Leaping the hurdles in developing regenerative treatments for the intervertebral disc from preclinical to clinical

Chronic back and neck pain is a prevalent disability, often caused by degeneration of the intervertebral disc. Because current treatments for this condition are less than satisfactory, a great deal of effort is being applied to develop new solutions, including regenerative strategies. However, the path from initial promising idea to clinical use is frought with many hurdles to overcome. Many of the keys to success are not necessarily linked to science or innovation. Successful translation to clinic will also rely on planning and awareness of the hurdles. It will be essential to plan your entire path to clinic from the outset and to do this with a multidisciplinary team. Take advise early on regulatory aspects and focus on generating the proof required to satisfy regulatory approval. Scientific demonstration and societal benefits are important, but translation cannot occur without involving commercial parties, which are instrumental to support expensive clinical trials. This will only be possible when intellectual property can be protected sufficiently to support a business model. In this manner, commercial, societal, medical, and scientific partners can work together to ultimately improve patient health. Based on literature surveys and experiences of the co‐authors, this opinion paper presents this pathway, highlights the most prominent issues and hopefully will aid in your own transational endeavors

Type IV Pili in Francisella tularensis: Roles of pilF and pilT in Fiber Assembly, Host Cell Adherence, and Virulence ▿

Francisella tularensis, a highly virulent facultative intracellular bacterium, is the causative agent of tularemia. Genome sequencing of all F. tularensis subspecies revealed the presence of genes that could encode type IV pili (Tfp). The live vaccine strain (LVS) expresses surface fibers resembling Tfp, but it was not established whether these fibers were indeed Tfp encoded by the pil genes. We show here that deletion of the pilF putative Tfp assembly ATPase in the LVS resulted in a complete loss of surface fibers. Disruption of the pilT putative disassembly ATPase also caused a complete loss of pili, indicating that pilT functions differently in F. tularensis than in model Tfp systems such as those found in Pseudomonas aeruginosa and Neisseria spp. The LVS pilF and pilT mutants were attenuated for virulence in a mouse model of tularemia by the intradermal route. Furthermore, although absence of pili had no effect on the ability of the LVS to replicate intracellularly, the pilF and pilT mutants were defective for adherence to macrophages, pneumocytes, and hepatocytes. This work confirms that the surface fibers expressed by the LVS are encoded by the pil genes and provides evidence that the Francisella pili contribute to host cell adhesion and virulence

Short-term Results of Platelet-Rich Plasma in the Treatment of Chronic Anal Fissure

Background: Anal fissure is one of the most common benign anal disorders, and medical treatments play an important role in its management.Objective: The purpose of this study was to investigate the short-term effects and success of platelet-rich-plasma in the treatment of chronic anal fissure.Design: The study is a two parallel-group, randomized, controlled clinical trial.Settings: The study was carried out in two tertiary university hospitals.Patients: Forty-four-patients with chronic anal fissure were randomly assigned to platelet-rich-plasma treatment or control group. Presenting symptoms and pain scores were recorded on enrolment. The control patient self-administered topical glyceryl trinitrate. Platelet-rich-plasma was injected locally in the intervention group followed by self-administered glyceryl trinitrate.Main outcome measures: Primary outcome measure is a reduction in pain scores.Results: On day 10 and one month after treatment, the mean pain score was significantly lower in the platelet-rich-plasma patients than in the controls (p=0.005 and p<0.005, respectively). By one month after treatment, mean pain score declined by 5.7 points in the platelet-rich plasma-treated group compared with a 4.1 mean pain score decline in the control group (mean difference:1.6 points [95% confidence interval;0.3-2.9]).According to the repeated-measures analyses, Pain scores decreased in both groups, but the decrease in the treatment group was statistically higher than the control group (p<0.001).Complete epithelialization and recovery rates were significantly higher in the platelet-rich-plasma group than in controls at all follow-up times, with p-values ranging from 0.034 to<0.001.The observed difference in complete epithelialization after two months of treatment between the platelet-rich-plasma group and the control group was 56.2% with a 95% confidence interval of 14.03% to 98.4%.Limitations: This study was limited by its small sample size and long-term follow-up of the patients was not given.Conclusions: Platelet-rich-plasma reduced complaints and accelerated epithelialization and healing in patients with chronic anal fissures

Deletion of TolC orthologs in Francisella tularensis identifies roles in multidrug resistance and virulence

The Gram-negative bacterium Francisella tularensis is the causative agent of tularemia. Interest in this zoonotic pathogen has increased due to its classification as a category A agent of bioterrorism, but little is known about the molecular mechanisms underlying its virulence, and especially what secretion systems and virulence factors are present. In this study, we characterized two genes in the F. tularensis genome, tolC and a gene we term ftlC, whose products have high homology with the Escherichia coli TolC protein. TolC functions as the outer membrane channel component for both type I secretion and multidrug efflux systems. We constructed deletion mutations of these genes in the F. tularensis live vaccine strain by allelic replacement. Deletion of either tolC or ftlC caused increased sensitivity to various antibiotics, detergents, and dyes, indicating both genes are involved in the multidrug resistance machinery of F. tularensis. Complementation of the deletion mutations in trans restored drug resistance. Neither tolC nor ftlC was required for replication of the live vaccine strain in murine bone marrow-derived macrophages. However, deletion of tolC, but not ftlC, caused a significant attenuation of virulence in a mouse model of tularemia that could be complemented by addition of tolC in trans. Thus, tolC is a critical virulence factor of F. tularensis in addition to its role in multidrug resistance, which suggests the presence of a functional type I secretion system