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Dental pulp stem cells immobilized in alginate microspheres for applications in bone tissue engineering
Author(s) -
Kanafi M. M.,
Ramesh A.,
Gupta P. K.,
Bhonde R. R.
Publication year - 2014
Publication title -
international endodontic journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.988
H-Index - 119
eISSN - 1365-2591
pISSN - 0143-2885
DOI - 10.1111/iej.12205
Subject(s) - dental pulp stem cells , osteocalcin , viability assay , chemistry , stem cell , alizarin red , mtt assay , alkaline phosphatase , microbiology and biotechnology , trypan blue , cell , biomedical engineering , biochemistry , staining , pathology , biology , medicine , enzyme
Aim To immobilize dental pulp stem cells ( DPSC ) in alginate microspheres and to determine cell viability, proliferation, stem cell characteristics and osteogenic potential of the immobilized DPSC s. Methodology Human DPSC s isolated from the dental pulp were immobilized in 1% w/v alginate microspheres. Viability and proliferation of immobilized DPSC s were determined by trypan blue and MTT assay, respectively. Stem cell characteristics of DPSC s post immobilization were verified by labelling the cells with CD 73 and CD 90. Osteogenic potential of immobilized DPSC s was assessed by the presence of osteocalcin. Alizarin red staining and O ‐cresolphthalein complexone method confirmed and quantified calcium deposition. A final reverse transcriptase PCR evaluated the expression of osteogenic markers – ALP , R unx‐2 and OCN . Results More than 80% of immobilized DPSC s were viable throughout the 3‐week study. Proliferation appeared controlled and consistent unlike DPSC s in the control group. Presence of CD 73 and CD 90 markers confirmed the stem cell nature of immobilized DPSC s. The presence of osteocalcin, an osteoblastic marker, was confirmed in the microspheres on day 21. Mineralization assays showed high calcium deposition indicating elevated osteogenic potential of immobilized DPSC s. Osteogenic genes‐ ALP , Runx‐2 and OCN were also upregulated in immobilized DPSC s. Surprisingly, immobilized DPSC s in the control group cultured in conventional stem cell media showed upregulation of osteogenic genes and expressed osteocalcin. Conclusion Dental pulp stem cells immobilized in alginate hydrogels exhibit enhanced osteogenic potential while maintaining high cell viability both of which are fundamental for bone tissue regeneration.