Name: biological compatibility profile on biomaterials for bone regeneration
Uploaded: 2018-11-16T13:00:00
Description: Watch this Scientific Journal Video about Biological Compatibility Profile on Biomaterials for Bone Regeneration at JoVE.com

This research project has received funding from the European Union&#39;s Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 263363.

The procedures were done with BALB/c mice following all guidelines for the Care and Use of Laboratory Animals of the Austrian Ministry of Education, Science and Research and were approved by the Committee on the Ethics of the Austrian Ministry of Education, Science and Research.
1. Primary Mouse OB Culture
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	<li>OB isolation from neonatal mouse calvaria using enzymatic digestion
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		<li>Euthanize 1&#8211;2 day old neonatal pups (60 in total) by decapitation and place...

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Here, we show a multidisciplinary approach for the preclinical assessment of biocompatibility for representative biomaterials developed for bone regeneration and repair. We tested the responses of OBs, OCs, and the in vivo healing response in a critical bone defect model in mice as well as in vitro and in vivo immune responses. We aimed to demonstrate how the assays work and summarize the data and conclusions derived from the examination of the biomaterials. We show that our strategy generates ...

Bone repair is a complex process that begins with hematoma formation, inflammation, callus formation and then remodeling1,2. However, bone regeneration potential is limited to the size of the bone fracture1,3. For instance, large bone fractures caused by trauma, cancer or osteoporosis may not heal and are termed non-union bone fractures. These bone lesions often require treatment to promote healthy physiological bone repair and regeneration. Currently, autograft and allograft bone transplantation is the approach of choice4 with 2.2 million bone replacement procedures annually5. Though these procedures have a high success rate, there may be complications, for example, limited availability of bone, infection, donor site morbidity, and rejection4. New alternatives for bone tissue engineering are being sought to address these challenges.
The design of biomaterials based on natural or synthetic polymers, bioceramics or metals in combination with cells and bioactive molecules is on the rise6. Our current understanding of physiological bone healing and healing in the context of biomaterials depends on multiple factors such as mechanical properties and multiple local and systemic factors including cells from the circulation and fracture site7,8,9. Biomaterials for bone regeneration aim to promote osteogenicity and osseointegration10 and are ideally biocompatible, biodegradable, and porous (promoting cell migration, oxygen, and nutrients). They also need to be sufficiently strong to support the fracture site to relieve pain. Additionally, inflammatory factors are required to initiate the healing process. However, if the biomaterial induces excessive inflammation and allergic responses, this might limit or inhibit bone healing11,12. Thus, an interdisciplinary approach is necessary to evaluate biomaterials developed for bone repair.
In this study, we present a pre-clinical evaluation of representative materials, 1) Orthovita Vitoss foam which is a commercially available cancellous bone graft substitute consisting of tricalcium phosphate composed of nanometer-sized pure &#946;-tricalcium phosphate (&#946;-TCP) particles and Type 1 bovine collagen (C) (&#946;-TCP/C foam) and 2) &#946;-TCP disks. Here, we illustrate biocompatibility testing of these biomaterials using primary osteoblast (OB) and osteoclast (OC) assays, an in vivo model of bone repair, an immunological assessment comprising in vitro T lymphocyte proliferation and cytokine production, and in vivo immunogenicity and allergenicity, as previously reported13.

biological compatibility profile on biomaterials for bone regeneration

Large non-union bone fractures are a significant challenge in orthopedic surgery. Although auto- and allogeneic bone grafts are excellent for healing such lesions, there are potential complications with their use. Thus, material scientists are developing synthetic, biocompatible biomaterials to overcome these problems. In this study, we present a multidisciplinary platform for evaluating biomaterials for bone repair. We combined expertise from bone biology and immunology to develop a platform including in vitro osteoclast (OC) and osteoblast (OB) assays and in vivo mouse models of bone repair, immunogenicity, and allergenicity. We demonstrate how to perform the experiments, summarize the results, and report on biomaterial biocompatibility. In particular, we tested OB viability, differentiation, and mineralization and OC viability and differentiation in the context of &#946;-tricalcium phosphate (&#946;-TCP) disks. We also tested a &#946;-TCP/Collagen (&#946;-TCP/C) foam which is a commercially available material used clinically for bone repair in a critical-sized calvarial bone defect mouse model to determine the effects on the early phase of bone healing. In parallel experiments, we evaluated immune and allergic responses in mice. Our approach generates a biological compatibility profile of a bone biomaterial with a range of parameters necessary for predicting the biocompatibility of biomaterials used for bone healing and repair in patients.

To assess &#946;-TCP for its effectiveness as a biomaterial for bone repair, we used in vitro and in vivo screening methods. Firstly, we measured the OB responses to the &#946;-TCP disks compared with baseline medium alone controls. Figure 2 demonstrates the OB viability in response to &#946;-TCP disks at 7 and 14 days of culture. Cell viability measured from metabolically active cells in the culture wells was the same for OBs with medium in...

Watch this Scientific Journal Video about Biological Compatibility Profile on Biomaterials for Bone Regeneration at JoVE.com

Biological Compatibility Profile on Biomaterials for Bone Regeneration

The number of novel biomaterials engineered for repairing large bone lesions is continuously expanding with the aim to enhance bone healing and overcome the complications associated with bone transplantation. Here, we present a multidisciplinary strategy for pre-clinical biocompatibility testing of biomaterials for bone repair.

Large non-union bone fractures are a significant challenge in orthopedic surgery. Although auto- and allogeneic bone grafts are excellent for healing such ...

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