Isolation and characterization of muscle stem cells, fibro-adipogenic progenitors, and macrophages from human skeletal muscle biopsies.

Animal models clearly illustrate that the maintenance of skeletal muscle mass depends on the function and interaction of a heterogeneous population of resident and infiltrating mononuclear cells. Several lines of evidence suggest that mononuclear cells also play a role in muscle wasting in humans, and targeting these cells may open new treatment options for intervention or prevention in sarcopenia. Methodological and ethical constraints have perturbed exploration of the cellular characteristics and function of mononuclear cells in human skeletal muscle. Thus, investigations of cellular phenotypes often depend on immunohistochemical analysis of small tissue samples obtained by needle biopsies, which do not match the deep phenotyping of mononuclear cells obtained from animal models. Here, we have developed a protocol for fluorescence-activated cell sorting (FACS), based on single-cell RNA-sequencing data, for quantifying and characterizing mononuclear cell populations in human skeletal muscle. Muscle stem cells, fibro-adipogenic progenitors, and two subsets of macrophages (CD11c+/-) are present in needle biopsies in comparable quantities per milligram tissue to open surgical biopsies. We find that direct cell isolation is preferable due to a substantial shift in transcriptome when using preculture before the FACS procedure. Finally, in vitro validation of the cellular phenotype of muscle stem cells, fibro-adipogenic progenitors, and macrophages confirms population-specific traits. This study demonstrates that mononuclear cell populations can be quantified and subsequently analyzed from needle biopsy material and opens the perspective for future clinical studies of cellular mechanisms in muscle wasting.

Local concentrations of gentamicin obtained by microdialysis after a controlled application of a GentaColl sponge in a porcine model.

Local treatment with gentamicin may be an important tool in the prevention and treatment of surgical site infections in high-risk procedures and patients. The aim of this study was to evaluate the pharmacokinetic profile of gentamicin in bone and surrounding tissue, released from a controlled application of a GentaColl sponge in a porcine model. In eight female pigs, a GentaColl sponge of 10 × 10 cm (1.3 mg gentamicin/cm2 ) was placed in a cancellous bone cavity in the proximal tibia. Microdialysis was used for sampling of gentamicin concentrations over 48 hours from the cavity with the implanted GentaColl sponge, cancellous bone parallel to the cavity over and under the epiphyseal plate, cortical bone, the intramedullary canal, subcutaneous tissue, and the joint cavity of the knee. Venous blood samples were obtained as reference. The main finding was a mean peak drug concentration (95% CI) of gentamicin in the cancellous bone cavity containing the implanted GentaColl sponge of 11 315 (9049-13 581) µg/mL, persisting above 1000 µg/mL until approximately 40 hours after application. Moreover, the concentrations were low (<1 µg/mL) in the surrounding tissues as well as in plasma. The mean peak gentamicin concentration from the cancellous bone cavity after a controlled application of a GentaColl sponge was high and may be adequate for the prevention of biofilm formation. However, high MIC strains and uncontrolled application of the GentaColl sponge may jeopardize this conclusion.

Life-threatening MRSA sepsis with bilateral pneumonia, osteomyelitis, and septic arthritis of the knee in a previously healthy 13-year-old boy: a case report.

The incidence and severity of methicillin resistant Staphylococcus aureus (MRSA) infections are increasing and cause high mortality and morbidity. We describe the first pediatric case in Scandinavia with Panton-Valentine leucocidin (PVL) positive MRSA septicemia who developed bilateral pneumonia, arthritis of the knee, and osteomyelitis of the tibia. Radiological investigation and interpretation directed the treatment, especially the surgical debridement, and combined with clinical and biochemical findings lead to close interdisciplinary treatment with frequent surgical interventions and antimicrobial combination therapy. The outcome was a healthy patient without sequelae, a favorable course unlike those previously described in the literature. This case underlines the necessity of a close interdisciplinary cooperation in children with severe MRSA infection encompassing pneumonia, septic arthritis, and osteomyelitis, using different imaging modalities to guide the surgical and antibiotic treatment.