clay and Prostatic-Neoplasms

Nanocomposite scaffolds show extensive applications in regenerative medicine and have shown promise as in vitro analogues of human tissue that can be used for the study of diseases. The complex nature of cancer metastasis is recently investigated using several 3D scaffold models. Herein, we report a polymer-nanoclay-based in vitro tumour model that recapitulates early stage of prostate cancer (PCa) colonization during skeletal metastasis on bone mimetic scaffolds. A unique cell culture system termed as "sequential culture (SC)" has been applied to create a bone-mimetic niche for colonization of PCa cells. Human mesenchymal stem cells (MSCs) were seeded on the bone-mimetic scaffolds, where they differentiated into bone cells and then formed mineralized bone matrix without osteogenic supplements. Further, PCa was seeded on MSCs-seeded scaffolds. Sequentially cultured PCa cells with MSCs formed self-organized multicellular tumoroids with distinct tight cellular junctions and hypoxic core regions. Extensive quantitative reverse transcription-polymerase chain reaction experiments were performed to evaluate the expressions of genes related to osteotropic bone metastasis of PCa. On the nanoclay scaffolds, the MSCs differentiated to mature osteoblasts and epithelial to mesenchymal transition was inhibited whereas mesenchymal to epithelial transition was enhanced, as also the hypoxia increased angiogenesis, and finally, PCa cells initiated osteoblastic lesion. Further, the SC technique has significant effects on expression of key metastasis-related genes. Therefore, the SC-based tumour model can be applied to recapitulate more consistent osteotropic cancer cell behavior in understanding tumour biology. This model also can be implemented for drug screening to target colonization stage of PCa cells in the bone microenvironment.

Topics: Biomimetic Materials; Cell Differentiation; Cell Hypoxia; Cell Line, Tumor; Cell Shape; Clay; Core Binding Factor Alpha 1 Subunit; Epithelial-Mesenchymal Transition; Gene Expression Regulation; Humans; Male; Mesenchymal Stem Cells; Nanoparticles; Neoplasm Metastasis; Neovascularization, Pathologic; Osteoblasts; Osteogenesis; Prostate-Specific Antigen; Prostatic Neoplasms; Spheroids, Cellular; Tissue Scaffolds

In recent times, the limitation of two-dimensional cultures and complexity of in vivo models has paved the way for the development of three-dimensional models for studying cancer. Here we report the development of a new tumor model using PCL/HAPClay scaffolds seeded with a sequential culture of human mesenchymal stem cells (hMSCs) followed by human prostate cancer cells (HPCCs). This nanocomposite system is used as a test-bed for studying cancer metastasis and efficacy of anti-cancer drugs using a polymersome delivery method. A novel sequential cell culture system in three-dimensional in vitro bone model provides a unique bone mimetic environment. The hMSCs seeded scaffolds are seeded with prostate cancer cells after the hMSCs have differentiated into osteoblasts. Sequential culture on the scaffolds has shown formation of tumoroids or microtissue consisting of organized, densely packed round cells with hypoxic core regions similar to in vivo tumors. Such tumoroids are not observed on HPCC seeded scaffolds or when HPCCs sequentially cultured with human osteoblast cells. Clearly, the newly differentiated hMSCs play a vital role in the ability of cancer cells to grow into tumoroids and cause disease. The PCL/HAPclay scaffold system seeded with the sequential culture of hMSCs, and HPCCs presents a good model system for study of the interactions between prostate cancer cells and bone microenvironment. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1591-1602, 2016.

Topics: Alkaline Phosphatase; Aluminum Silicates; Biomimetic Materials; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cell Survival; Clay; Drug Delivery Systems; Folic Acid; Humans; Male; Mesenchymal Stem Cells; Nanoparticles; Osteoblasts; Osteogenesis; Prostatic Neoplasms; Spheroids, Cellular; Tissue Scaffolds; Tumor Cells, Cultured

Circulating tumor cells (CTCs) can be used clinically to treat cancer. As a diagnostic tool, the CTC count can be used to follow disease progression, and as a treatment tool, CTCs can be used to rapidly develop personalized therapeutic strategies. To be effectively used, however, CTCs must be isolated at high purity without inflicting cellular damage.. We designed a microscale flow device with a functionalized surface of E-selectin and antibody molecules against epithelial markers. The device was additionally enhanced with a halloysite nanotube coating. We created model samples in which a known number of labeled cancer cells were suspended in healthy whole blood to determine device capture efficiency. We then isolated and cultured primary CTCs from buffy coat samples of patients diagnosed with metastatic cancer.. Approximately 50% of CTCs were captured from model samples. Samples from 12 metastatic cancer patients and 8 healthy participants were processed in nanotube-coated or smooth devices to isolate CTCs. We isolated 20-704 viable CTCs per 3.75-mL sample, achieving purities of 18%-80% CTCs. The nanotube-coated surface significantly improved capture purities (P = 0.0004). Experiments suggested that this increase in purity was due to suppression of leukocyte spreading.. The device successfully isolates viable CTCs from both blood and buffy coat samples. The approximately 50% capture rate with purities >50% with the nanotube coating demonstrates the functionality of this device in a clinical setting and opens the door for personalized cancer therapies.

Topics: Aluminum Silicates; Antibodies; Antigens, Neoplasm; Antigens, Surface; Blood Buffy Coat; Breast Neoplasms; Cell Adhesion; Cell Adhesion Molecules; Cell Count; Cell Separation; Clay; E-Selectin; Epithelial Cell Adhesion Molecule; Female; Glutamate Carboxypeptidase II; Humans; Leukocytes; Lung Neoplasms; Male; Nanotubes; Neoplasm Metastasis; Neoplastic Cells, Circulating; Ovarian Neoplasms; Polyurethanes; Prostatic Neoplasms