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Strengths and weaknesses Waleed and Sarah
The Study <ul><li>Original idea </li></ul><ul><li>GFP: </li></ul><ul><ul><li>Readily detected </li></ul></ul><ul><ul><li>N...
The rats! <ul><li>Sample size </li></ul><ul><li>Bone chimeric rats: 7 weeks old </li></ul><ul><li>Tendon chimeric rats: 12...
Bone chimeric rats <ul><li>Proportion of GFP-positive cells  </li></ul><ul><li>measured after transplantation. </li></ul><...
Tendon chimeric rats <ul><li>Same tendon used. </li></ul><ul><li>Absence of GFP signals in </li></ul><ul><li>bone marrow a...
Methods <ul><li>Left leg controls, confirmed GFP </li></ul><ul><li>positivity </li></ul><ul><li>Negative control: GFP-posi...
Immunohistochemistry  <ul><li>Monoclonal antibody </li></ul><ul><li>Fixation and quick freezing </li></ul><ul><li>damages ...
Tendon type <ul><li>Only patellar used  </li></ul><ul><li>What about intrasynovial tendons? </li></ul>
Subjective Analysis of Morphology <ul><li>Lack of markers for MCS and fibroblasts </li></ul><ul><li>‘Fibroblast like cells...
Functional Tissue Engineering <ul><li>Managed to create constructs </li></ul><ul><li>that can withstand large forces </li>...
Final thoughts <ul><li>Could there be other cells apart from fibroblasts involved? </li></ul><ul><li>Is it really biphasic...
 
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Strengths and Weakness

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  • Transcript of "Strengths and Weakness"

    1. 1. Strengths and weaknesses Waleed and Sarah
    2. 2. The Study <ul><li>Original idea </li></ul><ul><li>GFP: </li></ul><ul><ul><li>Readily detected </li></ul></ul><ul><ul><li>Not rejected by hosts </li></ul></ul><ul><ul><li>Expression sustained after cell division </li></ul></ul><ul><ul><li>Not expressed in RBC </li></ul></ul><ul><ul><li>Does it interfere with cellular interaction? </li></ul></ul>
    3. 3. The rats! <ul><li>Sample size </li></ul><ul><li>Bone chimeric rats: 7 weeks old </li></ul><ul><li>Tendon chimeric rats: 12 weeks old </li></ul><ul><li>Any difference in healing ability? </li></ul>
    4. 4. Bone chimeric rats <ul><li>Proportion of GFP-positive cells </li></ul><ul><li>measured after transplantation. </li></ul><ul><li>Absence of GFP signal in other tissues </li></ul><ul><li>confirmed using laser SEM. </li></ul><ul><li>Why were 60 rats irradiated? </li></ul><ul><li>Low proportion of GFP-positive cells </li></ul><ul><li>after transplantation, and there was no </li></ul><ul><li>means of assessing host bone marrow </li></ul><ul><li>cells. </li></ul>
    5. 5. Tendon chimeric rats <ul><li>Same tendon used. </li></ul><ul><li>Absence of GFP signals in </li></ul><ul><li>bone marrow and blood confirmed. </li></ul><ul><li>Histology and fluorescence of </li></ul><ul><li>patellar tendons observed to determine </li></ul><ul><li>survival of transplanted tendons. </li></ul><ul><li>Trauma of harvesting tendons from </li></ul><ul><li>GFP rats and adding nylon suture… </li></ul><ul><li>does it interfere with healing? </li></ul><ul><li>Only ≈65% GFP-positivity before injury </li></ul>
    6. 6. Methods <ul><li>Left leg controls, confirmed GFP </li></ul><ul><li>positivity </li></ul><ul><li>Negative control: GFP-positivity </li></ul><ul><li>measured before injury. </li></ul><ul><li>Standardized </li></ul><ul><li>7 days </li></ul><ul><li>Incision 1 wk after transplantation </li></ul><ul><li>in bone chimeric, but after 2 wks </li></ul><ul><li>in tendon chimeric. </li></ul>
    7. 7. Immunohistochemistry <ul><li>Monoclonal antibody </li></ul><ul><li>Fixation and quick freezing </li></ul><ul><li>damages tendon </li></ul><ul><li>Control for BrdU </li></ul>
    8. 8. Tendon type <ul><li>Only patellar used </li></ul><ul><li>What about intrasynovial tendons? </li></ul>
    9. 9. Subjective Analysis of Morphology <ul><li>Lack of markers for MCS and fibroblasts </li></ul><ul><li>‘Fibroblast like cells’-subjective </li></ul>
    10. 10. Functional Tissue Engineering <ul><li>Managed to create constructs </li></ul><ul><li>that can withstand large forces </li></ul><ul><li>Forces measured in vivo using </li></ul><ul><li>transducers. </li></ul><ul><li>In vitro v.s. in vivo environments </li></ul><ul><li>effects on construct? Long-term? </li></ul><ul><li>How accurate are tensile strength measurements? Variables: direction </li></ul><ul><li>of forces, long term, in vivo conditions? </li></ul>
    11. 11. Final thoughts <ul><li>Could there be other cells apart from fibroblasts involved? </li></ul><ul><li>Is it really biphasic? </li></ul><ul><li>Are rat tendons comparable to human? </li></ul>
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