In conclusion, the authors suggest that the decellularization of animal corneas with 1.5 M NaCl represents a useful method for the development of human bioengineered corneas with therapeutic potential.
Advances in the development of cornea substitutes by tissue engineering techniques have focused on the use of decellularized tissue scaffolds. In this work, we evaluated different chemical and physical decellularization methods on small intestine tissues to determine the most appropriate decellularization protocols for corneal applications. Our results revealed that the most efficient decellularization agents were the SDS and triton X-100 detergents, which were able to efficiently remove most cell nuclei and residual DNA. Histological and histochemical analyses revealed that collagen fibers were preserved upon decellularization with triton X-100, NaCl and sonication, whereas reticular fibers were properly preserved by decellularization with UV exposure. Extracellular matrix glycoproteins were preserved after decellularization with SDS, triton X-100 and sonication, whereas proteoglycans were not affected by any of the decellularization protocols. Tissue transparency was significantly higher than control non-decellularized tissues for all protocols, although the best light transmittance results were found in tissues decellularized with SDS and triton X-100. In conclusion, our results suggest that decellularized intestinal grafts could be used as biological scaffolds for cornea tissue engineering. Decellularization with triton X-100 was able to efficiently remove all cells from the tissues while preserving tissue structure and most fibrillar and non-fibrillar extracellular matrix components, suggesting that this specific decellularization agent could be safely used for efficient decellularization of SI tissues for cornea TE applications.
Ideally, biomaterials designed to play specific physical and physiological roles in vivo should comprise components and microarchitectures analogous to those of the native tissues they intend to replace. For that, implantable biomaterials need to be carefully designed to have the correct structural and compositional properties, which consequently impart their bio-function. In this study, we showed that the control of such properties can be defined from the bottom-up, using smart surface templates to modulate the structure, composition, and bio-mechanics of human transplantable tissues. Using multi-functional peptide amphiphile-coated surfaces with different anisotropies, we were able to control the phenotype of corneal stromal cells and instruct them to fabricate self-lifting tissues that closely emulated the native stromal lamellae of the human cornea. The type and arrangement of the extracellular matrix comprising these corneal stromal Self-Lifting Analogous Tissue Equivalents (SLATEs) were then evaluated in detail, and was shown to correlate with tissue function. Specifically, SLATEs comprising aligned collagen fibrils were shown to be significantly thicker, denser, and more resistant to proteolytic degradation compared to SLATEs formed with randomly-oriented constituents. In addition, SLATEs were highly transparent while providing increased absorption to near-UV radiation. Importantly, corneal stromal SLATEs were capable of constituting tissues with a higher-order complexity, either by creating thicker tissues through stacking or by serving as substrate to support a fully-differentiated, stratified corneal epithelium. SLATEs were also deemed safe as implants in a rabbit corneal model, being capable of integrating with the surrounding host tissue without provoking inflammation, neo-vascularization, or any other signs of rejection after a 9-months follow-up. This work thus paves the way for the de novo bio-fabrication of easy-retrievable, scaffold-free human tissues with controlled structural, compositional, and functional properties to replace corneal, as well as other, tissues.
There was a low prevalence of myopia in these African children, astigmatism being the most frequent refractive error. The mean refractive errors found were low, and therefore visual acuity was high in these children.
SUMMARY
Bleaching can cause perceptible color changes on resin-based composite (RBC) restorations that may not be stable with aging. The objective of this study was to evaluate color stability and whiteness variations of RBCs after bleaching and aging procedures. Discs (10 mm in diameter and 1 mm thick) of shades A2 and A3 were fabricated from two RBCs (Filtek Z250 and Filtek Z350 XT) and divided into three subgroups (for each composite and shade) (n=5) as follows: control (no bleaching), at-home bleaching, and in-office bleaching. All specimens underwent an accelerated artificial aging up to 450 KJ/m2 and 900 KJ/m2 in an aging chamber (Suntest XXL+). A spectroradiometer (SpectraScan PR-670) was used to obtain CIE L*a*b* coordinates. CIEDE2000 color difference (ΔE00) and whiteness index for dentistry (WID) were used to evaluate color stability. Color and whiteness differences data were analyzed considering the 50:50% visual color difference thresholds (perceptibility [PT] and acceptability [AT]) and 50:50% whiteness thresholds (whiteness perceptibility [WPT] and whiteness acceptability [WAT]). Analysis of variance and Tukey tests (α=0.05) were used to statistically analyze the data. After bleaching, all specimens showed ΔE00 and ΔWID values below their corresponding acceptability thresholds (AT and WAT, respectively). After aging, L* and WID values decreased while b* values increased (p≤0.05), resulting in ΔE00 and ΔWID values above AT and WAT, respectively. Color changes after bleaching RBCs were clinically acceptable, while aging provoked clinically perceptible color changes.
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