Abstract
BackgroundMesenchymal stem cells (MSCs) can regenerate missing tissues and treat diseases. Hence, the current work aimed to compare the proliferation rate and the osteogenic differentiation potential of bone marrow MSCs (BMSCs), gingival MSCs (GMSCs) and submandibular MSCs (SMSCs).Material and MethodsMSCs derived from bone marrow, gingiva and submandibular salivary gland were isolated and cultured from rats. The proliferation capacity was judged by MTT proliferation Assay. Osteogenic differentiation was assessed by Alzarin red stain and quantitative RT-PCR was performed for Runx-2 and MMP-13.ResultsThe highest significant proliferation was estimated in the BMSCs compared to GMSCs and SMSCs (p-value was < 0.01). All studied cell types formed mineralized nodules as stained with Alizarin Red stain at the 3rd passage of differentiation. However, BMSCs seemed to generate the highest level of mineralization compared to GMSCs and SMSCs. RT-PCR revealed that the expression of Runx-2 and MMP-13 mRNAs was significantly increased in the BMSCs compared to GMSCs and SMSCs (p-value was < 0.01).ConclusionsBMSCs displayed maximum osteogenesis results followed by the GMSCs and lastly by the SGSCs. Thus, it could be recommended that GMSCs can be used as a second choice after BMSCs when bone tissue reconstruction is needed. Key words:Mesenchymal stem cells, osteogenic differentiation, Runx-2, MMP-13.
Highlights
Mesenchymal stem cells (MSCs) are the most promising stem cells for clinical applications; they were originally found in the bone marrow, and have been isolated from many other tissues as skin, adipose tissue and various dental tissues [1,2]
bone marrow MSCs (BMSCs) can be isolated from the bone marrow of iliac crest by physicians, but the bone marrow aspiration procedure is invasive for the donors
The stem cells most commonly used to date for bone regeneration in dental patients are BMSCs obtained from the iliac crest, owing to their great potential for bone regeneration [13]
Summary
Mesenchymal stem cells (MSCs) are the most promising stem cells for clinical applications; they were originally found in the bone marrow, and have been isolated from many other tissues as skin, adipose tissue and various dental tissues [1,2]. We investigated and compared the proliferation rate and the osteogenic differentiation potential of different adult mesenchymal stems from rat bone marrow, gingival and salivary glands. Cells were plated in 25 ml culture flasks and proliferated in minimal essential medium (MEM) (Gibco-Life Technologies) supplemented with 15% fetal bovine serum (FBS), 1× Pen/Strep antibiotics (Invitrogen) and incubated at 37°C and 5% CO2. Cell Culture media were changed twice a week; with supplementation of LAscorbic Acid 2-Phosphate (50 μg/mL; Sigma-Aldrich) and they were monitored for 70 to 80% confluence by using inverted microscope software. -MTT proliferation Assay Measuring the MTT color absorbance among the three studied groups revealed that the highest significant proliferation was observed at two weeks culture in BMSCs (2.416±0.744), followed by GMSCs (1.281±0.577) and SMSCs (0.226±0.0225) (Table 2). At 7&14 days culture there was a statistically highly significant difference between all the studied groups as the p-value was < 0.01. -Osteogenic differentiation Osteogenic differentiation and mineralization were evidenced by calcium deposits which formed orange red fa-
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