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Classical approaches to engineer skeletal muscle tissue based on current regenerative and surgical procedures still do not meet the desired outcome for patient applications. Besides the evident need to create functional skeletal muscle tissue for the repair of volumetric muscle defects, there is also growing demand for platforms to study muscle-related diseases, such as muscular dystrophies or sarcopenia. Currently, numerous studies exist that have employed a variety of biomaterials, cell types and strategies for maturation of skeletal muscle tissue in 2D and 3D environments. However, researchers are just at the beginning of understanding the impact of different culture settings and their biochemical (growth factors and chemical changes) and biophysical cues (mechanical properties) on myogenesis. With this review we intend to emphasize the need for new in vitro skeletal muscle (disease) models to better recapitulate important structural and functional aspects of muscle development. We highlight the importance of choosing appropriate system components, e.g., cell and biomaterial type, structural and mechanical matrix properties or culture format, and how understanding their interplay will enable researchers to create optimized platforms to investigate myogenesis in healthy and diseased tissue. Thus, we aim to deliver guidelines for experimental designs to allow estimation of the potential influence of the selected skeletal muscle tissue engineering setup on the myogenic outcome prior to their implementation. Moreover, we offer a workflow to facilitate identifying and selecting different analytical tools to demonstrate the successful creation of functional skeletal muscle tissue. Ultimately, a refinement of existing strategies will lead to further progression in understanding important aspects of muscle diseases, muscle aging and muscle regeneration to improve quality of life of patients and enable the establishment of new treatment options.
Glyphosate-based herbicides were used as plant protection product globally for several decades. However, glyphosate is discussed as showing genotoxicity and many other side-effects such as inhibiting the mitochondrial succinate dehydrogenase, leading to a decreased ATP production. Therefore, finding alternative active substances is necessary. Pelargonic acid (nonanoic acid) and its ammonium salt (saponified form), which are used as alternatives to glyphosate-based herbicides, are biological derived substances considered as environmentally friendly herbicides. To test the effects of pelargonic acid in its acidic form (active substance in TopGun) and its saponified form (in Finalsan Plus) on the aquatic ecosystem, students of the master study program “environmental management and ecotoxicology” compared the toxicity levels of these substances using zebrafish embryos within the scope of a student research course. The project was developed by applying the 7-step problem-based learning method which allowed the students to design their experiments independently with the guidance and feedback of the lecturers. Acute toxicity was determined according to OECD test guideline 236 in D. rerio which revealed a LC50 of 1.55 mg/L of pelargonic acid, a LC50 value of 0.93 mg/L pelargonic acid in TopGun and a LC50 of 36.37 mg/L of Finalsan Plus. Neutral Red Uptake assays were performed on the rainbow trout-derived gill cell-line RTgill-W1 to determine the acute toxicity according to the OECD test guideline 129 which revealed the IC50 value of 12.4 mg/L pelargonic acid in TG. Due to solubility limits, no reliable IC50 could be obtained. The results of the tests indicate differences in the acute toxicity of the pelargonic acid-based formulations TopGun and Finalsan Plus. However, more investigations have to be done in order to analyse if the acidic form is more toxic than the saponified form. Furthermore, detailed ecotoxicological risk assessments and an evaluation of the genotoxicity of both herbicides has to be performed before any conclusion can be drawn. Financial support from the City of Vienna project PBL in Molecular Life Science (21-06) is gratefully acknowledged.
