Sperm preparation after freezing improves motile sperm count, motility, and viability in frozen-thawed sperm compared with sperm preparation before freezing-thawing process.

PURPOSE: The aim of this study is to evaluate which cryopreservation protocol, freezing before or after swim-up, optimizes cryopreservation outcomes in terms of motile sperm count, motility, morphology, and viability, and also to establish whether sperm viability could be assessed based on sperm motility. METHODS: Fifty-three fresh and 53 swim-up prepared samples were considered for the first experiment. In parallel, total motility evaluation by CASA system (computer-assisted sperm analyzer) and hypoosmotic swelling test (HOS-test) was performed in each sample to compare the viability results of both methods. In the second experiment, 21 normozoospermic semen samples and 20 semen samples from male factor patients were included. After fresh ejaculate evaluation, the semen sample of each patient was divided into two aliquots, one of them was frozen before swim-up and the other was frozen after swim-up. Motility, sperm count, morphology, and viability were evaluated after thawing. RESULTS: A linear regression model allows prediction of HOS-test viability results based on total motility: HOS = 1.38 + 0.97 · TM (R 2 = 99.10, residual mean squares = 9.51). Freezing before sperm selection leads to higher total and progressive motility, total motile sperm count, and viability rates than when sperm selection is performed before freezing (P < 0.005 in all cases). In fact, sperm selection prior to freezing reaches critical values when subfertile patients are considered. CONCLUSIONS: To conclude, total motility evaluation can predict HOS-test viability results, resulting in a more objective and less time-consuming method to assess viability. In addition, sperm freezing prior to swim-up selection must be considered in order to achieve better outcomes after thawing, especially in patients presenting poor sperm baseline.

Meropenem dosing requirements against Enterobacteriaceae in critically ill patients: influence of renal function, geographical area and presence of extended-spectrum ß-lactamases.

The aim of this study was the evaluation of the influence of the susceptibility patterns of Escherichia coli and Klebsiella pneumoniae isolates, specifically the presence of extended-spectrum ß-lactamases and the geographical area (Europe and USA), on the meropenem dosing requirements in critically ill patients with different degrees of renal function by estimation of the probability of pharmacokinetic/pharmacodynamic (PK/PD) target attainment. Additionally, estimation of the PK/PD breakpoints according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) approach was also an objective. Six dosing regimens were evaluated: 0.5 g, 1 g and 2 g every 8 h given as 0.5-h or 3-h infusions. Pharmacokinetic data were obtained from the literature, and susceptibility data to meropenem for E. coli and K. pneumoniae were collected from the Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) surveillance study. For the same dose level, the 3-h infusion provided a probability of target attainment (PTA) ≥90 % for minimum inhibitory concentration (MIC) values up to two-fold dilution higher than those obtained with the 0.5-h infusion. For E. coli, the cumulative fraction of response (CFR) was 100 % in most cases, and neither the dose nor the infusion length nor the geographical area significantly affected the probability to reach the target. With regards to K. pneumoniae, the CFR increased when increasing the dose and decreasing the creatinine clearance (CLCR). The CFR for Spanish and USA strains was higher than that calculated for European strains. Meropenem PK/PD breakpoints are dependent on the dose, infusion length and CLCR, ranging from 2 to 32 mg/L. Based on our results, meropenem administered as a extended infusion is the best option to treat infections due to E. coli and K. pneumoniae.

Pharmacokinetic-pharmacodynamic evaluation of daptomycin, tigecycline, and linezolid versus vancomycin for the treatment of MRSA infections in four western European countries.

PURPOSE: To evaluate the usefulness of daptomycin, tigecycline, and linezolid for the treatment of MRSA infection compared with vancomycin in Belgium, the United Kingdom/Ireland, and Spain. METHODS: The methodology included the following steps: acquisition of microbiological and pharmacokinetic data, Monte Carlo simulation, estimation of the probability of target attainment (PTA), and calculation of the cumulative fraction of response (CFR). RESULTS: We showed that differences in the susceptibility of MRSA strains among countries may justify differences in the antibiotic dose selection. Two, 3, and 4 g daily of vancomycin seem be adequate in Belgium, Spain, and United Kingdom/Ireland respectively. The CFR obtained with 50 mg tigecycline every 12 h was higher in Spain than in Belgium and the United Kingdom/Ireland, but with the highest dose (100 mg q12h) the CFR was always 100%. At least 8 mg/kg daptomycin is necessary in United Kingdom/Ireland, but 4 mg/kg may be sufficient in Spain, and probably in Belgium. Six hundred mg q12h linezolid may be adequate in the four countries. CONCLUSION: Our study reinforces the idea that the local MIC distribution must be considered in order to increase the probability of success of empirical treatment and must be periodically updated.

Deciphering pharmacokinetics and pharmacodynamics of fosfomycin.

Fosfomycin, a low molecular weight and hydrophilic drug with negligible protein binding, is eliminated almost exclusively by glomerular filtration, whose clearance is subject to patient renal function. The volume of distribution approximates to the extracellular body water (about 0.3 L/Kg) in healthy volunteers, but it is increased in critically ill patients with bacterial infections. Fosfomycin presents a high ability to distribute into many tissues, including inflamed tissues and abscess fluids. Based on PK/PD analysis and Monte Carlo simulations, we have evaluated different fosfomycin dosing regimen to optimize the treatment of septic patients due to Enterobacterales and Pseudomonas aeruginosa. As PK/PD targets, we selected %T>MIC > 70% for all pathogens, and AUC24/MIC > 24 and AUC24/MIC > 15 for net stasis of Enterobacterales and P. aeruginosa, respectively. Pharmacokinetic parameters in critically ill patients were obtained from the literature. Several dosing regimens were studied in patients with normal renal function: fosfomycin 2-8 g given every 6-12 hours, infused over 30 minutes- 24 hours. At the susceptibility EUCAST breakpoint for Enterobacterales and Staphylococcus spp. (MIC ≤ 32 mg/L), fosfomycin 4 g/8h or higher infused over 30 minutes achieved a probability of target attainment (PTA) > 90%, based in both %T>MIC and AUC24/MIC. For MIC of 64 mg/L, fosfomycin 6 g/6h in 30-minute infusion and 8 g/ 8h in 30-minute and 6 hours infusions also achieved PTA values higher than 90%. No fosfomycin monotherapy regimen was able to achieve PK/PD targets related to antimicrobial efficacy for P. aeruginosa with MICs of 256-512 mg/L.

Application of pharmacokinetic/pharmacodynamic analysis to evaluate the adequacy of antimicrobial therapy for pediatric acute otitis media in Spain before and after the introduction of the PCV7 vaccine.

OBJECTIVE: To evaluate, by applying pharmacokinetic/pharmacodynamic (PK/PD) analysis, if the change in antibiotic susceptibility after the introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) in Spain had any influence on the usefulness of the antimicrobials more frequently used as empirical treatment of pediatric acute otitis media (AOM). METHODS: PK parameters and susceptibility of Streptococcus pneumoniae and Haemophilus influenzae were obtained from bibliography. Monte Carlo simulation was used to estimate the cumulative fraction of response (CFR), understood as the expected probability of therapy success. For amoxicillin and amoxicillin/clavulanate, the target was free antibiotic concentration remaining above the minimum inhibitory concentration (MIC) for ≥50% of the dosing interval (fT>MIC≥50%), whereas for cefuroxime axetil and cefotaxime, the target was fT>MIC≥60%. CFR values ≥90% were considered successful. RESULTS: When all serotypes of S. pneumoniae are considered, amoxicillin and cefotaxime turned out to reach a high probability of success, and difference before and after vaccination was scarce. For H. influenzae, CFR values were higher with amoxicillin/clavulanate than with amoxicillin. For both microorganisms, cefuroxime axetil resulted in low probability of success in the two periods of study. CONCLUSIONS: We have shown that the introduction of the PCV7 vaccination did not lead to changes in the probability of success of the current empiric treatments of the AOM. Integrated PK/PD analysis has demonstrated to be a useful tool to identify changes in antimicrobial activity after the implantation of a vaccination program, providing complementary information to the simple assessment of MIC values.

Solid lipid nanoparticles for retinal gene therapy: transfection and intracellular trafficking in RPE cells.

Retinal pigment epithelial (RPE) cells are usually employed to study DNA systems for diseases related to problems in the retina. Solid lipid nanoparticles (SLNs) have been shown to be useful non-viral vectors for gene therapy. The objective of this work was to evaluate the transfection capacity of SLNs in the human retinal pigment epithelial established cell line (ARPE-19) in order to elucidate the potential application of this vector in the treatment of retinal diseases. Results showed a lower transfection level of SLNs in ARPE-19 cells than in HEK293 (2.5% vs. 14.9% EGFP positive cells at 72h post-transfection). Trafficking studies revealed a delay in cell uptake of the vectors in ARPE-19 cells. Differences in internalization process into the two cell lines studied explain, in part, the difference in the gene expression. The clathrin-mediated endocytosis in ARPE-19 cells directs the solid lipid nanoparticles to lysosomes; moreover, the low division rate of this cell line hampers the entrance of DNA into the nucleus. The knowledge of intracellular trafficking is very useful in order to design more efficient vectors taking into account the characteristics of the specific cell line to be transfected.

Population pharmacokinetics of daptomycin in critically ill patients.

Daptomycin has shown activity against a wide range of Gram-positive bacteria; however, the approved dosages usually seem insufficient for critically ill patients. The aim of this study was to develop a population pharmacokinetic model for daptomycin in critically ill patients and to estimate the success of the therapy by applying pharmacokinetic/pharmacodynamic (PK/PD) criteria. Sixteen intensive care unit patients were included, four of whom underwent continuous renal replacement therapies (CRRT). Blood and, when necessary, effluent samples were drawn after daptomycin administration at previously defined time points. A population approach using NONMEM 7.3 was performed to analyse data. Monte Carlo simulations were executed to evaluate the suitability of different dosage regimens. The probabilities of achieving the PK/PD target value associated with treatment success (ratio of the area under the plasma concentration-time curve over 24 h divided by the minimum inhibitory concentration (AUC24/MIC ≥ 666)) and to reach daptomycin concentrations linked to toxicity (minimum concentration at steady-state (Cminss) ≥ 24.3 mg/L) were calculated. The pharmacokinetics of daptomycin was best described by a one-compartment model. Elimination was conditioned by the creatinine clearance (Clcr) and also by the extra-corporeal clearance when patients were subjected to continuous renal replacement therapy (CRRT). The PK/PD analysis confirmed that 280- and 420-mg/d dosages would not be enough to achieve high probabilities of target attainment for MIC values ≥ 1 mg/L in patients with Clcr ≥ 60 mL/min or in subjects with lower Clcrs but receiving CRRT. In these patients, higher dosages (560-840 mg/d) should be needed. When treating infections due to MIC values ≥ 4 mg/L, even the highest dose would be insufficient.

Interleukin (IL) -1 ß, IL-6 and tumor necrosis factor in patients with seasonal flu.

OBJECTIVE: The role of tumor necrosis factor (TNF), interleukin (IL)-1ß and IL-6 in the pathogenicity of seasonal flu is unknown. METHODS: We analyzed the profiles of these cytokines in 77 flu patients and 17 controls with non-flu respiratory infection, using molecular biology techniques (real-time polymerase chain reaction). RESULTS: Flu patients had lower monocyte counts (p=0.029) and a slightly lower median level of IL-6 (P=0.05) than the control group. Twenty-four flu patients (31.2%) had pneumonia; this group had higher C-reactive proteins (p=0.01) and monocyte levels (p=0.009). Pro-inflammatory cytokines levels did not rise in patients with pneumonia complicating seasonal influenza. CONCLUSIONS: IL-6 levels were lower in adults with influenza.

Solid lipid nanoparticles: formulation factors affecting cell transfection capacity.

Since solid lipid nanoparticles (SLNs) were introduced as non-viral transfection systems, very few reports of their use for gene delivery have been published. In this work different formulations based on SLN-DNA complexes were formulated in order to evaluate the influence of the formulation components on the "in vitro" transfection capacity. SLNs composed by the solid lipid Precirol ATO 5, the cationic lipid DOTAP and the surfactant Tween 80, and SLN-DNA complexes prepared at different DOTAP/DNA ratios were characterized by studying their size, surface charge, DNA protection capacity, transfection and cell viability in HEK293 cultured cells. The incorporation of Tween 80 allowed for the reduction of the cationic lipid concentration. The formulations prepared at DOTAP/DNA ratios 7/1, 5/1 and 4/1 provided almost the same transfection levels (around 15% transfected cells), without significant differences between them (p>0.05). Other assayed formulations presented lower transfection. Transfection activity was dependent on the DOTAP/DNA ratio since it influences the DNA condensation into the SLNs. DNA condensation is a crucial factor which conditions the transfection capacity of SLNs, because it influences DNA delivery from nanoparticles, gene protection from external agents and DNA topology.

[Cellular immune response to 3 anesthetic techniques for simple abdominal hysterectomy]. / Estudio de la respuesta celular inmune de tres técnicas de anestesia para histerectomía abdominal simple.

BACKGROUND AND OBJECTIVE: The effect of surgery and anesthesia on the immune response may have a significant effect on perioperative tumor surveillance. The aim of this study was to characterize the cellular immune response of patients undergoing simple abdominal hysterectomy under 3 types of anesthesia. PATIENTS AND METHODS: ASA 1-2 patients undergoing simple abdominal hysterectomy were enrolled prospectively after they gave informed consent; the patients were randomized to 3 groups of 20 each to receive balanced anesthesia (group A), remifentanil-based anesthesia and analgesia (group B), or combined general-epidural anesthesia (group C). Postoperative analgesia was provided in accordance with group assignment. Four and 24 hours after surgery, 20 mL of blood was drawn from each patient for analysis of leukocyte populations and lymphocyte subpopulations. Statistics were calculated with the SPSS software package, version 12.0. RESULTS: All groups had elevated neutrophil counts after surgery; the lowest levels were in group C (P<.05). Patients in all 3 groups developed lymphopenia, which was still evident 24 hours after surgery (P<.05). CD3 cell counts at 4 hours were lowest in patients who had received combined anesthesia (group C), CD19 cell counts were highest in group A, and CD16 cell counts were lowest in group C; this last difference was maintained at 24 hours (P<.05 for all these comparisons). CONCLUSION: Combined general-epidural anesthesia seems to lower the counts of natural killer cells that are involved in tumor surveillance and destruction.

In vivo evaluation of EPO-secreting cells immobilized in different alginate-PLL microcapsules.

Alginates are the most employed biomaterials for cell encapsulation due to their abundance, easy gelling properties and apparent biocompatibility. However, as natural polymers different impurities including endotoxins, proteins and polyphenols can be found in their composition. Several purification protocols as well as different batteries of assays to prove the biocompatibility of the alginates in vitro have been recently developed. However, little is known about how the use of alginates with different purity grade may affect the host immune response after their implantation in vivo. The present paper investigates the long-term functionality and biocompatibility of murine erythropoietin (EPO) secreting C2C12 cells entrapped in microcapsules elaborated with alginates with different properties (purity, composition and viscosity). Results showed that independently of the alginate type employed, the animals presented elevated hematocrit levels until day 130, remaining at values between 70-87%. However, histological analysis of the explanted devices showed higher overgrowth around non-biomedical grade alginate microcapsules which could be directly related with higher impurity content of this type of alginate. Although EPO delivery may be limited by the formation of a fibrotic layer around non-biomedical grade alginate microcapsules, the high EPO secretion of the encapsulated cells together with the pharmacodynamic behaviour and the angiogenic and immune-modulatory properties of EPO result in no direct correlation between the biocompatibility of the alginate and the therapeutic response obtained.

Evaluation of human serum albumin as a substitute of foetal bovine serum for cell culture.

Cell microencapsulation requires clinically approved materials for their use in pharmaceutical and/or biomedical applications. The overwhelming majority of the literature has used the classical alginate-poly-l-lysine-alginate (APA) capsules for cell immobilization. Although alginate is granted with the medical approval, some of the remaining components such as foetal bovine serum (FBS), an essential ingredient of cell culture media, are not in accordance with the guidelines affirmed by the American Society for Testing and Materials (ASTM) and Food and Drug Administration (FDA). In this paper, human serum albumin (HSA), a medically approved substance, was evaluated as a potential substitute of FBS. The effect of different percentages of FBS and HSA was studied on the proliferation rate, viability and protein production of two different cell lines (C2C12 and baby hamster kidney (BHK) cells), maintained in culture and immobilized in APA microcapsules. Results show that substitution of FBS by HSA reduced the functionality of both non-encapsulated and encapsulated BHK cells. However, immobilized C2C12 cells presented the highest level of viability and a reduction in protein production of 25% when 1% HSA was used. It can be concluded that HSA might be a possible substitute of FBS in order to maintain or transport encapsulated C2C12 cells for short periods of time before implantation.

Structural recovery of the retina in a retinoschisin-deficient mouse after gene replacement therapy by solid lipid nanoparticles.

X-linked juvenile retinoschisis (XLRS) is a retinal degenerative disorder caused by mutations in the RS1 gene encoding a protein termed retinoschisin. The disease is an excellent candidate for gene replacement therapy as the majority of mutations have been shown to lead to a complete deficiency of the secreted protein in the retinal structures. In this work, we have studied the ability of non-viral vectors based on solid lipid nanoparticles (SLN) to induce the expression of retinoschisin in photoreceptors (PR) after intravitreal administration to Rs1h-deficient mice. We designed two vectors prepared with SLN, protamine, and dextran (DX) or hyaluronic acid (HA), bearing a plasmid containing the human RS1 gene under the control of the murin opsin promoter (mOPS). In vitro, the nanocarriers were able to induce the expression of retinoschisin in a PR cell line. After injection into the murine vitreous, the formulation prepared with HA induced a higher transfection level in PR than the formulation prepared with DX. Moreover, the level of retinoschisin in the inner nuclear layer (INL), where bipolar cells are located, was also higher. Two weeks after vitreal administration into Rs1h-deficient mice, both formulations showed significant improvement of the retinal structure by inducing a decrease of cavities and PR loss, and an increase of retinal and outer nuclear layer (ONL) thickness. HA-SLN resulted in a significant higher increase in the thickness of both retina and ONL, which can be explained by the higher transfection level of PR. In conclusion, we have shown the structural improvement of the retina of Rs1h-deficient mice with PR specific expression of the RS1 gene driven by the specific promoter mOPS, after successful delivery via SLN-based non-viral vectors.

Seasonal influenza in octogenarians and nonagenarians admitted to a general hospital: epidemiology, clinical presentation and prognostic factors.

OBJECTIVE: Seasonal influenza is responsible for high annual morbidity and mortality worldwide, especially in elderly patients. The aim of the study was to analyse the epidemiological, clinical and prognostic features of influenza in octogenarians and nonagenarians admitted to a general hospital, as well as risk factors associated with mortality. METHODS: Retrospective, cross-sectional, descriptive study in patients admitted and diagnosed with influenza by molecular biology in the General University Hospital of Alicante from 1 January to 31 April 2015. RESULTS: A total of 219 patients were diagnosed with influenza in the study period: 55 (25.1%) were ≤64 years-old; 77 (35.2%) were aged 65-79; 67 (30.6%) were aged 80-89 years; and 20 (9.1%) were aged ≥90 years. Most flu episodes were caused by influenza A (n=181, 82.6%). Patients aged 80 years or older had lower glomerular filtration rate (mean: 49.7 mL/min vs. 62.2 mL/min; p=0.006), a greater need for non-invasive mechanical ventilation (22% vs 9.3%; p=0.02), greater co-morbidity due to cardiac insufficiency (40.5% vs. 16.4%; p<0.001) and chronic renal disease (32.9 vs. 20%, p=0.03), and greater mortality (19% vs. 2.9%; p<0.001). In a multivariate analysis, mortality was higher in those aged 80 or over (adjusted odds ratio [ORa] 9.2, 95% confidence interval [CI] 1.65-51.1), those who had acquired the flu in a long-term care facility (ORa 11.9, 95% CI 1.06-134), and those with hyperlactataemia (ORa 1.89, 95% CI 1.20-3.00). CONCLUSIONS: Seasonal influenza is a serious problem leading to elevated mortality in octogenarian and nonagenarian patients admitted to a general hospital.

Cloning and sequencing of a tpr-met oncogene cDNA isolated from MNNG-transformed human XP fibroblasts.

A rearranged tpr-met oncogene was identified in a MNNG-transformed human Xeroderma pigmentosum (XP) cell line (ASKMN). A 2016 bp cDNA was cloned and sequenced, disclosing an ORF with a coding capacity for a 523 aa protein. The sequence of this tpr-met cDNA was very similar to that previously reported in another human MNNG-transformed cell line (MNNG-HOS).

Microcapsules prepared with different biomaterials to immobilize GDNF secreting 3T3 fibroblasts.

Cell microencapsulation represents a promising tool for the treatment of many central nervous system (CNS) diseases such as Parkinson's disease. In this technology, cells are surrounded by a semipermeable membrane which protects them from mechanical stress and isolates them from host's immune response. However, if the future clinical application of this strategy is wanted, many challenges remain including the improvement of the mechanical resistance of the microcapsules and the optimization of the intracapsular microenvironment conditions. In this way, the selection of the matrix is essential because the morphological and the physiological behavior of the cells depend on the interactions between the matrix and the enclosed cells. Assuming these considerations, three types of microcapsules elaborated with four different polymers: alginate, cellulose sulfate, agarose and pectin have been fabricated and compared in order to evaluate some key properties such as morphology, size and mechanical stability. Furthermore, GDNF secreting Fischer rat 3T3 fibroblasts were immobilized in each type of capsule and the viability and neurotrophic factor release was determinated. Results showed that the alginate and pectin microcapsules were the most resistant devices, maintaining an adequate microenvironment for the enclosed cells. In contrast, cells entrapped in alginate-cellulose sulfate matrices presented the lowest mechanical resistance, cell viability and GDNF production.

Determination of ceftazidime and cefepime in plasma and dialysate-ultrafiltrate from patients undergoing continuous veno-venous hemodiafiltration by HPLC.

We have developed and validated a new, rapid and reproducible HPLC method for the determination of cefepime and ceftazidime in plasma and dialysate-ultrafiltrate samples obtained from intensive care unit (ICU) patients undergoing continuous veno-venous hemodiafiltration (CVVHDF). The method for plasma samples involved protein precipitation with acetonitrile, followed by washing with dichloromethane to remove apolar lipophilic compounds. Dialysate-ultrafiltrate samples did not require any preparation. Separation was performed on a muBondapak C18 (30 cm x 3.9 mm x 10 microm) with UV detection. The mobile phase contained acetate buffer: ACN and was delivered at 2 ml/min. The coefficients of determination of the calibration curves were always > or = 0.998 and R.S.D.% of the response factors <10%. The intra and inter-assay precision and accuracy of the quality controls (QC) and limit of quantification (LOQ) were satisfactory in all cases. Plasma and dialysate-ultrafiltrate samples were stable at -20 and -80 degrees C for 2 months and also after three freeze/thaw cycles. Dialysate-ultrafiltrate samples were stable in the chromatographic rack for 24h at room temperature, but we recommend storing processed plasma samples at 4 degrees C until the analysis. The described method has proved to be useful to give accurate measurements of ceftazidime and cefepime in samples obtained from patients undergoing CVVHDF.

Evaluation of antimicrobial treatments in children with acute otitis media in Spain: a pharmacokinetic-pharmacodynamic (PK/PD) approach.

Pharmacokinetic/pharmacodynamic (PK/PD) principles are priceless tools for evaluating the effectiveness of different antimicrobial treatments for different infections. However, very few studies deal with pediatric dosages and take into account the unbound drug serum levels. Our study is focused on the most frequent antibiotic dosing schedules used in Spain for the treatment of acute otitis media (AOM) in children, where high rates of penicillin and macrolide resistance exist among pneumococcal isolates. Pharmacokinetic parameters of antibiotics in children where obtained from the literature. The minimum inhibitory concentrations (MIC90) of antibiotics for pediatric strains of Streptococcus pneumoniae and Haemophilus influenzae were obtained from the SAUCE 2 project. Only ceftriaxone (50 mg/kg single intramuscular dose) and high doses of co-amoxiclav (27-33 mg/kg q8h) provided adequate efficacy indexes (tss(%)>MIC) for both S. pneumoniae and H. influenzae in AOM in children. These results are consistent with MEF (medium ear fluid) levels obtained from the literature. Our results confirm the utility of serum unbound levels to predict efficacy of antibiotics in children with AOM.

Biocompatible oligochitosans as cationic modifiers of alginate/Ca microcapsules.

In the past, it has been proven that by properly adjusting the molecular mass of the oligochitosan samples, it is possible to optimize the formation of rigid, biocompatible capsules with semipermeable membranes under physiological conditions. In this study, the feasibility of four oligochitosan samples, with varying molar masses (M(n) in range 3-5 kDa), as biocompatible coatings of alginate/Ca capsules was investigated. By selection of appropriate depolymerization and purification methods we obtained oligochitosan samples that appeared to be noncytotoxic for C(2)C(12) myoblasts and did not influence the mammalian cell metabolism especially in relative short time during the process of capsule formation. Furthermore, oligochitosans can be used as a tool to reduce the membrane cut-off of the alginate capsules. However, such reduction, as well as mechanical resistance of formed microcapsules, depend on MM of the cationic polysaccharide and the chemical composition of the alginate (mannuronic/guluronic acid ratio). Here, we address that the use of low molar mass chitosan (< 5000 g/mol) permits the formation of mechanical stable capsules at physiological pH, which represents a strong advantage over other chitosan-based chemistries.