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Dear Hani
In response to your initial message (see below):
I stored my osteochondral plugs in Ringer's solution at -20°C while I was
conducting research in this area. Before the day of testing I would defrost
them overnight at 0-4°C. I never performed testing on cartilage specimens
that had been kept frozen for longer than 3 months (just an arbitrary length
of time that seemed appropriate). Also you may wish to store each plug
separately. If you store several in one container on defreezing you're
committed to using all of them for testing that day. I wouldn't recommend
refreezing for later use or extended storage at 0-4°C (unless you are using
a specialised storage protocol such as you've already referenced).
Throughout my testing program I always conducted one or two tests using
fresh (never frozen) cartilage specimens just to make sure the results
weren't different to those from the defrozen cartilage specimens. That's
the ultimate test. Once you've set up a storage protocol I'd advise you to
conduct a quick statistical analysis, comparing your defrozen cartilage
specimens to fresh cartilage specimens, according to whatever biomechanical
test you are conducting. If there's no significant difference you've
succeeded. i'd also recommend conducting the occasional fresh cartilage
specimen testing throughout your research just to maintain confidence in
your protocol.
I'd be interested to hear of advice you received from others.
Hamish
---------------------------------------------------
Hamish Forster PhD CEng
Smith+Nephew Orthopaedics
Memphis
901-399-5476
-----Original Message-----
From: Hani Awad [SMTP:awadhi@visto.com]
Sent: Thursday, January 06, 2000 8:52 AM
To: BIOMCH-L@NIC.SURFNET.NL
Subject: Cartilage Storage
Dear All,
I am looking for a protocol/recipe for storing osteochondral
cartilage specimens prior to compressive testing. The storage conditions
should be such that they do not affect the intrinsic material properties
(modulus and permeability) of cartilage.
I am aware of the procedures utilized by Athanasiou's group in which
they used saline solution with protease inhibitors to store specimens at
-80°C [Athanasiou et al, JOR, 12:340-349, 1994; Athanasiou et al, JOR,
9:330-340, 1991]. I am also aware of the study by Kwan et al [J Biomech
Eng, 114: 149-153, 1992] in which they used supplemented Earl MEM media to
store the specimens at 4°C. These studies claim that the methods they
describe did not affect the biomechanical properties of cartilage in
compression.
Is any body aware of more recent studies that challenge that claim
and recommend alternative procedures? Does any body recommend an alternative
protocol that he/she does not mind sharing??
Thanks a lot,
Hani Awad, PhD
Cincinnati, OH
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In response to your initial message (see below):
I stored my osteochondral plugs in Ringer's solution at -20°C while I was
conducting research in this area. Before the day of testing I would defrost
them overnight at 0-4°C. I never performed testing on cartilage specimens
that had been kept frozen for longer than 3 months (just an arbitrary length
of time that seemed appropriate). Also you may wish to store each plug
separately. If you store several in one container on defreezing you're
committed to using all of them for testing that day. I wouldn't recommend
refreezing for later use or extended storage at 0-4°C (unless you are using
a specialised storage protocol such as you've already referenced).
Throughout my testing program I always conducted one or two tests using
fresh (never frozen) cartilage specimens just to make sure the results
weren't different to those from the defrozen cartilage specimens. That's
the ultimate test. Once you've set up a storage protocol I'd advise you to
conduct a quick statistical analysis, comparing your defrozen cartilage
specimens to fresh cartilage specimens, according to whatever biomechanical
test you are conducting. If there's no significant difference you've
succeeded. i'd also recommend conducting the occasional fresh cartilage
specimen testing throughout your research just to maintain confidence in
your protocol.
I'd be interested to hear of advice you received from others.
Hamish
---------------------------------------------------
Hamish Forster PhD CEng
Smith+Nephew Orthopaedics
Memphis
901-399-5476
-----Original Message-----
From: Hani Awad [SMTP:awadhi@visto.com]
Sent: Thursday, January 06, 2000 8:52 AM
To: BIOMCH-L@NIC.SURFNET.NL
Subject: Cartilage Storage
Dear All,
I am looking for a protocol/recipe for storing osteochondral
cartilage specimens prior to compressive testing. The storage conditions
should be such that they do not affect the intrinsic material properties
(modulus and permeability) of cartilage.
I am aware of the procedures utilized by Athanasiou's group in which
they used saline solution with protease inhibitors to store specimens at
-80°C [Athanasiou et al, JOR, 12:340-349, 1994; Athanasiou et al, JOR,
9:330-340, 1991]. I am also aware of the study by Kwan et al [J Biomech
Eng, 114: 149-153, 1992] in which they used supplemented Earl MEM media to
store the specimens at 4°C. These studies claim that the methods they
describe did not affect the biomechanical properties of cartilage in
compression.
Is any body aware of more recent studies that challenge that claim
and recommend alternative procedures? Does any body recommend an alternative
protocol that he/she does not mind sharing??
Thanks a lot,
Hani Awad, PhD
Cincinnati, OH
---------------------------------------------------------------
To unsubscribe send SIGNOFF BIOMCH-L to LISTSERV@nic.surfnet.nl
For information and archives: http://isb.ri.ccf.org/biomch-l
---------------------------------------------------------------