compressive strain.

Animals↗
PubMed2003

Accuracy of peripheral quantitative computed tomography (pQCT) for assessing area and density of mouse cortical bone.

Peripheral quantitative computed tomography (pQCT) is increasingly used for measurement of cortical bone geometry and density in mice. We evaluated the accuracy of pQCT for area and density measurements of thin-walled aluminum phantoms and mouse femora. Aluminum tubes with varying wall thicknesses and femora from 1- to 6-month-old C3H/HeJ (C3H) and C57B1/6J (B6) mice (average cortical thickness 0.14-0.29 mm) were scanned at 70- or 90-microm resolution. pQCT values of area were compared to optical values determined after sectioning, while pQCT density (vBMD) was compared to solid aluminum density or correlated to bone ash content. For the aluminum phantoms, the error in pQCT area and density depended strongly on wall thickness, and density was consistently underestimated. For mouse femora, threshold values were found that produced zero error in bone area for each strain and age group, although the optimal threshold differed between groups. pQCT vBMD correlated strongly with ash content (r2=0.7), although the regression equations differed between strains and the magnitude of the inter-strain difference in vBMD was fourfold greater than the difference in ash content. This finding suggests that pQCT can overestimate the differences in volumetric mineral density between inbred mouse strains whose bones are of different thickness (e.g., C3H vs. B6). In conclusion, both area and density values obtained by pQCT depend strongly on specimen thickness, consistent with a partial volume averaging artifact. Investigators using pQCT to assess cortical bones in mice should be aware of the potential for cortical thickness-dependent errors.

Aluminum↗
PubMed2002

Low frequency noise and whole-body vibration cause increased levels of sister chromatid exchange in splenocytes of exposed mice.

Chronic exposure to low frequency (LF) noise and whole-body vibration (WBV) induces both physiological and psychological alterations in man. Recently, we have shown that long-term occupational exposure to LF noise and WBV produces genotoxic effects in man expressed as an increase in sister chromatid exchange (SCE) levels in lymphocytes. The objectives of the present study were to investigate whether the observed effect could be reproduced in a murine model and, if so, which of the agents, LF noise alone or in combination with WBV, would be instrumental in the SCE induction. SCEs were analyzed in spleen lymphocytes of mice exposed to LF noise alone and in combination with WBV for 300 and 600 hr. An effect at the cell cycle kinetics level was also investigated. The results revealed significant increases in the mean SCE number per cell and in the proportion of cells with high frequency of SCEs (HFCs) in lymphocytes of mice submitted to combined noise and WBV over controls. No significant differences were found between single noise-exposed and control mice. A cell cycle delay was observed exclusively in the noise and WBV exposure groups. In conclusion, we demonstrated that, as in exposed workers, prolonged exposure to the combination of LF noise and WBV determines an increase in SCE level in mice while LF noise alone is not effective in SCE induction.

Animals↗
PubMed2002

Haplotype study of microsatellites flanking the t(15;17) breakpoint in acute promyelocytic leukemia patients from North Portugal.

A higher frequency of acute promyelocytic leukemia (APL) has been noted in countries of Southern Europe and among 'Latino' patients of the United States with acute myeloid leukemia (AML). In order to discover whether there is any genetic predisposition to the disease, we analyzed microsatellites flanking PML and RARalpha genes in 29 t(15;17) APL patients from North Portugal and compared them with a control group of 123 healthy individuals. Fluorescent PCR products were analyzed using an automated capillary electrophoresis system and allele and haplotype frequencies of the two populations were determined. No significant differences were found, suggesting the same genetic origin of patients and healthy individuals. As suggested by the four microsatellites screened, MSI (microsatellite instability) does not explain the increased incidence of t(15;17) APL in this Portuguese population. These results intend to be a new approach to the study of APL, reflecting the particularity of the disease.

Chromosomes, Human, Pair 15↗
PubMed2002

[Determination of tissue optical parameters using HF modulation spectroscopy].

Radio frequency modulation spectroscopy is a capable method to determine tissue optical parameters in-vivo. For the eventual purpose of clinical measurements we have developed and tested an rf laser spectroscopy device which enables a measurement of the spatial amplitude and phase shift profiles of backscattered modulated laser light. Spectral absorption and scattering coefficients are computed by inverse formulas derived from analytical solutions of the diffusion model of light transport in a semi-infinite geometry.

Humans↗
PubMed2001

The effects of repetitive physiologic loading on bone turnover and mechanical properties in adult female and male rats.

Repetitive physiologic loading is widely believed to be beneficial in maintaining skeletal integrity. However, repetitive loading is also associated with bone injuries, including stress fractures and osteoporotic fractures, indicating that under certain conditions repetitive physiologic loading decreases the functional capacity of bone. Our objective was to identify the response of bone to excessive repetitive loading in adult rats. Male and female rats (8-9 months old) were exposed to 2 hours of treadmill running each day for 10 or 30 consecutive days. We examined bone response using biochemical, densitometric, and monotonic, relaxation, and cyclic mechanical outcomes. Urinary deoxypyridinoline, a marker of bone resorption, was not significantly affected by running nor were tibial or femoral bone mineral density (BMD) (P > 0.05). Tibial mechanical properties following running were not decreased (P > 0.05). We did observe a slight decrease in displacement to failure (P < 0.05) and energy to failure (P = 0.10) of the proximal femur. These findings indicate that 14,000 physiologic loading cycles per day did not increase systemic bone resorption levels or substantially degrade the mechanical properties of long bone in adult rats. The lack of response to low magnitude, high cycle number physiologic loading is consistent with the view that a metabolic bone disturbance, in addition to repetitive loading, may be necessary for the development of the stress injury in the adult skeleton.

Absorptiometry, Photon↗
PubMed2001

Raman spectroscopic detection of changes in bioapatite in mouse femora as a function of age and in vitro fluoride treatment.

Laser Raman microprobe spectroscopy, which characterizes the molecular structure of a mineral, was used to analyze microscopically small regions of bioapatite in mouse femora in order to study the effect of mouse age and in vitro fluoride treatment on the bone mineral (i.e., mineral identity and degree of crystallinity). Both femora that had and those that had not undergone in vitro NaF treatment underwent point analysis of 1 micron spots in the center of the compact bone's cross-section. The Raman spectra of bones treated with fluoride showed a peak up-shift of the PO4 vibration mode from 961 to 964 delta cm-1 indicating a conversion from a carbonated hydroxylapatitic to a carbonated fluorapatitic mineral phase. The spectral band width of the 961 delta cm-1 PO4 vibration in femora of 4-, 10-, and 24-week-old mice showed that aging, as well as in vitro treatment with 1.5 M NaF for 12 hours, significantly increases the degree of crystallinity of the bioapatite. In vitro fluoridation of 10-week-old mouse femora increased the bioapatite's degree of crystallinity to about the same degree as did aging to 24 weeks. Four-point bending tests indicated that the age-related increase in crystallinity of untreated bones was associated with decreased deformation to failure, i.e., increased brittleness. In contrast, the increased crystallinity following fluoridation of 10-week-old bones was associated with increased deformation, i.e., increased ductility, perhaps due to the altered mineral composition. This study shows that the laser Raman microprobe readily detects the conversion of carbonated hydroxylapatite to carbonated fluorapatite, as well as changes in crystallinity of either mineral phase, in microscopically small regions of a bone sample.

Aging↗
PubMed2001

BMP-12 gene transfer augmentation of lacerated tendon repair.

Bone morphogenetic protein (BMP) 12 is a recently discovered member of the human BMP family. It is the human homolog of mouse growth/differentiation factor (GDF)-7. Previously we reported that injection of mesenchymal progenitor cells transferred with the BMP-12 gene into the muscles of nude mice induced tendon-like tissue formation. In this study, we further investigated the effect of BMP-12 gene transfer on tendon cells. We observed that adenovirus mediated in vitro BMP-12 gene transfer into chicken tendon cells increased type I collagen synthesis. No change in alkaline phosphatase activity was observed following BMP-12 gene transfer. We also determined that BMP-12 gene transfer into a complete tendon laceration chicken model resulted in a two-fold increase of tensile strength and stiffness of repaired tendons, indicating improved tendon healing in vivo. We conclude that BMP-12 gene transfer is a promising procedure for improving the tendon repair process.

Alkaline Phosphatase↗
PubMed2001

The effect of variations in applied rehabilitation force on collagen concentration and maturation at the intrasynovial flexor tendon repair site.

The biochemical means by which accelerated rehabilitation alters intrasynovial flexor tendon repair site collagen synthesis and extracellular matrix maturation are not fully understood. We hypothesized that an increased level of applied rehabilitative force in a clinically relevant animal model would hasten the maturation of the repair site extracellular matrix as demonstrated by total collagen and collagen cross-link assessment. Twenty-eight flexor digitorum profundus tendons from 14 adult dogs were transected and repaired. The animals received either low- or high-force rehabilitation and were killed 10, 21, and 42 days after surgery. A 10-mm segment of tendon surrounding the repair site was obtained. Biochemical analysis showed that total collagen concentration was significantly reduced at each time point, that the reducible cross-link ratio of dihydroxylysinonorleucine to hydroxylysinonorleucine was significantly increased at each time point, and that the nonreducible pyridinoline cross-link content was significantly decreased at 10 days in both rehabilitative groups. Total collagen content did not vary to a statistically significant degree with either time or as a function of rehabilitation type. Based on these findings several clinically relevant observations can be made. Increasing collagen concentration and repair site maturation do not explain the previously demonstrated increased tensile properties of tendon that occur between 3 and 6 weeks after repair. Higher force rehabilitation does not alter the biochemical composition of the healing tendon through 6 weeks. Coupled with other recent data these findings suggest that high-force rehabilitation does not stimulate accelerated healing after intrasynovial flexor tendon repair.

Animals↗
PubMed2001

The influence of cross-sectional area on the tensile properties of flexor tendons.

Clinicians have long noted substantial variation in the cross-sectional size of flexor tendons in the hand; however, data indicating that surgical repair techniques of lacerated flexor tendons should be altered according to size are unavailable. Our objectives were to evaluate the cross-sectional size differences among tendons within the same hand and to correlate tendon size with tensile mechanical properties after suture repair. Fifty human cadaver flexor digitorum profundus tendons were measured with digital calipers to determine radioulnar and volardorsal diameters. Twenty tendons were used to measure resistance to suture pull-through; tendons were transected at the A2 pulley, and a transverse double-stranded 4-0 Supramid suture (S. Jackson, Inc, Alexandria, VA) was passed through the radioulnar plane of the tendon 1 cm from the transection site. The remaining tendons were transected and repaired by using a modified Kessler repair with double-stranded 4-0 Supramid suture. Both tendon repairs and tendon-suture pull-through specimens were tested to failure in tension by using a material testing machine. Dorsovolar tendon height and tendon cross-sectional area varied significantly between digits, with an average difference of approximately 40% between the values of the smallest (fifth) and largest (third) fingers. Yield and ultimate force determined by pull-through tests of the simple transverse suture correlated positively with tendon radioulnar width. Tensile properties of tendons repaired with a double-stranded modified Kessler repair, however, did not depend significantly on tendon size. These results indicate that the strength of the commonly used Kessler suture technique is not dependent on tendon cross-sectional size within the clinically relevant range of tendons evaluated.

Humans↗