Perfusion Flow Enhances Osteogenic Gene Expression and the Infiltration of Osteoblasts and Endothelial Cells into Three-Dimensional Calcium Phosphate Scaffolds

Table 1

Primer sequences and roles for genes measured in three-dimensional perfusion studies.

Gene name	Function	Primer sequence
Osteocalcin	Osteoblast differentiation	F: 5′-GAGTCTGACAAAGCCTTCATGTCC-3′;
		R: 5′-TGATAGCTCGTCACAAGCAGGGTTA-3′
Osteopontin	Osteoblast differentiation	F: 5′-CAGCTGGATGAACCAAGTCTGGAA-3′;
		R: 5′-ACTAGCTTGTCCTTGTGGCTGTGA-3′
Alkaline phosphatase	Osteoblast differentiation	F: 5′-GCCCTCTCCAAGACATATA-3′;
		R: 5′-CCATGATCACGTCGATATCC-3′
RUNX-2	Osteoblast differentiation	F: 5′-AGAGTCAGATTACAGATCCCAGGC-3′;
		R: 5′-GTCAGAGGTGGCAGTGTCATCAT-3′
Collagen type 1	Matrix protein	F: 5′-TGGTTTGGAGAGAGCATGACCGAT-3′;
		R: 5′-TGTAGGCTACGCTGTTCTTGCAGT-3′
Vascular endothelial growth factor A	Angiogenesis	F: 5′-ACAGAAGGAGAGCAGAAGTCCCAT-3′;
		R: 5′-ATGTGCTGGCTTTGGTGAGGTTTG-3′
Basic fibroblast growth factor 2	Angiogenesis	F: 5′-AGCGGCTCTACTGCAAGAAC-3′;
		R 5′-TGGCACACACTCCCTTGATA-3′
Macrophage colony stimulating factor	Angiogenesis	F: 5′-ATGGACACCTGAAGGTCCTG-3′;
		R: 5′-GCTGGAGAGGAGTCTCATGG-3′
COX-2	Inflammatory/osteogenic	F: 5′-TCAATACTGGAAGCCGAGCACCTT-3′;
		R: 5′-GCACTTGCATTGATGGTGGCTGTT-3′
HIF-1α	Hypoxia	F: 5′-AAACTTCTGGATGCCGGTGGTCTA-3′;
		R: 5′-TCTCACTGGGCCATTTCTGTGTGT-3′
GBE-1	Hypoxia	F: 5′-GCAGGTATAAGAAGTTTAGCCAGG-3′;
		R: 5′-GAGAAAATGGATTCCAACCACTGAA-3′
MIF	Hypoxia	F: 5′-CGCACAGTACATCGCAGTG-3′;
		R: 5′-CAGCGGTGCAGGTAAGTG-3′
Cyclophilin	Housekeeping	F: 5′-TCATGTGCCAGGGTGGTGACTTTA-3′;
		R: 5′-ATGCTTGCCATCCAGCCATTCAGT-3′
Beta-actin	Housekeeping	F: 5′-ATCACTATTGGCAACGAGCGGTTC-3′;
		R: 5′-TCTCCTTCTGCATCCTGTCAGCAA-3′
Ubiquitin	Housekeeping	F: 5′-CGTCGAGCCCAGTGTTACCACCAAGAAGG-3′;
		R: 5′-CCCCCATCACACCCAAGAACAAGCACAAG-3′