Dataset Information

Quantitative assessment of ultrastructure and light scatter in mouse corneal debridement wounds.

ABSTRACT:

Purpose

The mouse has become an important wound healing model with which to study corneal fibrosis, a frequent complication of refractive surgery. The aim of the current study was to quantify changes in stromal ultrastructure and light scatter that characterize fibrosis in mouse corneal debridement wounds.

Methods

Epithelial debridement wounds, with and without removal of basement membrane, were produced in C57BL/6 mice. Corneal opacity was measured using optical coherence tomography, and collagen diameter and matrix order were quantified by x-ray scattering. Electron microscopy was used to visualize proteoglycans. Quantitative PCR (Q-PCR) measured mRNA transcript levels for several quiescent and fibrotic markers.

Results

Epithelial debridement without basement membrane disruption produced a significant increase in matrix disorder at 8 weeks, but minimal corneal opacity. In contrast, basement membrane penetration led to increases in light scatter, matrix disorder, and collagen diameter, accompanied by the appearance of abnormally large proteoglycans in the subepithelial stroma. This group also demonstrated upregulation of several quiescent and fibrotic markers 2 to 4 weeks after wounding.

Conclusions

Fibrotic corneal wound healing in mice involves extensive changes to collagen and proteoglycan ultrastructure, consistent with deposition of opaque scar tissue. Epithelial basement membrane penetration is a deciding factor determining the degree of ultrastructural changes and resulting opacity.

SUBMITTER: Boote C

PROVIDER: S-EPMC3367468 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature

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Publications

Quantitative assessment of ultrastructure and light scatter in mouse corneal debridement wounds.

Boote Craig C Du Yiqin Y Morgan Sian S Harris Jonathan J Kamma-Lorger Christina S CS Hayes Sally S Lathrop Kira L KL Roh Danny S DS Burrow Michael K MK Hiller Jennifer J Terrill Nicholas J NJ Funderburgh James L JL Meek Keith M KM

Investigative ophthalmology & visual science 20120514 6

<h4>Purpose</h4>The mouse has become an important wound healing model with which to study corneal fibrosis, a frequent complication of refractive surgery. The aim of the current study was to quantify changes in stromal ultrastructure and light scatter that characterize fibrosis in mouse corneal debridement wounds.<h4>Methods</h4>Epithelial debridement wounds, with and without removal of basement membrane, were produced in C57BL/6 mice. Corneal opacity was measured using optical coherence tomogra ...[more]

PMID: 22467580

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Project description:Although sensory reinnervation occurs after injury in the peripheral nervous system, poor reinnervation in the elderly and those with diabetes often leads to pathology. Here we quantify sub-basal axon density in the central and peripheral mouse cornea over time after three different types of injury. The mouse cornea is highly innervated with a dense array of sub-basal nerves that form a spiral called the vortex at the corneal center or apex; these nerves are readily detected within flat mounted corneas. After anesthesia, corneal epithelial cells were removed using either a dulled blade or a rotating burr within an area demarcated centrally with a 1.5 mm trephine. A third wound type, superficial trephination, involved demarcating the area with the 1.5 mm trephine but not removing cells. By 7 days after superficial trephination, sub-basal axon density returns to control levels; by 28 days the vortex reforms. Although axon density is similar to control 14 days after dulled blade and rotating burr wounding, defects in axon morphology at the corneal apex remain. After 14 days, axons retract from the center leaving the sub-basal axon density reduced by 37.2 and 36.8% at 28 days after dulled blade and rotating burr wounding, respectively, compared with control. Assessment of inflammation using flow cytometry shows that persistent inflammation is not a factor in the incomplete reinnervation. Expression of mRNAs encoding 22 regeneration-associated genes involved in axon targeting assessed by QPCR reveals that netrin-1 and ephrin signaling are altered after wounding. Subpopulations of corneal epithelial basal cells at the corneal apex stop expressing ki67 as early as 7 days after injury and by 14 and 28 days after wounding, many of these basal cells undergo apoptosis and die. Although sub-basal axons are restored to their normal density and morphology after superficial trephination, sub-basal axon recovery is partial after debridement wounds. The increase in corneal epithelial basal cell apoptosis at the apex observed at 14 days after corneal debridement may destabilize newly reinnervated sub-basal axons and lead to their retraction toward the periphery.

Project description:PurposeA major challenge for accurate quantitative SPECT imaging of some radionuclides is the inadequacy of simple energy window-based scatter estimation methods, widely available on clinic systems. A deep learning approach for SPECT/CT scatter estimation is investigated as an alternative to computationally expensive Monte Carlo (MC) methods for challenging SPECT radionuclides, such as 90Y.MethodsA deep convolutional neural network (DCNN) was trained to separately estimate each scatter projection from the measured 90Y bremsstrahlung SPECT emission projection and CT attenuation projection that form the network inputs. The 13-layer deep architecture consisted of separate paths for the emission and attenuation projection that are concatenated before the final convolution steps. The training label consisted of MC-generated "true" scatter projections in phantoms (MC is needed only for training) with the mean square difference relative to the model output serving as the loss function. The test data set included a simulated sphere phantom with a lung insert, measurements of a liver phantom, and patients after 90Y radioembolization. OS-EM SPECT reconstruction without scatter correction (NO-SC), with the true scatter (TRUE-SC) (available for simulated data only), with the DCNN estimated scatter (DCNN-SC), and with a previously developed MC scatter model (MC-SC) were compared, including with 90Y PET when available.ResultsThe contrast recovery (CR) vs. noise and lung insert residual error vs. noise curves for images reconstructed with DCNN-SC and MC-SC estimates were similar. At the same noise level of 10% (across multiple realizations), the average sphere CR was 24%, 52%, 55%, and 67% for NO-SC, MC-SC, DCNN-SC, and TRUE-SC, respectively. For the liver phantom, the average CR for liver inserts were 32%, 73%, and 65% for NO-SC, MC-SC, and DCNN-SC, respectively while the corresponding values for average contrast-to-noise ratio (visibility index) in low-concentration extra-hepatic inserts were 2, 19, and 61, respectively. In patients, there was high concordance between lesion-to-liver uptake ratios for SPECT reconstruction with DCNN-SC (median 4.8, range 0.02-13.8) compared with MC-SC (median 4.0, range 0.13-12.1; CCC = 0.98) and with 90Y PET (median 4.9, range 0.02-11.2; CCC = 0.96) while the concordance with NO-SC was poor (median 2.8, range 0.3-7.2; CCC = 0.59). The trained DCNN took ~ 40 s (using a single i5 processor on a desktop computer) to generate the scatter estimates for all 128 views in a patient scan, compared to ~ 80 min for the MC scatter model using 12 processors.ConclusionsFor diverse 90Y test data that included patient studies, we demonstrated comparable performance between images reconstructed with deep learning and MC-based scatter estimates using metrics relevant for dosimetry and for safety. This approach that can be generalized to other radionuclides by changing the training data is well suited for real-time clinical use because of the high speed, orders of magnitude faster than MC, while maintaining high accuracy.

Dataset Information

Quantitative assessment of ultrastructure and light scatter in mouse corneal debridement wounds.

Purpose

Methods

Results

Conclusions

Publications

Quantitative assessment of ultrastructure and light scatter in mouse corneal debridement wounds.

Similar Datasets

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