A muscarinic receptor antagonist reverses multiple indices of diabetic peripheral neuropathy: preclinical and clinical studies using oxybutynin.

Preclinical studies indicate that diverse muscarinic receptor antagonists, acting via the M1 sub-type, promote neuritogenesis from sensory neurons in vitro and prevent and/or reverse both structural and functional indices of neuropathy in rodent models of diabetes. We sought to translate this as a potential therapeutic approach against structural and functional indices of diabetic neuropathy using oxybutynin, a muscarinic antagonist approved for clinical use against overactive bladder. Studies were performed using sensory neurons maintained in vitro, rodent models of type 1 or type 2 diabetes and human subjects with type 2 diabetes and confirmed neuropathy. Oxybutynin promoted significant neurite outgrowth in sensory neuron cultures derived from adult normal rats and STZ-diabetic mice, with maximal efficacy in the 1-100 nmol/l range. This was accompanied by a significantly enhanced mitochondrial energetic profile as reflected by increased basal and maximal respiration and spare respiratory capacity. Systemic (3-10 mg/kg/day s.c.) and topical (3% gel daily) oxybutynin reversed paw heat hypoalgesia in the STZ and db/db mouse models of diabetes and reversed paw tactile allodynia in STZ-diabetic rats. Loss of nerve profiles in the skin and cornea of db/db mice was also prevented by daily topical delivery of 3% oxybutynin for 8 weeks. A randomized, double-blind, placebo-controlled interventional trial was performed in subjects with type 2 diabetes and established peripheral neuropathy. Subjects received daily topical treatment with 3% oxybutynin gel or placebo for 6 months. The a priori designated primary endpoint, significant change in intra-epidermal nerve fibre density (IENFD) in skin biopsies taken before and after 20 weeks of treatments, was met by oxybutynin but not placebo. Secondary endpoints showing significant improvement with oxybutynin treatment included scores on clinical neuropathy, pain and quality of life scales. This proof-of-concept study indicates that muscarinic antagonists suitable for long-term use may offer a novel therapeutic opportunity for treatment of diabetic neuropathy. Trial registry number: NCT03050827.

Methods to Characterize Electrospun Scaffold Morphology: A Critical Review.

Electrospun scaffolds can imitate the hierarchical structures present in the extracellular matrix, representing one of the main concerns of modern tissue engineering. They are characterized in order to evaluate their capability to support cells or to provide guidelines for reproducibility. The issues with widely used methods for morphological characterization are discussed in order to provide insight into a desirable methodology for electrospun scaffold characterization. Reported methods include imaging and physical measurements. Characterization methods harbor inherent limitations and benefits, and these are discussed and presented in a comprehensive selection matrix to provide researchers with the adequate tools and insights required to characterize their electrospun scaffolds. It is shown that imaging methods present the most benefits, with drawbacks being limited to required costs and expertise. By making use of more appropriate characterization, researchers will avoid measurements that do not represent their scaffolds and perhaps might discover that they can extract more characteristics from their scaffold at no further cost.

How Fiber Surface Topography Affects Interactions between Cells and Electrospun Scaffolds: A Systematic Review.

Electrospun scaffolds have a 3D fibrous structure that attempts to imitate the extracellular matrix in order to be able to host cells. It has been reported in the literature that controlling fiber surface topography produces varying results regarding cell-scaffold interactions. This review analyzes the relevant literature concerning in vitro studies to provide a better understanding of the effect that controlling fiber surface topography has on cell-scaffold interactions. A systematic approach following PRISMA, GRADE, PICO, and other standard methodological frameworks for systematic reviews was used. Different topographic interventions and their effects on cell-scaffold interactions were analyzed. Results indicate that nanopores and roughness on fiber surfaces seem to improve proliferation and adhesion of cells. The quality of the evidence is different for each studied cell-scaffold interaction, and for each studied morphological attribute. The evidence points to improvements in cell-scaffold interactions on most morphologically complex fiber surfaces. The discussion includes an in-depth evaluation of the indirectness of the evidence, as well as the potentially involved publication bias. Insights and suggestions about dose-dependency relationship, as well as the effect on particular cell and polymer types, are presented. It is concluded that topographical alterations to the fiber surface should be further studied, since results so far are promising.

Main Morphological Characteristics of Tubular Polymeric Scaffolds to Promote Peripheral Nerve Regeneration-A Scoping Review.

The "nerve guide conduits" (NGC) used in nerve regeneration must mimic the natural environment for proper cell behavior. OBJECTIVE: To describe the main morphological characteristics of polymeric NGC to promote nerve regeneration. METHODS: A scoping review was performed following the Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) criteria in the PubMed, Web of Science, Science Direct, and Scientific Electronic Library Online (SciELO) databases. Primary studies that considered/evaluated morphological characteristics of NGC to promote nerve regeneration were included. RESULT: A total of 704 studies were found, of which 52 were selected. The NGC main morphological characteristics found in the literature were: (I) NGC diameter affects the mechanical properties of the scaffold. (II) Wall thickness of NGC determines the exchange of nutrients, molecules, and neurotrophins between the internal and external environment; and influences the mechanical properties and biodegradation, similarly to NGC (III) porosity, (IV) pore size, and (V) pore distribution. The (VI) alignment of the NGC fibers influences the phenotype of cells involved in nerve regeneration. In addition, the (VII) thickness of the polymeric fiber influences neurite extension and orientation. CONCLUSIONS: An NGC should have its diameter adjusted to the nerve with wall thickness, porosity, pore size, and distribution of pores, to favor vascularization, permeability, and exchange of nutrients, and retention of neurotrophic factors, also favoring its mechanical properties and biodegradability.

Peripheral Neuropathy in Mouse Models of Diabetes.

Peripheral neuropathy is a frequent complication of chronic diabetes that most commonly presents as a distal degenerative polyneuropathy with sensory loss. Around 20% to 30% of such patients may also experience neuropathic pain. The underlying pathogenic mechanisms are uncertain, and therapeutic options are limited. Rodent models of diabetes have been used for more than 40 years to study neuropathy and evaluate potential therapies. For much of this period, streptozotocin-diabetic rats were the model of choice. The emergence of new technologies that allow relatively cheap and routine manipulations of the mouse genome has prompted increased use of mouse models of diabetes to study neuropathy. In this article, we describe the commonly used mouse models of type 1 and type 2 diabetes, and provide protocols to phenotype the structural, functional, and behavioral indices of peripheral neuropathy, with a particular emphasis on assays pertinent to the human condition. © 2016 by John Wiley & Sons, Inc.