Effects of normobaric cyclic hypoxia exposure on mesenchymal stem-cell differentiation–pilot study on bone parameters in elderly

Table 1 Primer sequences and product sizes

Gene	Primer sequence (5’ —> 3’)	Product size (bp)
Runt-related transcription factor 2 (RUNX2)	TGGTTAATCTCCGCAGGTCAC; ACTGTGCTGAAGAGGCTGTTTG	143
Osterix (SP7)	AGCCAGAAGCTGTGAAACCTC; AGCTGCAAGCTCTCCATAACC	163
Collagen, type I, alpha 1 (COL1A1)	CGCTGGCCCCAAAGGATCTCCTG; GGGGTCCGGGAACACCTCGCTC	263
Integrin-binding sialoprotein (IBSP)	AGGGCAGTAGTGACTCATCCG; CGTCCTCTCCATAGCCCAGTGTTG	171
Osteocalcin (BGLAP)	CCATGAGAGCCCTCACACTCC; GGTCAGCCAACTCGTCACAGTC	258
Vascular endothelial growth factor A (VEGFA)	GCGAGCAGCGAAAGCGACAGG; CGTCAGCGCGACTGGTC	114
Peroxisome proliferator-activated receptor gamma 2 (PPARG2)	GCGATTCCTTCACTGATACACTG; GAGTGGGAGTGGTCTTCCATTAC	136
Fatty acid synthase (FASN)	AAGCTGAAGGACCTGTCTAGG; CGGAGTGAATCTGGGTTGATG	146
Adipose triglyceride lipase (ATGL) or patatin-like phospholipase domain-containing protein 2 (PNPLA2)	CCAACACCAGCATCCAGTTCA; ATCCCTGCTTGCACATCTCTC	102
Glycerol-3-Phosphate Dehydrogenase 1 (GPD1)	ATACAGCATCCTCCAGCACAAG; GGATGATTCTGCAGGCAGTG	120
Peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PPARGC1A)	CTGGTTGCCTGCATGAGTGTG; GGGCAAAGAGGCTGGTCTTCAC	173
Osteoprotegerin (OPG)	GGCGCTACCTTGAGATAGAGTTCTG; TGTTTTCTACAGGGTGCTTTAGATGAC	160
Receptor activator for nuclear factor kappa B ligand (RANKL)	CGTCGCCCTGTTCTTCTATTTC; AAATGCAGTGAGTGCCATCTTC	74
Low-density lipoprotein receptor related protein 5 (LRP5)	TACTGGACAGACTGGCAGACC; GTGTAGAAAGGCTCGCTTGG	209
Low-density lipoprotein receptor related protein 6 (LRP6)	TACTGGCCAAATGGACTGACT; TGTTGCAAGCCAAAATGGAGT	211
Dickkopf Wnt signaling pathway inhibitor 1 (DKK1)	ATGCGTCACGCTATGTGCT; GGAATACCCATCCAAGGTGCTA	144
Polymerase RNA II polypeptide A (POLR2A)	TTTTGGTGACGACTTGAACTGC; CCATCTTGTCCACCACCTCTTC	125

Table 2 Baseline characteristics of participants

Parameter	HYP (n = 5), mean ± SD	CON (n = 5), mean ± SD	P value
Age (yr)	83.00 ± 5.79	80.00 ± 4.06	0.371
Height (m)	1.61 ± 0.06	1.61 ± 0.06	0.960
Weight (kg)	62.00 ± 7.35	65.50 ± 2.87	0.350
BMI (kg ∙ m-2)	23.82 ± 2.68	25.26 ± 1.72	0.340
TUGT (s)	9.25 ± 0.87	10.47 ± 1.61	0.174
Diseases (n)	1.60 ± 1.52	2.20 ± 0.84	0.461
Medication (n)	5.60 ± 5.98	1.80 ± 0.45	0.194

HYP: Hypoxic exposure group; CON: Control group; BMI: Body mass index; TUGT: Timed up and go test.

Table 3 Dual-energy X-ray absorptiometry measurements at baseline (PRE) and after 18 wk (POST) for hypoxia exposure (n = 5) and control (n = 5) groups

Parameter		PRE, (mean ± SD)	%	POST, (mean ± SD)	Cohen’s d measure	MDC (%)	Wilcoxon’s test	Man Whitney U’s test
Fat mass (%)
Whole body	HYP	31.74 ± 10.08	–5.23	30.08 ± 12.47	0.15	12.38	0.225	0.841
	CON	36.39 ± 6.32	–1.37	35.89 ± 5.87	0.08	4.18	0.225
Trunk	HYP	25.13 ± 6.08	8.40	27.24 ± 12.27	0.23	34.74	0.500	0.841
	CON	33.45 ± 7.66	7.23	35.87 ± 9.08	0.29	33.27	0.893
Bone mineral density (g ∙ cm–2)
Whole body	HYP	0.880 ± 0.145	7.04$	0.942 ± 0.205	0.35	11.42	0.043	0.008
	CON	0.986 ± 0.053	–6.29	0.924 ± 0.052	1.18	5.37	0.042
Right leg	HYP	0.908 ± 0.149	0.44	0.912 ± 0.186	0.02	9.55	0.892	0.310
	CON	0.994 ± 0.071	–3.22	0.962 ± 0.068	0.46	4.98	0.109
Femoral	HYP	0.744 ± 0.118	0.81	0.750 ± 0.126	0.05	3.79	0.705	0.095
	CON	0.844 ± 0.042	–1.18	0.834 ± 0.051	0.22	2.06	0.129
Femoral neck	HYP	0.686 ± 0.106	–2.92	0.666 ± 0.108	0.19	1.96	0.041	1.000
	CON	0.752 ± 0.067	–2.39	0.734 ± 0.068	0.27	2.11	0.059
Trochanter	HYP	0.568 ± 0.097	3.17	0.586 ± 0.093	0.19	5.20	0.109	0.056
	CON	0.654 ± 0.064	–1.22	0.646 ± 0.069	0.12	4.66	0.461

HYP: Hypoxic exposure group; CON: Control group; %: Percentage of change between PRE and POST measurement outcomes; MDC: Minimum detectable change.