In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells.

BACKGROUND: The World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020. The retinal degenerative diseases affect the macula, which is responsible for central and detailed vision. Most macular degeneration, i.e., age-related macular degeneration (AMD) develops in the elderly; however, certain hereditary diseases, such as the Stargardt disease, also affect young people. This degeneration begins with loss of retinal pigmented epithelium (RPE) due to formation of drusen (atrophic) or abnormal vessels (exudative). In wet AMD, numerous drugs are available to successful treat the disease; however, no proven therapy currently is available to treat dry AMD or Stargardt. Since its discovery, human embryonic stem cells (hESCs) have been considered a valuable therapeutic tool. Some evidence has shown that transplantation of RPEs differentiated from hESCs cells can result in recovery of both RPE and photoreceptors and prevent visual loss. METHODS: The human embryonic WA-09 stem cell lineage was cultured under current Good Manufacturing Practices (cGMP) conditions using serum-free media and supplements. The colonies were isolated manually and allowed to spontaneously differentiate into RPE cells. RESULTS: This simple and effective protocol required minimal manipulation and yielded more than 10e8 RPE cells by the end of the differentiation and enrichment processes, with cells exhibiting a cobblestone morphology and displaying cellular markers and a gene expression profile typical of mature RPE cells. Moreover, the differentiated cells displayed phagocytic activity and only a small percentage of the total cells remained positive for the Octamer-binding transcriptions factor 4 (OCT-4) pluripotency cell marker. CONCLUSIONS: These results showed that functional RPE cells can be produced efficiently and suggested the possibility of scaling-up to aim at therapeutic protocols for retinal diseases associated with RPE degeneration.

Biomechanical Comparison of Vertical Mattress and Cross-stitch Suture Techniques and Single- and Double-Row Configurations for the Treatment of Bucket-Handle Medial Meniscal Tears.

BACKGROUND: Given the variety of suturing techniques for bucket-handle meniscal repair, it is important to assess which suturing technique best restores native biomechanics. PURPOSE/HYPOTHESIS: To biomechanically compare vertical mattress and cross-stitch suture techniques, in single- and double-row configurations, in their ability to restore native knee kinematics in a bucket-handle medial meniscal tear model. The hypothesis was that there would be no difference between the vertical mattress and cross-stitch double-row suture techniques but that the double-row technique would provide significantly improved biomechanical parameters versus the single-row technique. STUDY DESIGN: Controlled laboratory study. METHODS: Ten matched pairs of human cadaver knees were randomly assigned to the vertical mattress (n = 10) or cross-stitch (n = 10) repair group. Each knee underwent 4 consecutive testing conditions: (1) intact, (2) displaced bucket-handle tear, (3) single-row suture configuration on the femoral meniscus surface, and (4) double-row suture configuration (repair of femoral and tibial meniscus surfaces). Knees were loaded with a 1000-N axial compressive force at 0°, 30°, 60°, 90°, and 120° of flexion for each condition. Resultant medial compartment contact area, average contact pressure, and peak contact pressure data were recorded. RESULTS: Intact state contact area was not restored at 0° ( P = .027) for the vertical double-row configuration and at 0° ( P = .032), 60° ( P < .001), and 90° ( P = .007) of flexion for the cross-stitch double-row configuration. No significant differences were found in the average contact pressure and peak contact pressure between the intact state and the vertical mattress and cross-stitch repairs with single- and double-row configurations at any flexion angles. When the vertical and cross-stich repairs were compared across all flexion angles, no significant differences were observed in single-row configurations, but in double-row configurations, cross-stitch repair resulted in a significantly decreased contact area, average contact pressure, and peak contact pressure (all P < .001). CONCLUSION: Single- and double-row configurations of the vertical mattress and cross-stitch inside-out meniscal repair techniques restored native tibiofemoral pressure after a medial meniscal bucket-handle tear at all assessed knee flexion angles. Despite decreased contact area with a double-row configuration, mainly related to the cross-stitch repair, in comparison with the intact state, the cross-stitch double-row repair led to decreased pressure as compared with the vertical double-row repair. These findings are applicable only at the time of the surgery, as the biological effects of healing were not considered. CLINICAL RELEVANCE: Medial meniscal bucket-handle tears may be repaired with the single- or double-row configuration of vertical mattress or cross-stitch sutures.

Effects of Training and Overtraining on Intervertebral Disc Proteoglycans.

STUDY DESIGN: Animal experimental study. OBJECTIVE: Evaluate the effect of physical activity and overtraining condition on glycosaminoglycan concentration on the intervertebral disc (IVD) using a rat running model. SUMMARY OF BACKGROUND DATA: Some guidelines recommend the implementation of a physical exercise program as treatment for low back pain; however, cyclic loading impact on the health of the IVD and whether there is a dose-response relationship is still incompletely understood. METHODS: Thirty-two rats ages 8 weeks were divided into four groups with eight animals each. The first 8 weeks were the adaptive phase, the overtraining phase was from the ninth to the eleventh week, which consisted of increasing the number of daily training sessions from 1 to 4 and the recovery phase was represented by the 12th and 13th weeks without training. Control group 1 (CG1) did not undergo any kind of training. Control group 2 (CG2) completed just the adaptive phase. Overtraining group 1 (OT1) completed the overtraining phase. Overtraining group 2 (OT2) completed the recovery phase. Running performance tests were used to assess the "overtraining" status of the animals. IVD glycosaminoglycans were extracted and quantified, and identified by electrophoresis. RESULTS: Glycosaminoglycans showed a distribution between chondroitin sulfate and dermatan sulfate. Glycosaminoglycans quantification showed decreasing concentration at the following order: OT1 > CG2 > OT2 > CG1. Increased expression of dermatan sulfate was verified at the groups submitted to any training. CONCLUSION: Overtraining condition, as assessed by muscle and cardiovascular endurance did not lessen glycosaminoglycan concentration in the IVD. In fact, physical exercise increased glycosaminoglycan concentration in the IVD in proportion to the training load, even at overtraining condition, returning to normal levels after the recovery phase and glycosaminoglycan production is a reversible acute positive response for mechanical stimulation of the IVD. LEVEL OF EVIDENCE: N/A.

Bioabsorbable versus metallic interference screws for graft fixation in anterior cruciate ligament reconstruction.

BACKGROUND: Anterior cruciate ligament (ACL) tears are frequently treated with surgical reconstruction with grafts, frequently patella tendon or hamstrings. Interference screws are often used to secure the graft in bone tunnels in the femur and tibia. This review examines whether bioabsorbable interference screws give better results than metal interference screws when used for graft fixation in ACL reconstruction. OBJECTIVES: To assess the effects (benefits and harms) of bioabsorbable versus metallic interference screws for graft fixation in ACL reconstruction. SEARCH METHODS: We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register, CENTRAL (the Cochrane Library), MEDLINE, Embase, LILACS, trial registers and reference lists of articles. Date of search: January 2016. SELECTION CRITERIA: We included randomised controlled trials and quasi-randomised trials comparing bioabsorbable with metallic interferences screws in ACL reconstruction. The main outcomes sought were subjective-rated knee function, failure of treatment, and activity level. DATA COLLECTION AND ANALYSIS: At least two review authors selected eligible trials, independently assessed risk of bias, and cross-checked data. Data were pooled whenever relevant and possible. Requests for further information were sent to the original study authors. MAIN RESULTS: We included 12 trials (11 randomised and one quasi-randomised) involving a total of 944 participants, and reporting follow-up results for 774. Participants in the 12 trials underwent ACL reconstruction with either hamstring tendon grafts (five trials) or patellar tendon grafts (seven trials). Trials participants were randomly allocated to bioabsorbable or metallic interference screws for graft fixation in both femur and tibia (seven trials); femur only (three trials); tibia only (one trial); location was not reported in the remaining trial. A variety of materials was used for the bioabsorbable screws, Poly-L-lactic acid (PLLA) being the most common. The metallic screws, where reported, were titanium.All trials were at high risk of bias, which invariably included performance bias. Seven trials were at high risk of attrition bias and eight at high risk of reporting bias. The quasi-randomised trial was assessed as being at high risk for selection bias. Based on these study limitations and insufficiency of the available data, we judged the quality of evidence for all outcomes was very low.The majority of the available data for patient-reported knee function was presented as Lysholm scores (0 to 100; higher scores = better function). There was very low quality but consistent evidence of no clinically important differences between the two groups in Lysholm scores at 12 months follow-up (mean difference (MD) -0.08, 95% confidence interval (CI) -1.48 to 1.32; three trials, 168 participants); 24 months (MD 0.35, 95% CI -1.27 to 1.98; three trials, 113 participants) or five or more years follow-up (MD 1.23, 95% CI -2.00 to 4.47; two trials, 71 participants). This lack of between-group differences was also reported for Lysholm scores in several trials that did not provide sufficient data for pooling as well as for other self-reported knee function scores reported in several trials.Treatment failure was represented by the summed data for implant breakage during surgery and major postoperative complications (implant failure, graft rupture, symptomatic foreign body reactions, effusion and treated arthrofibrosis and related conditions) that were usually described in the trial reports as requiring further substantive treatment. There is very low-quality evidence of greater treatment failure in the bioabsorbable screw group (60/451 versus 29/434; risk ratio (RR) 1.94 favouring metallic screw fixation, 95% CI 1.29 to 2.93; 885 participants, 11 studies). In a population with an assumed risk (based on the median control group risk) of 56 participants per 1000 having treatment failure after metallic screw fixation, this equates to 53 more (95% CI 17 to 108 more) per 1000 participants having treatment failure after bioabsorbable screw fixation. All 16 intraoperative complications in the bioabsorbable screw group were implant breakages upon screw insertion. Treatment failure defined as postoperative complications only still favoured the metallic screw group but the 95% CI also included the potential for a greater risk of treatment failure after metallic screw fixation: 44/451 versus 29/434; RR 1.44, 95% CI 0.93 to 2.23. Based on the assumed risk of 56 participants per 1000 having postoperative treatment failure after metallic screw fixation, this equates to 25 more (95% CI 4 fewer and 69 more) per 1000 participants having this outcome after bioabsorbable screw fixation.There was very low-quality evidence of very similar activity levels in the two groups at 12 and 24 months follow-up measured via the Tegner score (0 to 10; higher scores = greater activity): 12 months (MD 0.08, 95% CI -0.39 to 0.55; 122 participants, two studies); 24 months (MD 0.01, 95% CI -0.54 to 0.57; 72 participants, two studies). AUTHORS' CONCLUSIONS: There is very low-quality evidence of no difference in self-reported knee function and levels of activity between bioabsorbable and metallic interference screws for graft fixation in ACL reconstruction. There is very low-quality evidence that bioabsorbable screws may be associated with more overall treatment failures, including implant breakage during surgery. Further research does not appear to be a priority, but if undertaken, should also examine costs.