Stromal Vascular Fraction and Amniotic Epithelial Cells: Preclinical and Clinical Relevance in Musculoskeletal Regenerative Medicine
Table 4
In vitro, in vivo, and clinical studies on AECs in bone regeneration.
Experimental model
Treatment groups
Evaluations
Results
Ref
In vitro: hAECs Purchased hADSCs
Group 1: hAECs () cultured in OM Group 2: hADSCs () cultured in OM Group 3: hAECs ( per pellet) cultured in CM Group 4: hADSCs ( per pellet) cultured in CM
RT-PCR (RUNX2, SOX9) Alizarin red staining Alcian blue staining IHC (OCN, AGG, and COLL II)
3, 7, and 14 days Group 1: ↑ RUNX2 gene expression, OCN protein than group 2 Group 3:↑ AGG and COLL II proteins, SOX9 gene expression, than group 4 2 and 4 wks Group 1: ↑ mineralization during time
In vitro: hAECs , 1 mT for 30 min each time, 2 for days with an interval of 12 hours
Group 1: hAECs in NM Group 2: hAECs in OM Group 3: hAECs in NM+PEMF Group 4: hAECs in OM+PEMF
RT-PCR (ALP, OCN, BMP2, RUNX2, NRF2, and KEAP1) ALP activity IHC (OCN) Calcium deposition
3, 7, and 11 days Group 4: ↑ ALP, OCN gene expression than groups 1-3 Group 3: ↑ ALP, OCN than groups 1 and 2 Group 2: ↑ ALP, OCN than group 1 7 days Group 3: ↑ ALP activity, calcium deposition than group 1 7, 11, and 21 days Group 2: ↑ ALP activity than groups 1 and 3 Group 4: ↑ ALP activity, OCN protein, calcium deposition than groups 1-3 Group 3: ↑ OCN protein than group 1 21 days Group 2: ↑ OCN protein, calcium deposition than groups 1 and 3 3, 11, and 21 days Group 4: ↑ BMP2, RUNX2, NRF2, and KEAP1 gene expression than groups 1-3
In vitro: Conditioned medium from hAECs after 24 hrs of culture Purchased hFOB1.19 ()
Group 1: hFOB1.19 in NM Group 2: hFOB1.19 in conditioned medium Group 3: hFOB1.19 in conditioned medium+TGFβ1 antibody (5 μg/ml)
Cell proliferation Cell migration ALP activity RT-PCR (ALP, OCN, OPN, and RUNX2)
2 hrs Group 2: ↑ hFOB1.19 migration than group 1 1, 2, and 3 days Group 2: ↑ hFOB1.19 proliferation than group 1 6 days Group 2: ↑ hFOB1.19 ALP activity, ALP, OCN, OPN, RUNX2 gene expression than group 1 Group 3: ↓ hFOB1.19 ALP activity, ALP, OCN gene expression than group 2 6 hrs Group 3: ↓ hFOB1.19 migration than group 2
In vitro: hAECs (3rd-5th passage) from 6 patients ()
Group 1: hAECs in NM Group 2: hAECs+mechanical stretch (maximum uniaxial stretched length (7.35 cm) for 2, 6, 12, and 24 hrs with 5% elongation at a frequency of 0.5 Hz Group 3: hAECs in OM for 21 days Group 4: hAECs+OM for 21 days+mechanical stretch for 2, 6, 12, and 24 hrs Group 5: hAECs+Runx2 shRNA+OM for 24 hrs Group 6: hAECs+Runx2 shRNA+mechanical stretch for 24 hrs Group 7: hAECs+Runx2 shRNA+OM+mechanical stretch for 24 hrs
RT-PCR (ALP, OCN, RUNX2, β-catenin, and Cyclin D1) WB (ALP, OCN, RUNX2, β-catenin, and Cyclin D1)
12 hrs Groups 3 and 4: ↑ OCN protein than group 1 6, 12, and 24 hrs Group 3: ↑ RUNX2, ALP, and OCN gene expression than group 1 6 and 12 hrs Group 4: ↑ RUNX2, ALP, OCN gene expression and protein than groups 2 and 3 12 hrs Group 3: ↑ β-catenin, RUNX2, Cyclin D1 gene expression and protein than group 1 Group 4: ↑ β-catenin, RUNX2, Cyclin D1 gene expression and protein than groups 2 and 3 24 hrs Groups 5-7: ↓ RUNX2, OCN gene expression, and protein than groups 2-4
In vivo: 6 adult sheep (2 yrs) that need bilateral sinus augmentation Allogenic oAECs () from 3 slaughtered sheep (3 mo of pregnancy) CaP (HA-βTCP: 30/60) scaffold
Group 1: scaffold Group 2: scaffold+AECs
Micro-CT Histology Histomorphometry IHC (VEGF)
45 days Group 2: ↑ VEGF, VA; ↑ BA than group 1 3 mo Group 2: ↓ VEGF, VA; ↑ BA than group 1
