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. 2021 May 11;9:661332. doi: 10.3389/fbioe.2021.661332

Table 5.

Pre-clinical studies using amniotic membrane derivated stromal cells for bone regeneration.

References Animal (No. per condition and per time) Model: Defect localization and size Amniotic cells characteristics (n = number of cells seeded on scaffold) Scaffold (t = culture duration before implantation) Treatments Evaluation methodology Results
Mattioli et al. (2012) Sheep (No. = 2) Tibial defectDiameter: 3 mm oAECs (2 × 106 cells) Fibrin glue (Tissuecol) (no culture) 1) Tissuecol
2) Tissuecol + oAECs
Histology Bone deposition was only observed in oAECs-transplanted defects after 45 days.
Tsuno et al. (2012) Rat (No. = 3) Calvarial defectDiameter: 5 mm hAMSCs (1 × 107 cells/mL) β-TCP (t = NS) 1) β-TCP
2) β-TCP + hAMSCs
Histology hAMSCs seeded scaffold showed immature bone deposition at 6 weeks and mature bone areas at 12 weeks.
Barboni et al. (2013) Sheep (No. = 3) Sinus augmentation oAECs (1 × 106 cells) HA/β-TCP (t = 3 days) 1) HA/β-TCP
2) HA/β-TCP + oAECs
Micro-CTHistology oAECs seeded scaffold displayed significant earlier bone formation and maturation at 45 days and induced significantly more bone deposition at 90 days.
Chen et al. (2014) Mice (No. = 6) Subcutaneous hAMSCs (5 × 104 cells/mL) CultiSpher S: Porcine gelatin microcarriers (t = 28–42 days) 1) CultiSpher S+ hAMSCs (no perfusion)
2) CultiSpher S+ hAMSCs (1 week perfusion)
3) CultiSpher + hAMSCs (2 weeks perfusion)
Micro-CTHistology Perfusion culture system increased mineralized matrix after 6 and 12 weeks Perfusion significantly enhanced vessel density.
Jiawen et al. (2014) Rat (No. = 4) Alveolar defectSize: NS hAECs (1.5 × 106) β-TCP (t = 3 days) 1) β-TCP
2) β-TCP + hAECs
Micro-CTHistology hAECs seeded scaffold significantly increased bone formation at 4 and 8 weeks postoperatively. hAECs seeded scaffold showed a significantly delayed macrophage response.
Si et al. (2015) Mice (No. = 3) Subcutaneous hAECs (1.5 × 106) β-TCP (t = 3 days) 1) β-TCP
2) β-TCP + hBMSCs
3) β-TCP + hAFMSCs
4) β-TCP + hAECs
HistologyImmunohisto-chemistry No sign of mineralization in all groups 1 month after implantation. OPN and OCN were expressed at a higher level with the seeded scaffolds.
Rameshbabu et al. (2016) Rabbit (No. = 5) Osteochondral defectDiameter: 4 mmDeep: 5 mm hAMSCs (NS) PEMS (7 days) 1) Empty defect
2) PEMS
3) PEMS + hAMSCs
Histology hAMSCs seeded scaffold seemed to induce higher bone formation and osteochondral regeneration 60 days post-implantation.
Jiang et al. (2020) Rabbit (No. = 3) Calvarial defectDiameter: 10 mm hAMSCs (5 × 106 cells/mL) Fibrinogen solution (NS) 1) Fibrin gel
2) Bio-oss
3) hAMSCs
4) Bio-oss + hAMSCs
Micro-CTHistologySequential fluorescent labeling Bone regeneration was significantly higher in groups 3 and 4 after 4 and 12weeks. Fluorescent labeling was significantly higher in group 4 after 3, 6, and 9 weeks. Vessels-like structure are significantly higher in presence of hAMSCs.
Li et al. (2020) Rat (No. = 3) Calvarial defectDiameter: 3 mm hAMSCs (1 × 106 cells/mL) Fibrin (NS) 1) Fibrin
2) Fibrin + hAMSCs
Micro-CTHistology hAMSCs seeded scaffold induced significantly higher bone formation after 8 weeks.
Datta et al. (2021) Rabbit (No. = NS) Tibial defectDiameter: 2.5 mmDeep: 2 mm hAMSCs (1 × 106 cells/mL) Hydrogel hybride (DBM + chitosan) 1) Empty defect
2) Hydrogel
3) Hydrogel + hAMSCs
Micro-CTHistology Bone regeneration was significantly higher using hydrogel + hAMSCs after 4 and 8 weeks. hAMSCs seeded scaffold showed more vascular structure after 4 weeks compared to the two other groups.

β-TCP, β-tricalcium phosphate; DBM, demineralized bone matrix; HA, hydroxyapatite; hAECs, human amniotic epithelial cells; hAMSCs, human amniotic mesenchymal stromal cells; NS, Not specified; oAECs, ovine amniotic epithelial cells; OCN, osteocalcin; OPN, osteopontin; PEMS, Placenta-derived Extracellular Matrix Sponge.