Purmorphamine increased adhesion, proliferation and expression of osteoblast phenotype markers of human dental pulp stem cells cultured on beta-tricalcium phosphate

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• 作者：Maryam Rezia Rad^a ; ¹ ; ^{m.rezai.rad@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; Moein Khojaste^a ; ¹ ; ^{moein.khojaste@yahoo.com" class="auth_mail" title="E-mail the corresponding author} ; Mehrnoosh Hasan Shahriari^a ; ¹ ; ^{shahriarim@yahoo.com" class="auth_mail" title="E-mail the corresponding author} ; Saeed Asgary^b ; ^c ; ¹ ; ^{saasgar@yahoo.com" class="auth_mail" title="E-mail the corresponding author} ; Arash Khojasteh^a ; ^d ; ^e ; ¹ ; ^{arashkhojasteh@yahoo.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Purmorphamine ; Osteoinductive small molecule ; Adhesion ; Proliferation ; Differentiation ; Human dental pulp stem cells ; β-TCP
• 刊名：Biomedicine & Pharmacotherapy
• 出版年：2016
• 出版时间：August 2016
• 年：2016
• 卷：82
• 期：Complete
• 页码：432-438
• 全文大小：1909 K

文摘

Growth factors play a significant role in cell proliferation and differentiation during different stages of the bone repair. However, several limitations have been brought researchers attention to an osteoinductive small molecule including Purmorphamine. In this study, we aimed to evaluate the effect of Purmorphamine on adhesion, proliferation and differentiation of human dental pulp stem cells (hDPSCs) seaded on beta-tricalcium phosphate (β-TCP) granules.

Methods

hDPSCs were established from extracted wisdom teeth of healthy volenteers. Cells at passage 3 were seeded on β-TCP in the presence or absence of Purmorphamine. Cell adhesion and proliferation were assessed using scanning electeron microscopy (SEM) and DNA counting assay, respectively, after 1, 3 and 5 days. Then, hDPSCs seeded on β-TCP were subjected to osteogenic medium with or without Purmorphamine. After 7 and 14 days osteogenic diffrentiation capability of hDPSCs were determined using real-time RT-PCR and alkaline phosphatase (ALP) activity assay.

Results

The significant increase in amount of DNA was observed at day 3 and 5 in the presence of Purmorphamine. SEM imaging also was confirmed the DNA counting assay; in all given time points, hDPSC attachment and growth was significantly higher in the presence of Purmorphamine. ALP activity was increased by Purmorphamine at both 7 and 14 days of induction. Purmorphamine showed to effect on osteopontin expression at earlier stage of osteogenic differentiation, whereas for osteocalcin expression, this effect was more evident at later stage of differentiation.

Conclusion

Purmorphamine had a promotive effect on adhesion, proliferation and osteogenic differentiation of hDPSCs cultured on β-TCP. The outcome of the current study would help in development of in vitro culture conditions for better osteogenic differentiation of hDPSCs prior to transplantation.