Role of Insulin-Transferrin-Selenium in Auricular Chondrocyte Proliferation and Engineered Cartilage Formation in Vitro

Abstract

The goal of this study is to determine the effects of Insulin-Transferrin-Selenium (ITS) on proliferation of auricular chondrocytes and formation of engineered cartilage in vitro. Pig auricular monolayer chondrocytes and chondrocyte pellets were cultured in media containing 1% ITS at different concentrations of fetal bovine serum (FBS, 10%, 6%, 2%, 0%), or 10% FBS alone as a control for four weeks. Parameters including cell proliferation in monolayer, wet weight, collagen type I/II/X (Col I, II, X) and glycosaminoglycan (GAG) expression, GAG content of pellets and gene expression associated with cartilage formation/dedifferentiation (lost cartilage phenotype)/hypertrophy within the chondrocyte pellets were assessed. The results showed that chondrocytes proliferation rates increased when FBS concentrations increased (2%, 6%, 10% FBS) in ITS supplemented groups. In addition, 1% ITS plus 10% FBS significantly promoted cell proliferation than 10% FBS alone. No chondrocytes grew in ITS alone medium. 1% ITS plus 10% FBS enhanced cartilage formation in terms of size, wet weight, cartilage specific matrices, and homogeneity, compared to 10% FBS alone group. Furthermore, ITS prevented engineered cartilage from dedifferentiation (i.e., higher index of Col II/Col I mRNA expression and expression of aggrecan) and hypertrophy (i.e., lower mRNA expression of Col X and MMP13). In conclusion, our results indicated that ITS efficiently enhanced auricular chondrocytes proliferation, retained chondrogenic phenotypes, and promoted engineered cartilage formation when combined with FBS, which is potentially used as key supplementation in auricular chondrocytes and engineered cartilage culture.

Role of Calcium Bio-Minerals in Regenerative Medicine and Tissue Engineering

“…Osteoclast differentiation from human blood precursors can be enhanced by using biomimetic calcium-phosphate substrates [44]. Insulintransferrin-selenium is also play important role in auricular chondrocyte proliferation and engineered cartilage formation in vitro [45]. It helps to retain chondrogenic phenotypes, and promoted engineered cartilage formation when combined with FBS, which is potentially used as key supplementation in auricular chondrocytes and engineered cartilage culture.…”

“…These composites are osteoinductive for human bone marrow mesenchymal stem cells (HBMS) and have wider application in tissue engineering [53] (Table 1). Composite scaffolds of nanohydroxyapatite and silk fibroin are used to enhance mesenchymal stem cell-based bone regeneration via the interleukin 1 alpha autocrine/paracrine signaling loop [54]. These scaffolds promote bone regeneration mainly through cell signaling pathway associated with cell-biomaterial interaction.…”

“…These scaffolds promote bone regeneration mainly through cell signaling pathway associated with cell-biomaterial interaction. Though, some other soluble factors also play a role in osteoinduction with nHAp-SF [54]. Superparamagnetic iron oxide nanoparticles (SPIONs) are also used in bone regeneration.…”

Effect of Insulin Transferrin Selenium Administration on Rat's Cultured In Vitro Embryo Post Warming After being Frozen using Vitrification Method

“…Insulin Transferrin Selenium is complex media supplements consisting of compounds of insulin, transferrin, dan selenium (Liu et al, 2014). Insulin Transferrin Selenium is complex protein which is able to stimulate cell growth, prevent cell damage due to anti oxidant role in it so that it can maintain embryo viability post warming.…”

The Role of MicroRNA-381 in Chondrogenesis and Interleukin-1-β Induced Chondrocyte Responses

“…PMCs were grown in M199 medium (Gibco, 11150-059) supplemented with 10% FBS, 1% penicillin and streptomycin, bFGF (Peprotech, Rocky Hill, NJ, USA), EGF (Peprotech), and insulin (Sigma, St. Louis, MO, USA) at 37°C in a humidified 5% CO 2 atmosphere. PHCs were cultured in DMEM/F12 supplemented with 5% FBS, 1% penicillin and streptomycin, and ITS (insulin, transferrin, and selenous acid) + Premix (BD Biosciences, San Jose, CA, USA catalog: 354352) as previously described [25,26,27,28,29] at 37°C in a humidified 5% CO 2 atmosphere. Chondrogenic ATDC5 cells, PHCs, and PMCs were treated with recombinant IL-1β (Peprotech, 200-01B) using various doses and times [30,31,32].…”