Matrix Production in Chondrocytes Transfected with Sex Determining Region Y-Box 9 and Telomerase Reverse Transcriptase Genes: An In Vitro Evaluation from Monolayer Culture to Three-Dimensional Culture

Abstract

Background:This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)box 9 (SOX9)-and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes fromWe use cookies to personalise content and ads, to provide social media features and to analyse our traffic. We also share information about your use of our site with our social media, advertising and analytics partners in accordance with our Privacy Statement. You can manage your preferences in Manage Cookies. ✓ OK Manage Cookies ❯ 3/20/2020 Matrix Production in Chondrocytes Transfected with Sex Determining Region Y-Box 9 and Telomerase Reverse Transcriptase Genes: An… https://link.springer.com/article/10.1007/s13770-019-00191-1 2/25monolayer to three-dimensional (3D) culture. Methods:The genes were transferred into chondrocytes at passage-1 (P1) via lipofection. The post-transfected chondrocytes (SOX9-,/TERT) were analysed at P1, P2 and P3. The non-transfected group was used as control. The 3D culture was established using the chondrocytes seeded in a discshaped PLGA/fibrin and PLGA scaffolds. The resulting 3D "cells-scaffolds" constructs were analysed at week-1, -2 and -3. The histoarchitecture was evaluated using haematoxylin and eosin, alcian blue and safranin o stains. The quantitative sulphated glycosaminoglycan (sGAG) content was measured using biochemical assay. The cartilage-specific markers expression were analysed via real-time polymerase chain reaction.

Does Quranic Recitation Work on a Non-Audible Cell? An Experimental Study in a Monolayer Cells Culture Model

“…This result supported the previous morphological result as the cells could sustain the shape of the cells. It is a normal phenomenon in cartilage research and documented in earlier studies (N. Md Nazir et al 2019;Tew et al 2005). The expression of collagen I and II in the cell culture showed the ability to maintain the phenotypic characteristics of the chondrocytes.…”

“…The descriptions for primers used (1st BASE, Singapore), i.e., collagen type II (COL2A1), SRY (sex-determining region Y)-box 9 (SOX9), and aggrecan-core protein (ACAN), collagen type I (COL1A2), and a housekeeping gene, hypoxanthine phosphoribosyltransferase 1 (HPRT1), were summarized in Table 2. The stepwise protocols, including total RNA extraction (TRIzol, Gibco, Life Technologies, USA), cDNA synthesis (Superscript™ II Reverse Transcriptase, Gibco and Gradient thermal cycler, TC-512, Techne, Inc., USA), qPCR, and gel electrophoresis (EC 300 XL, Thermo Scientific, USA and Alpha Imager HP System, Alpha Innotech, USA), were accomplished following the previous studies' methodology (Mohamad et al 2017;Nazir et al, 2019;Nazir et al, 2020).…”

“…Sulfated Glycosaminoglycan (sGAG) Production: All samples were first digested with papain digestion solution (125 µg/ml of papain, 5 mM L-cysteine, 100 mM Na2HPO4, 5 mM EDTA, pH 6.8) at 60°C for 16 hours (N. Md Nazir et al 2019;Noorhidayah Md Nazir et al 2020). Based on the manufacturer's protocol, the sGAG content was quantitatively analyzed using the sGAG Assay Kit (K-Assay®, Kamiya Biomedical Company, USA).…”

“…The insufficient presence of the proteoglycan in monolayer cultured chondrocytes is no more uncommon in many cartilage studies (Statham et al, 2021). It was suggested that culturing the cells in biomaterial scaffolds or introducing them to the three-dimensional environment can help the chondrocytes reinstate their properties to produce the proteoglycan (Munirah et al, 2008;Nazir et al, 2019;Nazir et al, 2020).…”

Comparison of Scaffolds Fabricated via 3D Printing and Salt Leaching: In Vivo Imaging, Biodegradation, and Inflammation

“…Biocompatible materials that are currently used to fabricate 3D scaffolds via the salt-leaching method include natural biomaterials, such as collagen, hyaluronic acid, and gelatin, as well as synthetic materials, such as polyester and polyethylene glycol (PEG) [ 6 , 7 , 8 ]. Synthetic polyesters can be precisely designed to have specific molecular weights and compositions [ 9 , 10 , 11 , 12 ]. In addition, synthetic polyesters can be produced in large quantities with little or no batch-to-batch variation.…”