Biomolecular evaluation of cryopreserved amniotic membranes for ophthalmological use by ELISA and RT-PCR at one and eighteen months.

PURPOSE: To study the presence of certain proteins - EGF (epidermal growth factor), KGF (keratinocyte growth factor), IL-10 (interleukin 10), HGF (hepatocyte growth factor), Alpha2-macroglobulin and IL-1RA (interleukin 1 receptor antagonist) in cryopreserved amniotic membranes at 1 and 18 months and, as a secondary objective, to detect mRNA corresponding to KGF, IL-1Ra, Alpha2-macroglobulin, Fas Ligand, TGF beta (transforming growth factor beta) and Lumican by RT-PCR in membranes preserved at 1 and 18 months. MATERIAL AND METHODS: Four samples of amniotic membrane were divided into 2 groups: the first group (N=2) cryopreserved for 1 month and the second group (N=2) cryopreserved for 18 months, in order to be studied by RT-PCR and ELISA. RESULTS: RT-PCR detected KGF, IL-1Ra, Alpha2-macroglobulin, Fas Ligand, and Lumican. Of these, FAS Ligand mRNA was found in samples preserved for 1and 18 months. KGF, Lumican, and alpha2-microglobulin mRNA were found only at 1 month, and IL-1Ra mRNA was absent in both sample groups. RT-PCR for TGF-beta was inconclusive. ELISA was performed for detection and quantification of 6 proteins (EGF, KGF, IL-10, HGF, Alpha2-macroglobulin and IL-1Ra) in both amniotic membrane groups. All 6 proteins were found in all samples, with a lower concentration at 18 months compared to 1 month of preservation. CONCLUSION: This study shows that membranes cryopreserved in 50% glycerol for 18 months do retain the proteins necessary for regeneration of the corneal surface, giving these membranes their biochemical properties.

Long-term follow up of intracorneal ring segment implantation in 932 keratoconus eyes.

PURPOSE: To evaluate the progression of keratoconus in 932 eyes of 659 patients through visual, refractive and topographic data after intracorneal ring segment (ICRS) implantation. METHODS: Retrospective review of 659 patients who underwent ICRS (Intacs®) implantation for keratoconus between September 1997 and November 2017. Demographics, preoperative and postoperative uncorrected visual acuity (UCVA) and best-corrected visual acuity (BCVA) in LogMAR, corneal topography parameters (thinnest pachymetry, Kmax), central corneal pachymetry and total follow-up time were evaluated. Statistical analysis was performed using IBM SPSS Statistics 24.0 for windows. RESULTS: Nine hundred and thirty-two eyes of 659 patients, with a mean age of 30.41 years (range 11-76 years), were evaluated. Mean total follow up time was 3.02 years. Forty-one eyes had a total follow up of over 10 years. Both UCVA and BCVA improved significantly after ICRS implantation (P<0.01). Only 18 eyes (2.66% of eyes of patients under 35 years of age) were found to have progression of keratoconus based on postoperative topographic data (Mean age 23.00 years, 55.6% female, total follow-up 2 to 10 years) CONCLUSION: ICRS implantation showed long-term improvement and stability in visual and topographic results in a large case series of patients with keratoconus. Only a minimal rate of progression was detected in young patients. However, further prospective studies need to be conducted to better define predictability of postoperative visual outcomes and progression.

Effects of cross-linkage on fatigue life and failure modes of stainless steel posterior spinal constructs.

This study tested the effects of cross-linkage on the fatigue performance of posterior spinal constructs (i.e., AcroMed stainless steel Isola systems). The failure modes encountered during fatigue were also examined. The results of this study confirmed earlier findings that the use of cross-linkage does not significantly affect the stability of posterior constructs during axial loading. Their influence in torsion loading is much more pronounced. During the fatigue tests, posterior stainless steel spinal implants instrumented without cross-linkage reached 1 million cycles at 500- and 750-N loads. When the load was increased to 1,000 N, the number of cycles to failure dropped by two-thirds. These findings demonstrate that the endurance limit was between 750 N and 1,000 N for spinal constructs without cross-linkage, with the limit being closer to 750 N. Devices equipped with one or two cross-linkages reached 1 million cycles at 500 N. The number of cycles to failure dropped dramatically as the load was increased to 750 and 1,000 N. It appears that the endurance limits for spinal devices using cross-linkage should be 500 and 750 N, with the limit closer to the 500-N load. All rod fractures occurred near the junction between the longitudinal and transverse rods. Stress concentration was greatly in the vicinity of that contact point. These results should provide a basis for future improvement in endurance limits of spinal implants equipped with cross-linkage. Higher endurance limits will reduce the toxic effects encountered during fracture modes. The implants will also be better able to withstand the high physiologic loads experienced by obese individuals.

An experimental study of lumbar destabilization. Restabilization and bone density.

STUDY DESIGN: An investigation of the effects of bone density on lumbar spine stability using destabilizing and restabilizing procedures. OBJECTIVES: To measure cadaveric vertebral bone densities computed tomographic scans and to correlate the measured densities with lumbar spine stability in the intact and during sequential destabilization and restabilization. SUMMARY OF BACKGROUND DATA: The stabilizing effects of lumbar pedicle screw fixation have been widely described. Numerous construct failure mechanisms have been observed, including screw loosening in osteoporosis. Although previous studies have analyzed the effect of bone density on the compression strength of bone similar to that used in interbody fusion and the relationship of pedicle screw pull-out strength to vertebral bone density, a combined study of bone density and construct stability using an interbody bone spacer with pedicle fixation has not been performed. METHODS: Bone densities were measured in 20 human cadaveric lumbar spines using computed tomography scans and a hydroxyapatite phantom. After the specimens were mounted in a testing frame, the L4-L5 motion segments were subjected to cyclic axial compression-torsional loads, and axial and rotational intervertebral displacements were monitored. Laminectomy, facetectomy, and pedicle screw-plate fixation were performed sequentially in three specimens. Ten others underwent these procedures with an additional destabilization procedure, discectomy, after facetectomy. Seven others underwent the same sequence as the previous group, followed by the insertion of interbody bone. Cyclic testing was resumed after each procedure. RESULTS: Average bone densities varied widely among the specimens. Average bone densities of the pedicle and of the vertebral body for individual specimens were well-correlated (r = 0.897). Displacements were recorded as a percentage of the intact state before destabilization; average percentages are reported as follows: axial displacements increased after facetectomy (145%) and subsequent discectomy (251%), and rotational displacements increased after facetectomy (295%) and discectomy (390%). Instrumentation without interbody bone resulted in specimens with decreased axial (126%) and rotational (156%) displacements. The addition of interbody bone further decreased axial (111%) and rotational (117%) displacements. The rotational stabilization provided by instrumentation was well-correlated with vertebral bone density (r = 0.804). This correlation was enhanced by the use of interbody bone (r = 0.939). CONCLUSION: The unstable lumbar spine can be partially stabilized using fixation. Interbody bone provides additional stability. The immediate stability provided by pedicle screws is greater in lumbar vertebrae with higher bone density.

Biomechanical assessment of titanium and stainless steel posterior spinal constructs: effects of absolute/relative loading and frequency on fatigue life and determination of failure modes.

The goal of this study was to examine the effects of absolute/relative loads and frequency on the fatigue life of titanium and stainless steel posterior spinal constructs, and to determine the failure fracture modes. The stainless steel constructs had higher stiffness and yield strength than the titanium constructs, but the ultimate static strength was almost equal for both types of constructs. Titanium constructs, however, exhibited higher variability than the stainless steel constructs. In fatigue tests, the stainless steel constructs were significantly affected by the external load and were frequency independent. It appears from fatigue curves that 500 N can be approximated as the endurance limit for the stainless steel constructs. Titanium constructs were load-frequency dependent, and their endurance limit was somewhere between the 500 and 750 N load levels. There were no differences in performance between the stainless steel and titanium constructs at 16 Hz. At 4 Hz, titanium constructs performed as well or better than stainless steel constructs. Also, the titanium constructs resulted in better performance than the stainless steel constructs in the elastic region, and with smaller differences in the plastic region. Most of the failure modes for stainless steel constructs were in screw bending at 16 Hz with a smaller percentage of rod fractures at high loads, with a higher percentage of rod fractures observed for the stainless steel constructs at 4 Hz. Most of the failure modes for titanium constructs occurred in screw bending or fracture.