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1.
J Biomed Mater Res A ; 100(4): 918-28, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22275163

ABSTRACT

Revision surgery for particle-induced implant loosening in total joint replacement is expected to increase dramatically over the next few decades. This study was designed to investigate if local tissue and serum markers of bone remodeling reflect implant fixation following administration of lipopolysaccharide (LPS)-doped polyethylene (PE) particles in a rat model. Twenty-four rats received bilateral implantation of intramedullary titanium rods in the distal femur, followed by weekly bilateral intra-articular injection of either LPS-doped PE particles (n = 12) or vehicle that contained no particles (n = 12) for 12 weeks. The group in which the particles were injected had increased serum C-terminal telopeptide of type I collagen (CTX-I), decreased serum osteocalcin (OC), increased peri-implant eroded surface, decreased peri-implant bone volume, and decreased mechanical pull-out strength compared to the controls. Implant fixation strength was positively correlated with peri-implant bone volume and serum OC and inversely correlated with serum CTX-I, while energy to yield was positively correlated with serum OC and inversely correlated with the number of tartrate-resistant acid phosphatase positive cells at the interface and the amount of peri-implant eroded surface. There was no effect on trabecular bone volume at a remote site. Thus, the particle-induced impaired fixation in this rat model was directly associated with local and serum markers of elevated bone resorption and depressed bone formation, supporting the rationale of exploring both anticatabolic and anabolic strategies to treat and prevent particle-related implant osteolysis and loosening, and indicating that serum markers may prove useful in tracking implant fixation.


Subject(s)
Biomarkers , Bone Remodeling , Prostheses and Implants , Animals , Male , Models, Animal , Rats , Rats, Sprague-Dawley
2.
PLoS One ; 5(10)2010 Oct 01.
Article in English | MEDLINE | ID: mdl-20957030

ABSTRACT

Enhanced understanding of differential gene expression and biological pathways associated with distinct phases of intramembranous bone regeneration following femoral marrow ablation surgery will improve future advancements regarding osseointegration of joint replacement implants, biomaterials design, and bone tissue engineering. A rat femoral marrow ablation model was performed and genome-wide microarray data were obtained from samples at 1, 3, 5, 7, 10, 14, 28, and 56 days post-ablation, with intact bones serving as controls at Day 0. Bayesian model-based clustering produced eight distinct groups amongst 9,062 significant gene probe sets based on similar temporal expression profiles, which were further categorized into three major temporal classes of increased, variable, and decreased expression. Osteoblastic- and osteoclastic-associated genes were found to be significantly expressed within the increased expression groups. Chondrogenesis was not detected histologically. Adipogenic marker genes were found within variable/decreased expression groups, emphasizing that adipogenesis was inhibited during osteogenesis. Differential biological processes and pathways associated with each major temporal group were identified, and significantly expressed genes involved were visually represented by heat maps. It was determined that the increased expression group exclusively contains genes involved in pathways for matrix metalloproteinases (MMPs), Wnt signaling, TGF-ß signaling, and inflammatory pathways. Only the variable expression group contains genes associated with glycolysis and gluconeogenesis, the notch signaling pathway, natural killer cell mediated cytotoxicity, and the B cell receptor signaling pathway. The decreased group exclusively consists of genes involved in heme biosynthesis, the p53 signaling pathway, and the hematopoietic cell lineage. Significant biological pathways and transcription factors expressed at each time point post-ablation were also identified. These data present the first temporal gene expression profiling analysis of the rat genome during intramembranous bone regeneration induced by femoral marrow ablation.


Subject(s)
Bone Marrow , Bone Regeneration , Femur , Gene Expression Profiling , Animals , Bayes Theorem , Male , Matrix Metalloproteinases/metabolism , Oligonucleotide Array Sequence Analysis , Rats , Rats, Sprague-Dawley , Transforming Growth Factor beta/metabolism , Wnt Proteins/metabolism
3.
Anat Rec (Hoboken) ; 292(2): 214-24, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19089906

ABSTRACT

The early events associated with axonal growth into 10-mm nerve gaps were studied histologically in the rat sciatic nerve model to determine if the outgrowth of blood vessels, Schwann cells, and axons could be enhanced. In the first two experimental groups, collagen nerve guides were filled with either saline or Matrigel. Marrow-derived mesenchymal stem cells (MSCs) were added to Matrigel in two other groups, one of which contained cells transfected with VEGF (MSC/VEGF). After 21 days, the injury site was exposed, fixed, sectioned, and volume fractions of the conduit contents were determined by point counting. The bioresorbable collagen conduits appropriately guided the axons and vessels in a longitudinal direction. The volume fraction of axons was significantly greater in the group with saline when compared with all three groups with Matrigel. This measure had a significant positive correlation with actual counts of myelinated axons. The blood vessel volume fraction in the Matrigel group decreased compared with the saline group, but was restored in the MSC/VEGF group. All Matrigel groups had comparable cellularity and showed a distribution of residual Matrigel in acellular zones. The saline group, by contrast, sustained a network of delicate fibroblastic processes that compartmentalized the nerve and its natural matrix as it became infiltrated by axons as minifascicles. In conclusion, the reduction of axonal outgrowth in the Matrigel groups, when compared with the saline group, suggests that Matrigel may impede the early regenerative process even when enriched by the addition of MSCs or VEGF-transfected cells.


Subject(s)
Axons/physiology , Guided Tissue Regeneration/methods , Mesenchymal Stem Cells/physiology , Nerve Regeneration/physiology , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/physiology , Animals , Axons/ultrastructure , Collagen , Drug Combinations , Laminin , Mesenchymal Stem Cells/ultrastructure , Mice , Proteoglycans , Rats , Rats, Inbred F344 , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Tissue Culture Techniques , Tissue Engineering , Tissue Scaffolds , Transfection
4.
J Neurosci Methods ; 172(2): 236-44, 2008 Jul 30.
Article in English | MEDLINE | ID: mdl-18562013

ABSTRACT

A new partial nerve lesion (PNL) model is needed to better simulate traumatic lesions seen clinically that result in both dysfunction and neuropathic pain. We assessed surgical variability and several outcome measures including histology during the acute postoperative period. A surgical lesion was created in the rat tibial nerve by removing a segment, later confirmed by myelinated axon counts. Variability in the model was assessed with four different outcome measures during the first postoperative week (n=24), with additional histological outcomes at 7 days (n=13) and pain testing at 21 days (n=9). At 7 days postoperative, the PNL resulted in a tibial functional index (TFI) of -41.3% distinct from a percent motor deficit (PMD) of -76.3%. However, the respective deficits from 2 to 7 days were similar. Either test could detect outliers, but PMD measurements had a lower coefficient of variation and were easier to perform and analyze. The deleted segment contained 26% of the myelinated axons and resulted in distal degeneration that was either 46% based on axon counts or 54% based on area. Replicated experiments confirmed the PMD, muscle atrophy, and formation of neuropathic pain. In conclusion, our partial lesion histologically progresses twofold during the first postoperative week with profound behavioral deficits involving both motor and sensory loss. These results based on sensitive and correlative outcome measures support the application of this novel model in experimental nerve lesion studies.


Subject(s)
Neurosurgical Procedures/methods , Peripheral Nerve Injuries , Peripheral Nerves/physiopathology , Peripheral Nervous System Diseases/physiopathology , Wounds and Injuries/physiopathology , Animals , Axons/pathology , Behavior, Animal/physiology , Denervation/adverse effects , Disability Evaluation , Disease Models, Animal , Disease Progression , Female , Muscle, Skeletal/innervation , Muscle, Skeletal/physiopathology , Muscular Atrophy/pathology , Muscular Atrophy/physiopathology , Paralysis/etiology , Paralysis/pathology , Paralysis/physiopathology , Peripheral Nerves/surgery , Peripheral Nervous System Diseases/pathology , Rats , Rats, Inbred F344 , Sensation Disorders/etiology , Sensation Disorders/pathology , Sensation Disorders/physiopathology , Tibial Nerve/injuries , Tibial Nerve/physiopathology , Tibial Nerve/surgery , Time Factors , Wallerian Degeneration/pathology , Wallerian Degeneration/physiopathology , Wounds and Injuries/pathology
5.
Anat Rec ; 266(1): 21-9, 2002 01 01.
Article in English | MEDLINE | ID: mdl-11748568

ABSTRACT

The aim of our study was to provide quantitative data on pudendal motor neuron cell bodies and axons in the female rat. To confirm earlier studies, fluorescent retrograde tracers were used to label the motor neurons for correlation with myelinated axon counts along the length of the motor pudendal nerve. The external urethral sphincter of female rats was injected with diamidino yellow and the external anal sphincter with fast blue. The L(6) spinal cord revealed labeled motor neurons. Those in the dorsolateral column (60.8 +/- 10.6) had nuclei labeled yellow from the external urethral sphincter and those in the dorsomedial column (31.7 +/- 8.5) had cytoplasm labeled blue from the external anal sphincter. Double labeling was not present, suggesting that pudendal motor neurons in each column innervate separate sphincters. The motor pudendal nerve in the ischiorectal fossa was also characterized by light microscopy. The mean myelinated axon count (151.4 +/- 17.0) was highly correlated (r = 0.995) in the proximal fascicles and the sum of distal fascicles. This indicated that myelinated axons do not branch at the point where the main motor pudendal nerve branches into separate fascicles. Axon counts between sides were not as well correlated (r = 0.883). The ratio of motor neurons to myelinated axons is 56%, suggesting that some myelinated axons either innervate other muscles or are sensory. This reproducible characterization of the normal pudendal nerve anatomy provides an excellent basis for experimental studies associated with pudendal nerve denervation as a model for neurogenic incontinence.


Subject(s)
Motor Neurons/cytology , Perineum/innervation , Spinal Cord/anatomy & histology , Anal Canal/innervation , Animals , Axonal Transport/physiology , Female , Fluorescent Dyes , Lumbosacral Plexus/anatomy & histology , Microscopy, Fluorescence , Motor Neurons/physiology , Nerve Fibers, Myelinated , Neural Pathways/anatomy & histology , Neural Pathways/physiology , Rats , Rats, Sprague-Dawley , Urethra/innervation
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