Screening of Mineralogenic and Osteogenic Compounds in Zebrafish—Tools to Improve Assay Throughput and Data Accuracy

Abstract

Bone disorders affect millions of people worldwide and treatments currently available often produce undesirable secondary effects or have limited efficacy. It is therefore of the utmost interest for patients to develop more efficient drugs with reduced off-target activities. In the long process of drug development, screening and preclinical validation have recently gained momentum with the increased use of zebrafish as a model organism to study pathological processes related to human bone disorders, and the development of zebrafish high-throughput screening assays to identify bone anabolic compounds. In this review, we provided a comprehensive overview of the literature on zebrafish bone-related assays and evaluated their performance towards an integration into screening pipelines for the discovery of mineralogenic/osteogenic compounds. Tools available to standardize fish housing and feeding procedures, synchronize embryo production, and automatize specimen sorting and image acquisition/analysis toward faster and more accurate screening outputs were also presented.

Contribution of Signal Transducer and Activator of Transcription 3 (STAT3) to Bone Development and Repair

“…The Stat3 KO mutants displayed significantly reduced stat3 expression ( Supplementary Figure S3 ) and similar defects in growth and skeletal development as observed with the other zebrafish Stat3 KO mutants, which together provide support that this reflects a hypomorphic mutant. Bone formation was explored through analysis of dermal bones as key indicators of bone development in zebrafish [ 38 , 39 , 40 ]. The cleithrum, operculum, ceratobranchial, and ceratohyal bones were all significantly smaller in Stat3 KO mutants, along with reduced expression of the key osteoblast markers runx2b , col10a , and spp1 .…”

Persistent mesodermal differentiation capability of bone marrow MSCs isolated from aging patients with low energy traumatic hip fracture and osteoporosis: clinical evidence

“…Nevertheless, the detailed differences are not yet investigated comprehensively. Therefore, our BMSCs bank collected from aging osteoporotic patients could be one of the high throughputs in vitro models for the osteogenic and anabolic dual-functioned drug screening before the complex in vivo selection; it is an expensive and time-consuming experiment, even utilizing a facile zebra sh model (33). Furthermore, such as previous publications using MSCs high throughput screening models successfully identi ed potent chondrogenic drugs (9,34).…”

Metabolic bone disorders and the promise of marine osteoactive compounds

“…These small teleosts offer several technical advantages that make them well suited for drug screening, e.g., smaller size, cost-effectiveness, shorter life span, and higher fecundity when compared to mammalian models. Moreover, the translucency of embryonic stages throughout development and the amenability to gene editing has enabled the generation of a vast array of transgenic and mutant lines that can be used for in vivo-cell tracking and disease modeling [ 296 ]. Furthermore, teleost ability to regenerate bone and cartilage tissues offer a different approach for evaluating the osteoactivity of drugs and compounds [ 297 ].…”