The Effect of Injection Needle Gauge Size on the Viability of Equine Mesenchymal Stem Cells
Aims Performance horses are at high risk of injury to the superficial digital flexor tendon (SDFT). Studies have shown that autologous mesenchymal stem cells (MSCs) injected into SDFT lesions subsequently result in a significant reduction of re‐injury in National Hunt racehorses; however, recent stu...
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| Veröffentlicht in: | Equine veterinary journal 2013-09, Vol.45 (S44), p.4-4 |
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| creator | Cree, S.J. Smith, R.K. Dudhia, J. |
| description | Aims
Performance horses are at high risk of injury to the superficial digital flexor tendon (SDFT). Studies have shown that autologous mesenchymal stem cells (MSCs) injected into SDFT lesions subsequently result in a significant reduction of re‐injury in National Hunt racehorses; however, recent studies show that only 25% of implanted MSCs survive 24 h post injection. The reason for this loss is unclear but may relate to cell quiescence or injection‐related mortality. We hypothesised that cell viability and mortality is increased with needle gauge.
Methods
Equine MSCs cultured in vitro were resuspended to a final suspension density of 5 x 105 cells/ml to mimic that used for implantation in clinic. The cell suspension was injected through a 19 gauge, 21 gauge (current practice in clinic) or 23 gauge needle. Mesenchymal stem cells viability and mortality was analysed over a 24 h period post injection using alamarBlue® and Annexin V (apoptosis) assays, respectively.
Results
There was a 25% reduction in viability (P |
| doi_str_mv | 10.1111/evj.12145_9 |
| format | Article |
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Performance horses are at high risk of injury to the superficial digital flexor tendon (SDFT). Studies have shown that autologous mesenchymal stem cells (MSCs) injected into SDFT lesions subsequently result in a significant reduction of re‐injury in National Hunt racehorses; however, recent studies show that only 25% of implanted MSCs survive 24 h post injection. The reason for this loss is unclear but may relate to cell quiescence or injection‐related mortality. We hypothesised that cell viability and mortality is increased with needle gauge.
Methods
Equine MSCs cultured in vitro were resuspended to a final suspension density of 5 x 105 cells/ml to mimic that used for implantation in clinic. The cell suspension was injected through a 19 gauge, 21 gauge (current practice in clinic) or 23 gauge needle. Mesenchymal stem cells viability and mortality was analysed over a 24 h period post injection using alamarBlue® and Annexin V (apoptosis) assays, respectively.
Results
There was a 25% reduction in viability (P<0.01) and mortality (P<0.01) compared with noninjected MSCs over the 24 h period post injection. All needle gauges also induced a decrease in cell metabolic activity immediately post injection but with recovery by 2 h post injection. Furthermore, 21 gauge and 23 gauge needles increased early apoptotic cells immediately post injection, whereas the 19 gauge needle showed a delayed increase in apoptosis until 2 h post injection.
Conclusions
The delayed apoptosis may correlate with a subpopulation of quiescent cells subsequently becoming apoptotic. The proportion of early and late apoptotic MSCs, while significant, does not account for the total cell loss reported after intra‐lesional injection.
Practical significance
Needle gauge selection has significant implications for the survival of MSC intra‐lesional injections but it may not be the sole factor implicated in post injection cell mortality. We therefore recommend a minimum diameter of 19 gauge should be used for implantation.
Ethical animal research
Stem cell lines were obtained from bone marrow with approval of the Institutional Ethical Committee and under UK Home Office licence. Sources of funding: Funded by the Medical Research Council and The Royal Veterinary College. Competing interests: None.</description><identifier>ISSN: 0425-1644</identifier><identifier>EISSN: 2042-3306</identifier><identifier>DOI: 10.1111/evj.12145_9</identifier><identifier>CODEN: EQVJAI</identifier><language>eng</language><publisher>Fordham: Blackwell Publishing Ltd</publisher><subject>Apoptosis ; Bone marrow ; Mortality ; Stem cells</subject><ispartof>Equine veterinary journal, 2013-09, Vol.45 (S44), p.4-4</ispartof><rights>2013 The Author(s). Equine Veterinary Journal © 2013 EVJ Ltd</rights><rights>2013 EVJ Ltd</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2259-e606d4326cac86cd157508c8157d3f0ada5c4316971c7dd96ea0d36c5d935f7f3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fevj.12145_9$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fevj.12145_9$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,777,781,1412,27905,27906,45555,45556</link.rule.ids></links><search><creatorcontrib>Cree, S.J.</creatorcontrib><creatorcontrib>Smith, R.K.</creatorcontrib><creatorcontrib>Dudhia, J.</creatorcontrib><title>The Effect of Injection Needle Gauge Size on the Viability of Equine Mesenchymal Stem Cells</title><title>Equine veterinary journal</title><addtitle>Equine Vet J</addtitle><description>Aims
Performance horses are at high risk of injury to the superficial digital flexor tendon (SDFT). Studies have shown that autologous mesenchymal stem cells (MSCs) injected into SDFT lesions subsequently result in a significant reduction of re‐injury in National Hunt racehorses; however, recent studies show that only 25% of implanted MSCs survive 24 h post injection. The reason for this loss is unclear but may relate to cell quiescence or injection‐related mortality. We hypothesised that cell viability and mortality is increased with needle gauge.
Methods
Equine MSCs cultured in vitro were resuspended to a final suspension density of 5 x 105 cells/ml to mimic that used for implantation in clinic. The cell suspension was injected through a 19 gauge, 21 gauge (current practice in clinic) or 23 gauge needle. Mesenchymal stem cells viability and mortality was analysed over a 24 h period post injection using alamarBlue® and Annexin V (apoptosis) assays, respectively.
Results
There was a 25% reduction in viability (P<0.01) and mortality (P<0.01) compared with noninjected MSCs over the 24 h period post injection. All needle gauges also induced a decrease in cell metabolic activity immediately post injection but with recovery by 2 h post injection. Furthermore, 21 gauge and 23 gauge needles increased early apoptotic cells immediately post injection, whereas the 19 gauge needle showed a delayed increase in apoptosis until 2 h post injection.
Conclusions
The delayed apoptosis may correlate with a subpopulation of quiescent cells subsequently becoming apoptotic. The proportion of early and late apoptotic MSCs, while significant, does not account for the total cell loss reported after intra‐lesional injection.
Practical significance
Needle gauge selection has significant implications for the survival of MSC intra‐lesional injections but it may not be the sole factor implicated in post injection cell mortality. We therefore recommend a minimum diameter of 19 gauge should be used for implantation.
Ethical animal research
Stem cell lines were obtained from bone marrow with approval of the Institutional Ethical Committee and under UK Home Office licence. Sources of funding: Funded by the Medical Research Council and The Royal Veterinary College. Competing interests: None.</description><subject>Apoptosis</subject><subject>Bone marrow</subject><subject>Mortality</subject><subject>Stem cells</subject><issn>0425-1644</issn><issn>2042-3306</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><recordid>eNp9kEtPwzAQhC0EEuVx4g9Y4ogCfic5olLKoxSJAkXiYBl7Ay5pAnEKlF-Pq1ZwYy8zWn2zKw1Ce5Qc0jhH8DE5pIwKqfM11GFEsIRzotZRJ1qZUCXEJtoKYUII50ywDnq8fQHcKwqwLa4LfF5NovN1hYcArgTcN7NnwCP_DTgu2wjfe_PkS9_OF3zvfeYrwFcQoLIv86kp8aiFKe5CWYYdtFGYMsDuSrfR3WnvtnuWDK77593jQWIZk3kCiignOFPW2ExZR2UqSWazqI4XxDgjreBU5Sm1qXO5AkMcV1a6nMsiLfg22l_efWvq9xmEVk_qWVPFl5oKTjJCM8EidbCkbFOH0ECh3xo_Nc1cU6IX7enYnl61F2m-pD99CfP_UN27v_hLJcuUDy18_aZM86pVylOpx8O-5vxinD1c3ugT_gM1xYBO</recordid><startdate>201309</startdate><enddate>201309</enddate><creator>Cree, S.J.</creator><creator>Smith, R.K.</creator><creator>Dudhia, J.</creator><general>Blackwell Publishing Ltd</general><general>Wiley Subscription Services, Inc</general><scope>BSCLL</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>K9.</scope></search><sort><creationdate>201309</creationdate><title>The Effect of Injection Needle Gauge Size on the Viability of Equine Mesenchymal Stem Cells</title><author>Cree, S.J. ; Smith, R.K. ; Dudhia, J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2259-e606d4326cac86cd157508c8157d3f0ada5c4316971c7dd96ea0d36c5d935f7f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Apoptosis</topic><topic>Bone marrow</topic><topic>Mortality</topic><topic>Stem cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Cree, S.J.</creatorcontrib><creatorcontrib>Smith, R.K.</creatorcontrib><creatorcontrib>Dudhia, J.</creatorcontrib><collection>Istex</collection><collection>CrossRef</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><jtitle>Equine veterinary journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cree, S.J.</au><au>Smith, R.K.</au><au>Dudhia, J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Effect of Injection Needle Gauge Size on the Viability of Equine Mesenchymal Stem Cells</atitle><jtitle>Equine veterinary journal</jtitle><addtitle>Equine Vet J</addtitle><date>2013-09</date><risdate>2013</risdate><volume>45</volume><issue>S44</issue><spage>4</spage><epage>4</epage><pages>4-4</pages><issn>0425-1644</issn><eissn>2042-3306</eissn><coden>EQVJAI</coden><abstract>Aims
Performance horses are at high risk of injury to the superficial digital flexor tendon (SDFT). Studies have shown that autologous mesenchymal stem cells (MSCs) injected into SDFT lesions subsequently result in a significant reduction of re‐injury in National Hunt racehorses; however, recent studies show that only 25% of implanted MSCs survive 24 h post injection. The reason for this loss is unclear but may relate to cell quiescence or injection‐related mortality. We hypothesised that cell viability and mortality is increased with needle gauge.
Methods
Equine MSCs cultured in vitro were resuspended to a final suspension density of 5 x 105 cells/ml to mimic that used for implantation in clinic. The cell suspension was injected through a 19 gauge, 21 gauge (current practice in clinic) or 23 gauge needle. Mesenchymal stem cells viability and mortality was analysed over a 24 h period post injection using alamarBlue® and Annexin V (apoptosis) assays, respectively.
Results
There was a 25% reduction in viability (P<0.01) and mortality (P<0.01) compared with noninjected MSCs over the 24 h period post injection. All needle gauges also induced a decrease in cell metabolic activity immediately post injection but with recovery by 2 h post injection. Furthermore, 21 gauge and 23 gauge needles increased early apoptotic cells immediately post injection, whereas the 19 gauge needle showed a delayed increase in apoptosis until 2 h post injection.
Conclusions
The delayed apoptosis may correlate with a subpopulation of quiescent cells subsequently becoming apoptotic. The proportion of early and late apoptotic MSCs, while significant, does not account for the total cell loss reported after intra‐lesional injection.
Practical significance
Needle gauge selection has significant implications for the survival of MSC intra‐lesional injections but it may not be the sole factor implicated in post injection cell mortality. We therefore recommend a minimum diameter of 19 gauge should be used for implantation.
Ethical animal research
Stem cell lines were obtained from bone marrow with approval of the Institutional Ethical Committee and under UK Home Office licence. Sources of funding: Funded by the Medical Research Council and The Royal Veterinary College. Competing interests: None.</abstract><cop>Fordham</cop><pub>Blackwell Publishing Ltd</pub><doi>10.1111/evj.12145_9</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| source | Wiley Online Library Journals Frontfile Complete |
| subjects | Apoptosis Bone marrow Mortality Stem cells |
| title | The Effect of Injection Needle Gauge Size on the Viability of Equine Mesenchymal Stem Cells |
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