Semi-synthetic degradable notochordal cell-derived matrix hydrogel for use in degenerated intervertebral discs: Initial in vitro characterization.

Low back pain is the leading cause of disability worldwide, but current therapeutic interventions are palliative or surgical in nature. Loss of notochordal cells (NCs) and degradation of the healthy matrix in the nucleus pulposus (NP), the central tissue of intervertebral discs (IVDs), has been associated with onset of degenerative disc changes. Recently, we established a protocol for decellularization of notochordal cell derived matrix (NCM) and found that it can provide regenerative cues to nucleus pulposus cells of the IVD. Here, we combined the biologically regenerative properties of decellularized NCM with the mechanical tunability of a poly(ethylene glycol) hydrogel to additionally address biomechanics in the degenerate IVD. We further introduced a hydrolysable PEG-diurethane crosslinker for slow degradation of the gels in vivo. The resulting hydrogels were tunable over a broad range of stiffness's (0.2 to 4.5 kPa), matching that of NC-rich and -poor NP tissues, respectively. Gels formed within 30 min, giving ample time for handling, and remained shear-thinning post-polymerization. Gels also slowly released dNCM over 28 days as measured by GAG effusion. Viability of encapsulated bone marrow stromal cells after extrusion through a needle remained high. Although encapsulated NCs stayed viable over two weeks, their metabolic activity decreased, and their phenotype was lost in physiological medium conditions in vitro. Overall, the obtained gels hold promise for application in degenerated IVDs but require further tuning for combined use with NCs.

Neo-cartilage formation using human nondegenerate versus osteoarthritic chondrocyte-derived cartilage organoids in a viscoelastic hydrogel.

Current regenerative cartilage therapies are associated with several drawbacks such as dedifferentiation of chondrocytes during expansion and the formation of fibrocartilage. Optimized chondrocyte expansion and tissue formation could lead to better clinical results of these therapies. In this study, a novel chondrocyte suspension expansion protocol that includes the addition of porcine notochordal cell-derived matrix was used to self-assemble human chondrocytes from osteoarthritic (OA) and nondegenerate (ND) origin into cartilage organoids containing collagen type II and proteoglycans. Proliferation rate and viability were similar for OA and ND chondrocytes and organoids formed had a similar histologic appearance and gene expression profile. Organoids were then encapsulated in viscoelastic alginate hydrogels to form larger tissues. Chondrocytes on the outer bounds of the organoids produced a proteoglycan-rich matrix to bridge the space between organoids. In hydrogels containing ND organoids some collagen type I was observed between the organoids. Surrounding the bulk of organoids in the center of the gels, in both OA and ND gels a continuous tissue containing cells, proteoglycans and collagen type II had been produced. No difference was observed in sulphated glycosaminoglycan and hydroxyproline content between gels containing organoids from OA or ND origin after 28 days. It was concluded that OA chondrocytes, which can be harvested from leftover surgery tissue, perform similar to ND chondrocytes in terms of human cartilage organoid formation and matrix production in alginate gels. This opens possibilities for their potential to serve as a platform for cartilage regeneration but also as an in vitro model to study pathways, pathology, or drug development.

Evaluating Initial Integration of Cell-Based Chondrogenic Constructs in Human Osteochondral Explants.

Integration of an implant with the surrounding tissue is a major challenge in cartilage regeneration. It is usually assessed with in vivo animal studies at the end-stage of implant development. To reduce animal experimentation and at the same time increase screening throughput and speed up implant development, this study examined whether integration of allogeneic cell-based implants with the surrounding native cartilage could be demonstrated in an ex vivo human osteochondral culture model. Chondrocytes were isolated from smooth cartilage tissue of fresh human tibial plateaus and condyles. They were expanded for 12 days either in three-dimensional spinner flask cultures to generate organoids, or in two-dimensional culture flasks for standard cell expansion. Three implant groups were created (fibrin+organoids, fibrin+cells, and fibrin only) and used to fill a Ø 6 mm full-depth chondral defect created in human osteochondral explants (Ø 10 mm, bone length cut to 4 mm) harvested from a second set of fresh human tibial plateaus. Explants were cultured for 1 or 28 days in a double-chamber culture platform. Histology showed that after 28 days the organoids on the interface of the defect remodeled and merged, and cells migrated through the fibrin glue bridging the space between the organoids and between the organoids and the native cartilage. For both conditions, newly formed tissue rich in proteoglycans and collagen type II was present mainly on the edges and in the corners of the defect. In these matrix-rich areas, cells resided in lacunae and the newly formed tissue integrated with the surrounding native cartilage. Biochemical analysis revealed a statistically significant effect of culture time on glycosaminoglycan (GAG) content, and showed a higher hydroxyproline (HYP) content for organoid-filled implants compared with cell-filled implants at both timepoints. This ex vivo human osteochondral culture system provides possibilities for exploration and identification of promising implant strategies based on evaluation of integration and matrix production under more controlled experimental conditions than possible in vivo.

Self-Oxygenation of Tissues Orchestrates Full-Thickness Vascularization of Living Implants.

Bioengineering of tissues and organs has the potential to generate functional replacement organs. However, achieving the full-thickness vascularization that is required for long-term survival of living implants has remained a grand challenge, especially for clinically sized implants. During the pre-vascular phase, implanted engineered tissues are forced to metabolically rely on the diffusion of nutrients from adjacent host-tissue, which for larger living implants results in anoxia, cell death, and ultimately implant failure. Here it is reported that this challenge can be addressed by engineering self-oxygenating tissues, which is achieved via the incorporation of hydrophobic oxygen-generating micromaterials into engineered tissues. Self-oxygenation of tissues transforms anoxic stresses into hypoxic stimulation in a homogenous and tissue size-independent manner. The in situ elevation of oxygen tension enables the sustained production of high quantities of angiogenic factors by implanted cells, which are offered a metabolically protected pro-angiogenic microenvironment. Numerical simulations predict that self-oxygenation of living tissues will effectively orchestrate rapid full-thickness vascularization of implanted tissues, which is empirically confirmed via in vivo experimentation. Self-oxygenation of tissues thus represents a novel, effective, and widely applicable strategy to enable the vascularization living implants, which is expected to advance organ transplantation and regenerative medicine applications.

Tethering Cells via Enzymatic Oxidative Crosslinking Enables Mechanotransduction in Non-Cell-Adhesive Materials.

Cell-matrix interactions govern cell behavior and tissue function by facilitating transduction of biomechanical cues. Engineered tissues often incorporate these interactions by employing cell-adhesive materials. However, using constitutively active cell-adhesive materials impedes control over cell fate and elicits inflammatory responses upon implantation. Here, an alternative cell-material interaction strategy that provides mechanotransducive properties via discrete inducible on-cell crosslinking (DOCKING) of materials, including those that are inherently non-cell-adhesive, is introduced. Specifically, tyramine-functionalized materials are tethered to tyrosines that are naturally present in extracellular protein domains via enzyme-mediated oxidative crosslinking. Temporal control over the stiffness of on-cell tethered 3D microniches reveals that DOCKING uniquely enables lineage programming of stem cells by targeting adhesome-related mechanotransduction pathways acting independently of cell volume changes and spreading. In short, DOCKING represents a bioinspired and cytocompatible cell-tethering strategy that offers new routes to study and engineer cell-material interactions, thereby advancing applications ranging from drug delivery, to cell-based therapy, and cultured meat.

The regenerative effect of different growth factors and platelet lysate on meniscus cells and mesenchymal stromal cells and proof of concept with a functionalized meniscus implant.

Meniscus regeneration could be enhanced by targeting meniscus cells and mesenchymal stromal cells (MSCs) with the right growth factors. Combining these growth factors with the Collagen Meniscus Implant (CMI®) could accelerate cell ingrowth and tissue formation in the implant and thereby improve clinical outcomes. Using a transwell migration assay and a micro-wound assay, the effect of insulin-like growth factor-1, platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), transforming growth factor beta 1 (TGF-ß1), fibroblast growth factor, and platelet lysate (PL) on migration and proliferation of meniscus cells and MSCs was assessed. The formation of extracellular matrix under influence of the above-mentioned growth factors was assessed after 28 days of culture of both MSCs and meniscus cells. As a proof of concept, the CMI® was functionalized with a VEGF binding peptide and coated with platelet-rich plasma (PRP) for clinical application. Our results demonstrate that PDGF, TGF-ß1, and PL stimulate migration, proliferation, and/or extracellular matrix production of meniscus cells and MSCs. Additionally, the CMI® was successfully functionalized with a VEGF binding peptide and PRP which increased migration of meniscus cell and MSC into the implant. This study demonstrates proof of concept of functionalizing the CMI® with growth factor binding peptides. A CMI® functionalized with the right growth factors holds great potential for meniscus replacement after partial meniscectomy.

De novo neo-hyaline-cartilage from bovine organoids in viscoelastic hydrogels.

Regenerative therapies for articular cartilage are currently clinically available. However, they are associated with several drawbacks that require resolution. Optimizing chondrocyte expansion and their assembly, can reduce the time and costs of these therapies and more importantly increase their clinical success. In this study, cartilage organoids were quickly mass produced from bovine chondrocytes with a new suspension expansion protocol. This new approach led to massive cell proliferation, high viability and the self-assembly of organoids. These organoids were composed of collagen type II, type VI, glycosaminoglycans, with Sox9 positive cells, embedded in a pericellular and interterritorial matrix similarly to hyaline cartilage. With the goal of producing large scale tissues, we then encapsulated these organoids into alginate hydrogels with different viscoelastic properties. Elastic hydrogels constrained the growth and fusion of the organoids inhibiting the formation of a tissue. In contrast, viscoelastic hydrogels allowed the growth and fusion of the organoids into a homogenous tissue that was rich in collagen type II and glycosaminoglycans. The encapsulation of organoids to produce in vitro neocartilage also proved to be superior to the conventional method of encapsulating 2D expanded chondrocytes. This study describes a multimodal approach that involves chondrocyte expansion, organoid formation and their assembly into neohyaline-cartilage which proved to be superior to the current standard approaches used in cartilage tissue engineering. STATEMENT OF SIGNIFICANCE: In this manuscript, we describe a new and simple methodology to quickly mass produce self-assembling cartilage organoids. Due to their matrix content and structure similarities with native cartilage, these organoids on their own have the potential to revolutionize cartilage research and the manner in which we study signaling pathways, disease progression, tissue engineering, drug development, etc. Furthermore, these organoids and their fast mass production were combined with a key relatively ignored hydrogel characteristic, viscoelasticity, to demonstrate their fusion into a neo-tissue. This has the potential to open the door for large scale cartilage regeneration such as for entire joint surfaces.

Characterization of biomaterials intended for use in the nucleus pulposus of degenerated intervertebral discs.

Biomaterials for regeneration of the intervertebral disc must meet complex requirements conforming to biological, mechanical and clinical demands. Currently no consensus on their characterization exists. It is crucial to identify parameters and their method of characterization for accurate assessment of their potential efficacy, keeping in mind the translation towards clinical application. This review systematically analyses the characterization techniques of biomaterial systems that have been used for nucleus pulposus (NP) restoration and regeneration. Substantial differences in the approach towards assessment became evident, hindering comparisons between different materials with respect to their suitability for NP restoration and regeneration. We have analysed the current approaches and identified parameters necessary for adequate biomaterial characterization, with the clinical goal of functional restoration and biological regeneration of the NP in mind. Further, we provide guidelines and goals for their measurement. STATEMENT OF SIGNIFICANCE: Biomaterials intended for restoration of regeneration of the nucleus pulposus within the intervertebral disc must meet biological, biomechanical and clinical demands. Many materials have been investigated, but a lack of consensus on which parameters to evaluate leads to difficulties in comparing materials as well as mostly partial characterization of the materials in question. A gap between current methodology and clinically relevant and meaningful characterization is prevalent. In this article, we identify necessary methods and their implementation for complete biomaterial characterization in the context of clinical applicability. This will allow for a more unified approach to NP-biomaterials research within the field as a whole and enable comparative analysis of novel materials yet to be developed.

Spatiotemporal material functionalization via competitive supramolecular complexation of avidin and biotin analogs.

Spatiotemporal control over engineered tissues is highly desirable for various biomedical applications as it emulates the dynamic behavior of natural tissues. Current spatiotemporal biomaterial functionalization approaches are based on cytotoxic, technically challenging, or non-scalable chemistries, which has hampered their widespread usage. Here we report a strategy to spatiotemporally functionalize (bio)materials based on competitive supramolecular complexation of avidin and biotin analogs. Specifically, an injectable hydrogel is orthogonally post-functionalized with desthiobiotinylated moieties using multivalent neutravidin. In situ exchange of desthiobiotin by biotin enables spatiotemporal material functionalization as demonstrated by the formation of long-range, conformal, and contra-directional biochemical gradients within complex-shaped 3D hydrogels. Temporal control over engineered tissue biochemistry is further demonstrated by timed presentation and sequestration of growth factors using desthiobiotinylated antibodies. The method's universality is confirmed by modifying hydrogels with biotinylated fluorophores, peptides, nanoparticles, enzymes, and antibodies. Overall, this work provides a facile, cytocompatible, and universal strategy to spatiotemporally functionalize materials.

From fiber curls to mesh waves: a platform for the fabrication of hierarchically structured nanofibers mimicking natural tissue formation.

Bioinstructive scaffolds for regenerative medicine are characterized by intrinsic properties capable of directing cell response and promoting wound healing. The design of such scaffolds requires the incorporation of well-defined physical properties that mimic the native extracellular matrix (ECM). Here, inspired by epithelial tissue morphogenesis, we present a novel approach to code nanofiber materials with controlled hierarchical wavy structures resembling the configurations of native EMC fibers through using thermally shrinking materials as substrates onto which the fibers are deposited. This approach could serve as a platform for fabricating functional scaffolds mimicking various tissues such as trachea, iris, artery wall and ciliary body. Modeling affirms that the mechanical properties of the fabricated wavy fibers could be regulated through varying their wavy patterns. The nanofibrous scaffolds coded with wavy patterns show an enhanced cellular infiltration. In addition, we further investigated whether the wavy patterns could regulate transforming growth factor-beta (TGF-ß) production, a key signalling pathway involved in connective tissue development. Our results demonstrated that nanofibrous scaffolds coded with wavy patterns could induce TGF-ß expression without the addition of a soluble growth factor. Our new approach could open up new avenues for fabricating bioinstructive scaffolds for regenerative medicine.

A Versatile Protocol for Studying Anterior Cruciate Ligament Reconstruction in a Rabbit Model.

Anterior cruciate ligament (ACL) injuries are frequent, as >200,000 injuries occur in the United States alone each year. Owing to the risks for associated meniscus and cartilage damage, ACL injuries are a significant source of both orthopedic care and research. Given the extended recovery course after ACL injury, which often lasts 1-2 years, and is associated with limited participation in sports and activities of daily living for patients, there is a critical need for the evolution of new and improved methods for ACL repair. Subsequently, animal models of ACL reconstruction (ACLR) play a key role in the development and initial trialing of novel ACL interventions. This article provides a clear operative description and associated illustrations for a validated, institutional animal care and use committee, and veterinarian approved and facile model of ACLR to serve researchers investigating ACLR.

Direct and Indirect Flap Measurements in Femtosecond Laser-Assisted In Situ Keratomileusis.

PURPOSE: To compare direct and indirect LASIK flap thickness measurements using ultrasound and Scheimpflug technology. METHODS: Eighty-two eyes treated with laser-assisted in situ keratomileusis refractive surgery using a femtosecond laser (IntraLase FS150) were prospectively included in the study. Flap thickness was set to 115 µm. Corneal flap thickness was measured using the direct method-ie, ultrasound pachymetry immediately after flap construction in the presence of cavitation bubbles-and indirect methods, with subtraction of intraoperative post-lift corneal thickness measured using ultrasound pachymetry (intrastroma) from preoperative central corneal thickness using ultrasound (Indirect-US) or Scheimpflug thinnest pachymetry (Indirect-Scheimpflug). RESULTS: Mean flap thickness was overestimated using the indirect methods, Indirect-US and Indirect-Scheimpflug (122.6 ± 24.5 µm and 128.1 ± 26.1 µm, respectively; P < 0.0060 and P < 0.0001, respectively). There were no significant correlations between the direct and indirect methods. Indirect-Scheimpflug was significantly higher (P = 0.0122) than Indirect-US. The closest average flap thickness compared with the set parameter of 115 µm was that of the direct method (115.6 ± 8.6 µm; 95% confidence interval: -1.3 to 2.5; P = 0.5163). The direct method provided the lowest SD of all groups (SD: 8.64). CONCLUSIONS: The direct method of flap thickness measurement was the most comparable to the set parameter compared with the indirect subtraction methods. Additional studies are needed to determine which method allows for the most accurate measurement of flap thickness.

Bioactive Tape With BMP-2 Binding Peptides Captures Endogenous Growth Factors and Accelerates Healing After Anterior Cruciate Ligament Reconstruction.

BACKGROUND: The anterior cruciate ligament (ACL) has poor regenerative capacity, and an injury leads to loss of function, limiting quality of life and increasing the incidence of osteoarthritis. Surgical interventions can stabilize the joint and improve functional recovery. The delivery of growth factors (GFs) enhances the healing process; however, this is complex in its regulation, is high in costs, has side effects, and can only be accomplished with supraphysiological concentrations and thus is currently not clinically feasible. However, the immobilization of a patient's endogenous GFs in biomaterials can overcome these problems. PURPOSE: To develop a method to capture endogenous bone morphogenetic protein-2 (BMP-2) and ultimately show enhanced ACL healing in vivo using this novel methodology. STUDY DESIGN: Controlled laboratory study. METHODS: BMP-2 binding peptides were synthetized, purified, and immobilized on polycaprolactone (PCL) films. The affinity between the peptide and human BMP-2 (hBMP-2) was confirmed with immunofluorescence and enzyme-linked immunosorbent assay. The C2C12 Luc reporter cell line was used to confirm the bioactivity of immobilized BMP-2. For in vivo experiments, the same functionalization technology was applied to the commercially available Polytape, and the functionalized tape was sutured together with the graft used for ACL reconstruction in rats. Each animal underwent reconstruction with either native Polytape (n = 3) or Polytape with BMP-2 binding peptides (n = 3). At 2 and 6 weeks after surgery, the graft was assessed by histology and micro-computed tomography. RESULTS: The covalent immobilization of the peptide in PCL was successful, allowing the peptide to capture hBMP-2, which remained bioactive and led to the osteogenic differentiation of C2C12. In vivo experiments confirmed the potential of the Polytape functionalized with the BMP-2 binding peptide to capture endogenous BMP-2, leading to enhanced bone formation inside the femoral and tibial tunnels and ultimately improving the graft's quality. CONCLUSION: The incorporation of BMP-2 binding peptides into materials used for ACL reconstruction can capture endogenous hBMP-2, which enhances the healing process inside the bone tunnels. CLINICAL RELEVANCE: These results demonstrate the potential of using synthetic peptides to endow biomaterials with novel biological functions, namely to capture and immobilize endogenous GFs.

Lens: Management of Cataract Surgery, Cataract Prevention, and Floppy Iris Syndrome.

According to the World Health Organization, cataract is the major cause of reversible visual impairment in the world. It is present as the cause of decreased visual acuity in 33% of the visual impaired citizens. With the increase of life expectancy in the last decades, the number of patients with cataract is expected to grow for the next 20 years. Nowadays, the only effective treatment for cataracts is surgery and its surgical outcomes have been increasingly satisfactory with the technological advancement.Pharmaceutical development has been also responsible for surgical outcomes enhancement. This includes the development of new ophthalmic viscoelastic devices (OVDs), intraocular dyes, mydriatics, miotics, anesthetics, irrigating solutions, and antibiotics. However, the increased costs and demand for cataract surgery may be hard to meet in the future unless clinical preventive and curative options are evaluated.In this chapter, we review the studies that addressed pharmacological applications in cataract.

Mesotelioma pleural benigno: apresentação de um caso singular / Benign pleural mesothelioma: presentation of a singular case

Os autores relatam um caso de mesotelioma pleural benigno. São abordados aspectos histológicos, etiopatogênicos, genéticos, epidemiológicos e clínicos, bem como a casuística estudada, a terapêutica instituída e o segmento obtido. Abordam os critérios para o diagnóstico, resultante da somação de vários fatores, dando ênfase ao quadro clínico compatível, confirmação histopatológica, imuno-histopatológicos compatíveis com resposta positiva clínico-radiográfica e eficaz após a cirurgia proposta.

Cases of benign mesothelioma of the pleura are reported by authors. Histological, etiopathogenic, genetic, epidemiological and clinical aspects are approached as well the casuistry that was studied, established therapy and the acquired segment. It approaches the criteria for diagnosis resulted from many factors emphasizing compatible clinical condition, histopathological confirmation, compatible immuno-histopathology, effective and positive clinical radiography answer after the proposed surgery.

Cross-Talk Between Human Tenocytes and Bone Marrow Stromal Cells Potentiates Extracellular Matrix Remodeling In Vitro.

Tendon and ligament (T/L) pathologies account for a significant portion of musculoskeletal injuries and disorders. Tissue engineering has emerged as a promising solution in the regeneration of both tissues. Specifically, the use of multipotent human mesenchymal stromal cells (hMSC) has shown great promise to serve as both a suitable cell source for tenogenic regeneration and a source of trophic factors to induce tenogenesis. Using four donor sets, we investigated the bidirectional paracrine tenogenic response between human hamstring tenocytes (hHT) and bone marrow-derived hMSC. Cell metabolic assays showed that only one hHT donor experienced sustained notable increases in cell metabolic activity during co-culture. Histological staining confirmed that co-culture induced elevated collagen protein levels in both cell types at varying time-points in two of four donor sets assessed. Gene expression analysis using qPCR showed the varied up-regulation of anabolic and catabolic markers involved in extracellular matrix maintenance for hMSC and hHT. Furthermore, analysis of hMSC/hHT co-culture secretome using a reporter cell line for TGF-ß, a potent inducer of tenogenesis, revealed a trend of higher TGF-ß bioactivity in hMSC secretome compared to hHT. Finally, hHT cytoskeletal immunostaining confirmed that both cell types released soluble factors capable of inducing favorable tenogenic morphology, comparable to control levels of soluble TGF-ß1. These results suggest a potential for TGF-ß-mediated signaling mechanism that is involved during the paracrine interplay between the two cell types that is reminiscent of T/L matrix remodeling/turnover. These findings have significant implications in the clinical use of hMSC for common T/L pathologies.

Image-guided femtosecond laser-assisted cataract surgery in Peters anomaly type 2.

UNLABELLED: We describe a technique for image-guided femtosecond laser-assisted cataract surgery in a case of Peters anomaly type 2. The femtosecond laser technology enabled reliable construction of a complete capsulotomy despite central corneal opacification and a tented anterior capsule. Use of an image-guided femtosecond laser for cataract extraction in Peters anomaly type 2 is a safe method for removing these complex cataracts while preserving capsular and corneal integrity. FINANCIAL DISCLOSURE: Dr. De la Cruz is a consultant to Alcon Surgical, Inc. No other author has a financial or proprietary interest in any material or method mentioned.

Refractive and Visual Outcomes of Different Intraocular Lenses with Femtosecond Laser Cataract Surgery: The Expectation of Independence from Spectacles.

PURPOSE: To evaluate the visual performance of different types of intraocular lens (IOL) in eyes submitted to femtosecond laser (FSL) cataract surgery and to analyze whether this technology could result in spectacles independence. DESIGN: Retrospective comparative case series. METHODS: Consecutive eyes that were scheduled for FSL cataract surgery were included in this study. After one month postoperative, patients were divided into two groups: cases that required prescription for eyeglasses and cases that did not require prescription. In addition, the patients were analyzed according to the type of IOL that was implanted (monofocal, monofocal toric, multifocal and multifocal toric). RESULTS: Thirty-five cataract surgeries were analyzed. Twenty-three eyes had a monofocal IOL implanted, and 12 had a multifocal IOL implanted. After 1 month, 12 cases required prescription (Group 1), and 23 did not require prescription (Group 2). The proportion of eyes that did not require corrective lenses was significantly greater (P = 0.02) in eyes that received multifocal IOL (91.67%) compared with those that received monofocal IOL (47.83%). Additionally, 100% of eyes that received multifocal toric IOL did not require eyeglasses at one month postoperatively. CONCLUSION: The FSL surgical precision associated with modern IOL technology may be an important factor related to vision satisfaction after cataract surgery.

Clinical experience with the first 40 cases with femtosecond laser cataract surgery technology: safety of the learning curve / Experiência clínica com os primeiros 40 casos de cirurgia de catarata com a tecnologia de laser de femtossegundo: segurança da curva de aprendizagem

Objective:To evaluate the introduction of the femtosecond laser (FSL) to perform the key steps of the traditional cataract surgery process and the operational difficulties and safety of this new technology during routine use in an operating room in Brazil.MethodsA retrospective study was conducted using the first cases operated on at a single center using the laser platform LenSx/Alcon with a soft contact lens patient interface.All patients underwent a detailed preoperative assessment.The anterior capsulotomy, nuclear fragmentation, and corneal incisions were created with the FSL; then, the surgery was completed following the standard phacoemulsification procedure. The main outcome measurements were difficulties and complications related to the learning curve and an analysis of postoperative uncorrected distance visual acuity (UDVA).Results:Of 31 patients (40 eyes), 9 patients had FSL cataract surgery in both eyes.The mean age was 64 ± 12 years (ranging from 42 to 82), the mean cataract nuclear sclerosis was grading 2 ± 0.6 (ranging from 1 to 4), and the preoperative mean UDVA in logMAR was 0.4 ± 0.2 (ranging from 0.1 to 1.3). Anterior capsulotomy was complete in all patients, and scissors were not needed to cut off any intact portion. The postoperative corneal incisions were not completely linear and showed some irregularities. Laser phaco-fragmentation was effective, with the division of the nucleus into smaller segments easily performed before phacoemulsification.After 1 month, the postoperative mean UDVA in logMAR was 0.1 ± 0.1 (ranging from 0.0 to 0.4) (P < 0.0001).Conclusion:With increasing surgical cases and experience, the phacoemulsification steps are performed precisely and effectively with FSL pretreatment, resulting in a safe learning curve.

Objetivo:Avaliar a introdução do laser de femtossegundo (FSL) para realizar etapas fundamentais da cirurgia de catarata tradicional e as dificuldades e segurança desta nova tecnologia na rotina de um centro cirúrgico no Brasil.Métodos:Um estudo retrospectivo foi realizado com os primeiros casos operados em um único centro usando a plataforma de laser LenSx/Alcon, sendo utilizado lentes de contato gelatinosa na interface do paciente. Todos os pacientes foram submetidos a uma avaliação pré-operatória detalhada. Capsulotomia anterior, fragmentação nuclear e incisões na córnea foram criados com a FSL, em seguida, a cirurgia foi concluída com procedimento de facoemulsificação padrão. As principais medidas de desfecho foram dificuldades e complicações relacionadas à curva de aprendizado e análise da acuidade visual pós-operatória não corrigida à distância (UDVA).Resultados:Dos 31 pacientes (40 olhos), 9 realizaram cirurgia de catarata em ambos os olhos com FSL. A média de idade foi de 64 ± 12 anos (variando de 42 a 82), a média de classificação da catarata através da esclerose nuclear foi 2 ± 0,6 (variando de 1 a 4) e UDVA pré-operatória em logMAR foi de 0,4 ± 0,2 (variando de 0,1 a 1,3). A capsulotomia anterior foi completa em todos os pacientes e não precisou usar da pinça para cortar qualquer porção intacta. Incisões corneanas pós-operatórias não foram lineares, mostrando ainda algumas irregularidades. A facofragmentação a laser foi eficaz, com a divisão do núcleo em segmentos menores facilmente realizados antes da facoemulsificação. Após 1 mês de pós-operatório, UDVA em logMAR foi de 0,1 ± 0,1 (variando de 0,0 a 0,4) (P < 0,0001).Conclusão:Com o aumento do volume dos casos cirúrgicos e ganho de experiência, passos precisos da facoemulsificação são realizados com o pré-tratamento com o FSL, resultando em uma segura curva de aprendizagem.

Sleep deprivation and accidental fall risk in children.

OBJECTIVES: To look for an association between sleep deprivation and risk of accidental falls (AF) in children. METHODS: A questionnaire was applied to two groups of children aged 1-14 years, encompassing children observed in an emergency room for AF (G1) and children attending health care visits (HV) (G2). Collected data included demographic characteristics, medical history, previous week's sleep pattern (PWSP), sleep duration and sleep pattern in the preceding 24 h, mechanism of fall, and injury severity. EXCLUSION CRITERIA: acute or chronic disease or exposure to drugs interfering with sleep. Statistical analyses included Fisher's exact test, Pearson Chi-square, Fisher-Freeman-Halton test, T and Mann-Whitney tests for independent samples, and multivariate logistic regression (α=5%). RESULTS: We obtained 1756 questionnaires in G1 and 277 in G2. Of those, 834 in G1 and 267 in G2 were analyzed. We found an increased risk of AF in boys (OR 1.6; 95% CI 1.2-2.4). After controlling for age, gender, summer holidays, parental education and profession, lack of naps and PWSP were associated with increased risk (OR 2.1; 95% CI 1.3-3.3 and OR 2.7; 95% CI 1.2-6.1). In 3-5 year-old children there was an association between AF and a shorter than usual sleep duration in the previous 24 h (p=0.02). CONCLUSIONS: To our knowledge, our study is the largest so far to assess the association between sleep deprivation and childhood injury. It evidences a protective effect of naps in children. Sleep duration of less than 8 h increases risk of AF. Pre-schoolers may be particularly susceptible to sleep deprivation.