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PubMed1994

Uncemented total shoulder arthroplasty. A review.

Clinicians have had much experience with uncemented humeral components. A press-fitted humeral component will usually remain stable in the absence of a glenoid component. From the information available, surgeons should not continue to use press-fitted humeral components for total shoulder arthroplasty. Tissue ingrowth humeral components offer promise; however, the reports to date have short or intermediate length followup, and radiographic results do not equal those of cemented components. Early clinical results with tissue ingrowth glenoid components are excellent; radiographically evident changes occur much less frequently than they do following cement fixation. However, their disadvantage is the possibility of accelerated polyethylene wear and subsequent metal-induced synovitis. As such, there are no clear cut indications for cement fixation versus tissue ingrowth fixation for the glenoid component other than those intuitively based on bone quality and quantity in the glenoid fossa. Accruing experience will help to define the indications better, but given the similarity of clinical outcomes to date, distinction between the options may be difficult.

Arthroplasty↗
PubMed1996

A simplified and rapid procedure for in situ hybridization on human, flash-frozen, post-mortem brain and its combination with immunohistochemistry.

A simplified and rapid method is described for in situ hybridization (ISHH) studies of human post-mortem brain. Brain tissue was dissected into slices and was flash-frozen at -70 degrees C for storage. ISHH was carried out on 12 microns cryostat sections, post-fixed in 4% paraformaldehyde. The histology of human brain tissue prepared by this technique rivalled that of formalin-fixed, wax-embedded tissue. In ISHH studies, flash-frozen tissue gave superior results to those obtained following long-term fixation of tissue in 10% formalin with subsequent wax-embedding, or short-term prefixation in 4% paraformaldehyde. A systematic evaluation of commonly employed preparative procedures for ISHH was carried out on flash-frozen brain and a simplified protocol, consisting only of fixation and dehydration, was developed as a result of these studies. Specific hybridization of probes to a number of mRNA species was demonstrable in neurons in different brain regions. Using 0.5% glutaraldehyde/4% paraformaldehyde post-fixation, immunohistochemical labelling of TH-positive cortical catecholaminergic neurons and striatal dopaminergic terminals was successfully demonstrated in flash-frozen tissue. The same fixation technique also allowed combination of ISHH and immunohistochemistry for the simultaneous demonstration of tyrosine hydroxylase mRNA and peptide in neurons of human brain stem and cortex. mRNA and peptides in flash-frozen tissue were found to be stable for more than 3 years. ISHH could be readily performed on relatively large brain structures. In addition to permitting excellent ISHH and immunohistochemistry, alone or in combination, flash-freezing allows the maximum versatility of tissue use and does not compromise its study by other neuroscience techniques.

Aged↗
PubMed1986

Aminoalkylsilane-treated glass slides as support for in situ hybridization of keratin cDNAs to frozen tissue sections under varying fixation and pretreatment conditions.

Attempts to investigate the cellular localization of keratin mRNAs by in situ hybridization with specific [35S]-labelled cDNA probes to mouse epithelia have been seriously impeded by the uncontrollable detachment of frozen tissue sections on conventionally coated glass slides (i.e. those coated with egg white, gelatin, collagen). Similarly, a variety of other coating and attachment devices have proved to be unsatisfactory or impracticable for large scale investigations. These difficulties were completely overcome and in situ hybridization was possible after a short immersion of the glass slides in a 2% solution of 3-aminopropyltriethoxysilane in acetone. This treatment provides the glass surface with aminoalkyl groups which are apparently able to react covalently with aldehyde or ketone functions of frozen tissue sections. The resulting firm adhesion of the sections enabled us to investigate the influence of different fixation and prehybridization procedures on the quality of the in situ hybridization. It was found that especially harsh prehybridization, involving hydrochloric acid, heat and proteinase K treatment, drastically reduces the morphological integrity of the sections, thus rendering a reliable assignment of the label difficult. In contrast, a mild prehybridization, consisting mainly of a rehydration of the sections in phosphate-buffered saline and equilibration in 0.1 M glycine, leaves the morphology intact and leads to a highly efficient and specific in situ hybridization.

Animals↗
PubMed1999

A high frequency of sequence alterations is due to formalin fixation of archival specimens.

Genomic analysis of archival tissues fixed in formalin is of fundamental importance in biomedical research, and numerous studies have used such material. Although the possibility of polymerase chain reaction (PCR)-introduced artifacts is known, the use of direct sequencing has been thought to overcome such problems. Here we report the results from a controlled study, performed in parallel on frozen and formalin-fixed material, where a high frequency of nonreproducible sequence alterations was detected with the use of formalin-fixed tissues. Defined numbers of well-characterized tumor cells were amplified and analyzed by direct DNA sequencing. No nonreproducible sequence alterations were found in frozen tissues. In formalin-fixed material up to one mutation artifact per 500 bases was recorded. The chance of such artificial mutations in formalin-fixed material was inversely correlated with the number of cells used in the PCR-the fewer cells, the more artifacts. A total of 28 artificial mutations were recorded, of which 27 were C-T or G-A transitions. Through confirmational sequencing of independent amplification products artifacts can be distinguished from true mutations. However, because this problem was not acknowledged earlier, the presence of artifacts may have profoundly influenced previously reported mutations in formalin-fixed material, including those inserted into mutation databases.

Base Sequence↗
PubMedDate not supplied

Fixation methods for implantable port chamber: comparative study using glue, self-stabilizing leg and suture fixations in rabbits.

OBJECTIVE: To evaluate the fixation strength and tissue reaction of the glue fixation and self-stabilizing leg fixation methods and to compare the results with those of the conventional tagging suture fixation method. MATERIALS AND METHODS: Twelve healthy rabbits were selected and three different methods of implanting the port chamber were employed on the back of each rabbit. A total of thirty six port chambers were implanted with these three different methods, viz. the glue fixation method using tissue adhesive, the self-stabilizing leg method using a self-expandable stabilizing leg, and the suture fixation method. The fixation strength and the gross and histopathologic changes of each fixation method were evaluated at three days, one week, two weeks and four weeks after port implantation. RESULTS: The glue fixation method showed a good fixation strength, which was similar to that of the tagging suture method (p = 0.3486). Five of the six ports (83%) implanted with the glue fixation method which were examined after two weeks showed cracks on the external surface, but this had no adverse effects on their function. A large amount of granulation tissue reaction was found at the bottom of the chamber (p = 0.0025). The fixation with the self-stabilizing leg showed relatively lower fixation strength (p = 0.0043), but no turning-over of the chamber occurred. The fixation strength improved with time after the first week, and minimal granulation tissue reaction was observed with this method. CONCLUSION: The glue fixation method exhibited equal fixation strength compared to the suture fixation, but showed cracking and a large amount of granulation tissue, whereas the fixation with a self-stabilizing leg showed weaker fixation strength.

Alloys↗
PubMed1991

Differential antigen preservation during tissue autolysis.

Immediate fixation or snap freezing of tissue is ordinarily done to maximize antigen preservation for immunocytochemistry; however, delay in tissue allocation or spontaneous lymph node infarction can render tissue suboptimal for immunostaining. To test the effects of tissue autolysis/necrosis on the preservation of various lymphoid, epithelial, and mesenchymal markers, two lymph nodes (one with reactive lymphoid hyperplasia and one with metastatic ductal breast carcinoma) were evaluated for immunocytochemically demonstrated antigen preservation at 0-, 4-, 8-, 12-, 24-, 48-, and 72-hour intervals of autolysis at 37 degrees C. All specimens were stained by frozen section and formalin-fixed paraffin section immunocytochemical reactions with antibodies against CLA (CD45), UCHL-1 (CD45RO), L-26, kappa, lambda, anti-epithelial keratins (AE-1 and AE-3), epithelial membrane antigen, and vimentin. Frozen sections were additionally stained for Leu-1 (CD5), Leu-2a (CD8), Leu-3a+b (CD4), Leu-4 (CD3), and Leu-14 (CD22). The most resilient lymphoid antigen preservation was observed with CLA and UCHL-1, both exhibiting immunoreactivity at 72 hours in both frozen and fixed preparations. L-26 showed similar reactivity in frozen sections, but detectable antigen was observed only up to 24 hours in formalin-fixed tissue. Leu-2a proved to be the most labile antigen, persisting for only 12 hours in frozen sections. The epithelial markers epithelial membrane antigen and AE-1 exhibited excellent antigenic preservation in both frozen and fixed preparations; AE-3 persisted well in frozen section but was not demonstrated in fixed tissue. Vimentin immunoreactivity was vastly superior in frozen, as compared with fixed, tissue sections. Most antigens showed remarkable preservation despite morphologic degradation; however, differential antigenic resilience was demonstrated. Knowledge of this variation in antigen decay is critical for evaluation of immunoperoxidase phenotypic studies of autolyzed or necrotic tissue.

Antigens↗
PubMed1991

Preparation of inner ear sensory hair bundles for high resolution scanning electron microscopy.

Chemical fixation techniques for preservation of sensory hair bundles in the mammalian inner ear for scanning electron microscopy (SEM) are reviewed. Fixatives employed were glutaraldehyde, glutaraldehyde-picrate, glutaraldehyde-tannic acid, glutaraldehyde-formaldehyde, glutaraldehyde followed by postfixation with osmium tetroxide and the osmium thiocarbohydrazide (OTOTO) method. Dehydration was routinely accomplished with ascending grades of acetone followed by critical point drying with liquid CO2 or fluorocarbon sublimation. Specimens other than those prepared by the OTOTO method were metal coated with gold, gold-palladium or platinum. Material was viewed at high resolution (2-3 nm) in a transmission electron microscope (TEM) fitted with a scanning system and an LaB6 filament. A few specimens, which were either coated with platinum, carbon or uncoated, were examined in a field emission SEM. We have concluded that glutaraldehyde fixation followed by critical point drying with CO2 and coating with platinum gives the best general preservation of stereocilia and their cross-links for routine high resolution SEM, but that carbon-coated or uncoated specimens offer potentially better results free from metal coating artifacts when viewed with field emission SEM. These methods have enabled us to make novel observations upon the surface detail and cross-links of stereocilia which have helped considerably in understanding the mechanical properties of hair bundles particularly in relation to sensory transduction. We have found that stereocilial surface detail and cross-links are sensitive to fixation regimens. In particular they are degraded by exposure to osmium tetroxide; they are also highly labile since deleterious changes in their appearance can be detected as early as 15 minutes following death.

Animals↗
PubMed2003

Removal of mercuric chloride deposits from B5-fixed tissue will affect the performance of immunoperoxidase staining of selected antibodies.

B5 clearing is a step used before immunoperoxidase staining to remove the precipitated mercuric chloride deposits caused by B5 fixation of tissue. In the B5 clearing procedure, the slides are treated with Lugol's iodine and 5% sodium thiosulfate before antigen retrieval and the application of the primary antibody. The goal of this project was to study the effect of the B5 clearing protocol on immunoperoxidase staining on paraffin-embedded tissue, which has not been previously reported in a series of antibodies. We evaluated 75 antibodies using the 2-step clearing protocol and performed paired immunoperoxidase staining on the Ventana ES instrument, with and without the clearing protocol. We found that among 75 antibodies studied, 3 (CD5, CD30, and synaptophysin) showed total obliteration of reactivity, and 3 (ALK, Ulex, and GFAP) showed partial reduction of the staining compared with the controls. Pathologists must be aware of the possible false-negative staining effect caused by the routinely used B5 clearing protocol. Control tissues must receive the same clearing protocol (i.e., placed on case slides) to ensure detection of this effect.

Antibodies↗
PubMed1973

Cyclic adenosine monphosphate, metabolites, and phosphorylase in neural tissue: a comparison a methods of fixation.

Fixation of rat brain tissue by freeze-blowing, microwave irradiation, immersion of whole rats in liquid nitrogen, and decapitation into liquid nitrogen indicates that postmortem changes in metabolites and enzyme forms are minimal in freeze-blown brains. Cyclic adenosine monophosphate levels are lowest in microwave-irradiated brains, which has been interpreted by some investigators to indicate rapid fixation and minimal anoxia. However, the changes in phosphocreatine, adenosine triphosphate, lactate, and phosphorylase clearly demonstrate that fixation by freeze-blowing or immersion in liquid nitrogen more closely approximate the state in vivo.

Adenosine Triphosphate↗
PubMed2006

Experimental results of the tracheoesophageal tissue connector for improved fixation of shunt valves in laryngectomized patients.

BACKGROUND: After total laryngectomy and voice rehabilitation using a tracheoesophageal shunt valve, patients often have valve-related complications such as leakage. To solve these problems, a tracheoesophageal tissue connector (TE-TC) was devised to serve as an interface between the patient's tissue (trachea and esophagus) and the shunt valve. METHODS: The TE-TC is a permucosal connection constructed from a titanium ring (filled with a silicon rubber plug) combined with polypropylene or titanium mesh. After implantation in adult goats for 12 weeks the implants were submitted to histologic investigation. RESULTS: Firm implant fixation was achieved. In nearly all (18/19), no signs of infection of the implant were seen; 11 of 19 animals died before the end of the experiment owing to complications not related to the implant. CONCLUSIONS: The TE-TC is a new device with potential in the solution for fixation-related problems in tracheoesophageal voice rehabilitation.

Animals↗
PubMed1997

[Reduction of the weight of prostatic tissue during transurethral resection].

OBJECTIVE: To study the weight reduction of prostatic tissue removed during transurethral resection. METHOD: 22 hyperplastic prostate glands enucleated via a transvesical approach were resected in vitro, under conditions as close as possible to in vivo resection in 11 cases. The authors compared the weight of the enucleated prostates with the weight of fresh resection chips and the weight of chips fixed in Bouin solution. RESULTS: The weight of the chips was 28% lower than that of the prostates, while the weight of chips fixed in Bouin solution was 8% higher than that of the fresh chips. CONCLUSION: Transurethral resection causes a 28% loss of the weight of resected prostatic tissue and fixation in Bouin solution of these same chips increases their weight by an average of 8%.

Acetic Acid↗
PubMed2006

Hybrid femoral fixation of soft-tissue grafts in anterior cruciate ligament reconstruction using the EndoButton CL and bioabsorbable interference screws: a biomechanical study.

PURPOSE: The purpose of this study was to evaluate the effect of hybrid femoral fixation with bioabsorbable interference screws (BioRCI; Smith & Nephew Endoscopy, Andover, MA) and EndoButton CL (Smith & Nephew Endoscopy) fixation. METHODS: Biomechanical testing of 3 different fixation techniques was performed by use of porcine hind-limb distal femurs and mature bovine extremity common extensor tendons. Two independent testing sessions were examined. The first testing session (group A) compared femoral fixation via the EndoButton CL device (n = 6) with femoral fixation via the EndoButton CL device with the addition of a BioRCI screw (n = 6). The second testing session (group B) compared femoral fixation via BioRCI screws alone (n = 6) with femoral fixation via the EndoButton CL device with the addition of a BioRCI screw (n = 6). The femur-graft complex was cyclically loaded between 50 and 250 N at 1 Hz for 1,000 cycles. After cycling, the amount of graft slippage was determined by measuring the change in grip-to-grip distance. The complex was then loaded to failure at 1 mm/s, and the ultimate tensile strength, stiffness, and mode of failure were determined. RESULTS: In group A the addition of an interference screw to the EndoButton CL fixation increased the ultimate tensile strength (1,364.7 +/- 102.4 N for EndoButton CL alone v 1,449.3 +/- 94.4 N for combined technique, P = .035) and stiffness (195.5 +/- 12.1 N/mm for EndoButton CL alone v 307.3 +/- 54.9 N/mm for combined technique, P = .004) and decreased the amount of graft slippage (2.6 +/- 0.5 mm for EndoButton CL alone v 2.0 +/- 0.3 mm for combined technique, P = .017). In group B the addition of the EndoButton CL device to interference screw fixation significantly increased the ultimate tensile strength (643.5 +/- 148.4 N for BioRCI screws alone v 1,290.3 +/- 254.4 N for combined technique, P = .004) but had no effect on stiffness (315.7 +/- 38.9 N/mm for BioRCI screws alone v 341.5 +/- 64.0 N/mm for combined technique, P = .267) or graft slippage (2.7 +/- 1.0 mm for BioRCI screws alone v 2.0 +/- 0.6 mm for combined technique, P = .087). CONCLUSIONS: Our study shows that hybrid femoral fixation of double-looped gracilis-semitendinosus grafts via the EndoButton CL device and a bioabsorbable interference screw is stronger than interference or EndoButton CL fixation alone with respect to ultimate tensile strength, stiffness, and slippage. The addition of an interference screw to suspensory fixation via the EndoButton CL device increased the ultimate tensile strength from 1,360 N to 1,450 N, improved reconstruction stiffness from 200 N/mm to 300 N/mm, and decreased the amount of graft slippage resulting from cyclic loading from 2.6 mm to 2.0 mm. CLINICAL RELEVANCE: The hybrid fixation of the EndoButton CL device and an interference screw is a stronger and stiffer construct than either device alone and allows for aperture fixation, which may translate into better clinical results.

Absorbable Implants↗
PubMed1978

Ultrastructural localization of cationic proteins in human polymorphonuclear leukocytes.

The present investigation is concerned with the use of the post-formalin ammoniacal silver reaction to detect the arginine-rich cationic proteins in human polymorphonuclear leukocytes at the ultrastructural level. These proteins appear to function as neutral proteases in antibacterial action and as mediators of inflammation. Originally, the ammoniacal silver reaction relied upon primary fixation in dilute formalin which prevented optimum fixation of tissues. This study shows that by using the proper sequence of glutaraldehyde fixation and the ammoniacal silver solution in conjunction with osmium tetroxide treatment, better fixation of the tissue and localization of the ammoniacal silver reaction can be achieved. Also, the ammoniacal silver reaction in human polymorphonuclear leukocytes is exclusively located in the large, crystalline cytoplasmic granules of eosiniphils. All other cytoplasmic granules of neutrophils, eosinophils, and basophils were found to be devoid of the ammoniacal silver reaction product. These results are contrary to previously published experimental data. Possible explanations for this discrepancy are discussed.

Ammonia↗
PubMed2005

Compaction of a bone dowel in the tibial tunnel improves the fixation stiffness of a soft tissue anterior cruciate ligament graft: an in vitro study in calf tibia.

BACKGROUND: Despite increasing attention on fixation of a soft tissue anterior cruciate ligament graft in the tibia, there have been no studies on the use of a bone dowel as a joint line fixation device for promoting fixation properties, especially stiffness at the time of implantation. PURPOSE: To determine whether compacting a bone dowel into the tibial tunnel improves fixation stiffness, yield load, and resistance to slippage of a soft tissue anterior cruciate ligament graft. STUDY DESIGN: Controlled laboratory study. METHODS: A double-looped tendon graft was fixed at the distal end of the tibial tunnel with a WasherLoc in 24 calf tibias. The tibial tunnels were treated with or without a dowel of cancellous bone. The bone dowel was harvested from the tibial tunnel and then compacted into a tapered space anterior to the anterior cruciate ligament graft as a joint line fixation device. A cyclic load and measurement test was administered to determine fixation stiffness, yield load, slippage, and failure mode. RESULTS: The specimens with the bone dowel had 58 N/mm more stiffness (P = .04); however, the yield load and resistance to slippage were similar in specimens with and without the bone dowel. CONCLUSIONS: A bone dowel harvested from the tibial tunnel can be used as a joint line fixation device in series with a distal fixation method to improve initial fixation stiffness and increase the fit, which is known to enhance tendon graft-to-bone healing in the tibia.

Animals↗
PubMed1975

[Intramedullary fixation of fragments in soft-tissue injuries (author's transl)].

In cases of open fractures, careful reconstruction of the soft tissue and stable internal fixation are generally regarded as the best means of preventing infection. With this in mind, some of the author's own cases of open fractures of the upper and lower leg-stabilized either with conventional intramedullary nails or with interlocking nails-are demonstrated. The infection rate in the especially vulnerable open fracture of the tibia was 7.89 percent.

Adolescent↗