Effects of insulin-like growth factor 1 and basic fibroblast growth factor on the morphology and proliferation of chondrocytes embedded in Matrigel in a microfluidic platform

Abstract

Abstract. An integrated microfluidic device was utilized in the present study to investigate the morphology and proliferation of rabbit articular chondrocytes embedded in Matrigel in the presence of insulin-like growth factor 1 (IGF-1) and/or basic fibroblast growth factor (bFGF). The microfluidic device was composed of two parallel channels and a central perfusion-based three-dimensional cell culture module. The rabbit chondrocytes were cultured for 2 weeks at series of concentration gradients of IGF-1 and/or bFGF, which were generated through a diffusion process. At the end of the experiment, the morphology and quantity of cells were measured. Since high expression of collagen II is essential to the function of hyaline cartilage, immunofluorescent images of collagen II expression prior to and after the experiments were gathered for each group. The mean fluorescence intensity ratio (MIR) of collagen II in each group was calculated. The MIRs of collagen II in chondrocytes treated with IGF-1 ranged from 0.6-0.81, those in the cells treated with bFGF ranged from 0.47-0.52, and those in cells treated with a combination of IGF-1 and bFGF ranged from 0.63-0.83. Chondrocyte aggregations were observed in the group treated with 75-100 ng/ml IGF-1 (3.46-fold proliferation ratio). Similarly, a 3.83-fold proliferation ratio was identified in chondrocytes treated with 2.5-5.0 ng/ml bFGF. The group treated with 50-75 ng/ml IGF-1 and 2.5-5.0 ng/ml bFGF exhibited the optimum increase in proliferation (4.83-fold proliferation ratio). The microfluidic device used in the present study can be easily adapted to investigate other growth factors at any concentration gradient. In addition, parallel experiments can be performed simultaneously with a small quantity of cells, making it an attractive platform for the high-throughput screening of cell culture parameters. This platform will aid in the optimization of culture conditions for the in vitro expansion of chondrocytes while maintaining their in vivo morphology, which will improve autologous chondrocyte implantation capabilities for the treatment of cartilage injury.

Engineered adipose-derived stem cells with IGF-1-modified mRNA ameliorates osteoarthritis development

“…In addition, IGF-1 also plays important roles in cartilage repair that can promote chondrocyte survival, proliferation, and production of cartilage matrix [25]. Due to these characteristics, IGF-1 has garnered considerable interest for OA therapy and showed great potential for clinical translation [26][27][28]. Herein, we transfected ADSCs with IGF-1 modRNA, characterized the expression pattern of IGF-1-ADSC and explored its potential therapeutic effect in a murine OA model.…”

Two-Dimensional and Three-Dimensional Cartilage Model Platforms for Drug Evaluation and High-Throughput Screening Assays

“…Microfluidic devices have been used to culture hydrogels in dynamic conditions, thus creating shear forces and concentration gradients which help to recapitulate the endogenous environment. Li et al 101 reported the use of such a device to screen the combinatorial effects of two growth factors on type II collagen production in Matrigel-encapsulated chondrocytes. Immunostaining of the entire polydimethylsiloxane (PDMS) chip, with the aid of image analysis software, allowed for rapid data acquisition.…”

A Progress Report and Roadmap for Microphysiological Systems and Organ-On-A-Chip Technologies to Be More Predictive Models in Human (Knee) Osteoarthritis

“…As the most recent advancement in the palette of 3D culture techniques, organ-on-a-chip (OOAC) and microphysiological systems (MPS) aim for an even better recapitulation of a native tissue-like environment, tissue architecture and cell-specific responses relevant to OA modeling by combining stem cell or patient-derived primary cell-based models for cartilage and synovium ( Fomby et al, 2010 ; Smeriglio et al, 2015 ; Tian et al, 2016 ; Li et al, 2017 ; Lin et al, 2019 ) as well as adipose ( Loskill et al, 2017 ) and bone-like tissues ( Mansoorifar et al, 2021 ; Nasello et al, 2021 ). In this context, microfluidic technologies can create an even more dynamic yet more controllable musculoskeletal disease microenvironment.…”