Human amnion-derived mesenchymal stem cells promote osteogenic differentiation of human bone marrow mesenchymal stem cells via H19/miR-675/APC axis

Abstract

Bone volume inadequacy is an emerging clinical problem impairing the feasibility and longevity of dental implants. Human bone marrow mesenchymal stem cells (HBMSCs) have been widely used in bone remodeling and regeneration. This study examined the effect of long noncoding RNAs (lncRNAs)-H19 on the human amnion-derived mesenchymal stem cells (HAMSCs)-droved osteogenesis in HBMSCs. HAMSCs and HBMSCs were isolated from abandoned amniotic membrane samples and bone marrow. The coculture system was conducted using transwells, and H19 level was measured by quantitative real-time reverse transcriptionpolymerase chain reaction (RT-PCR). The mechanism was further verified. We here discovered that osteogenesis of HBMSCs was induced by HAMSCs, while H19 level in HAMSCs was increased during coculturing. H19 had no significant effect on the proliferative behaviors of HBMSCs, while its overexpression of H19 in HAMSCs led to the upregulated osteogenesis of HBMSCs in vivo and in vitro; whereas its knockdown reversed these effects. Mechanistically, H19 promoted miR-675 expression and contributed to the competitively bounding of miR-675 and Adenomatous polyposis coli (APC), thus significantly activating the Wnt/β-catenin pathway. The results suggested that HAMSCs promote osteogenic differentiation of HBMSCs via H19/miR-675/APC pathway, and supply a potential target for the therapeutic treatment of bone-destructive diseases.

Human Amniotic Mesenchymal Stem Cells Promote Endogenous Bone Regeneration

“…Therefore, we proposed to introduce hAMSCs to bone defects' microenvironment and stimulate and accelerate the endogenous vascularized bone regeneration. Several studies have shown that hAMSCs enhance the osteogenic differentiation of AdSCs, BMSCs, and promote the tube-formation of human umbilical vein endothelial cells (HUVECs) ( 84 – 89 ). When hAMSCs are co-cultured with BMSCs in a transwell system, ALP activity of BMSCs and the expression of osteogenic markers, including OCN and Runx2, were upregulated ( 90 ).…”

Human amnion-derived mesenchymal stem cells promote osteogenic differentiation of lipopolysaccharide-induced human bone marrow mesenchymal stem cells via ANRIL/miR-125a/APC axis

“…Mandible samples were harvested and analyzed by hematoxylin and eosin (H&E), Masson trichrome and immunohistochemistry. Primary antibodies against RUNX2 (1:300 dilution) were used for immunohistochemistry as reported (23). Positive areas were observed under the microscope.…”

“…Primary antibody [ -Catenin (D10A8) XP® Rabbit mAb #8480(1:100), Cell Signaling Technology, Danvers, MA, USA] and DAPI were used to perform immuno uorescence staining as reported (23). Images were captured under the inverted uorescence microscope (Olympus, Japan).…”

Human amnion-derived mesenchymal stem cells promote osteogenic differentiation of lipopolysaccharide-induced human bone marrow mesenchymal stem cells via ANRIL/miR-125a/APC axis

“…After 7 days of osteogenic induction, ALP staining was used to detect the activity with an NBT/BCIP staining kit (CoWin Biotech, Beijing, China) and an ALP assay kit (Jiancheng Corp, Nanjing, China) as previously reported (21,22). Mineralized matrix formation was determined after 14 days of osteogenic induction as previously reported (23).…”

“…In vivo bone formation assay Approximately 10 5 cells (5 × 10 HAMSCs and 5 × 10 4 HBMSCs NC /HBMSCs ANRIL / HBMSCs shNC /HBMSCs shANRIL pretreated with LPS) were attached to each HA/TCP biomaterial (Φ 5 × H 2 mm, Sichuan University, Chengdu, Sichuan, China). After 12 hours, the complexes were subcutaneously implanted into the rat mandibular defect area designed as previously reported (four female nude rats per group, with an average weight of 280 g) (23). All animal experiments were conducted in compliance with the regulations and guidelines of Nanjing Medical University institutional animal care.…”