MRI findings of traumatic and degenerative rotator cuff tears and introduction of the "cobra sign".

Background: A rotator cuff tear (RCT) is a common shoulder diagnosis and its etiology may be acute, traumatic, or chronic degenerative. Differentiation between the 2 etiologies may be important for multiple reasons, but remains difficult based on imaging. Further knowledge about radiographic and magnetic resonance findings to distinguish traumatic from degenerative RCT is needed. Methods: We analyzed magnetic resonance arthrograms (MRAs) of 96 patients with traumatic or degenerative superior RCT, which were matched according their age and the affected rotator cuff muscle into the 2 groups. Patients older than 66 years of age were excluded from the study to avoid including cases with pre-existing degeneration. In the case of traumatic RCT, the time between the trauma and MRA had to be less than 3 months. Various parameters of the supraspinatus (SSP) muscle-tendon unit were assessed (tendon thickness, presence of a remaining tendon stump at the greater tubercle, magnitude of retraction, layer appearance). The retraction of the 2 SSP layers were individually measured to determine the difference of retraction. Additionally, edema of the tendon and muscle, the tangent- and kinking-sign as well as the newly introduced Cobra-sign (bulging of the distal part of the ruptured tendon with slim configuration of the medial part of the tendon) were analyzed. Results: Edema within the SSP muscle (sensitivity 13%, specificity 100%, P = .011) or the tendon (sensitivity 86%, specificity 36%, P = .014) are more frequent in traumatic RCT. The same association was found for the kinking-sign (sensitivity 53%, specificity 71%, P = .018) and the Cobra sign (sensitivity 47%, specificity 84%, P = .001). Even though not statistically significant, tendencies were observed toward thicker tendon stumps in traumatic RCT, and greater difference in retraction between the 2 SSP layers in the degenerative group. The cohorts had no difference in the presence of a tendon stump at the greater tuberosity. Conclusion: Muscle and tendon edema, as well as tendon kinking appearance and the newly introduced cobra-sign are suitable MRA parameters to distinguish between traumatic and degenerative etiology of a superior RTC.

Quantification of Malalignment and Corrective Osteotomies in Patients With Malunion After Elastic Stable Intramedullary Nailing of Pediatric Forearm Fractures.

Purpose: This study looked at postoperative malunion with restricted function after elastic stable intramedullary nailing (ESIN) in pediatric patients in their childhood or adolescence. The primary objective was to compare the magnitude of the osseous malposition to the healthy opposite side. Second, these individuals were treated with patient-specific surgical instrumentation, and functional outcomes were documented. Methods: Patients under the age of 18 at the time of a corrective osteotomy due to a forearm malunion after initial ESIN treatment were included in this study. The healthy contralateral side was used as a reference for preoperative analysis and planning of the osteotomy. Osteotomies were performed using patient-specific guides and the direction and extent of the malunion were compared to the change in range of motion (ROM) after the operation. Results: Fifteen patients met the inclusion criteria at three years after initial ESIN placement, with the most pronounced malposition in the rotational axis. The postoperative function significantly improved by 12° (pre-op: 60° ± 17; post-op: 72° ± 10) of pronation and 33° (pre-op: 43° ± 26; post-op: 76° ± 13) of supination. There was no correlation between the amount and direction of malformation and the change in ROM. Conclusions: The most noticeable malunion after forearm fractures treated with the ESIN technique is in the rotational direction. Patient-specific corrective osteotomy of pediatric forearm malunion following forearm fracture fixation with ESIN achieves significant improvement in forearm range of motion. Clinical relevance: The findings are clinically relevant since forearm fractures are the most common pediatric fracture, affecting a large number of patients who can benefit from the findings of this study. It has the potential to raise awareness of the significance of the accurate rotational component of intraoperative bone alignment in the ESIN procedure.

Compliance with wearing a thoracolumbar orthosis in nonoperative treatment of osteoporotic vertebral fractures: a prospective sensor-controlled study.

BACKGROUND CONTEXT: Hyperextension orthoses (HOs) for nonoperative treatment of osteoporotic vertebral fractures (OVFs) are widely prescribed. However, the compliance, how much an HO is worn after it has been prescribed, is widely unknown. PURPOSE: This study was performed to assess the wearing time of HOs for OVFs in a prospective blinded, sensor-controlled manner. STUDY DESIGN / SETTING: A prospective, single blinded observational study was performed. PATIENT SAMPLE: This study prospectively included 18 patients who were treated nonoperatively with an HO for OVFs. OUTCOME MEASURES: The true wearing time was measured using a hidden temperature-based sensor. The patients were invited to return for regular follow-up every 2 weeks for 6 weeks, at which time clinical evaluation (including a visual analog scale for pain and the Oswestry disability questionnaire) and radiographs of the spine were performed. METHODS: Full compliance was defined as a wearing time of 15 hours per day. Correlation between compliance and demographic differences, patient reported outcomes and radiographic changes of the vertebral structures were calculated. RESULTS: The mean HO wearing time was 5.5±3.3 hours (37%±22% compliance). Female patients used the HO significantly longer per day than did male patients (6.5±3.2 vs 2.9±2.0 hours, p=.039). Age and body mass index had no influence on wearing behavior. CONCLUSIONS: Overall, compliance with wearing HOs is poor and shows great variability with significant gender-dependency but not associated with BMI, age, or pain-level. Further studies are required to confirm our results that the wearing time does not have an influence on kyphotic progression of the osteoporotic fractured segment, nor on clinical outcome at short term. LEVEL OF EVIDENCE: I.

Endoscopic Treatment of Stump Infection of the Residual Synovial Cavity After Through-the-Knee Amputation: A Case Report.

CASE: We report the case of a 35-year-old patient who presented with a septic residual synovial cavity infection 8 weeks after a through-the-knee amputation because of a parosteal sarcoma. An endoscopic evacuation of the turbid fluid and synovial debridement through parapatellar portals as in a standard knee arthroscopy was performed, in conjunction with appropriate antibiotic therapy. One year postoperatively, there were no signs of residual infection. CONCLUSION: Endoscopic treatment of a septic stump infection of the residual synovial cavity after through-the-knee amputation is feasible. In our case, this approach resulted in rapid wound healing and early prosthesis mobility.

3D planning and patient specific instrumentation for intraarticular corrective osteotomy of trapeziometacarpal-, metacarpal and finger joints.

BACKGROUND: Intra-articular malunions of the finger can lead to deformity and loss of function and can be treated with intra-articular corrective osteotomies. The aim of this study was to evaluate radiographic joint congruency, feasibility and functional outcome of three-dimensional (3D) printed patient-specific instrumentation (PSI) for corrective osteotomies at the trapeziometacarpal and finger joints. METHODS: Computer-tomography (CT) scans were acquired preoperatively for standard 3D planning, which was followed by calculation of cutting planes and the design of individualized bone surface contact drilling, sawing and reposition guides. Follow-up CT scans and clinical examinations (range of motion, grip strength) were performed. Postoperative complications were documented and patient-reported outcome measurements were assessed (Single Assessment Numeric Evaluation (SANE) score, brief Michigan Hand Questionnaire (MHQ)). RESULTS: Ten patients (mean age 28.4 ± 12.8,range 13.8-51.3) years) were included with a mean follow-up of 21 ± 18 (3-59) months including seven osteotomies at the trapeziometacarpal or metacarpophalangeal joints and three at the proximal interphalangeal joint (PIP). All radiographic follow-up examinations showed the planned correction with good joint congruency and regular osseous consolidation. At the latest follow-up, the range of motion (ROM) increased and the average grip strength recovered to the level of the contralateral side. No postoperative complication was detected. The mean SANE score improved from 44 ± 23 (0-70) to 82 ± 12 (60-90) after a mean of 72 ± 20 (44-114) months. The mean postoperative brief MHQ was 92 ± 8 (71-98). CONCLUSION: The use of 3D PSI in treating intra-articular malunions at the trapeziometacarpal and finger joints restored articular congruency accurately. ROM and grip strength improved postoperatively comparable to the healthy contralateral side and patient-reported outcome measures improved after medium-term follow-up.

3D analysis of the distal ulna with regard to the design of a new ulnar head prosthesis.

STUDY DESIGN: A retrospective, single center, data analysis. OBJECTIVE: Persistent pain and instability are common complications after distal ulnar head arthroplasty. One main reason may be the insufficient representation of the anatomical structures with the prosthesis. Some anatomical structures are neglected such as the ulnar head offset and the ulnar torsion which consequently influences the wrist biomechanics. METHODS: CT scans of the ulnae of forty healthy and asymptomatic patients were analyzed in a three-dimensional surface calculation program. In the best fit principle, cylinders were fitted into the medullary canal of the distal ulna and the ulnar head to determine their size. The distance between the central axes of the two cylinders was measured, which corresponds to the ulnar offset, and also their rotational orientation was measured, which corresponds to the ulnar torsion. RESULTS: The mean medullary canal diameter was 5.8 mm (±0.8), and the ulnar head diameter was 15.8 mm (±1.5). The distance between the two cylinder axes was 3.89 mm (±0.78). The orientation of this offset was at an average of 8.63° (±15.28) of supination, reaching from 23° pronation to 32° supination. CONCLUSION: With these findings, a novel ulnar head prosthesis should have different available stem and head sizes but also have an existing but variable offset between these two elements. A preoperative three-dimensional analysis is due to the high variation of offset orientation highly recommended. These findings might help to better represent the patients natural wrist anatomy in the case of an ulnar head arthroplasty. LEVEL OF EVIDENCE: III.

Musculoskeletal biomechanics of patients with or without adjacent segment degeneration after spinal fusion.

STUDY DESIGN: A retrospective, single center, case-control study was performed. OBJECTIVE: The present study employed patient-specific biomechanical modeling to find potential biomechanical differences after spinal fusion at L4/5 in patients with and without subsequent development of adjacent segment disease (ASD). METHODS: The study population comprised patients who underwent primary spinal fusion at L4/5 and were either asymptomatic during > 4 years of follow-up (CTRL; n = 18) or underwent revision surgery for ASD at L3/4 (n = 20). Landmarks were annotated on preoperative and follow-up lateral radiographs, and specific musculoskeletal models were created using a custom-built modeling pipeline. Simulated spinal muscle activation and lumbar intervertebral shear loads in unfused segments were analyzed in upright standing and forward flexion. Differences between the pre- and postoperative conditions were computed for each patient. RESULTS: The average postoperative muscle activity in the upright standing posture was 88.4% of the preoperative activity in the CTRL group (p <  0.0001), but did not significantly change from pre- to postoperatively in the ASD group (98.0%). The average shear load magnitude at the epifusional joint L3/4 during upright standing increased from pre- to postoperatively in the ASD group (+ 3.9 N, +/- 17.4 (n = 18)), but decreased in the CTRL group (- 4.6 N, +/- 23.3 (n = 20); p <  0.001). CONCLUSION: Patient-specific biomechanical simulation revealed that spinal fusion surgery resulted in greater shear load magnitude and muscle activation and therefore greater forces at the epifusional segment in those with ASD compared with those without ASD. This is a first report of patient-specific disc load and muscle force calculation with predictive merits for ASD.

Patient-specific statistical shape modeling for optimal spinal sagittal alignment in lumbar spinal fusion.

PURPOSE: The present study compared patients developing ASD after L4/5 spinal fusion with a control group using a patient-specific statistical shape model (SSM) to find alignment-differences between the groups. METHODS: This study included patients who had undergone spinal fusion at L4/5 and either remained asymptomatic (control group; n = 25, follow-up of > 4 years) or required revision surgery for epifusional ASD (n = 22). Landmarks on preoperative and postoperative lateral radiographs were annotated, and the optimal spinal sagittal alignment was calculated for each patient. The two-dimensional distance from the SSM-calculated optimum to the actual positions before and after fusion surgery was compared. RESULTS: Postoperatively, the additive mean distance from the SSM-calculated optimum was 86.8 mm in the ASD group and 67.7 mm in the control group (p = 0.119). Greater differences were observed between the groups with a larger distance to the ideal in patients with ASD at more cranial levels. Significant difference between the groups was seen postoperatively in the vertical distance of the operated segment L4. The patients with ASD (5.69 ± 3.0 mm) had a significant greater distance from the SSM as the control group (3.58 ± 3.5 mm, p = 0.034). CONCLUSION: Patients with ASD requiring revision after lumbar spinal fusion have greater differences from the optimal spinal sagittal alignment as an asymptomatic control group calculated by patient-specific statistical shape modeling. Further research might help to understand the value of SSM, in conjunction with already established indexes, for preoperative planning with the aim of reducing the risk of ASD. LEVEL OF EVIDENCE I: Diagnostic: individual cross-sectional studies with consistently applied reference standard and blinding.

Unplanned Return Visits to a Pediatric Emergency Department.

OBJECTIVES: Unplanned return visits (URVs) to emergency departments (EDs) account internationally for 2.5% to 5.2% of all consultations. ED crowding is an increasing challenge, and URVs seem to contribute to this problem. This study aimed to assess factors for URVs at the ED of a tertiary children's hospital to analyze if they are jointly responsible for the steadily rising amount of treated patients. METHODS: All patients with an URV to a pediatric ED in Switzerland between January and December 2013 were included in the study. Data were taken retrospectively from the electronic patient files, and different variables were defined and analyzed. RESULTS: URVs occurred at an incidence of 4.6%, and mostly concerned infants and toddlers (46%). URVs were independent of weekdays and mostly occurred between 10 am and 10 pm. In 84.2% of the cases, the URVs were judged as unnecessary, and in 15.8%, a hospitalization was indicated, mainly for children with a worsening respiratory illness. CONCLUSIONS: The occurrence of URVs in our ED was within the incidence reported in the literature. While URVs lead to hospitalization in some patients, the majority of URVs were unnecessary from a medical point of view. These results indicate that a correct evaluation of the child's health state by parents is often challenging and requires repeated medical attendance following a first ED visit, especially in infants with airway diseases and infections. Intensive counseling and scheduled short-term follow-up consultation at the pediatrician's office could prevent URVs to the ED.

Antileishmanial Activity of Dimeric Flavonoids Isolated from Arrabidaea brachypoda.

Leishmaniasis are diseases caused by parasites belonging to Leishmania genus. The treatment with pentavalent antimonials present high toxicity. Secondary line drugs, such as amphotericin B and miltefosine also have a narrow therapeutic index. Therefore, there is an urgent need to develop new drugs to treat leishmaniasis. Here, we present the in vitro anti-leishmanial activity of unusual dimeric flavonoids purified from Arrabidaea brachypoda. Three compounds were tested against Leishmana sp. Compound 2 was the most active against promastigotes. Quantifying the in vitro infected macrophages revealed that compound 2 was also the most active against intracellular amastigotes of L. amazonensis, without displaying host cell toxicity. Drug combinations presented an additive effect, suggesting the absence of interaction between amphotericin B and compound 2. Amastigotes treated with compound 2 demonstrated alterations in the Golgi and accumulation of vesicles inside the flagellar pocket. Compound 2-treated amastigotes presented a high accumulation of cytoplasmic vesicles and a myelin-like structure. When administered in L. amazonensis-infected mice, neither the oral nor the topical treatments were effective against the parasite. Based on the high in vitro activity, dimeric flavonoids can be used as a lead structure for the development of new molecules that could be useful for structure-active studies against Leishmania.

Intranasal administration of resveratrol successfully prevents lung cancer in A/J mice.

Lung cancer is the most lethal cancer in the world. About 80% of lung cancer deaths are linked to tobacco use. As a complement to tobacco control, efficient chemoprevention strategies are needed to tackle lung cancer epidemic. Resveratrol is one of the most studied natural products, notably for its cancer chemoprevention properties. However, its low oral bioavailability has often limited the translation of in vitro activities to in vivo effects. While oral administration of resveratrol effectively inhibited colorectal carcinogenesis, it failed to protect mice from chemically-induced lung carcinogenesis. Therefore, non-invasive parenteral routes must be considered to bring resveratrol to the lungs. In the present study, intranasal administration of a concentrated formulation proved to be a valid method to expose the lungs to a sufficient amount of resveratrol. This formulation was administered three times a week for 25 weeks to A/J mice having 4-[methyl(nitroso)amino]-1-(3-pyridinyl)-1-butanone-induced lung carcinogenesis. Resveratrol-treated mice showed a 27% decrease in tumour multiplicity, with smaller tumours, resulting in 45% decrease in tumour volume/mouse. In vitro investigations highlighted apoptosis as a potential mechanism of action. This study presents an effective way to overcome resveratrol low oral bioavailability, encouraging a reevaluation of its use in future clinical trials.

Identification and Mode of Action of a Plant Natural Product Targeting Human Fungal Pathogens.

Candida albicans is a major cause of fungal diseases in humans, and its resistance to available drugs is of concern. In an attempt to identify novel antifungal agents, we initiated a small-scale screening of a library of 199 natural plant compounds (i.e., natural products [NPs]). In vitro susceptibility profiling experiments identified 33 NPs with activity against C. albicans (MIC50s ≤ 32 µg/ml). Among the selected NPs, the sterol alkaloid tomatidine was further investigated. Tomatidine originates from the tomato (Solanum lycopersicum) and exhibited high levels of fungistatic activity against Candida species (MIC50s ≤ 1 µg/ml) but no cytotoxicity against mammalian cells. Genome-wide transcriptional analysis of tomatidine-treated C. albicans cells revealed a major alteration (upregulation) in the expression of ergosterol genes, suggesting that the ergosterol pathway is targeted by this NP. Consistent with this transcriptional response, analysis of the sterol content of tomatidine-treated cells showed not only inhibition of Erg6 (C-24 sterol methyltransferase) activity but also of Erg4 (C-24 sterol reductase) activity. A forward genetic approach in Saccharomyces cerevisiae coupled with whole-genome sequencing identified 2 nonsynonymous mutations in ERG6 (amino acids D249G and G132D) responsible for tomatidine resistance. Our results therefore unambiguously identified Erg6, a C-24 sterol methyltransferase absent in mammals, to be the main direct target of tomatidine. We tested the in vivo efficacy of tomatidine in a mouse model of C. albicans systemic infection. Treatment with a nanocrystal pharmacological formulation successfully decreased the fungal burden in infected kidneys compared to the fungal burden achieved by the use of placebo and thus confirmed the potential of tomatidine as a therapeutic agent.

A novel cyclosporin a aqueous formulation for dry eye treatment: in vitro and in vivo evaluation.

PURPOSE: The aim of the present study was the in vitro and in vivo evaluation of a novel aqueous formulation based on polymeric micelles for the topical delivery of cyclosporine A for dry eye treatment. METHODS: In vitro experiments were carried out on primary rabbit corneal cells, which were characterized by immunocytochemistry using fluorescein-labeled lectin I/isolectin B4 for the endothelial cells and mouse monoclonal antibody to cytokeratin 3+12 for the epithelial ones. Living cells were incubated for 1 hour or 24 hours with a fluorescently labeled micelle formulation and analyzed by fluorescence microscopy. In vivo evaluations were done by Schirmer test, osmolarity measurement, CyA kinetics in tears, and CyA ocular distribution after topical instillation. A 0.05% CyA micelle formulation was compared to a marketed emulsion (Restasis). RESULTS: The in vitro experiments showed the internalization of micelles in the living cells. The Schirmer test and osmolarity measurements demonstrated that micelles did not alter the ocular surface properties. The evaluation of the tear fluid gave similar CyA kinetics values: AUC = 2339 ± 1032 min*µg/mL and 2321 ± 881.63; Cmax = 478 ± 111 µg/mL and 451 ± 74; half-life = 36 ± 9 min and 28 ± 9 for the micelle formulation and Restasis, respectively. The ocular distribution investigation revealed that the novel formulation delivered 1540 ± 400 ng CyA/g tissue to the cornea. CONCLUSIONS: The micelle formulation delivered active CyA into the cornea without evident negative influence on the ocular surface properties. This formulation could be applied for immune-related ocular surface diseases.

Ocular biocompatibility of novel Cyclosporin A formulations based on methoxy poly(ethylene glycol)-hexylsubstituted poly(lactide) micelle carriers.

Topical ocular drug delivery has always been a challenge for pharmaceutical technology scientists. In the last two decades, many nano-systems have been studied to find ways to overcome the typical problems of topical ocular therapy, such as difficult corneal penetration and poor drug availability. In this study, methoxy poly(ethylene glycol)-hexylsubstituted poly(lactides) (MPEG-hexPLA) micelle formulations, which are promising nanocarriers for poorly water soluble drugs, were investigated for the delivery of Cyclosporin A (CsA) to the eye. As a new possible pharmaceutical excipient, the ocular compatibility of MPEG-hexPLA micelle formulations was evaluated. An in vitro biocompatibility assessment on human corneal epithelial cells was carried out using different tests. Cytotoxicity was studied by using the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] (MTT), and clonogenic tests and revealed that the CsA formulations and copolymer solutions were not toxic. After incubation with MPEG-hexPLA micelle formulations, the activation of caspase-dependent and -independent apoptosis as well as autophagy was evaluated using immunohistochemistry by analyzing the localization of four antibodies: (1) anti-caspase 3; (2) anti-apoptotic inducing factor (AIF); (3) anti-IL-Dnase II and (4) anti-microtubule-associated protein 1 light chain 3 (LC3). No apoptosis was induced when the cells were treated with the micelle solutions that were either unloaded or loaded with CsA. The ocular tolerance was assessed in vivo on rabbit eyes by Confocal Laser Scanning Ophthalmoscopy (CLSO), and very good tolerability was seen. The observed corneal surface was comparable to a control surface that was treated with a 0.9% NaCl solution. In conclusion, these results demonstrate that MPEG-hexPLA micelles are promising drug carriers for ocular diseases involving the activation of cytokines, such as dry eye syndrome and autoimmune uveitis, or for the prevention of corneal graft rejection.

Recent advances in confocal microscopy for studying drug delivery to the eye: concepts and pharmaceutical applications.

Since its seminal introduction 50years ago, confocal microscopy has been applied in numerous fields in life sciences. This review presents the different key elements of confocal microscopes, in particular scanning techniques, light sources and especially laser sources are described in this review. Furthermore, an overview of the different image processing systems coupled with confocal microscopy is provided. The chapter closes with the applications of confocal microscopy in drug delivery to the eye.

Comparative studies of various hyaluronic acids produced by microbial fermentation for potential topical ophthalmic applications.

This work presents a comparative study of various hyaluronic acids (HA) produced by fermentation of either Bacillus subtilis or Streptococcus towards the selection of an optimal molecular weight (MW) HA for the preparation of topical ophthalmic formulations. The influence of HA MW on water binding capacity, sterile filtration, rheological properties, precorneal residence time and ocular tolerance of ophthalmic solutions was investigated. Molecular weight did not affect hydration of hyaluronic acid according to differential scanning calorimetry (DSC). In general, medium MW HA (0.6-1 MDa) resulted in solutions that were superior in terms of sterile filtration and kinematic viscosity requirements compared to high MW HA (>1 MDa). Moreover, all HA-based solutions exhibited well-defined viscoelastic properties that depend on MW. Gamma scintigraphic data indicated that HA MW at 0.1% concentration (w/v) and HA origin did not significantly affect the corneal residence time on rabbit eyes. A 0.3% solution of high MW HA had a prolonged residence time in the precorneal area compared to a medium MW HA at the same concentration. Finally, an in vivo ocular irritation test based on confocal laser scanning ophthalmoscopy (CLSO) conclusively showed the excellent tolerance of both Bacillus-derived HA and Streptococcus-derived HA after topical instillation onto the corneal surface. Overall, this comprehensive work highlights the superiority of medium MW hyaluronic acid for topical ophthalmic formulations based on their physico-chemical and biological properties, tolerance and handling. Such solutions are expected to enhance tear film stability, to allow for maximum comfort, and to exhibit high residence times, while being biocompatible and easy to sterile filter.

New surface-active polymers for ophthalmic formulations: evaluation of ocular tolerance.

Two n-octenylsuccinate starch (AS) types of unknown molecular weights were assessed for ocular tolerance. Irritation potential of different solutions (containing 2 and 15% (w/w) AS) and AS stabilized emulsions (containing 15% (w/w) AS) was evaluated in vivo in rabbit eyes, using a confocal laser scanning microscope, and in vitro on treated excised pig corneas by light microscopy of histological cross sections. Both AS types were previously characterized by viscosity, osmolality and surface tension measurements. All tested solutions and emulsions showed good eye tolerance regardless of concentration and emulsifying properties suggesting AS to be a good alternative to commonly used solubilizing or emulsifying agents in ophthalmic formulations.

Ocular tolerance of absorption enhancers in ophthalmic preparations.

The use of absorption promoters is a way to improve the bioavailability and therapeutic response of topically applied ophthalmic drugs. The ocular tolerance of 9 potential absorption promoters was investigated as well as the influence of the enhancers' concentration on the ocular tolerance. The substances tested were instillated repetitively (4 times per day, during 3 days, and once just before examination) as aqueous solutions onto rabbit corneas. Fluorescein dyeing enabled us to specifically mark corneal damage that was observed by confocal microscopy. The degree of corneal injury was assessed with an image-processing system that calculated the total fluorescent areas. Confocal microscopy results showed the relatively good tolerance of permeation enhancers like dimethyl sulfoxide (DMSO), decamethonium, edetate, glycocholate, and cholate in contrast to the poorly tolerated saponin and fusidate. Increasing the promoters' concentration led generally to an increase in corneal lesions.

Ocular tolerance of preservatives and alternatives.

Eye drops are multiple dosage forms protected against microbial contamination by means of preservatives. However, the ocular tolerance of these chemicals can vary and this may result in adverse toxic or allergic reactions. This overview presents the pharmacopoeial requirements for the preservation of eye drops, the factors affecting ocular tolerance as well as the adverse external ocular effects induced by preservatives. The alternatives to the use of preservatives are also discussed, including the recent progress in eye drops packaging.