Study of effects of metformin on osteogenic differentiation of mesenchymal stem cells and bone tissue regeneration in type I osteogenesis imperfecta mice

doi:10.11958/20221341

Abstract

Abstract:

Objective To evaluate the effect of metformin on the osteogenic differentiation of adipose derived mesenchymal stem cells of type I osteogenesis imperfecta mice (oimADSCs), and the therapeutic effect of metformin on bone tissue regeneration in oim mice. Methods The oimADSCs were isolated and cultured. Cell morphology observation, alkaline phosphatase staining and oil red O staining were used to validate the multiple differentiation potential of oimADSCs．After osteogenic induction，ALP staining and ALP activity were compared between groups. qPCR was applied to analyze the expression of osteogenic differentiation associated genes. Micro-CT was used to detect the femoral microstructure and analyze effects of metformin on bone regeneration in oim mice. Results Compared with the control group, ALP staining was more deeper and the ALP activity level was obviously increased in the osteogenesis+metformin group. The transcription levels of Bglap, Col1a1, Runx2 and BMP2 were significantly up-regulated. The micro-CT results revealed that after metformin treatment, BV/TV, trabecular number and Tb.Th of oim mice were increased. Conclusion Metformin can enhance osteogenic differentiation of oimADSCs in vitro and improve bone microstructure of oim mice in vivo．

Key words: osteogenesis imperfecta, mesenchymal stem cells, metformin, type I osteogenesis imperfecta, adipose derived mesenchymal stem cells, osteogenic differentiation

CLC Number:

R681.1

Figures/Tables 7

References 22

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基因名称	引物序列（5′→3′）	产物大小（bp）
Bglap	上游：CTGACCTCACAGATGCCAAGCC	191
	下游：CATACTGGTCTGATAGCTCGTCACAAG	191
Col1a1	上游：GACAGGCGAACAAGGTGACAGAG	85
	下游：CAGGAGAACCAGGAGAACCAGGAG	85
Runx2	上游：GCAGCAGCAGCAGCAGGAG	181
	下游：GCACGGAGCACAGGAAGTTGG	181
BMP2	上游：AACACCGTGCGCAGCTTCCATC	142
	下游：CGGAAGATCTGGAGTTCTGCAG	142
GAPDH	上游：CATCACTGCCACCCAGAAGACTG	152
	下游：ATGCCAGTGAGCTTCCCGTTCAG	152

基因名称	引物序列（5′→3′）	产物大小（bp）
Bglap	上游：CTGACCTCACAGATGCCAAGCC	191
	下游：CATACTGGTCTGATAGCTCGTCACAAG	191
Col1a1	上游：GACAGGCGAACAAGGTGACAGAG	85
	下游：CAGGAGAACCAGGAGAACCAGGAG	85
Runx2	上游：GCAGCAGCAGCAGCAGGAG	181
	下游：GCACGGAGCACAGGAAGTTGG	181
BMP2	上游：AACACCGTGCGCAGCTTCCATC	142
	下游：CGGAAGATCTGGAGTTCTGCAG	142
GAPDH	上游：CATCACTGCCACCCAGAAGACTG	152
	下游：ATGCCAGTGAGCTTCCCGTTCAG	152

组别	BMP2	Runx2	Col1a1	Bglap
成骨诱导组	1.10±0.10	0.93±0.12	1.10±0.10	1.06±0.05
实验组	2.53±0.42	1.68±0.04	1.82±0.11	2.01±0.36
t	5.798^＊＊	10.583^＊＊	8.519^＊＊	4.531^＊＊

组别	BMP2	Runx2	Col1a1	Bglap
成骨诱导组	1.10±0.10	0.93±0.12	1.10±0.10	1.06±0.05
实验组	2.53±0.42	1.68±0.04	1.82±0.11	2.01±0.36
t	5.798^＊＊	10.583^＊＊	8.519^＊＊	4.531^＊＊

组别	BV/TV	Tb. N/mm^-1	Tb. Th/mm
对照组	2.06±0.31	0.55±0.05	0.042±0.003
二甲双胍组	6.17±1.00	1.48±0.15	0.051±0.002
t	9.595^＊＊	14.461^＊＊	6.320^＊＊