An improved intrafemoral injection with minimized leakage as an orthotopic mouse model of osteosarcoma

Abstract

Osteosarcoma, the most common type of primary bone cancer, is the second highest cause of cancer-related death in pediatric patients. To understand the mechanisms behind osteosarcoma progression and to discover novel therapeutic strategies for this disease, a reliable and appropriate mouse model is essential. For this purpose, osteosarcoma cells need to be injected into the bone marrow. Previously, the intratibial and intrafemoral injection methods were reported; however, the major drawback of these methods is the potential leakage of tumor cells from the injection site during or after these procedures. To overcome this, we have established an improved method to minimize leakage in an orthotopic mouse model of osteosarcoma. By taking advantage of the anatomical benefits of the femur with less bowing and larger medullary cavity than those of the tibia, osteosarcoma cells are injected directly into the femoral cavity following reaming of its intramedullary space. To prevent potential leakage of tumor cells during and after the surgery, the injection site is sealed with bone wax. This method requires a minor surgery of approximately 15 minutes under anesthesia. Our established orthotopic osteosarcoma model could serve as a valuable and reliable tool for examining tumor progression of various types of bone tumors.

Engineering 2D Mesoporous Silica@MXene‐Integrated 3D‐Printing Scaffolds for Combinatory Osteosarcoma Therapy and NO‐Augmented Bone Regeneration

“…The orthotopic osteosarcoma in the mouse model was not suitable for current multifunctional research because of several critical challenges and technical pitfalls. [ 52,53 ] First of all, the major concern is leakage, as an inevitable complication during injection procedure, which might induce the alteration of tumor formation and inconsistent experimental results. [ 53 ] Second, based on clinical therapeutics, local tumor removal was critical followed by scaffold implantation.…”

“…[ 52,53 ] First of all, the major concern is leakage, as an inevitable complication during injection procedure, which might induce the alteration of tumor formation and inconsistent experimental results. [ 53 ] Second, based on clinical therapeutics, local tumor removal was critical followed by scaffold implantation. However, such surgical procedures including limb‐sparing maneuvers might not be applicable for nude mice with xenograft osteosarcoma.…”

“…However, such surgical procedures including limb‐sparing maneuvers might not be applicable for nude mice with xenograft osteosarcoma. Although some orthotopic mouse osteosarcoma models have been established with several advantages, such as minimized leakage and pathological similarity, [ 53–55 ] the ecotopic osteosarcoma model might be more suitable for validating dual function of tumor ablation and bone regeneration with straightforward biological performance. Therefore, tumor ablation and bone‐regeneration capacity were validated by ectopic osteosarcoma and cranial defect model separately in this study.…”

Genome-wide RNAi screening identifies TMIGD3 isoform1 as a suppressor of NF-κB and osteosarcoma progression

“…2a). We further performed orthotopic tumour transplantation assays by injecting SJSA-1 and Saos2 cells with or without knockdown of TMIGD3 into the femurs of immunocompromised NOD-scid IL2Rγ null mice22. Knockdown of TMIGD3 significantly enhanced primary tumour growth in femurs and metastasis formation in the lungs (SJSA-1) or the livers (Saos2, Fig.…”

“…Mice were monitored for laboured breathing and the numbers of pulmonary tumour nodules were evaluated 6 weeks after injections. For orthotopic injections, 15 μl of cell suspension (1 × 10 5 ) was injected into femoral bone marrow space of anaesthetized NOD-scid IL2Rγ null mice (The Jackson Laboratories)22. When the tumours reached ∼1 cm in thigh diameter, the mice were killed.…”