Determination of Total Protein and Calcium in Gingival Mesenchymal Stem Cell-Conditioned Medium

2021 ◽  
Vol 10 (2) ◽  
pp. 65-70
Author(s):  
Banun Kusumawardani ◽  
Intan Julita Purnamasari ◽  
Dea Ajeng Pravita Suendi

Gingival mesenchymal stem cell-conditioned medium (GMSC-CM) is a conditioned medium obtained from cultured gingival mesenchymal stem cells. GMSCs are easily isolated from gingival tissue, whereas gingival tissue is easily obtained by minimally invasive techniques. On the other hand, CM contains proteins, cytokines, chemokines and growth factors that play an important role in osteogenic differentiation.This study aims to determine the total protein and calcium levels in GMSC-CM.GMSCs were grown in culture media with 10% of FBS. CM of GMSC was obtained from the media collection process with a 0.22 µm filter and concentrated with a centrifugal filter to obtain a concentrated GMSC-CM. The concentration of total protein was performed by bicinchoninic acid protein assay on GMSC-CM and concentrated GMSC-CM. Calcium level was performed by atomic absorption spectroscopy method. The results arethat GMSC-CM had a total protein concentration of 2502±0.06 µg/ml, and the concentratedGMSC-CM was 1.912±0.08 μg/ml.The calcium level of GMSC-CM was 0.009% and concentratedGMSC-CM was 0.009%. It can be concluded thatGMSC-CM had a high concentration of total protein and calcium. These parameters were used to develop bioprocesses to enhance the production of GMSC-CM, which will support the implementation of cell-free therapy for tissue regeneration.

2021 ◽  
Vol 20 (2) ◽  
pp. 426-430
Author(s):  
Nurhasan Agung Prabowo ◽  
Zainal Arifin Adnan ◽  
Arief Nurudhin ◽  
Yulyani Werdiningsih ◽  
Kukuh Prasetyo

Background: Systemic lupus erythematosus is a chronic autoimmune disease that affects target organs. mesenchymal stem cell conditioned medium has immunosuppressive, anti-inflammatory, and immunoregulatory properties in lupus. Methyl prednisolone is a standard drug for lupus with immunosuppressive and anti-inflammatory properties. This study aims to compare the therapeutic effect of mesenchymal stem cell conditioned medium administration compared to methyl prednisolone on interleukin 10 levels and the degree of pulmonary vasculitis of lupus mice. Methods: The subjects were 24 female mice of Mus musculus Balb/C strain, which were categorized into 4 groups of 8 mice, i.e. the control group receiving 0.5 cc of 0.9% NaCl injection and placebo, the lupus group receiving 0.5 cc of pristane injection and placebo, and the treatment mesenchymal stem cell conditioned medium group receiving 0.5 cc pristane injection and mesenchymal stem cell conditioned medium 0,5 cc, and methylprednisolone group receiving 0,5 cc pristiane injection and methylprednisolone p.o 1,5 mg/kgbodyweight. After 24 days the mice were terminated and kidney and blood samples were taken. Statistical analysis was performed using ANOVA test followed by independent T-test. The p value was considered significant when the p < 0.05. Results: The study showed that there was no difference on the levels of interleukin level10 among mesenchymal stem cell conditioned medium goup and methyl prednisolone group (CM = 5,94 ± 2,49 pg/mL, mp = 5,86+1,73 pg/mL; p = 1) and the degree of pulmonary vasculitis (CM= 1,94 ± 0,25, MP=1,89+ 0,11 pg/ml; p = 0.667). Mesenchymal stem cell conditioned medium as good as methyl prednisolone in decreasing levels of interleukin 10 and the degree of pulmonary vasculitis in lupus mice. Conclusion: Mesenchymal stem cell conditioned medium as good as methyl prednisolone in decreasing levels of interleukin 10 and the degree of pulmonary vasculitis in lupus mice Bangladesh Journal of Medical Science Vol.20(2) 2021 p.426-430


2020 ◽  
Vol 10 (8) ◽  
pp. 2646
Author(s):  
Keng-Liang Ou ◽  
Yun-Wen Kuo ◽  
Chia-Yu Wu ◽  
Bai-Hung Huang ◽  
Fang-Tzu Pai ◽  
...  

The study elucidated the wound healing and hair regeneration properties of a conditioned medium prepared from the culture of human hair follicle mesenchymal stem cells (HFMSCs). The wound-healing effects of mesenchymal stem cell-conditioned medium (MSC-CM) were tested in vitro using scratch assays co-cultured with HaCaT keratinocyte and monitored through optical microscopy. The cell proliferation of HFMSCs and the HaCaT keratinocyte were observed in the presence of different kinds of drugs including UK5099, sodium L-lactate, lactate dehydrogenase-A, MSC-CM, caffeine, and caffeic acid. The hair regeneration properties were investigated in vivo by administrating the MSC-CM solutions to adult B6 mouse models. For quantification, hematoxylin and eosin staining were performed following euthanasia. In vitro results revealed that MSC-CM promotes dermal cell migrations and enhances proliferation of HFMSCs and HaCaT keratinocytes, demonstrating wound-healing properties. Moreover, when the MSC-CM solutions were applied to the shaved mouse skin, a dark area that expanded overtime was seen. Although no hair growth was found, histological analysis proved that a fat layer thickness increment was found under the mouse’s skin, ultimately projecting the formation of new hair growth. MSC-CM promotes the migration and proliferation of dermal keratinocytes that are beneficial for wound healing and hair growth. It is believed that MSC-CM can potentially serve as the basis of alternative therapeutic applications for wound closure and skin regeneration as well as hair growth stimulation and hair loss prevention in alopecia.


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