Research On The Key Technology Of The Fabrication Of In Vitro Bioactive Muscular Models For Biohybrid Robots

Biohybrid robots are constructed by the integration of living cells and bioactive materials,which can simulate the shape and movement of real organisms.Compared with biomimetic robots,biobots exhibit superiority in terms of miniaturization,bio-syncretic,and self-repair.Therefore,they have a wide application prospect in military and medical fields such as building micro-nano reconnaissance robots and vascular robots.Meanwhile,the fabrication of in vitro bioactive muscular models is a key step to realizing the muscle drive of biobots,aiming to model real biological muscle tissue.Their principle is to generate in vitro muscle tissue by assembling muscle cells with bioactive materials that harness biologically equivalent properties.It can be concluded that the fabrication of in vitro bioactive muscular models is an important basis for the realization of muscle-driven functions of biobots.Therefore,this thesis will conduct relevant theoretical and technological research on key technologies such as the structural design of bioactive muscular in vitro models,muscle cell culture,bioactive material synthesization,3D bioprinting and printing quality evaluation,and fabrication of bioactive muscular in vitro models to support and promote the developments of biobots and muscle-driven.The specific research contents are as follows:(1)Based on the analysis of the fabrication mechanism of skeletal muscle tissue,the structure of bioactive muscular was designed and the 3D bioprinting method was established.C2C12 myoblasts,Matrigel,and other materials were selected as the main components of the in vitro bioactive muscle models.Meanwhile,the properties of these components were analyzed,and the configuration process of the materials was summarized in detail.(2)Myoblasts were cultured in vitro and detailed culture processes were established.The four main factors affecting the performance and printability of the in vitro bioactive muscle models were studied as well as the range of cell concentration and material ratio.(3)The synthesization method of bioactive muscle models material in vitro was proposed,and the deficiency of Matrigel which was difficult to 3D bioprinting was overcome.The rheological properties of bioactive materials were tested via temperature sweeps,time sweeps,amplitude sweeps,frequency sweeps,flow curve sweeps.As a result,the yield characteristics,gelling time,and other performance affecting parameters were obtained.(4)Based on the understanding of the 3D bioprinting principle,an easamatic version of 3D bioprinter is refitted,and a semi-quantitative(RN)evaluation method was proposed to evaluate the printing quality of in vitro bioactive muscle models to ensure that cells survivability and printing parameters were optimized.(5)3D bioprinting and culture the in vitro bioactive muscle models based on the proposed biomaterials synthesization method and optimized bioprinting parameters,the muscle cells can survive more than 5 days.In summary,this thesis established a detailed 3D bioprinting process and evaluation method for assessing printing quality for the fabrication of in vitro bioactive muscle model,which lay an important foundation for biohybrid robot muscle driving in the future.