Journal article
Effects of calcium concentration on nonviral gene delivery to bone marrow‐derived stem cells
Journal of tissue engineering and regenerative medicine, Vol.13(12), pp.2256-2265
12/2019
DOI: 10.1002/term.2971
PMID: 31677246
Abstract
Background
Calcium ions (Ca2+) influence natural bone healing, and calcium is frequently used in bone tissue engineering scaffolds and cements. Scaffolds can also incorporate gene delivery systems to further promote osteoblast differentiation. Thus, our goal was to identify if Ca2+ concentration affects the transfection of bone marrow stromal cells because these cells play a major role in bone healing and can infiltrate gene‐activated scaffolds designed to promote bone growth.
Methods
Bone marrow‐derived mesenchymal stem cells (BMSCs) were cultured in media with Ca2+ concentrations ranging from 0 to 20 mM and transfected with polyethyleneimine‐plasmid DNA (PEI‐pDNA) complexes. Cell viability and transfection efficiency were determined using MTS assays and flow cytometry, respectively. PEI‐pDNA complex localization in BMSCs was assessed using fluorescence microscopy. To determine BMSC differentiation, messenger RNA (mRNA) for osteocalcin and CBFA1 was quantified using real time‐polymerase chain reaction (PCR). Calcium deposition was qualitatively assessed after three and 14 days using Alizarin Red staining.
Result
Our results indicate that Ca2+ levels between 8 and 12 mM positively impacted transfection of BMSCs with PEI‐pDNA complexes in terms of cell viability and transfection efficiency. A Ca2+ concentration of 10 mM also increased the expression of an osteogenic gene, osteocalcin, when the cells were transfected with plasmid DNA encoding bone morphogenetic protein 2 (BMP‐2).
Conclusion
Ca2+ at a 10 mM concentration can significantly reduce toxicity and enhance transfection efficiency when combined with PEI‐pDNA complexes, and this combination can be specifically applied to further enhance the differentiation of BMSCs by using the combination of polyethyleneimine‐plasmid bone morphogenetic protein 2 (PEI‐pBMP‐2) and 10 mM Ca2+ as compared with PEI‐pBMP‐2 alone.
Details
- Title: Subtitle
- Effects of calcium concentration on nonviral gene delivery to bone marrow‐derived stem cells
- Creators
- Timothy M Acri - College of Pharmacy University of IowaNoah Z Laird - College of Pharmacy University of IowaSean M Geary - College of Pharmacy University of IowaAliasger K Salem - College of Pharmacy University of IowaKyungsup Shin - College of Dentistry and Dental Clinics University of Iowa
- Resource Type
- Journal article
- Publication Details
- Journal of tissue engineering and regenerative medicine, Vol.13(12), pp.2256-2265
- DOI
- 10.1002/term.2971
- PMID
- 31677246
- NLM abbreviation
- J Tissue Eng Regen Med
- ISSN
- 1932-6254
- eISSN
- 1932-7005
- Number of pages
- 10
- Grant note
- American Association of Orthodontists Foundation (Martin "Bud" Schulman Postdoctoral Fellowship Awar)
- Language
- English
- Date published
- 12/2019
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Orthodontics; Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Dental Research; Chemical and Biochemical Engineering
- Record Identifier
- 9984216704602771
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