Low‐intensity Ultrasound Stimulation Enhances Chondrogenic Differentiation in Alginate Culture of Mesenchymal Stem Cells

Abstract

Mesenchymal stem cells (MSCs) are regarded as a potential autologous source for cartilage repair, because they can differentiate into chondrocytes by transforming growth factor‐beta (TGF‐β) treatment under the 3‐dimensional (3‐D) culture condition. However, more efficient and versatile methods for chondrogenic differentiation of MSCs are still in demand for its clinical application. Recently, low‐intensity ultrasound (LIUS) was shown to enhance fracture healing in vitro and induce chondrogenesis of MSCs in vitro. In this study, we investigated the effects of LIUS on the chondrogenesis of rabbit MSCs (rMSCs) in a 3‐D alginate culture and on the maintenance of chondrogenic phenotypes after replating them on a monolayer culture. The LIUS treatment of rMSCs increased: (i) the matrix formation; (ii) the expression of chondrogenic markers such as collagen type II, aggrecan, and Sox‐9; (iii) the expression of tissue inhibitor of metalloprotease‐2 implicated in the integrity of cartilage matrix; and (iv) the capacity to maintain the chondrogenic phenotypes in a monolayer culture. Notably, LIUS effects were clearly shown even without TGF‐β treatment. These results suggest that LIUS treatment could be an efficient and cost‐effective method to induce chondrogenic differentiation of MSCs in vitro for cartilage tissue engineering.

Effects of Low Intensity Ultrasound on the Chondrogenic Differentiation of Adult Stem Cells From Adipose Tissue

“…The results of this study are in agreement with the study that they applied ultrasound parameters close to parameters of our study [38]. LIUS parameters of this study and frequency (1.5 MHz) but they showed chondrogenic differentiation without focus on hypertrophic markers evaluations.…”

“…The low intensity ultrasound (LIUS) device (Novin, Iran) with continuous wave at 1 MHz and intensity of 200 mW/cm 2 was applied for 10 minutes per day in ultrasound cultures [35,38]. The distance between transducer and cultures was determined by the Sarvazyan method [39][40][41].…”

Purinergic and Store-Operated Ca2+ Signaling Mechanisms in Mesenchymal Stem Cells and Their Roles in ATP-Induced Stimulation of Cell Migration

“…Cells were fixed with 4% paraformaldehyde for 30 for 48 hours by gentle shaking and then in BM or chondrogenic medium (0.1 mM dexamethasone, 10 ng/ml Transforming growth factor (TGF)-b3, 50 mg/ml ascorbic acid 2-phosphate, 1.0 mg/ml recombinant human insulin, 0.55 mg/ml human transferrin, and 0.5 lg/ml sodium selenite in BM) for 3 weeks with the media changed every 3 days. Cell pellets were paraffin embedded, sectioned, and stained with Alcian blue and Sirius red [29].…”

Low-Intensity Ultrasound (LIUS) as an Innovative Tool for Chondrogenesis of Mesenchymal Stem Cells (MSCs)

“…[9][10][11][12][13][14][15][16] Among these mechanical stimulations, LIUS is of particular interest because it is simple, cost-effective and already proven to activate chondrocyte phenotypes in vitro [17][18][19][20][21][22] and improve cartilage repair in animal models. 23,24 This review will introduce findings by other groups and ours on the LIUS effect on MSCs during the chondrogenic differentiation and will discuss some of the issues regarding its mechanism and future applications.…”