Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Bone Enhancement with BMP-2 for Safe Clinical Translation
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Polymer Chemistry.
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Bone morphogenetic protein-2 (BMP-2) is considered a promising adjuvant for the treatment of bone regeneration. However, BMP-2 delivery in a conventional collagen scaffold needs a high dose to achieve an effective outcome. Moreover, such dosage may lead to serious side effects. The aim of the following thesis was to find clinically acceptable strategies reducing the required dose of BMP-2 by improving the delivery and optimizing the preclinical testing of the new approaches. In all the studies hyaluronic acid (HA) hydrogels was used as a carrier for BMP-2.

The HA hydrogel/BMP-2 construct was modified with bioactive matrix components in order to obtain an effective release of BMP-2 and an enhanced bone formation. The most promising were two strategies. In the first one, BMP-2, precomplexed with the glycosaminoglycans dermatan sulfate or heparin prior to loading it into HA hydrogel, protected and prolonged the delivery of the protein, resulting in twofold larger bone formation in comparison to non-complexed BMP-2. In the second strategy, the fibronectin fragment integrin-binding domain (FN) was covalently incorporated into HA hydrogel. The FN remarkably improved the capacity of the material to support the cells attachment and spreading, providing the formation of twice as much bone in comparison to non-functionalized HA hydrogel/BMP-2.

Furthermore, the importance of a proper design of the preclinical study for BMP-2 delivery systems was highlighted. Firstly, proper physicochemical handling of BMP-2 showed the improvement in further in vivo activity.  The use of glass storage vials and an acidic formulation buffer was superior to plastic surfaces and physiological pH. Secondly, while regenerative medicine strategy testing required the use of animal models that matched the research questions related to clinical translation, two new animal models were developed. The subperiosteal mandibular and calvarial models in rats were found to be minimally invasive, convenient and rapid solution for the evaluation of a broad range of approaches including bone augmentation, replacement and regeneration. Both models are primarily relevant for the initial testing of the injectable bone engineering constructs. 

Those clinically translatable approaches presented here could prove to be a powerful platform for a wider use of BMP-2 in orthopedic, plastic surgery and regenerative medicine research.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2013. , 74 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1009
Keyword [en]
Bone repair, Bone healing, Bone morhogenetic protein-2, Osteogenesis, Extracelular matrix, Hyaluronan, Animal model
National Category
Orthopedics Biomaterials Science
Research subject
Orthopaedics; Engineering Science with specialization in Materials Science
Identifiers
URN: urn:nbn:se:uu:diva-188027ISBN: 978-91-554-8572-6 (print)OAI: oai:DiVA.org:uu-188027DiVA: diva2:577093
Public defence
2013-02-08, Museum Gustavianum - Auditorium Minus, Akademigatan 3, Uppsala, 13:15 (English)
Opponent
Supervisors
Available from: 2013-01-18 Created: 2012-12-12 Last updated: 2013-02-11Bibliographically approved
List of papers
1. Critical assessment of rhBMP-2 mediated bone induction: An in vitro and in vivo evaluation.
Open this publication in new window or tab >>Critical assessment of rhBMP-2 mediated bone induction: An in vitro and in vivo evaluation.
Show others...
2012 (English)In: Journal of Controlled Release, ISSN 0168-3659, E-ISSN 1873-4995, Vol. 162, no 3, 646-653 p.Article in journal (Refereed) Published
Abstract [en]

Understanding the influence of formulation and storage conditions on rhBMP-2 bioactivity is extremely important for its clinical application. Reports in the literature show that different research groups employ different parameters such as formulation conditions, storage, doses for in vivo applications etc. that makes it difficult to correlate results from different experiments. We therefore decided to rationalize these anomalies by performing a basic study on such parameters using two commercially available BMPs. Our in vitro experiments suggest that BMPs from different sources have significant differences in their bioactivity. The clinically approved rhBMP-2 (InductOs®; BMP-P) showed superior stability, compared to rhBMP-2 from R&D Systems (BMP-R) at physiological pH (determined by ALP assay). This BMP-P also showed lower binding to polypropylene Eppendorf tube. The BMP-R almost lost its bioactivity within 30min at physiological pH and also shows more adhesion to plastic surfaces. This aggregation behavior was unequivocally ascertained by performing light scattering studies of the two BMPs, which revealed linear aggregation with time for BMP-R unlike BMP-P. The in vitro results were also reflected in the in vivo experiments, in a rat ectopic model with injectable hyaluronic acid (HA) hydrogel as BMP carrier. After 7weeks post-implantation we observed larger bone volume with oriented collagen in the BMP-P group but a smaller bone with disoriented collagen in the BMP-R case. Our results highlight the large difference in activity between seemingly identical substances and also the importance of proper handling of such sensitive proteins.

National Category
Natural Sciences Polymer Chemistry
Research subject
Chemistry with specialization in Polymer Chemistry
Identifiers
urn:nbn:se:uu:diva-181153 (URN)10.1016/j.jconrel.2012.08.004 (DOI)000310506900021 ()22902595 (PubMedID)
Available from: 2012-09-17 Created: 2012-09-17 Last updated: 2017-12-07Bibliographically approved
2. Smart design of stable hydrazone crosslinked extracellular matrix mimetic hydrogel for tissue engineering application
Open this publication in new window or tab >>Smart design of stable hydrazone crosslinked extracellular matrix mimetic hydrogel for tissue engineering application
Show others...
2012 (English)In: Journal of Tissue Engineering and Regenerative Medicine, ISSN 1932-6254, Vol. 6, no suppl 1, 192-192 p.Article in journal, Meeting abstract (Other academic) Published
Abstract [en]

Injectable hydrogels are important biomaterials with enormous applications. They are used for various biomedical applications such as diagnostics, 3D cell culture matrix, drug reservoir, encapsulation of bioactive compounds and growth factors, scaffold for tissue engineering etc. We here present our recent development in our efforts to develop hydrogel scaffolds with enhanced rigidity, stability, swelling characteristics. Hydrazone crosslinked gels are attractive due to its simplicity and versatility which could be formed by mixing appropriate aldehyde and hydrazide functionalized hyaluronan. By fine-tuning the electronic character around the hydrazone linkage, we succeeded in developing extremely stable hydrazone bond and utilized it for developing hyaluronan (HA) based synthetic extracellular matrix (ECM) hydrogel. Among the different hydrazides tested, we identified carbonyldihydrazide (CDH) as the best candidate to deliver stable hydrazone linkage. This stability is presumably due to extensive delocalization of the positive charge across neighboring amino groups of CDH. The hydrolytic stability imparted by this group was found to be several folds under acidic, basic and physiological pH when compared to other hydrazones. This tailored hydrogel with CDH also exhibited superior swelling and mechanical properties and enzymatic stability which makes it ideal for tissue engineering application.

National Category
Medical and Health Sciences Polymer Chemistry
Research subject
Chemistry with specialization in Polymer Chemistry
Identifiers
urn:nbn:se:uu:diva-182412 (URN)10.1002/term.1586 (DOI)000308313001311 ()
Available from: 2012-10-12 Created: 2012-10-10 Last updated: 2013-02-11Bibliographically approved
3. Complexation and sequestration of BMP-2 from an ECM mimetic hyaluronan gel for improved bone formation
Open this publication in new window or tab >>Complexation and sequestration of BMP-2 from an ECM mimetic hyaluronan gel for improved bone formation
Show others...
2013 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 8, no 10, e78551Article in journal (Refereed) Published
Abstract [en]

Bone morphogenetic protein-2 (BMP-2) is considered a promising adjuvant for the treatment of skeletal non-union and spinal fusion. However, BMP-2 delivery in a conventional collagen scaffold necessitates a high dose to achieve an efficacious outcome. To lower its effective dose, we precomplexed BMP-2 with the glycosaminoglycans (GAGs) dermatan sulfate (DS) or heparin (HP), prior to loading it into a hyaluronic acid (HA) hydrogel. In vitro release studies showed that BMP-2 precomplexed with DS or HP had a prolonged delivery compared to without GAG. BMP-2-DS complexes achieved a slightly faster release in the first 24 h than HP; however, both delivered BMP-2 for an equal duration. Analysis of the kinetic interaction between BMP-2 and DS or HP showed that HP had approximately 10 times higher affinity for BMP-2 than DS, yet it equally stabilized the protein, as determined by alkaline phosphatase activity. Ectopic bone formation assays at subcutaneous sites in rats demonstrated that HA hydrogel-delivered BMP-2 precomplexed with GAG induced twice the volume of bone compared with BMP-2 delivered uncomplexed to GAG.

Keyword
Bone morphogenetic protein-2, Bone repair, BMP-2 release, Heparin, Dermatan sulfate
National Category
Orthopedics Biomaterials Science
Research subject
Orthopaedics; Engineering Science with specialization in Materials Science
Identifiers
urn:nbn:se:uu:diva-188175 (URN)10.1371/journal.pone.0078551 (DOI)000326034500093 ()
Available from: 2012-12-13 Created: 2012-12-13 Last updated: 2017-12-06Bibliographically approved
4. Improving the osteogenic potential of BMP-2 with hyaluronic acid hydrogel modified with integrin-specific fibronectin fragment
Open this publication in new window or tab >>Improving the osteogenic potential of BMP-2 with hyaluronic acid hydrogel modified with integrin-specific fibronectin fragment
Show others...
2013 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 34, no 3, 704-712 p.Article in journal (Refereed) Published
Abstract [en]

While human bone morphogenetic protein-2 (rhBMP-2) is a promising growth factor for bone regeneration, its clinical efficacy has recently shown to be below expectation. In order to improve the clinical translation of rhBMP-2, there exists strong motivation to engineer better delivery systems. Hyaluronic acid (HA) hydrogel is a suitable carrier for the delivery of rhBMP-2, but a major limitation of this scaffold is its low cell adhesive properties. In this study, we have determined whether covalent grafting of an integrin-specific ligands into HA hydrogel could improve cell attachment and further enhance the osteogenic potential of rhBMP-2. A structurally stabilized fibronectin (FN) fragment containing the major integrin-binding domain of full-length FN (FN III9 *-10) was engineered, in order to be incorporated into HA hydrogel. Compared to non-functionalized HA hydrogel, HA-FN hydrogel remarkably improved the capacity of the material to support mesenchymal stem cell attachment and spreading. In an ectopic bone formation model in the rat, delivery of rhBMP-2 with HA-FN hydrogel resulted in the formation of twice as much bone with better organization of collagen fibers compared to delivering the growth factor in non-functionalized HA hydrogel. This engineered hydrogel carrier for rhBMP-2 can be relevant in clinical bone repair.

Keyword
Bone regeneration, Cell adhesion, Fibronectin, Hyaluronic acid hydrogel, Integrins, RhBMP-2
National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-184869 (URN)10.1016/j.biomaterials.2012.10.015 (DOI)000312759800011 ()
Available from: 2012-11-21 Created: 2012-11-15 Last updated: 2017-12-07Bibliographically approved
5. Bone Engineering by Biomimetic Injectable Hydrogel
Open this publication in new window or tab >>Bone Engineering by Biomimetic Injectable Hydrogel
Show others...
2012 (English)In: Molecular Crystals and Liquid Crystals, ISSN 1542-1406, Vol. 555, no 1, 177-188 p.Article in journal (Refereed) Published
Abstract [en]

Osteoporosis is a multifactorial bone disease characterized by low bone mineral density (BMD) and deterioration of micro-architecture of cancellous bone leading to bone fragility and risk of fractures. In the current work, a novel tissue engineering strategy was experimented to enhance bone architecture in the risk areas via local injection of a biomimetic/osteoinductive injectable hyaluronan based hydrogel loaded with nano-hydroxyapatite crystals (Hya/HA) with/without bone morphogenetic protein (BMP-2), in distal femur of normal and ovariectomized New Zealand white rabbits. Our results revealed the osteoinductive effect of the Hya/HA composite that enhanced bone density and architecture of the rabbit distal femur.

Keyword
Bone engineering, hyaluronic acid, hydroxyapatite, injectable hydrogels, osteoinduction, osteoporosis
National Category
Medical and Health Sciences Polymer Chemistry
Research subject
Chemistry with specialization in Polymer Chemistry
Identifiers
urn:nbn:se:uu:diva-174215 (URN)10.1080/15421406.2012.635530 (DOI)000302300400019 ()
Conference
11th International Conference on Frontiers of Polymers and Advanced Materials (ICFPAM 2011)
Available from: 2012-05-15 Created: 2012-05-14 Last updated: 2013-02-11Bibliographically approved
6. Minimally invasive mandibular bone augmentation using injectable hydrogels
Open this publication in new window or tab >>Minimally invasive mandibular bone augmentation using injectable hydrogels
Show others...
2012 (English)In: Journal of Tissue Engineering and Regenerative Medicine, ISSN 1932-6254, Vol. 6, no S3, s15-s23 p.Article in journal (Refereed) Published
Abstract [en]

Hyaluronic acid-based hydrogels are proven biocompatible materials and excellent carriers of bone morphogenetic protein-2 (BMP-2) that have been successfully tested for bone generation in vivo. Different formulations, with or without nanohydroxyapatite, have shown promise for craniofacial applications. In this study, 28 rats were used to investigate whether it is possible to achieve mandibular bone augmentation upon injection of novel hyaluronic acid-based hydrogels containing nanohydroxyapatite and different concentrations of BMP-2 (0, 5 and 150ÎŒg/ml). The biomaterials were injected subperiosteally through fine needles into the innate mandibular diastema, imitating a clinical procedure for resorbed mandibles. No incisions, flaps or sutures were necessary. After 8weeks the mandibles were evaluated by peripheral quantitative computed tomography (pQCT), micro-computed tomography (ÎŒCT), histology, immunohistochemistry and fluorochrome labelling. As a result, engineered bone was observed in all treated mandibles, with a statistically significant increase in mandibular bone volume correlated with the amount of BMP-2 loaded in the hydrogel formula. We therefore demonstrated that minimally invasive mandibular bone augmentation is possible upon injection in rats, when using the appropriate injectable scaffolds. This represents an attractive clinical alternative for oral implantology patients.

Keyword
Bone morphogenetic protein-2, Bone tissue engineering, Hyaluronic acid, Hydroxyapatite, Injectable hydrogel, Mandible, Minimally invasive, Rat animal model
National Category
Natural Sciences Polymer Chemistry
Research subject
Chemistry with specialization in Polymer Chemistry
Identifiers
urn:nbn:se:uu:diva-181021 (URN)10.1002/term.1593 (DOI)000313431100003 ()
Note

Correspondence Address: Hilborn, J.; Polymer Chemistry, Department of Chemistry - Ņngström Laboratory, Science for Life Laboratory email: hilborn@mkem.uu.se

Available from: 2012-09-14 Created: 2012-09-14 Last updated: 2013-02-19Bibliographically approved
7. Evaluation of injectable constructs for bone repair with a subperiosteal cranial model in the rat
Open this publication in new window or tab >>Evaluation of injectable constructs for bone repair with a subperiosteal cranial model in the rat
Show others...
2013 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 8, no 8, 140 p.e71683- p.Article in journal (Refereed) Published
Abstract [en]

While testing regenerative medicine strategies, the use of animal models that match the research questions and that are related to clinical translation is crucial. During the initial stage of evaluating new strategies for bone repair, the main goal is to state whether the strategies efficiently induce the formation of new bone tissue at an orthotopic site. Here, we present a subperiosteal model in rat calavria that allow the evaluation of a broad range of approaches including bone augmentation, replacement and regeneration. Easy and fast to perform, the model is minimally invasive and no defect are created. The procedure enables to evaluate the outcomes quantitatively using micro-computed tomography and qualitatively by histology and immunohistochemistry. For establishing the model, we used bone morphogenetic protein-2 as an osteoinductive factor and hyaluronic acid hydrogel as injectable biomaterial. We showed that this subperiosteal cranial model offers a minimally invasive and promising solution for a rapid evaluation of bone tissue engineering strategies, even for investigator with limited experience in orthopedic surgery. We believe that this approach could be a powerful platform for orthopedic research and regenerative medicine.

Publisher
140 p.
Keyword
Subperiosteal rat model, Minimally invasive surgery, Regenerative medicine, Bone repairr matrix, Hyaluronan, Animal model
National Category
Biomaterials Science Orthopedics
Research subject
Orthopaedics; Surgery; Engineering Science; Engineering Science
Identifiers
urn:nbn:se:uu:diva-188001 (URN)10.1371/journal.pone.0071683 (DOI)000323115800075 ()
Available from: 2012-12-13 Created: 2012-12-12 Last updated: 2017-12-06Bibliographically approved

Open Access in DiVA

fulltext(15943 kB)1507 downloads
File information
File name FULLTEXT01.pdfFile size 15943 kBChecksum SHA-512
80a63d94320f607f6ae1dac0abbda77e030855106926458be27f0f55fe93700487e5ea5c77bc0863b8f174ea97d63a3112288ce479fee4467045aa1858067041
Type fulltextMimetype application/pdf
Buy this publication >>

Search in DiVA

By author/editor
Kisiel, Marta
By organisation
Polymer Chemistry
OrthopedicsBiomaterials Science

Search outside of DiVA

GoogleGoogle Scholar
Total: 1507 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

isbn
urn-nbn

Altmetric score

isbn
urn-nbn
Total: 1360 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf