An in vitro 3D bone metastasis model by using a human bone tissue culture and human sex-related cancer cells

Abstract

One of the main limitations, when studying cancer-bone metastasis, is the complex nature of the native bone environment and the lack of reliable, simple, inexpensive models that closely mimic the biological processes occurring in patients and allowing the correct translation of results. To enhance the understanding of the mechanisms underlying human bone metastases and in order to find new therapies, we developed an in vitro three-dimensional (3D) cancer-bone metastasis model by culturing human breast or prostate cancer cells with human bone tissue isolated from female and male patients, respectively. Bone tissue discarded from total hip replacement surgery was cultured in a rolling apparatus system in a normoxic or hypoxic environment. Gene expression profile, protein levels, histological, immunohistochemical and four-dimensional (4D) micro-CT analyses showed a noticeable specificity of breast and prostate cancer cells for bone colonization and ingrowth, thus highlighting the species-specific and sex-specific osteotropism and the need to widen the current knowledge on cancer-bone metastasis spread in human bone tissues. The results of this study support the application of this model in preclinical studies on bone metastases and also follow the 3R principles, the guiding principles, aimed at replacing/reducing/refining (3R) animal use and their suffering for scientific purposes.

Prostate Cancer Phenotype Influences Bone Mineralization at Metastasis: A Study Using an In Vitro Prostate Cancer Metastasis Testbed

“…ex vivo studies have demonstrated the osteoblastic characteristic of PC‐3 cells in a 3D cancer–bone model using a co‐culture of live calvarial bones and cancer cells . A mixed (osteoblastic and osteolytic) characteristic of PC‐3 cells in an in vitro 3D bone model by using human bone tissue culture was also shown . The induction of osteoblastic activity of the PC‐3 cell by inhibiting DKK‐1 has also been reported .…”

“…(16) A mixed (osteoblastic and osteolytic) characteristic of PC-3 cells in an in vitro 3D bone model by using human bone tissue culture was also shown. (70) The induction of osteoblastic activity of the PC-3 cell by inhibiting DKK-1 has also been reported. (71) Bone matrix proteins, OCN and osteopontin play important roles in the growth and progression of metastatic prostate cancer.…”

Dynamic Collision Behavior Between Osteoblasts and Tumor Cells Regulates the Disordered Arrangement of Collagen Fiber/Apatite Crystals in Metastasized Bone

“…To date, various kinds of biomimetic bone metastasis models involving spheroid cultures [ 12 ], microfluidic cell culture models [ 13 ], or 3D organ culture systems [ 14 ] have been constructed. These tumor engineering techniques have achieved certain results for understanding tumor growth characteristics and chemotherapy resistance.…”

Evaluation of RNA from human trabecular bone and identification of stable reference genes

“…The study was approved by the Ethics Committee of the Rizzoli Orthopedic Institute (Protocol MET‐3D; approved May 22, 2014) and informed consent was obtained from all subjects. Bone specimens were obtained from patients undergoing total hip replacement as previously described; the patients were not exposed to pharmacological treatments that could affect bone metabolism (Salamanna et al, ).…”