CUSTOM IMPLANTS Research, Regenerative Medicine and Personalized Surgery at Rizzoli Orthopedic Institute PERSONALIZED SCAFFOLDS: In-vitro validation of magnetic microstructured 3D printed scaffolds In-vitro validation of microstructured scaffolds through 3D printing in bioabsorbable materials colonized with human cells. A magnetic manipulator of cells, magnetized by internalization or by coating of magnetic nanoparticles, is developing its prototype

1. Anna Sagnella
www.laboratoriomister.it

2. PERSONALIZED SCAFFOLDS
Dediu Alek - v.dediu@bo.ismn.cnr.it
IOR – RIT – Lab. RAMSES
Dott.ssa Grigolo Brunella - brunella.grigolo@ior.it
Ing. Petretta Mauro - mauro.petretta@ior.it Industrial Partner I+ Srl

3. PERSONALIZED SCAFFOLDS
Realization of biomaterials and/or cell-based
devices by 3D-printing and in-vitro
cellularization

4. CUSTOM IMPLANTS 4
Scope:
In-vitro study and validation of “mixed” systems realized by
magnetic cellularization of micro-structured scaffolds
produced by 3D-printing

5. CUSTOM IMPLANTS 5
Magnetic Scaffolds:
First Formulation:
PCL (PolyCaproLactone)
+ magnetic nanoparticles (Fe2O3)
Improvement of Osteogenic potential
Biomaterial-based
Nanocomposite: Polymer + Filler
New Formulation with
calcium phosfate or hydroxyapatite
(HA)

6. CUSTOM IMPLANTS 6
Analysis carried out on biomaterial-based nanocomposites:
➢ Presence of magnetic nanoparticles (in red)
➢ Presence of hydroxyapatite nanoparticles (in blu)
➢ Not formation of cluster
AFM Analisys:
Topografia Fase meccanica Fase magnetica
Topografic Mechanic phase Magnetic phase

7. CUSTOM IMPLANTS 7
Scaffolds Realization:
Different scaffolds for internal geometry and architecture
are projected and fabricated by Bioprinting «3D Discovery»
(regenHU) platform, with magnetic nanoparticles (Image A)
and without magnetic nanoparticles as reference (Image B)
(A) (B)

8. CUSTOM IMPLANTS 8
Biocompatibility:
Biocompatibility studies carried out on PCL/HA scaffolds, with and
without magnetic nanoparticles; stromal mesenchymal cells from
marrow (MSCs) are used.
B
MSCs
B
MSCs
PCL/HA
Microscopy (Day 21)
MSCs: mesenchymal cell
B: biomaterial
PCL/HA
+Fe2O3
MSCs
B B
MSCs
VITALITA' CELLULARE
GIORNO 1 GIORNO 7 GIORNO 14 GIORNO 21
0
2000
4000
6000
PCL - Fe2O3
PCL + Fe2O3
Assorbanzaa570nm
The presence of magnetic nanoparticle does not
affect negatively on cell vitality
Cell vitality

9. CUSTOM IMPLANTS 9
Osteogenic differentiation:
Evaluation of osteogenic differentiation potential by in-vitro
studies in presence of osteogenic factors (FO), days 1,7, 14 e 21
VITALITA' CELLULARE
GIORNO 1 GIORNO 7 GIORNO 14 GIORNO 21
0
2000
4000
6000
PCL + Fe2O3 - FO
PCL + Fe2O3 + FO
Assorbanzaa570nm
PCL + Fe2O3 + FO
PCL + Fe2O3 - FO
Cell vitality: Alamar-blue assay
PCL/HA/
Fe2O3
-FO
PCL/HA/
Fe2O3
+FO
day 14 day 21
MSCs
MSCs
MSCs
MSCs
B
B
B
B
➢ PCL/HA/Fe2O3 +/- Fe2O3 scaffolds show a good cell vitality
and they can promote cell proliferation
➢ Work in progress: specific analysis focused on
mineralization process
MSCs: mesenchymal cell
B: biomaterial
FO: Osteogenic factors

10. CUSTOM IMPLANTS 10
Scaffolds cellularization:
➢ By magnetic guide for cells
➢ Magnetized and no-magnetized cells
➢ Magnetized and no-magnetized scaffolds
➢ Magnetic nanoparticles low cytotoxicity
➢ Specific cell internalization
Magnetic cellularization of 12
scaffolds: 6 magnetic and 6 non
magnetic
Cellularization of scaffolds placed in
magnetic gradients is 1.5 more
efficient

11. CUSTOM IMPLANTS 11
Realization of a prototype:
magnetic manipulator
Study
Design
Assembly
An optimal magnetic guide to
move and collocate the cells
into scaffolds fabricated by 3D-
printing

12. CUSTOM IMPLANTS 12
Magnetic manipulator:
assembly and development
• 1 transparent tube with
perfusion system
• 8 pointed magnets and 8
pointed focusing in 3D
architecture controlled by
computer
• 2 magnetic manipulator high
precision (ca. 10 μm)
• Optical and fluorescence
microscopy
• 1 mathematic model
COMSOL for magnetic
topographic

13. CUSTOM IMPLANTS 13
Magnetic manipulator:
operation
• 3D-configuration
• The magnets produces
magnetic gradients of
different intensity and
geometry
• The cells are moved into
and outside the scaffolds
placed into perfused tube
• In situ observation

14. CUSTOM IMPLANTS 14
Magnetic porous
Scaffolds
Magnets
Distribution of
magnetic
gradients around
the scaffolds in
perpendicular
fields
Presence of a local
area of magnetic
attraction near the
circumscribed fiber
(60 ° angle) and
remaining
repulsion potential
➢ Improvement of precision for cell internalization
➢ Loading of different cells type without overlays
Magnetic manipulator:
in vitro cellularization

15. CUSTOM IMPLANTS 15
Magnetic cellularization of 12 scaffolds: 6
magnetic and 6 non magnetic
Cellularization of scaffolds placed in magnetic
gradients is 1.5 more efficient
Magnetic manipulator:
in vitro cellularization

16. CUSTOM IMPLANTS 16
Thanks
Dediu Alek - v.dediu@bo.ismn.cnr.itSagnella Anna - sagnella@laboratoriomister.it
IOR – RIT – Lab. RAMSES
Dott.ssa Grigolo Brunella - brunella.grigolo@ior.it
Ing. Petretta Mauro - mauro.petretta@ior.it
Dott.ssa Cavallo Carola - carola.cavallo@ior.it
Dott.ssa Desando Giovanna - giovanna.desando@ior.it
Partner industriale I+ Srl