Abstract
The capability to 3D print bespoke biologically receptive parts within short time periods has driven the growing prevalence of additive manufacture (AM) technology within biological settings, however limited research concerning cellular interaction with 3D printed polymers has been undertaken. In this work, we used skeletal muscle C2C12 cell line in order to ascertain critical evidence of cellular behaviour in response to multiple bio-receptive candidate polymers; polylactic acid (PLA), acrylonitrile butadiene styrene (ABS), polyethylene terephthalate (PET) and polycarbonate (PC) 3D printed via fused deposition modelling (FDM). The extrusion based nature of FDM elicited polymer specific topographies, within which C2C12 cells exhibited reduced metabolic activity when compared to optimised surfaces of tissue culture plastic, however assay viability readings remained high across polymers outlining viable phenotypes. C2C12 cells exhibited consistently high levels of morphological alignment across polymers, however differential myotube widths and levels of transcriptional myogenin expression appeared to demonstrate response specific thresholds at which varying polymer selection potentiates cellular differentiation, elicits pre-mature early myotube formation and directs subsequent morphological phenotype. Here we observed biocompatible AM polymers manufactured via FDM, which also appear to hold the potential to simultaneously manipulate the desired biological phenotype and enhance the biomimicry of skeletal muscle cells in vitro via AM polymer choice and careful selection of machine processing parameters. When considered in combination with the associated design freedom of AM, this may provide the opportunity to not only enhance the efficiency of creating biomimetic models, but also to precisely control the biological output within such scaffolds.
Highlights
The capability to 3D print bespoke biologically receptive parts within short time periods has driven the growing prevalence of additive manufacture (AM) technology within biological settings, limited research concerning cellular interaction with 3D printed polymers has been undertaken
Channel widths (Fig. 2E) observed in polylactic acid (PLA) had significantly reduced diameter than acrylonitrile butadiene styrene (ABS) (P ≤ 0.0005), polyethylene terephthalate (PET) (P ≤ 0.0005) and PC (P = 0.005), no variation was observed between ABS and PET (P = 0.270)
We observed polymer dependent viability of C2C12 cells when compared to respective controls, with decreases documented in PLA, PET and PC polymer conditions; it is important to outline that surfaces associated with 3D printing, will often yield lower viability when compared to the industrially optimised equivalent of tissue culture plastic
Summary
The capability to 3D print bespoke biologically receptive parts within short time periods has driven the growing prevalence of additive manufacture (AM) technology within biological settings, limited research concerning cellular interaction with 3D printed polymers has been undertaken. The design freedom associated with 3D printing technology has reduced the complexity and enhanced the efficiency of the engineering methods required to create biological models,[21,22,23] despite the growing prevalence of 3D printing, limited research regarding cellular compatibility has been undertaken As such, if these technologies are going to form the platforms within which complex cellular physiological processes are to be re-created, understanding and defining the compatibility between various mammalian cell types and the printed materials is of paramount importance.[24,25] Skeletal muscle is of specific interest due to its regenerative capacity via multipotent stem cells known as satellite cells,[26] predicating its use within in vitro models[27] as pre-clinical test beds to study the cellular and molecular mechanisms that are regulated in musculoskeletal and neuromuscular disease.[28] Cell lines such as C2C12 murine skeletal myoblasts[29] are frequently used within these models due to a lack of availability of primary human muscle cells, and their capacity to provide a biologically relevant, repeatable and reliable model of in vitro skeletal muscle
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