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Isolation of osteogenic progenitors from human amniotic fluid using a single step culture protocol.

Antonucci I, Iezzi I, Morizio E, Mastrangelo F, Pantalone A, Mattioli-Belmonte M, Gigante A, Salini V, Calabrese G, Tetè S, Palka G, Stuppia L - BMC Biotechnol. (2009)

Bottom Line: Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases.Electron microscopy analysis evidenced the best cell growth on this latter surface.The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Sciences, G, d'Annunzio University, Chieti-Pescara, Italy. antonucciivana@libero.it

ABSTRACT

Background: Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases. In the present study, we report a novel single step protocol for the osteoblastic differentiation of human amniotic fluid cells.

Results: The described protocol is able to provide osteoblastic cells producing nodules of calcium mineralization within 18 days from withdrawal of amniotic fluid samples. These cells display a complete expression of osteogenic markers (COL1, ONC, OPN, OCN, OPG, BSP, Runx2) within 30 days from withdrawal. In order to test the ability of these cells to proliferate on surfaces commonly used in oral osteointegrated implantology, we carried out cultures onto different test disks, namely smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium). Electron microscopy analysis evidenced the best cell growth on this latter surface.

Conclusion: The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology.

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Related in: MedlinePlus

Scanning Electron Microscope analysis of osteoblastic cells cultured on SLA titanium disks. a) 22×, b) 500×: adherent cells covering the whole surface of SLA titanium disk; c) 1250×, d) 1250×: evidence of philophodia surrounding cell surfaces.
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Figure 5: Scanning Electron Microscope analysis of osteoblastic cells cultured on SLA titanium disks. a) 22×, b) 500×: adherent cells covering the whole surface of SLA titanium disk; c) 1250×, d) 1250×: evidence of philophodia surrounding cell surfaces.

Mentions: In order to evaluate the growth ability of osteoblastic cells obtained by Protocol 2 on different surfaces commonly used in oral implantology, cultures were carried out on smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium) test disks, and evaluated using Electron Scanning Microscopy. Titanium is universally considered as the first-rate material for oral osseointegrated implantology. Additional treatments on commercially pure (c.p.) titanium surface provide further enhancement of bone-to-implant contact, thus reducing the osseointegration period, improving treatment outcome and increasing applicability to poor bone quality. The investigation of implants with different surface treatments, both in vitro and in vivo, is a crucial point in order to define the surface morphology which could permit a good osteoblastic cell proliferation and osseointegration around implant. In our experiments, adherent cells were not detected on smooth copper surface (negative control) at day 3, while different behaviour of osteoblastic-like cells were observed on machined titanium and SLA titanium surfaces. On machined titanium surfaces, few adherent cells were observed around the titanium disk. On the contrary, adherent cells were found to cover the whole surface of SLA titanium disk (Figure 5a). Cell aggregates were arranged almost uniformly and formed a single layer cell culture on the disk surface (Figure 5b). At high magnification philophodia surrounding cell surfaces were clearly visible (Figure 5c–d).


Isolation of osteogenic progenitors from human amniotic fluid using a single step culture protocol.

Antonucci I, Iezzi I, Morizio E, Mastrangelo F, Pantalone A, Mattioli-Belmonte M, Gigante A, Salini V, Calabrese G, Tetè S, Palka G, Stuppia L - BMC Biotechnol. (2009)

Scanning Electron Microscope analysis of osteoblastic cells cultured on SLA titanium disks. a) 22×, b) 500×: adherent cells covering the whole surface of SLA titanium disk; c) 1250×, d) 1250×: evidence of philophodia surrounding cell surfaces.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2654889&req=5

Figure 5: Scanning Electron Microscope analysis of osteoblastic cells cultured on SLA titanium disks. a) 22×, b) 500×: adherent cells covering the whole surface of SLA titanium disk; c) 1250×, d) 1250×: evidence of philophodia surrounding cell surfaces.
Mentions: In order to evaluate the growth ability of osteoblastic cells obtained by Protocol 2 on different surfaces commonly used in oral implantology, cultures were carried out on smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium) test disks, and evaluated using Electron Scanning Microscopy. Titanium is universally considered as the first-rate material for oral osseointegrated implantology. Additional treatments on commercially pure (c.p.) titanium surface provide further enhancement of bone-to-implant contact, thus reducing the osseointegration period, improving treatment outcome and increasing applicability to poor bone quality. The investigation of implants with different surface treatments, both in vitro and in vivo, is a crucial point in order to define the surface morphology which could permit a good osteoblastic cell proliferation and osseointegration around implant. In our experiments, adherent cells were not detected on smooth copper surface (negative control) at day 3, while different behaviour of osteoblastic-like cells were observed on machined titanium and SLA titanium surfaces. On machined titanium surfaces, few adherent cells were observed around the titanium disk. On the contrary, adherent cells were found to cover the whole surface of SLA titanium disk (Figure 5a). Cell aggregates were arranged almost uniformly and formed a single layer cell culture on the disk surface (Figure 5b). At high magnification philophodia surrounding cell surfaces were clearly visible (Figure 5c–d).

Bottom Line: Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases.Electron microscopy analysis evidenced the best cell growth on this latter surface.The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Sciences, G, d'Annunzio University, Chieti-Pescara, Italy. antonucciivana@libero.it

ABSTRACT

Background: Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases. In the present study, we report a novel single step protocol for the osteoblastic differentiation of human amniotic fluid cells.

Results: The described protocol is able to provide osteoblastic cells producing nodules of calcium mineralization within 18 days from withdrawal of amniotic fluid samples. These cells display a complete expression of osteogenic markers (COL1, ONC, OPN, OCN, OPG, BSP, Runx2) within 30 days from withdrawal. In order to test the ability of these cells to proliferate on surfaces commonly used in oral osteointegrated implantology, we carried out cultures onto different test disks, namely smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium). Electron microscopy analysis evidenced the best cell growth on this latter surface.

Conclusion: The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology.

Show MeSH
Related in: MedlinePlus