Fabrication of polycaprolactone-silanated β-tricalcium phosphate-heparan sulfate scaffolds for spinal fusion applications
Copyright © 2017 Elsevier Inc. All rights reserved..
BACKGROUND CONTEXT: Interbody spinal fusion relies on the use of external fixation and the placement of a fusion cage filled with graft materials (scaffolds) without regard for their mechanical performance. Stability at the fusion site is instead reliant on fixation hardware combined with a selected cage. Ideally, scaffolds placed into the cage should both support the formation of new bone and contribute to the mechanical stability at the fusion site.
PURPOSE: We recently developed a scaffold consisting of silane-modified PCL-TCP (PCL-siTCP) with mechanical properties that can withstand the higher loads generated in the spine. To ensure the scaffold more closely mimicked the bone matrix, we incorporated collagen (Col) and a heparan sulfate glycosaminoglycan sugar (HS3) with increased affinity for heparin-binding proteins such as bone morphogenetic protein-2 (BMP-2). The osteostimulatory characteristic of this novel device delivering exogenous BMP2 was assessed in vitro and in vivo as a prelude to future spinal fusion studies with this device.
STUDY DESIGN/SETTING: A combination of cell-free assays (BMP2 release), progenitor cell-based assays (BMP2 bioactivity, cell proliferation and differentiation), and rodent ectopic bone formation assays was used to assess the osteostimulatory characteristics of the PCL-siTCP-based scaffolds.
MATERIALS AND METHODS: Freshly prepared rat mesenchymal stem cells were used to determine reparative cell proliferation and differentiation on the PCL-siTCP-based scaffolds over a 28-day period in vitro. The bioactivity of BMP2 released from the scaffolds was assessed on progenitor cells over a 28-day period using ALP activity assays and release kinetics as determined by enzyme-linked immunosorbent assay. For ectopic bone formation, intramuscular placement of scaffolds into Sprague Dawley rats (female, 4 weeks old, 120-150 g) was achieved in five animals, each receiving four treatments randomized for location along the limb. The four groups tested were (1) PCL-siTCP/Col (5-mm diameter×1-mm thickness), PCL-siTCP/Col/BMP2 (5 µg), (3) PCL-siTCP/Col/HS3 (25 µg), and (4) PCL-siTCP/Col/HS3/BMP2 (25 and 5 µg, respectively). Bone formation was evaluated at 8 weeks post implantation by microcomputed tomography (µCT) and histology.
RESULTS: Progenitor cell-based assays (proliferation, mRNA transcripts, and ALP activity) confirmed that BMP2 released from PCL-siTCP/Col/HS3 scaffolds increased ALP expression and mRNA levels of the osteogenic biomarkers Runx2, Col1a2, ALP, and bone gla protein-osteocalcin compared with devices without HS3. When the PCL-siTCP/Col/HS3/BMP2 scaffolds were implanted into rat hamstring muscle, increased bone formation (as determined by two-dimensional and three-dimensional µCTs and histologic analyses) was observed compared with scaffolds lacking BMP2. More consistent increases in the amount of ectopic bone were observed for the PCL-siTCP/Col/HS3/BMP2 implants compared with PCL-siTCP/Col/BMP2. Also, increased mineralizing tissue within the pores of the scaffold was seen with modified-tetrachrome histology, a result confirmed by µCT, and a modest but detectable increase in both the number and the thickness of ectopic bone structures were observed with the PCL-siTCP/Col/HS3/BMP2 implants.
CONCLUSIONS: The combination of PCL-siTCP/Col/HS3/BMP2 thus represents a promising avenue for further development as a bone graft alternative for spinal fusion surgery.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2018 |
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| Erschienen: |
2018 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:18 |
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| Enthalten in: |
The spine journal : official journal of the North American Spine Society - 18(2018), 5 vom: 01. Mai, Seite 818-830 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Bhakta, Gajadhar [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 11.01.2019 Date Revised 11.01.2019 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.spinee.2017.12.002 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM279330294 |
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| 100 | 1 | |a Bhakta, Gajadhar |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Fabrication of polycaprolactone-silanated β-tricalcium phosphate-heparan sulfate scaffolds for spinal fusion applications |
| 264 | 1 | |c 2018 | |
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| 500 | |a Date Completed 11.01.2019 | ||
| 500 | |a Date Revised 11.01.2019 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2017 Elsevier Inc. All rights reserved. | ||
| 520 | |a BACKGROUND CONTEXT: Interbody spinal fusion relies on the use of external fixation and the placement of a fusion cage filled with graft materials (scaffolds) without regard for their mechanical performance. Stability at the fusion site is instead reliant on fixation hardware combined with a selected cage. Ideally, scaffolds placed into the cage should both support the formation of new bone and contribute to the mechanical stability at the fusion site | ||
| 520 | |a PURPOSE: We recently developed a scaffold consisting of silane-modified PCL-TCP (PCL-siTCP) with mechanical properties that can withstand the higher loads generated in the spine. To ensure the scaffold more closely mimicked the bone matrix, we incorporated collagen (Col) and a heparan sulfate glycosaminoglycan sugar (HS3) with increased affinity for heparin-binding proteins such as bone morphogenetic protein-2 (BMP-2). The osteostimulatory characteristic of this novel device delivering exogenous BMP2 was assessed in vitro and in vivo as a prelude to future spinal fusion studies with this device | ||
| 520 | |a STUDY DESIGN/SETTING: A combination of cell-free assays (BMP2 release), progenitor cell-based assays (BMP2 bioactivity, cell proliferation and differentiation), and rodent ectopic bone formation assays was used to assess the osteostimulatory characteristics of the PCL-siTCP-based scaffolds | ||
| 520 | |a MATERIALS AND METHODS: Freshly prepared rat mesenchymal stem cells were used to determine reparative cell proliferation and differentiation on the PCL-siTCP-based scaffolds over a 28-day period in vitro. The bioactivity of BMP2 released from the scaffolds was assessed on progenitor cells over a 28-day period using ALP activity assays and release kinetics as determined by enzyme-linked immunosorbent assay. For ectopic bone formation, intramuscular placement of scaffolds into Sprague Dawley rats (female, 4 weeks old, 120-150 g) was achieved in five animals, each receiving four treatments randomized for location along the limb. The four groups tested were (1) PCL-siTCP/Col (5-mm diameter×1-mm thickness), PCL-siTCP/Col/BMP2 (5 µg), (3) PCL-siTCP/Col/HS3 (25 µg), and (4) PCL-siTCP/Col/HS3/BMP2 (25 and 5 µg, respectively). Bone formation was evaluated at 8 weeks post implantation by microcomputed tomography (µCT) and histology | ||
| 520 | |a RESULTS: Progenitor cell-based assays (proliferation, mRNA transcripts, and ALP activity) confirmed that BMP2 released from PCL-siTCP/Col/HS3 scaffolds increased ALP expression and mRNA levels of the osteogenic biomarkers Runx2, Col1a2, ALP, and bone gla protein-osteocalcin compared with devices without HS3. When the PCL-siTCP/Col/HS3/BMP2 scaffolds were implanted into rat hamstring muscle, increased bone formation (as determined by two-dimensional and three-dimensional µCTs and histologic analyses) was observed compared with scaffolds lacking BMP2. More consistent increases in the amount of ectopic bone were observed for the PCL-siTCP/Col/HS3/BMP2 implants compared with PCL-siTCP/Col/BMP2. Also, increased mineralizing tissue within the pores of the scaffold was seen with modified-tetrachrome histology, a result confirmed by µCT, and a modest but detectable increase in both the number and the thickness of ectopic bone structures were observed with the PCL-siTCP/Col/HS3/BMP2 implants | ||
| 520 | |a CONCLUSIONS: The combination of PCL-siTCP/Col/HS3/BMP2 thus represents a promising avenue for further development as a bone graft alternative for spinal fusion surgery | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Bone morphogenetic protein-2 | |
| 650 | 4 | |a Collagen | |
| 650 | 4 | |a Ectopic bone | |
| 650 | 4 | |a Glycosaminoglycans | |
| 650 | 4 | |a PCL | |
| 650 | 4 | |a Spinal fusion | |
| 650 | 7 | |a BMP2 protein, human |2 NLM | |
| 650 | 7 | |a Bone Morphogenetic Protein 2 |2 NLM | |
| 650 | 7 | |a Calcium Phosphates |2 NLM | |
| 650 | 7 | |a Polyesters |2 NLM | |
| 650 | 7 | |a beta-tricalcium phosphate |2 NLM | |
| 650 | 7 | |a polycaprolactone |2 NLM | |
| 650 | 7 | |a 24980-41-4 |2 NLM | |
| 650 | 7 | |a Collagen |2 NLM | |
| 650 | 7 | |a 9007-34-5 |2 NLM | |
| 650 | 7 | |a Heparitin Sulfate |2 NLM | |
| 650 | 7 | |a 9050-30-0 |2 NLM | |
| 700 | 1 | |a Ekaputra, Andrew K |e verfasserin |4 aut | |
| 700 | 1 | |a Rai, Bina |e verfasserin |4 aut | |
| 700 | 1 | |a Abbah, Sunny A |e verfasserin |4 aut | |
| 700 | 1 | |a Tan, Tuan Chun |e verfasserin |4 aut | |
| 700 | 1 | |a Le, Bach Quang |e verfasserin |4 aut | |
| 700 | 1 | |a Chatterjea, Anindita |e verfasserin |4 aut | |
| 700 | 1 | |a Hu, Tao |e verfasserin |4 aut | |
| 700 | 1 | |a Lin, Tingxuan |e verfasserin |4 aut | |
| 700 | 1 | |a Arafat, M Tarik |e verfasserin |4 aut | |
| 700 | 1 | |a van Wijnen, Andre J |e verfasserin |4 aut | |
| 700 | 1 | |a Goh, James |e verfasserin |4 aut | |
| 700 | 1 | |a Nurcombe, Victor |e verfasserin |4 aut | |
| 700 | 1 | |a Bhakoo, Kishore |e verfasserin |4 aut | |
| 700 | 1 | |a Birch, William |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Li |e verfasserin |4 aut | |
| 700 | 1 | |a Gibson, Ian |e verfasserin |4 aut | |
| 700 | 1 | |a Wong, Hee-Kit |e verfasserin |4 aut | |
| 700 | 1 | |a Cool, Simon M |e verfasserin |4 aut | |
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