Surface Acoustic Wave 3D Printing Device Builds Colorectal Cancer Research Model For Clinical Adjuvant Therapy

3D bioprinting has been widely used in tissue engineering and biomedical engineering due to its excellent tissue structure construction and manipulability of biomaterials.Bioprinting has been oriented towards building structures and has less application in the study of biological mechanisms.SAW 3D printed devices enable acoustic droplet printing,single-cell manipulation,multi-material,high-viscosity liquid printing,and fine-scale construction.SAW 3D printing technology can be used to solve various pressing problems in biomedical engineering,such as the establishment of disease models,personalized treatment research.In this paper,the applications of surface acoustic wave(SAW)3D printing devices in biomedical engineering are explored based on the developed SAW 3D printing devices.In view of the problems existing in the current personalized cancer treatment,such as the lack of mechanism research tools,ignoring the impact of patients’ tumor invasion ability,specific cancer treatment methods,difficult to reproduce the real tumor environment,drug resistance analysis,etc.,this paper conducted a targeted study.Firstly,the feasibility of the device was verified.The tumor sphere-fibroblast(CAFs)invasion model was constructed by acoustic 3D printing,and the observation of the model proved that the acoustic 3D printing device could realize the fine operation of single cell spheres and the construction of biological model.Secondly,acoustic 3D printing of organoid fine arrangement combined with organoid culture technology was performed for the adjuvant treatment of low rectal cancer intersphincter resection(ISR).Finally,aiming at the shortcomings of the existing personalized cancer treatment,the acoustic 3D printing device is used to construct the microtissue model of normal organoids from the same patient surrounding the cancer organoids,reconstruct the real tumor structure,and make the drug test more accurate,which can be used for personalized treatment of colorectal cancer.Compared with the existing 3D printing technology,the surface acoustic wave 3D printing technology has more accurate manipulation ability of single-cell droplet(tumor ball or organoid),so as to meet the establishment of fine model,and be used for mechanism research and clinical adjuvant therapy.1.Using surface acoustic wave 3D printing to precisely control the characteristics of tumor spheres and cells,a single tumor sphere-fibroblast(CAFs)model was constructed to study the mechanism of CAFs and tumor invasion in colorectal cancer.The patient’s cancer tissue-derived cells were quickly(36 h)cultured into tumor spheres by the hanging drop method,and anti-Ep CAM immunomagnetic beads were used to complete the negative separation and culture of homologous patient CAFs.Using the open-cavity structure of acoustic printing devices,fine manipulation of trace amounts of liquids and non-blocking printing of high-viscosity materials can be achieved.A single tumor sphere was printed on the receiving plate,and then CAFs containing methacrylated gelatin(Gel MA)were printed around the tumor sphere to construct a colorectal tumor sphere-CAFs invasion model to observe the invasion process of tumor spheres and CAFs to determine the tumor invasive ability.Subsequently,the model was tested with the clinical drug 5-fluorouracil(5-FU),and the immunofluorescence changes of various proteins in the model were observed.Based on the results,recommendations are made for clinical prevention of the patient.As the modeling verification and invasion research of SAW 3D printing device,it proves the feasibility of SAW 3D printing invasion model and the invasion model has certain clinical guiding significance.2.Using the characteristics of surface acoustic wave 3D printing devices to build tumor models,combined with the developed organoid culture technology,the cancer organoids derived from the cancer tissue of patients with rectal cancer and the normal organoids derived from the tissue 1-2cm adjacent to the cancer of the patient were cultured,Using the advantages of acoustic 3D printing devices to build fine structures,we can print and arrange cancer organoids and normal organoids from the same patient,build an organoid invasion model,simulate the spatial layout of low rectal cancer tumors,and observe the invasion process of cancer organoids.To evaluate its invasive ability,combined with clinical Magnetic resonance imaging(MRI)and mismatch repair(MMR)proteins detection in patients with low rectal cancer,to determine the tumor location and infiltration of patients,and to help determine whether intersphincterectomy(ISR)is required for patients with organoids derived from surgical treatment.3.SAW 3D printing devices to construct organoid tumor microtissue models(PDMs)for personalized treatment of patients.We combined the previously developed acoustic printing technology with the organoid culture technology to precisely position the patient’s colon organoids around the tumor organoids to form a cancer organoid in the center and a normal organoid structure around it,which is called an organoid tumor microstructure tissues(PDMs),reproducing the real tumor structure.5-FU reagent and invasiveness studies were performed on PDMs,and tumor microtissue growth was analyzed by time-lapse imaging of tumor-colon organoid interaction kinetics.The sensitivity of PDMs to 5-FU was analyzed by protein immunofluorescence.In addition,the 5-FU-treated and control group PDMs were differentially studied by RNA transcriptome sequencing(RNA-SEQ)to analyze the sensitivity of the PDMs model to5-FU.DNA whole exome sequencing(DNA WES)was performed on the organoids.Combined with 70 patient samples,it is proved that the surface acoustic wave 3D printing device to construct organoid tumor microtissue has a wide range of applications in clinical personalized treatment.