miR-196a-5p Promoted the Osteogenic Differentiation and Calvarial Bone Defect Repair of Wharton’s Jelly Umbilical Cord Stem Cells

Abstract Background: To investigate the effect of miR‐196a-5p on the osteogenic differentiation and defected bone repair of Wharton’s jelly umbilical cord stem cells (WJCMSCs). Methods: miR‐196a-5p mimic or inhibitor was applied to overexpress or silence miR‐196a-5p expression in WJCMSCs. The alkaline phosphatase (ALP) activity, mineralization ability, and osteogenic markers expression were used to test WJCMSCs osteogenic potential in vitro. Calvarial bone defect model of rat was used to evaluate WJCMSCs bone regeneration ability in vivo. mRNA microarray was used to reveal the underling mechanisms that miR‐196a-5p regulated bone repair.Results: miR-196a-5p inhibition reduced the ALP activity, mineralization ability, and level of osteogenic markers OCN, DSPP, DMP1 and BSP, while miR-196a-5p overexpression enhanced the ALP activity, mineralization ability, and level of OCN, DSPP, DMP1 and BSP of WJCMSCs in vitro. Next, the micro-CT and histopathology results showed miR-196a-5p-overexpressed-WJCMSCs obviously promoted the new bone tissue regeneration and calvarial bone defect repair after MSCs transplanted for 12 weeks. Further, mRNA microarray of miR-196a-5p-overexpressed-WJCMSCs revealed totally 959 significantly differentially expressed genes (DEGs), among which 34 upregulated and 925 downregulated. Also, 241 miR-196a-5p targeted genes were predicted by using miRNA targeted websites and only 19 predicted genes were consistent with microarray results. On this basis, one significantly downregulated gene SERPINB2 was selected and revealed that SERPINB2 deletion obviously enhanced the ALP activity and mineralization ability of WJCMSCs in vitro.Conclusions: miR-196a-5p promoted the osteogenic differentiation potential and calvarial bone defect repair ability of WJCMSCs. And SERPINB2 acted as one key downstream gene to participate in the miR-196a-5p promoted MSCs osteogenic differentiation.

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Cryopreserved platelet-rich plasma enhancing calvarial bone defect repair in rabbits: an observation of outcome

Academic Journal of Second Military Medical University ◽

10.3724/sp.j.1008.2015.00391 ◽

2015 ◽

Vol 36 (4) ◽

pp. 391

Author(s):

Jun CHEN ◽

Yun-fu ZHAO ◽

Qing-fu ZHANG ◽

Guo-qin LIU ◽

Man-jiao LIU ◽

...

Keyword(s):

Bone Defect ◽

Platelet Rich Plasma ◽

Calvarial Bone ◽

Bone Defect Repair ◽

Defect Repair ◽

Calvarial Bone Defect

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The immunogenic reaction and bone defect repair function of ε-poly-L-lysine (EPL)-coated nanoscale PCL/HA scaffold in rabbit calvarial bone defect

Journal of Materials Science Materials in Medicine ◽

10.1007/s10856-021-06533-7 ◽

2021 ◽

Vol 32 (6) ◽

Author(s):

Bin Tian ◽

Na Wang ◽

Qingsong Jiang ◽

Lijiao Tian ◽

Lei Hu ◽

...

Keyword(s):

Tissue Engineering ◽

Bone Defect ◽

Macrophage Infiltration ◽

T Helper ◽

Calvarial Bone ◽

Bone Defect Repair ◽

Defect Repair ◽

Calvarial Bone Defect ◽

Helper Type

AbstractTissue engineering is a promising strategy for bone tissue defect reconstruction. Immunogenic reaction, which was induced by scaffolds degradation or contaminating microorganism, influence cellular activity, compromise the efficiency of tissue engineering, or eventually lead to the failure of regeneration. Inhibiting excessive immune response through modulating scaffold is critical important to promote tissue regeneration. Our previous study showed that ε-poly-L-lysine (EPL)-coated nanoscale polycaprolactone/hydroxyapatite (EPL/PCL/HA) composite scaffold has enhanced antibacterial and osteogenic properties in vitro. However, the bone defect repair function and immunogenic reaction of EPL/PCL/HA scaffolds in vivo remains unclear. In the present study, three nanoscale scaffolds (EPL/PCL/HA, PCL and PCL/HA) were transplanted into rabbit paraspinal muscle pouches, and T helper type 1 (Th1), T helper type 2 (Th2), T helper type 17 (Th17), and macrophage infiltration were analyzed after 1 week and 2 weeks to detect their immunogenic reaction. Then, the different scaffolds were transplanted into rabbit calvarial bone defect to compare the bone defect repair capacities. The results showed that EPL/PCL/HA composite scaffolds decreased pro-inflammatory Th1, Th17, and type I macrophage infiltration from 1 to 2 weeks, and increased anti-inflammatory Th2 infiltration into the regenerated area at 2 weeks in vivo, when compared to PCL and PCL/HA. In addition, EPL/PCL/HA showed an enhanced bone repair capacity compared to PCL and PCL/HA when transplanted into rabbit calvarial bone defects at both 4 and 8 weeks. Hence, our results suggest that EPL could regulate the immunogenic reaction and promote bone defect repair function of PCL/HA, which is a promising agent for tissue engineering scaffold modulation.

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In Vitro Prevascularization of Self-Assembled Human Bone-Like Tissues and Preclinical Assessment Using a Rat Calvarial Bone Defect Model

Materials ◽

10.3390/ma14082023 ◽

2021 ◽

Vol 14 (8) ◽

pp. 2023

Author(s):

Fabien Kawecki ◽

Todd Galbraith ◽

William P. Clafshenkel ◽

Michel Fortin ◽

François A. Auger ◽

...

Keyword(s):

Bone Defect ◽

Bone Repair ◽

Human Bone ◽

Male Rats ◽

Defect Model ◽

Calvarial Bone ◽

Calvarial Bone Defect ◽

Self Assembled

In vitro prevascularization has the potential to address the challenge of maintaining cell viability at the core of engineered constructs, such as bone substitutes, and to improve the survival of tissue grafts by allowing quicker anastomosis to the host microvasculature. The self-assembly approach of tissue engineering allows the production of biomimetic bone-like tissue constructs including extracellular matrix and living human adipose-derived stromal/stem cells (hASCs) induced towards osteogenic differentiation. We hypothesized that the addition of endothelial cells could improve osteogenesis and biomineralization during the production of self-assembled human bone-like tissues using hASCs. Additionally, we postulated that these prevascularized constructs would consequently improve graft survival and bone repair of rat calvarial bone defects. This study shows that a dense capillary network spontaneously formed in vitro during tissue biofabrication after two weeks of maturation. Despite reductions in osteocalcin levels and hydroxyapatite formation in vitro in prevascularized bone-like tissues (35 days of culture), in vivo imaging of prevascularized constructs showed an improvement in cell survival without impeding bone healing after 12 weeks of implantation in a calvarial bone defect model (immunocompromised male rats), compared to their stromal counterparts. Globally, these findings establish our ability to engineer prevascularized bone-like tissues with improved functional properties.

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Functional tissue-engineered bone-like graft made of a fibrin scaffold and TG2 gene-modified EMSCs for bone defect repair

NPG Asia Materials ◽

10.1038/s41427-021-00297-w ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Wentao Shi ◽

Yunduan Que ◽

Xuan Zhang ◽

Lu Bian ◽

Xuejian Yu ◽

...

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Bone Defect ◽

Osteogenic Medium ◽

Fibrin Gel ◽

Bone Defect Repair ◽

Functional Tissue ◽

Fibrin Scaffold ◽

Defect Repair

AbstractThe transplantation of tissue-engineered scaffolds with stem cells is a promising therapeutic approach for bone defect repair. To improve the therapeutic efficacy of this approach, in this study, a novel biofunctional live tissue-engineered bone-like graft was designed and constructed using a fibrin scaffold loaded with TG2 gene-modified ectomesenchymal stem cells (TG2-EMSCs) derived from nasal respiratory mucosa for bone defect repair. Autocalcification of the cell-free fibrin gel in osteogenic medium with additional alkaline phosphatase (ALP) and the osteogenic differentiation of TG2-EMSCs on the fibrin scaffold were assessed in vitro. The results indicated that the cell-free fibrin gel could autocalcify in the osteogenic medium with ALP and that the overexpression of TG2 by TG2-EMSCs could promote the osteogenic differentiation of these stem cells in the fibrin scaffold. Moreover, TG2 could enhance the deposition of extracellular matrix proteins in the fibrin scaffold, followed by calcification of the bone matrix in vitro. After transplantation into critical-sized cranial defects in rats, the functional tissue-engineered bone-like grafts improved bone regeneration. These results indicate that this tissue-engineered bone-like graft could improve the process of bone defect repair.

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