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Protein Enhanced Bone Recovery. By Amanda Walker Working with Ashley Aston Weiner Mentored by Prasad Shastri. Analysis- Primary Objective. Main goal- to optimize protein release from a bone repair biopolymer Goal components minimize burst effect by optimizing porosity

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Protein enhanced bone recovery

Protein Enhanced Bone Recovery

By Amanda Walker

Working with Ashley Aston Weiner

Mentored by Prasad Shastri


Analysis primary objective
Analysis- Primary Objective

  • Main goal- to optimize protein release from a bone repair biopolymer

  • Goal components

    • minimize burst effect by optimizing porosity

    • Obtain a linear release of protein over 3 to 4 weeks (life span of polymer)

  • Factors to vary

    • Porogen material

    • Porogen concentration

    • Size of gelatin

      microspheres

http://www.oplin.org/point/

http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T3D-438BNXR-1&_coverDate=06%2F15%2F2001&_alid=368044133&_rdoc=1&_fmt=&_orig=search&_qd=1&_cdi=4944&_sort=d&view=c&_acct=C000006878&_version=1&_urlVersion=0&_userid=86629&md5=d5be4d79321979caf317181be7c98482


Analysis problem statement
Analysis- Problem Statement

  • The data we have for the porosity effects are not sufficient.

  • Porogens mixed into polymer control protein release.

  • Previous protein release studies done from linear polymers.

http://members.tripod.com/sunfh/pol6.gif


Analysis polymer explanation
Analysis- Polymer explanation

How does it work?

  • Polyanhydride degrades over the course of a month

  • Porogens quickly dissolve creating holes

    • Exposes proteins

    • Proteins are growth factors

  • Bone grows in as material degrades

  • http://www.synergizedsolutions.com/simpsons/pictures/bart/scientist.gif

    http://www.nrc-cnrc.gc.ca/education/images/science/photos/victoria_02.jpg


    Market analysis
    Market Analysis

    • The market

      • Broken bones

        • 3 femur fractures per 10,000 population annually for young and old people

    • Bone disease

      • 50% of women 50+ have fractures related to osteoporosis

  • Spinal fusion

    • 290,000 surgeries in 2003

  • http://www.drrathresearch.org/clinical_studies/condition_bonefracture_print.html

    http://www.paralumun.com/osteoporosisstats.htm

    http://www.aaos.org/wordhtml/research/stats/spinefusion_recent.htm


    Market analysis1
    Market Analysis

    • Competition

      • ETEX corporation

        • Endothermic curing takes 15-20 minutes

        • No proteins

    • NovaBone

      • Molds to shape of injury, but not immediately

  • Benefits over competition

    • Less invasive

    • More comfortable

    • Custom fit

    • Instant curing

    • Enhancement by growth

      factors

    • Crosslinking=material

      strength

  • http://www.etexcorp.com/etex/content/products/index.shtml

    http://www.novabone.com/prod03.htm


    Analysis background
    Analysis- Background

    • Precedent- BCNU release study

      • Wafer placed in hole after glioblastoma removal

      • Release of BCNU kills leftover cancer cells

      • Improves lifespan

    • This device is currently being used in medicine.

    • Several other papers were researched.

    http://www.neuropat.dote.hu/jpeg/tumor/3gliobl1.jpg


    Analysis material components
    Analysis- Material Components

    +

    Porogens and Proteins

    • Monomers

      • MSA

      • MCPH

    • Porogen

      • Salt

      • Gelatin

    • Protein/sugar

      • HRP as test protein

    • Crosslinking initiator

    MCPH

    +

    MSA

    Photoinitiator/

    Blue light

    Crosslinked polymer


    Analysis polymerization
    Analysis- Polymerization

    • Initiator= camphorquinone

    • Forms free radicals when exposed to blue light

      • By hydrogen atom extraction

  • Radicals initiate crosslinking

  • http://www.bonartmed.com/upload/product/1/1096423878014_0.JPG

    http://www3.interscience.wiley.com/cgi-bin/abstract/109792870/ABSTRACT?CRETRY=1&SRETRY=0


    Synthesis of samples block diagram
    Synthesis of Samples- Block Diagram

    Mix protein and sugar

    Coomassie assay

    Granulate protein

    Mix monomers, protein, and porogen

    Make and/or quantify porogen

    Add initiator

    Mold and expose


    Hypothesis performance metrics
    Hypothesis- Performance Metrics

    How will improvement be measured?

    • Polymerized samples will be degraded in 37 degree PBS.

    • PBS will be changed daily.

    • The old PBS will be analyzed using a protein quantification assay.

    • This will determine how much protein was released over the last time period.


    Analysis performance criteria
    Analysis- Performance Criteria

    • Constraints

      • Release=3-4 weeks

      • Avoid burst effect

    • Limitations

      • Time- 4 months

      • Money- no real constraints

    • Exclusions- some porogens

      • FITC-dextran, for example

      • BSA as a protein- too much background in negative controls

    Burst Effect


    Validation current status
    Validation- Current Status

    • Accomplished:

      • Protein uniformity verification

      • Protein quantification

      • Protein and porogen granulation

      • Made the cylindrical samples for the first porogen (NaCl)

  • Current work

    • Had to change the protein used from BSA to horseradish peroxidase (HRP)

      • Due to this we are using a different assay to quantify release in the degradation studies

      • The Coomassie assay was still used for protein uniformity verification

    • Degradation study of the NaCl porogen samples

      • Slightly delayed (about 2 weeks) due to the necessity of changing the protein



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