Document XO8a7OKO0pNBr15eVJE50z8gR
AR226-3061
Study mio
TOP Biocfflnpatibility Testing of S-22072. S-22073, and H-22074
Study Completed on October 4, 1996
Contracting MmrfltffrY
North American Science Auoeiaf, Inc.
2261 Tracy Road Northwood, Ohio 43619-1397
for
Baskell
B. X. du Font de Neooure and Coapany
Laboratory for Toxicology and Industrial Elktoa Road, P.O. Box 50
Newark, Delaware 19714
Medicine
Study Monitor Susan A. MacKenzie, V.M.D., Ph.D., D.A.B.T.
Medical Research Project Ho.
Laboratory Pro-1 act ID Haskell Laboratory Report No. 907-96
Page 1 of 53
i i
Company Sanitized. Does not contain TSCA CBI
Substance Tested; Synonyms/Other Codes:
SBSSSSt-SSSBSSaSSX^. Tegt_J3ubBfcaiae_ftl
DuPont HLO S07-96
aamSA Lab. No.
Phys i cal Portn: Purity:
Composition:
Contaminants; CAS Registry Mo-
96T-12210-00
Blue fabric Not provided by sponsor
oropany Sanitized.
2 -
DuPont HLO 907-96
QgmgBM. niPQBMM-IOM (Continusd) TMfc Subatance 82
Subattmcc Teated: Synonvma/Other Codea:
H-22073
Haasa_rf>b. MQ. gta^Ags3UgsgB:
Purifcy: Congtoaition:
96T-12211-00
Blue fabric
Mot provided by epoaaaz
Contamj^ianta: CAS Reqiatrv Mo.;
gubafeance Tested: Svnonvms/Other Codee:
Teet Substance 3
3
'Sanitized. Does;. .':... ..
-
DUPont HLO 907-96
NAmSA Lah. No.
Phvaical Form: Purity: Composition:
96T-12212-00
Blue-green fabric
Contaminants: CAS Registry No.: Study Initiated-Completed:
08/27/96 - 10/04/96
-
4
-
'^BffiSpanySanitized. Does not contain TSCA CBS
DuPont Central Research and Development
Haskell Laboratory for Toxicology and Industrial Medicine
DuPont HLO 907-96 October 4, 1996
DuPont Mon-Woven Old Hickory, Tennessee
USP Biocompatibnitv Testing of H-22572. H-22073. and H-22074
Attached you will find reports from North American Science Associates, Inc.
The results for the H-22072. H-22073, and H-22074 samples are;
Test Substance H-22072 H-22073 H-22074
Cytotoxicity
(Aaarose Overlay)
Mon-toxic Kon-toxic Kon-toxic
Test Results
USP Muscle
USP Intracutaneous Implantation
SC& Pass
CSpb______(7 day)
Pass
KS"
Pass
Pass
NS
Pass
Pass
N5
a Sodium chloride extract
b Cotton Seed oil extract
c Not significant
^^n^v^^ ^ML^I 0..
1^/11%
Susan A. MacKenzie. V.M.D., Ph.D., D.A.B.T. Research Toxicologist
Company Sanitized. Does not contain TSCA CBI
5 -
World Leader in Teslinn Sm'irt"; for the Medical Device Industry
DuPONT HLO 907-96
2261 Tracy Road Northwood, OH 43619 Phone 419*666*94S5
FAX 419^66.2954
LAB NO. P.O. NO.
96T 12210 00 LGX 592565
E. I. du Font de Nemoun & Co. HASKELL LABORATORY
ELKTON RD. P.O. BOX 50 NEWARK, DE
19714-0050
ATTN: DR. SUSAN A. MACKENZIE
ID NO. H-22072
CYTOTOXICITY - AGAROSE OVERLAY, SOLID
Test Article: H-22072 (control item)
Test Article Description: Sheet - 1 cm2 piece
P-ocedure:
A monolayer of L-929 mouse flbroblast cells wu grown to confluency and
overlaid with Miaicaum Essential Medium supplemented with aenim, antibiotics, neutral red, and 1% agaroae. The test article, a 0.5 cm x 0.5 cm piece of 725871H as a positive control, and a 1.0 cm length piece of UP-1 aa a negative
control were placed on me solidified overlay surface. Following incubation for 24 hours, me culture was microscopically examined for evidence of cell
decolorizadon to determine me zone of cell lysis. Any decolorized zone preaent
was examined microscopically to confirm cell lysis.
Score
Nontoxic (N)
Toxic (D
Observations
Normal cell morphology in proximity to test sample.
Cellular death and degeneration associated with the area beneath the test sample and possibly extended beyond the perimeter of the test sample. Where a zone of lysis was observed, die distance from the edge
of the sample to the edge of me zone was measured
and reported in millimeters (mm).
<gompai%anitized.Does not contain TSCA CBI
^lu
Completed/?-I \ ~^C>
Approved irony Fh*fp8
MG030-110 Page 1 of 2
Authoiizalion (or duplication ol Ihis report except in whole, is reserved pending NAmSA's writtfln approval
N A i n S A World Lvadfr in Tallinn Srrvirvs for Ihe Mrdiral Drnrc Industry
DuPont HLO 907-96
226 ITracy Road Northwood, OH 43619 Phone 419*666*9455 FAX 419^66.2954
LAB NO. 96T 12210 00
CYTOTOXICTTY - AGAROSE OVERLAY, SOLID
Results:
Test/Control Articles
Test Article Negative Control: UP-1 NAmSA Positive Control: 725871H
Scofe
7wff]^mlvia)
Conclusion: Comments:
Results and conclusions cpply only to the tet article toted. No further evaluation of theae results is made by NAmSA. Any extrapolation of tfaeae data to other aamplea is me
respooaibuity of die sponsor.
The above teat article was nontoxic for L-929 mouse fibrobh
test parameter*.
' me above deacribod
Placed me 'bluest' side of fabric down on agarose. All raw data pertaining to this study and a copy of me final report are to be retained i
designated NAfflSA archive files.
Test Article Received: 9-4-96 Date Prepared: 9-10-96
Date Terminated: 9-11-96
ompany Sanitized. Does not contain TSCA CBI
I ^ A la*
Completed.
Approved_
-
7
(See page 1)
Page 2 of 2
I Aulhonzaii> n for duplication ol this report, except in whole, is reserved pending NAmSA's written approval
Confidential TU013-800
DuPont HLO 907-96
Lab No.
96T 12210 00
P.O. No. LGX 592565
STUDY TITLE: USP INTRACUTANEOUS TOXICITY STUDY IN THE RABBIT
(Extracts)
TEST ARTICLE: H-22072 (control item)
IDENTIFICATION NO.:
H-22072
SPONSOR:
DR. SUSAN A. MACKENZIE
E. I. du Font de Nemours & Co.
HASKELL LABORATORY
ELKTON RD. P.O. BOX 50
NEWARK. DE
19714-0050
8-
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W,HL.^.ri.T.,,iHlS.r.ic., far,l,.M.^c.H)..ic.l^.,,r,
2261 Ti*cy (toad
Northoood. Oh 43619
"""
FAX
41M66.84SS <19<6M954
Aufhonzjtion Iw dupautkm of Wi npofl. nfX In hon. l< r*Mrvd pwdlng NAn SA'i iriiun ippimol
TU013-800
DuPont HLO 907-96
Lab No. 96T 12210 00
TABLE OF CONTENTS
P9K V\m^!T
SUMMARY ................................................. 3 INTRODUCTION ............................................... 4
MATERIALS ................................................. 4 METHODS ................................................... 4
RESULTS .................................................... 6 CONCLUSION ................................................ 6
RECORD STORAGE ............................................. 6
TABLE
I - USP INTRACUTANEOUS TOXICITY OBSERVATIONS ..................
7
^mpany Sanded. Does not contain TSCA CBI
9 _
Page 2 of 7
NAinSA ,^_| ^_B_.
i
WorU U^ir l Ttill,, Stnirtt
'w lln MfUcl 0>(t Mitrf
2261 Tracy ROM
Nortturood. OH 4361
'*"
FAX
<lfr666.9455
< 'Ma<M4
AuttiortzjUan tot duplication n mr npon. <upl in wnoh. r fwnd ixndtng NAmSA't irHn ippfovl
TU013-800
DuPont HLO 907-96
Lab No. 96T 12210 00
SUMMARY
The test article, H-22072 (control item), H-22072. was extracted in 0.9% sodium chloride USP solution and cottonseed oil, NF. These extracts were evaluated for intracutaneous toxicuy in accordance with the guidelines of the current USP.
A 0.2 ml dose of the appropriate test article extract was injected by the imracuianeous romc into five separate sites on the right side of the back of each rabbit. Similarly, the corresponding blank vehicle was injected on the
left side of the back of each rabbit. Two rabbits were used for each pair of extracts. Observations for erythema and edema were conducted at 24, 48, and 72 hours after intracuianeous injection.
Under the conditions of this study, there was no evidence of significant irritation or toxicity from the extracts injected intracuianeously into rabbits. Each test article extract met the USP requirements.
Study and Supervisory Personnel:
Donald P. Barda, LAT Karen D. Cobum. ALAT Barbara M. Monroe Tina M. Cox, BS. LAT
Approved by:
l.tUL-JLQMJU.VU.^ Lit-
Timothy Jansen, BLS Section Manager, Testing Services
0,1 QJ^
Date Completed
/las
r^w
1
- 10 -
empany Sanitized. Doas not corrts^ yc^ A ^ Page 3 of 7
N A i n S A WorU U^if in Tiilliit Swim far lln UtUnl Dfficf iHfuiirj
226t Tftcy Red
Nonnwood. OH 43619
^'" 'F"A"X
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TUO 13-800
DuPont HLO 907-96 Lab No. 96T1221000
INTRODUCTION
The lest article identified below was extracted and the extracts were evaluated for bioconpatibility in accordance with the guidelines of the current USP. The purpose ot (he study was to determine whether leachables extracted from the material would cause local dermal irritant or toxic effects following injection into rabbit skin. The test anicle was received on September 4, 1996. The animals were injected on September 10. 1996, and the observations were concluded on September 13, 1996.
MATERIALS
The sample provided by (he sponsor was identified and handled as follows:
Test Anicle:
H-22072 (control item)
Identification No: Storage Conditions: Vehicles:
Preparation:
H-22072
Room temperature
0.9% sodium chloride USP solution (SC) cottonseed oil, NF (CSO)
A ratio of 120 cm':20 ml (surface area of test article to volume of vehicle) was used for each preparation. The test article was extracted in SC and CSO at 121 "C for 1 hour. The extraction vehicles without test article were similarly prepared to serve as control blanks.
Condition of Extracts: Sample Disposition:
SC: CSO:
Teat clear with test article particulates clear with test article particulates
Any remaining sample was discarded.
METHODS
Control clear clear
Test System:
Species: Breed:
Source: Sex:
Body Weight Range: Age: Acclimation Period: Number of Animals: Identification Method:
Rabbit fOrvctolayus cuniculus)
New Zealand White Mohican Valley Rabbiuy No particular gender was prescribed for this test No particular body weight range was prescribed for this test No particular age was prescribed for this test Minimum 2 days Two per pair of extracts
Ear tag or tattoo
Justification of Test System:
The intracutaneous injection test in rabbits is specified in the current USP and has been used historically to evaluate biomaterial extracts.
Husbandry:
Conditions conformed to Standard Operating Procedures which are based on the "Guide for the Care and Use of Laboratory Animals."
Food:
PROLAB* High Fiber Rabbit Diet was provided daily.
Company Sanitized. Does not contain TSCA CBI Page 4 of 7
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2261 Truy Red
Norttwood. OH 43619
Pfton* 419466.94JS
FAX
4taeeBS4
iKUng NAmSA't wrtnn approvil
TU013-800
DuPont HLO 907-96
Lab No. 96T 12210 00
Water: Contaminants: Housing:
Freely available, municipal (Toledo. OH) water was delivered through an automatic
watering system.
Reasonably expected contaminants in feed or water supplies did not have the potential to influence the outcome of this test.
Animals were individually housed in stainless steel suspended cages identified by card indicating the lab number, animal number, test code, sex, and date dosed.
Environmental:
Facility: Personnel: Selection:
The room temperature was nnitored daily. The temperature range for the rabbit was 64-74-F.
The room humidity was monitored daily. The humidity range for the rabbit was 30-70%.
The light cycle was controlled using an automatic timer (12 bows light, 12 hours dark).
NAmSA is an AAALAC accredited facility.
Associates involved were appropriately qualified and trained.
Only healthy, thin-skinned animals free of mechanical irritation or trauma that could
interfere with the test were selected.
Experimental Procedure:
Prior to treatment, each rabbit was clipped free of fur from the back and both sides of the spinal column to yield a sufficient injection area. Two rabbits were prepared per pair of extracts. A 0.2 ml dose of test article extract was injected intracuianeously into five separate sites on me right side of the back of each mbbit; 0.2 ml of the control blank was injected into five separate sites on the corresponding left side of the back. Injections were spaced approximately 2 cm apart. The animals were returned to their respective cages following the
procedure.
Observations for erythema and edema were conducted at 24, 48, and 72 hours after injection. Reactions were scored on a 0 to 4 basis. Any reaction at the injection sites was also noted. The reactions were evaluated according to the following subjective rating scale:
ERYTHEMA (ER)
0 No erythema 1 Very slisia (barely perceptible) 2 Well-defined (pink) 3 Moderate to severe (red) 4 Severe (beet redness) to slight eschar
formation (injuries in depth)
EDEMA (ED)
0 No edema 1 Very slight (barely pere-otible) 2 Slight (edges of area wcii-defirwl by
definite swelling)
3 Moderate (raised approxinuf'/ 1 a. a)
4 Severe (raised more than 1 sasa. may
extend beyond the area of exposure)
The cumulativt; average erythema and edema score for each test extract and corresponding control blenk was calculated. For each extract, a difference in average scores (test minus control blank) of 1.0 or less was
considered to be acceptable. A difference of 0.6 to 1.0 indicated a slight, but acceptable, reaction. A difference > 1.0 was considered to be unacceptable. Additionally, the average score for each test extract and
blank was calculated for each interval. Any adverse reaction noted in the test extract was compared to the
corresponding blank.
12
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2261 Tiacy Rod
NOHhwood. OH 43619
Phone 41948.94SS
FAX
41966<S4
iding NAmSA't wrtltr ipprovl
SiinTSCACBI ' Page 5 of 7
TU013-800
DuPont HLO 907-96
Lab No. 96T 12210 00
RESULTS
Results of scores for individual rabbits appear in Table I. The findings an summarized below:
Extract SC
CSO
Avenge Test (- Avenge Control = Difference == Assessment
0.0
0.0
0.0
Passed
1.5
1.7
0.0
PMMd
The difference in average scores was considered acceptable. The difference between test and control avenge scores did not exceed 1.0 at any observation interval.
Results and conclusions apply only to the test article tested. No further evaluation of these results is nude by NAmSA. Any extrapolation of these data to other samples is the responsibility of the sponsor.
CONCLUSION
Under the conditions of this study, there was no evidence of significant irritation or toxicity from me extracts injected intracutaneously fito rabbits. Each test article extract met the USP requuemenu.
RECORD STORAGE
All raw data pertaining to this study and a copy of the Rnal report are to be retained in designated NAmSA
archive flies.
Company Sanitized. Does not co';>te-i TSCA. C31
13 -
N A m S A WarU Ln4ir In Tnlll Strrlcti for Ikt Mifttlil Dflei liUsilrj
2261 Trcy Road
Nonhwood. OH 436l9
Ption. 4168-9455
FAX
41834
tUMwWWi loi duplcilkm ol ttu npoil. EJOI In itxita. li rnnd pending NAmSA'i rmw pproval
Page 6 of 7
TU013-800
DuPont HLO 907-9ft Lab No. 96T 12210 00 TABLE I
USP 1NTRACUTANEOUS TOX1CITY OBSERVATIONS
RABBIT NUMBER 97235
97209
97205 97209
EXTRACT
SC SC CSO CSO
TEST EXTRACT Scoring buefval (HOUR)
24
48
72
01
ED ER ED
ER ED
0
0
0
0
0
0
0
0
0
0
0
0
2
2
2
1
1
1
2
2
2
1
1
1
ER = Erythema ED = Edema
SC =0.9% sodium chloride USP solution
CSO = cottonrccd oil, NF
CONTROL BLANK Scorinf Interval (Houn)
24
48
72
ER
ED ER ED ER
ED
0
0
0
0
0
0
0
0
0
0
0
0
2
2
2
1
2
1
2
2
2
1
2
1
Offlipany Sanitized, Does not'
14
N A i n K A ICorW Ultfir In TflliKI Scrricti for tin Mftfica/ Dic Miiiir)
2261 Trcy Rwd
Northwood. OH 3619
Phone 41W6fl.94S5
PAX
41948649S4
AuttiaiaiMn tu dialcjikn of ttiH rapCTl. OIBMX m tor. ll f--rvd pfxflng NAmSA'i inmn pprwl
Page 7 of 7
Confidential TU014-807
DuPont HLO 907-96
Lab No.
96T 12210 00
P.O. No. LGX 592565
S-HJDY TITLE: USP MUSCLE IMPLANTATION STUDY IN THE RABBIT
(1 WEEK)
TEST ARTICLE: H-22072 (control itca)
IDENTIFICATION NO..
H-22072
SPONSOR:
DR. SUSAN A. MACKENZIE E. 1. du Pont de Ncmours & Co. HASKELL LABORATORY
ELKTON RD. P.O. BOX 50 NEWARK, DE 19714-0050
, c
_
Company Sanitized. Does n"? ----- Page 1 of 7
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Nonhxooa. OH 0619
FAX
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TU014-807
OuPont HLO 907-96
Lab No. 96T 12210 00
TABLE OF CONTENTS
SUMMARY .................................................. INTRODUCTION ............................................... MATERIALS ................................................. METHODS ................................................... RESULTS .................................................... CONCLUSION ................................................
RECORD STORAGE .............................................
fWfVwffF
3 4 4 4 6 7 7
Company Sanitized. Does not contain TSCA CBI
16 -
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22B1 Tie Rnd
Nonhmod. OH 3er
ph<)" 419-M6S5
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Page 2 of 7
TUO 14-807
DuPont HLO 907-96
Lab No. 96T 12210 00
SUMMARY
"M test article, H-22072 (control item). H-22072, was implanted in muscle tissue of the rabbit. The muscle tissue was evaluated for evidence of irritation or toxicity in accordance with the guidelines of the current USP.
Implant samples and USP negative control strips were loaded into needles and sterilized by steam. Rabbits were implanted and were then euthanatized 1 week later. Muscle tissues were excised and the implant sites were examined macroscopically.
Under the conditions of this study, the macroscopic reaction was not significant as compared to the USP negative control implant material. The implanted test article met the USP requirements.
Study and Supervisory Personnel:
Christina M. Tollcs
Jason A. Karstcn Paul A. Bell. HT(ASCP) Tina M. Cox, BS. LAT
/mm
fV}pAn
Company Sanitized. Does not contain TSCA CB8
- 17 -
NAmSA iU--
^
^^-
WarU Lia^ir ill TfUlHi Smicfi
fw llr MiMnl Diitf iHimltrj
2261 Tricy Roul
Northmad. OH 43619
''"'" MB.94S5
FAX
41 MSS-ZW
AulhorUftton hx durullon ef thu repon. siapt In lul*. ta mM<vd pMKftig NAmSA'i finn tpprovl
Page 3 of 7
TUO 14-807
DuPont HLO 907-96
Lab No. 96T 12210 00
INTRODUCTION
The test article identified below was evaluated for biocompatibility in accordance with the guidelines of the
current USP. The purpose of the study was to evaluate the potential for a local irritant or toxic response to
material implanted in direct contact with muscle tissue. The test article was received on September 4, 1996. The animals were implanted on September 6. 1996. and muscles were explaued on September 13, 1996.
MATERIALS
The sample provided by the sponsor was identified and handled as follows:
Test Article:
H-22072 (control item)
Identification No:
H-22072
Storage Conditions:
Room temperature
Control Article: Preparation:
USP plastic 1 mm diameter negative control reference strips purchased by NAmSA from the offices of the US Phannacopeial Convention and cut into 10 mm sections.
A n""TMTM of six sections of the test article per rabbit, each approximately 1 mm x 10 mm, were loaded into 16 gauge needles. For each rabbit, a minimum of four USP strips were loaded into the same size needles as used for the test article. Test and control articles were sterilized by steam.
Sample Disposition:
Any remaining sample was discarded.
METHODS
Test System:
Species:
Breed: Source: Sex: Body Weight Range: Age: Acclimation Period:
Number of Animals:
Identification Method:
Rabbit fOrvctolaeus cuniculus)
New Zealand White Mohican Valley Rabbitry No particular gender was prescribed for this test 2.7 kg to 2.8 kg at implantation No particular age was prescribed for this test Minimum 2 days
Two
Ear tag or tattoo
Justification of Test System:
The rabbit is suggested as an appropriate animal model for evaluating polymer materials by the current USP testing guidelines. The muscle tissue has been used historically because the response to implanted material is easily graded and compared to a known negative control material.
18
Company Sanitized. Does not con!
K A i n S^&AAfB WorU Lfdir in T*J*( Sirmti /or <*< Mi^lcal Dr( liifutlrj
22*1 Tricy Rod
Nonhoood. OH 43618
Ptioo* 41M66-B45S
FAX
418fr29S4
W Amhortmton ftx auptcnion rt mt upon. xc*(X In atlote, a r--nnd p*n*ig HfvnSA'i ortltn pprmgl.
TU014-807
DuPont HLO 907-96
Lab No. 96T 12210 00
Ar Husbandry:
Food: Water: Contaminants: Housing: Environmental:
Facility: Personnel:
Conditions conformed to Standard Operating Procedures which are based on the "Guide for the Care and Use of Laboratory Animals.*
PROLAB High Fiber Rabbit Diet was provided daily.
' Freely available, municipal (Toledo, OH) water was delivered through an automatic
watering system.
Reasonably expected contaminants in feed or water supplies did not have the potential to influence the outcome of this test.
Animals were individually housed in stainless steel suspended cages identified by a card indicating the lab number, animal number, test code, sex, and date implanted.
The room temperature was monitored daily. The temperature range for the rabbit was 64-74-F.
The room humidity was monitored daily. The humidity range for the rabbit was 30-70%.
The light cycle was controlled using an automatic timer (12 hours light, 12 noun daifc).
NAmSA is an AAALAC accredited facility.
Associates involved were appropriately qualified and trained.
Selection:
Healthy animals were selected. To reduce the number of animals used for testing, and to comply with the directives of the NAmSA IACUC, rabbits on this study may have been used previously in an unrelated test model. Any previously evaluated test or control articles did not cause a response in the animals. Complete history of animal usage is traceable in laboratory records. Animals used for previous evaluations are identified in
the report.
Experimental Procedure:
The fur was clipped from the area over the paravcrtcbral muscles and the rabbits were sedated with acepromazine maleate (0.2 ml/kg dose). After the muscle area was wiped clean with 70% alcohol, each paravenebral muscle was infiltrated with 1% lidocaine to achieve local anesthesia on both sides of the vertebral column.
Needles containing test article were inserted into Ac muscle at a 45 angle. A stylet was placed in the needle and the need!; was withdrawn, leaving the sample in the paravenebral muscle. Six test article sections were implanted in the right paravenebral muscle of each rabbit. In the opposite muscle, four USP negative control strips were similarly implanted.
Rabbits were returned to their respective cages following the procedure. Rabbits were observed daily for general health. Body weights were recorded prior to implantation and at termination.
19
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0empany saniti2ed Does not extern TSCA
2261 Tricy Road
Nonhwood. OH 436)9
Phoiw 41W66.M5S
FAX
4194664B64
idmg NAmSA'i illtMi appiovl
Page 5 of 7
TU014-807
DuPont HLO 907-96 Lab No. 96T1221000
At 1 week, the rabbits were weighed and euthanatized by an intravenous injection of a sodium pentobarbital based drug. The paravenebral muscles were dissected free and methodically cut to locate four of the test article sites and two of the USP negative control sites in each rabbit. Capsule formation or other evidence of irritation was scored using appropriate magnification aid an auxiliary 1'ght source. Scores were assigned and recorded as
follows:
0 No capsule, no adverse reaction (other than minimal hemorrhage) 1 Up to 0.5 mm capsule or reaction area 2 0.6 to 1.0 mm capsule or reaction area
3 I.I to 2.0 mm capsule or reaction area
4 > 2.0 mm capsule or reaction area
Average macroscopic scores for test implants were compared to average scores of the comrol sites. Calculations were rounded off to the nearest 0.1. A Reaction Index difference of 0.0 to 0.5 in scores (test
minus control) was regarded as "not significant,' 0.6 to 1.0 as a "trace,* I.I to 2.0 as "slight.* 2.1 to 3.0 as
"moderate* and S3.1 as "marked.* The requirements of the USP test were met if the difference between test
and control score averages (macroscopic) was not greater man 1.0. The requirements of the test were not met
if the difference between the test and control score for two (or more) implant sites exceeded 1 for any animal
implanted.
RESULTS
Clinical Observations: All animals appeared clinically normal throughout the duration of the study. Body weight data for individual rabbits were considered acceptable.
Macroscopic Observations: The findings for the macroscopic evaluation are shown below.
Weight (kg)
Rabbit
Number
DayO
Day 7
Site
1
96648*
2.7
2.8
2
3
4
1
96762*
2.8
2.8
2
3
4
Test
0 0 0 0
0 0 0 0
USP Negative Control
0 0
0 0
Average:
0.0
0.0
Previous use history traceable in laboratory records.
Macroscopically, there was no difference between the reactions of the test and control materials. This resulted in a macroscopic reaction of "not significant* tissue contact irritation.
Results and conclusions apply only to the test article tested. No further evaluation of these results is made by NAmSA. Any extrapolation of these data to other samples is the responsibility of the sponsor.
gSeinpany Sanitized. Does not contain TSCA CB1
KAmSA
-20-
WorH Lsadir In Torinf Simicit far Ihi Miticsl Dtfici Ifdutlrj
2261 Trcy Pud
Nonhwood. OH 43619
Phon* 410^66-9455
FAX
4iafi6<9f4
Aulhanulloi lor duplication ct ihu npot. xcx m whod, u reurved pending NAmSA'a MWn ippimil.
Page 6 of 7
FE
HED
OaTnHEaR LTT LDUERTE CE leeee Ee R
ky:
SEnotemmiieTSyr aio s RE s a NE
hd. Ge mE hades fol
i
ame
TREE Le
ifiol
jd
SCldidea llm See
LH
GE es cRA Ral
2
a
;
P Cha ees se . Se
TE
ah ei Bend a
SSEeraTh leE d
.
iis
onli of
Lp
=r :
SRR
y
sg deg
ciety Tg
!
}
3
Feit
Pipes od
:
`Company Santized.Doss not conta TSCA Gat 1
Cn
el Me]
:
mt
rdngnsw
LAB NO. P.O. NO.
96T 12211 00 LOX 592565
E. I. du loot de Nemoun & Co.
HASKELL LABORATORY
ELKTON RD. P.O. BOX 50
NEWARK, DE
19714-0050
ATTN: DR. SUSAN A. MACKENZIE
ID NO. H-22073
CYTOTOXICITY - AGAROSE OVERLAY, SOUD
Ten Article: H-22073 (teat item)
Procedure:
Test Article Description; Sheet -1 cm? piece
A mooolayer of Lr-929 mouse fibroUut cell* w grown to confluency ad overlaid with Mimmmn E--eatiil Medinm ipplemcoted with aenm, "^^j^^y. oeubu red, and 2ft (uow. Hie teat article, a 0.5 cm x 0.5 cm piece of 725871H -- poaidve control, iad a 1.0 cm length piece of UP-1 a Mftfhw cootrol were placed cm die Mlidifiedaveflaynnftce. Following ini-abtioa for 24 hcMn, the culture waa maciXMcopicaOyexumaed for evideoce of ceO decolorizatiantodetermiDetbezDoeofcellly--. Aay decolorized zaae preMBt
was nr-^n'M imcnMcopically to coafum cell lyu.
Score
Nontoxic (N) Toxic (T)
OlMervattooa
Nonnal cell morpholofy in proximity to tea --aple.
Cellular death and degeaentiaa anociated wim the area beaeath the tert mnple aad poaubly extended beyoodtheperimeter of dieted ample. Where B zone of lyais was obaerved, die dutance from the edge of the (ample to the edge of me aone wa< mcaured and reported m rnilliTnetcr* (mm).
^ n-l^-^L lu
Completed
Approved.
- 22 -
MG030-110
Page 1 of 2
Aulhofi/ation lor duplication ol this report, except in whole, is reserved pending NAmSA's written approval.
N A i n X A World Lradfr in Testing Srrvimhr Iht Mtdifal Devire IndliiSrv
DuPont HLO 907-96
2261 Tracy Road Northwood. OH 43619 Phone 419*666*9455 FAX 419*666*29S4
LAB NO. 96T 1221100
CYTOTOXICTTY - AGAROSE OVERLAY, SOLID
Rexults:
ZSBE^fJLlCfiLdDIB)
Tot Article
N
Negative Control: UP-1
N
NAmSA Poaitive Control: 725871H
T
Coaclunoa:
Renlta aad caacluaioo* apply ooly to die teat article (cited. Nofiirtha'entutfioaoftfaMe remlte u mile by NAmSA. AnyextnpotoioaofaMe&tetoodiBrnaDpteuthe iwpoaMMBty (n uie ipomor.
The above tort article wax noBtojdc for L-929 i
i fibmUnt celb under die above deicribed
Placed "bluef ode of bbric down 00 faroee. All raw data r**8""'f to ddi tudy and s copy of die filial report are to be i
deagnated NAmSA archive file*.
Tot Article Received: 9-4-96 Dte Prepared: 9-1046
Date Terminated: 9-11-96
eompany Sanitized. Does not contain TSCA CBI
l
Completed.
mrviQ
Approved. - 23 -
(Seepage I)
Page 2 of 2
[ Authorization for duplication ol this report, except in whole, is reserved pending NAmSA's written approval
Confidential TU013-800
DuPont HLO 907-96
Lib No.
96T 12211 00
P.O. No. LGX 592565
SrUPYTTTLE: USP INTRACUTANEOUS TOXICITY STUDY IN THE RABBIT
(Extracts)
TEST ARTICLE: H-22073 (test item)
ffiENTRWA-RONNSL:
H-22073
pesapasy Sanitized. Does not contain TSCA CBI
SPONSOR:
OR. SUSAN A. MACKENZIE E. I. du Font de Nemoura ft Co. HASKELL LABORATORY
ELKTON RD. P.O. BOX 50
NEWARK. DE
19714-0050
24 -
MmSA t irUU^rirntiiSmkn /r ( M*nJ Dnfa* l^mlry
2201 Trcy Rout
NOfOHKWd. OK 43618
^^j*"^ phon*
FAX
41B4M4BM
Autuxtullon tor *iptai d (hte npon. eoapl In hoi*, u i----vd pmfling NAnSA't iitMn (ipro--1
Page I of 7
TU013-800
DuPont HLO 907-96
Lab No. 96T 1221100
TABLE OF CONTENTS
Page Nun^ber
SUMMARY .................................................. 3 INTRODUCTION ............................................... 4
MATERIALS ................................................. 4 METHODS ................................................... 4 RESULTS .................................................... 6 CONCLUSION ................................................ 6
RECORD STORAGE ............................................. 6
TABLE
I - USP INTRACUTANEOUS TOXICITY OBSERVATIONS ..................
7
(Company Sanitized. Does not contain TSCA CBI
- 25 -
iVAinSA W,rULM^r^T.ltlvS,mc; 'fr lln UlMtit Dfif llUftUj
2261TrcyRoa
"wow* Nontloood. OH 43818
P;"* <;.MBW45i f--
AuftOrilMon for dilcallBn ol Ihll (WO^ W M irtnb. ! mwn^ pe(ng NAfflSA'l iiiin Wptonl.
Page2of7
TU013-800
DuPont HLO 907-96
Lab No.
96T 12211 00
WMMArUr
The test aniclc. H-22073 (test item), H-22073. was extracted in 0.9% sodium chloride USP solution and cottonseed oil. NF. These extracts were evaluated for imracutaneoua toxicity in accordance with the guidelines of the current USP.
A 0.2 ml dose of the appropriate test article extract was injected by the intracutaneous route into five separate sites on the right side of the back of each rabbit. Similariy, the corresponding blank vehicle was infected on the left side of the back of each rabbit. Two rabbits were used for each pair of extracts. Observations for erythema and edema were conducted at 24, 48, and 72 hours after imracutaneous injection.
Under the conditions of this study, there was no evidence of significant irritation or toxicity from the extracts injected intracuianeously into rabbits. Each test article extract met the USP requirements.
Study and Supervisory Personnel:
Donald P. Barda, LAT Karen D. Coburo. ALAT Barbara M. Monroe Tina M. Cox. BS. LAT
Approved by:
LHoJ^ sk^^J^ Ltyi.
Timothy Jansen, BLS Section Manager, Testing Services
c^(Q.-Q(o
Date Completed
/las
mm
- 26 -
gtorapany Sanitized. Doss not contain TSCA CBi Page 3 of 7
N A m S . K WrU Uaftr In Tattly Ssrvicci /r (* fff 0r<c lufsitrj
2281 1Ycy Ra*d
Nofthwood. OH 43619
"w
rAX
41frM5f 'frBs<
eutwtWm fw dupteltlon of BU* npon. wcw m txX*. h r--rvd pwng NAmSA'* inun ppimal
TU013-TOO
DuPont HLO 907-96 Lib No. 96T 1221100
INTRQPVCTON
The test aiticle identified below was extracted and the extracts were evaluated for bioconpxdbility in accordance with the guidelines of the current USP. The purpose of the study was to determine whether leactubles extracted from the material would cause local dermal irritant or toxic effects following injection into rabbit skin. The test article was received on September 4. 1996. The animals were injected on September 10,1996. and the observations were concluded on September 13, 1996.
MATERIALS
The sample provided by the sponsor was identified and handled as follows:
Test Article:
H-22073 (test item)
Identification No:
H-22073
Storage Conditions:
Room temperature
Vehicles: Preparation:
0.9% sodium chloride USP solution (SC) cottonseed oil, NF (CSO)
A ratio of 120 cm1^ ml (surface area of leal article to volume of vehicle)
was used for each preparation. The test article was extracted in SC and CSO at 12PC for 1 hour. The extraction vehicles without test article were similarly prepaied to serve as control blanks.
Condition of Extracts:
Sample Disposition:
SC: CSO:
dear with test article paniculates dear with test article paniculate*
Any remaining sample was discarded.
METHODS
dear dear
Test System:
Species: B*r-e_ e- d-:-
Source: Sex:
Bo&y Weight Range: Age: Acclimation Period: Number of Animals: Identification Method:
Rabbit fOrvctolams cuniculus^ New Zealand White Mohican Valley Rabbioy No particular gender was prescribed for this test No particular body weight range was prescribed for this test No particular age was prescribed for this test Minimum 2 deys Two per pair of extracts
Ear tag or tattoo
Justification of Test System:
The iniracutancous injection test in rabbits is specified in the current USP and has been used historically to evaluate biomaterial extracts.
Animal Management:
Husbandry:
Conditions conformed to Standard Operating Procedures which are based on the "Guide for the Care and Use of Laboratory Animals."
Food:
PROLAB* High Fiber Rabbit Diet was provided daily. Company Sanitized. Does not contain TSCA CBI
- 27 -
N A m X A W,rU Ushr ta Hlllf SinUft fw tin Mfnl Dntti liutsilry
aeiTrcyBod Nonhwooa. OH 3619
Ption* 4<SfrB4SS
FAX
4(ftftW4
AuftortaHai lar ljufMrsW d IN* npart. sasw In wtau. h nwvM CKSng NAmSA't <tMn ppnw.
Page 4 of 7
TU013-800
DuPont HLO 907-96
Lab No. 96T 12211 00
Water: Contaminants:
Freely available, municipal (Toledo, OH) water was delivered through an automatic
watering system.
Reasonably expected contaminantf in feed or water suppliesdid not have the potential to influence the outcome of this test.
Housing:
Animals were individually housed in stainless steel suspended cages identified by a card indicating the lab number, animal number, test code, sex, and date dosed.
Environmental:
The room temperature was monitored daily. The temperature range for the rabbit was 64-74-F.
The room humidity was monitored daily. The humidity range for the rabbit was 30-70%.
The light cycle was controlled using an automatic timer (12 hours light, 12 hours dark).
Facility:
NAmSA is an AAALAC accredited facility.
Personnel:
Associates involved were appropriately qualified and trained.
Selection:
Only healthy, thin-sldnned animals free of mechanical irritation or trauma that could interfere with the test were selected.
Experimental Procedure:
Prior to treatment, each rabbit was clipped free of fur from me back and bom sides of the pinal column to yield a sufficient injection area. Two rabbits were prepared per pair of extracts. A 0.2 ml k. ose of test aiticle extract was injected intracutaneously into five separate sites on the right side of the back of each rabbit; 0.2 ml of the control blank was injected into five separate sites on the corresponding left side of the back. Injections were spaced approximately 2 cm apart. The animals were returned to their respective cages following the
procedure.
Observations for erythema and edema were conducted at 24,48, and 72 hours after injection. Reactions were
scored on a 0 to 4 basis. Any reaction at the injection sites was also noted. The reactions were evaluated according 10 the following subjective rating scale:
ERYTHEMA (ER)
0 No erythema 1 Very slight (barely perceptible) 2 Well-defined (pink) 3 Moderate to severe (red) 4 Severe (beet redness) to slight eschar
formation (injuries in depth)
EDEMA (ED)
0 No edema 1 Very slight (barely perceptible) 2 Slight (edges of area well-defined by
definite swelling) 3 MoJeratc (raised approximately 1 mm) 4 Severe (raised more than 1 mm, may
extend beyond the area of exposure)
The cumulative average erythema am", edema score for each test extract and corresponding control blank was calculated. For each extract, a difference in average scores (test minus control blank) of 1.0 or less was considered to be acceptable. A difference of 0.6 to 1.0 indicated a slight, but acceptable, reaction. A difference > 1.0 was considered to be unacceptable. Additionally, the average score for each test extract and blank was calculated for each interval. Any adverse reaction noted in the test extract was compared to the
corresponding blank.
company Sanitized. Does not contain TSCA CBI
- 28 -
K A i n S A WarU Lfadir in Tillinf Simlctt for Ihi M^^lcai Dirici Iwlutlrj
2261 Tttcy Roul
NoKhMOOd, OH 43618
Phona <I<66-9<55
FAX
41M6ft^9M
AuOiortzjUon la ituptoiiKxi ai o" npox, >c*pl m who)*, it rcurvad pending N;nSA' wmwi (ppmii.
Page 5 of 7
TU013-800
DuPont HLO 907-96
Lab No. 96T 12211 00
RESULTS
Results of scores for individual rabbits appear in Table I. The findings are summarized below:
Extract SC
CSO
Average Test (- i Average Control = Difference =- Assessment
0.0
0.0
0.0
Passed
1.7
1.8
0.0
Passed
The difference in average scores was considered acceptable. The difference between test and control average scores did not exceed 1.0 at any observation interval.
Results and conclusions apply only to the test article tested. No further evaluation of these results is made by NAmSA. Any extrapolation of these data to other samples is the responsibility of me sponsor.
CONCLUSION
Under the conditions of this study, there was no evidence of significant irritation or toxicry from the extracts injected intracutancously into rabbits. Each test article extract met the USP requirements.
RECQRD STORAGE
All raw data pertaining to this study and a copy of the final report are to be retained in designated NAmSA
archive files.
Company Sanitized. Does not contain TSCA CBI
- 29
WorM Lnitr in TtHist Sintcii frr ihi Miiteat Df'lci lis^vilij
2261 Tracy RotD
Nonnwood. OH 43619
Phorx 41M66-845&
FAX
4194G6.29S4
Aulhortutnn lor duptcJIIon 1 thr rport. *xcjp< In otiola, a (MMVd puvunf NAmSA's wrun p(rol
Page 6 of 7
TUO 13-800
DuPont HLO 907-96
Lab No. 96T 1221100 TABLE I USP 1NTRACUTANEOUS TOXICITY OBSERVATIONS
TEST EXTRACT Scorinf Interval (Hour)
RABBIT NUMBER
97210 97/14 97210 97214
EXTRACT
sc sc cso cso
24
ER
ED
0
0
0
0
2
2
2
2
48
ER ED
0
0
0
0
2
2
2
2
.,,.
ER ED
0
0
0
0
1
1
1
1
ER = Erythema ED = Edema
SC = 0.9% sodium chloride USP solution CSO = cottonseed oil, NF
CONTROL BLANK Scoring Inieivil (Houn)
24
48
72
ER
ED ER ED ER ED
0
0
0
0
0
0
0
0
0
0
0
0
2
2
2
2
1
1
2
2
2
2
1
2
^"otcoma.-nTSCACB,
30
K A i n S A WorU Usfit in TflUni Swiecl far Ihi Mifical Dtfin St^sstrj
2261 Tf>cy Ro.d
Nonhwood. OH 43619
Phone 41466-84S5
FAX
41a649S4
Authoflulloi lot du(*c*!ton ol mil nwn. oxpi in ohor. u nwmd p*r>dig NAmSA'B rrnn appwal.
Page 7 of 7
Confidential TU014-807
DuPont HLO 907-96
Lab No.
96T 1221100
P.O. No. LGX 592565
STUDY TITLE: USP MUSCLE IMPLANTATION STUDY IN THE RABBIT
(1WEEK) TEST ABHCLE:
H 22073 (tett item)
IDENTIFICATION NO.:
H-22073
SPONSOR:
DR. SUSAN A. MACKENZIE
E. I. du Pom de Nemours & Co.
HASKELL LABORATORY
ELKTON Rl NEWARK. DE
l^TrS^Bo"11"-oes noi contain TSCA CBl
- 31 -
,
Page 1 of 7
K A m S j i ;WerU Lfa^ir in Tiiliiii Smien fft tin MiMcsl 0<ri MMIJ
2261 Tfcy ROW Northood. OH 43619
'>'n <'M55
rAX *' *"frS4
Authertutlon lor diwIutKin el thr npon. M|X In wtioM. k r----nd funOn) NAmSA't rffiw pprovil
TU014-807
DuPont HLO 907-96
Lab No. 96T 1221100
TABLE OF CONTENTS
SUMMARY .................................................. INTRODUCTION ...............................................
MATERIALS ................................................. METHODS ................................................... RESULTS .................................................... CONCLUSION ................................................
RECORD STORAGE .............................................
^B* ^Irfn
3 4 4 4 6 7 7
- 32 -
CiUOpwy Ssmtized. Does notcontaimTSCA CB1 Page 2 of 7
NAinSA! U---- ^
H'orU^aJrrfiirIfJfl(SfrJ far tin Mfca< 0<c irfr|>
I
2261 Trcy Hold
Nonnwood. OM 43619
"x1"*
rAX
'9<86-9455 '9<86<!9S4
Aulhonubal tor duBlcflon 01 thi* no. xcpl In whol*. r fmvd (Mntfng NAmSA'* wntti PCWI.
TU014-807
SUMMARY
DuPont HLO 907-96
Lab No. 96T 12:11 00
The test article, H-22073 (test item), H-22073. was implanted in muscle tissue of the rabbit. The muscle tissue was evaluated for evidence of irritation or toxicity in accordance with the guidelines of the current USP.
Implant samples and USP negative control strips were loaded into needles and sterilized toysteam. Rabbits were implanted and weie then euthanatized 1 week later. Muscle tissues were excised and the implant sites were examined macroscopically.
Under the conditions of this study, the macroscopic reaction was not significant as compared a the USP negative control implant material. The implanted test article met the USP requirements.
Study and Supervisory Personnel:
Christina M. Tollcs
Jason A. Karsten Paul A. BcU. HT(ASCP) Tina M. Cox. BS, LAT
/mm
fA^^j
I
- 33 -
@eenpany Sanitized. Does not contain TSCACBl Page 3 of 7
W,rU L.a^r in T.,,1,1 S,rric,,
for (* Mftflcaf Dnif lnSinlry
2261 Tctcy Rwa
Nwthwood. OH 3S19
phBr" 4'M869455
----
'68wg54
Auttwtnmn lor duplication of ihr neorl. iapl In luM. H r--rwa pending NAmSA'i wrtTUn pofwtl.
TU014-807
DuPont HLO 907-96
Lab No. 96T 1221100
INTRODUCTION
The test article identified below was evaluated for biocompatibility in accordance with the guidelines of die
current USP. The purpose of the study was to evaluate the potential for a local irritant or toxic response to
material implanted in direct contact with muscle tissue. The test article was received on September 4, 1996. The animals were implanted on September 6, 1996. and muscles were explained on September 13, 1996.
MATERIALS
The sar-ple provided by the sponsor was identified and handled as follows:
Test Article:
H-22073 (test item)
Identification No:
H-22073
Storage Conditions: Control Article:
Preparation:
Room temperature
USP plastic 1 mm diameter negative control reference strips purchased by NAmSA from the offices of the US Pharmacopeia! Convention and cut into 10 mm sections.
A minimum of six sections of the test article per rabbit, each approximately
1 mm x 10 mm, were loaded into 16 gauge needles. For each rabbit, a BMW""" of four USP strips were loaded into the same sue needles as used for the test article. Test and control articles were sterilized by steam.
Sample Disposition:
Any remaining sample was discarded.
METHODS
Test System:
Species:
Breed: Source: Sex: Body Weight Range: Age: Acclimation Period:
Number of Animals: Identification Method:
Rabbit fOivctolaeus cuniculus) New Zealand White Mohican Valley Rabbitry No particular gender was prescribed for this test 2.8 kg at implantation No particular age was prescribed for this test Minimum 2 days
Two
Ear tag or tattoo
Justification of Teal System:
The rabbit is suggested as an appropriate animal model for evaluating polymer materials by the current USP testing guidelines. The muscle tissue has been used historically because the response to implanted material is easily graded and compared to a known negative control material.
^^.o^-----"^'8'
34
N A i n S A WorU Liaiir is Tiitift Services fst tin MUltsl Drtci la^mitty
2261 Tricy Road
Northwood. OH 43610
Phons 419^66.9455
FAX
41>66.;954
Aulhoittabon lor dwkauon d Wv npvH. xc*ol in (rfiote. u nwncd pending NAmSA't fttgn tpproval
Page 4 of 7
TUO 14-807
DuP-nt HLO 907-96
Lab No. 96T 1221100
Animal Management:
Husbandry:
Conditions conformed to Standard Operating Procedures which are based on the "Guide for the Care and Use of Laboratory Animals."
Food:
PROLAB* High Fiber Rabbit Diet was provided daily.
Water:
Freely available, municipal (Toledo, OH) water was delivered through an automatic
watering system.
Contaminants:
Reasonably expected contaminants in feed or water supplies did not have the potential to influence the outcome of this test.
Housing:
Animals were individually housed in stainless steel suspended cages identified by a card indicating the lab number, animal number, test code, sex, and date implanted.
Environmental:
The room temperature was monitored daily. The temperature range for the rabbit was
64-74T.
The room humidity was monitorjd daily. The humidity range for the rabbit was 30-70%.
The light cycle was controlled using an automatic timer (12 hours light, 12 hours dark).
Facility:
NAmSA is an AAALAC accredited facility.
Personnel:
Associates involved were appropriately qualified and trained.
Selection:
Healthy animals were selected. To reduce the number of animals used for testing, and to comply with the directives of the NAmSA IACUL, rabbits on this study may have been used previously in an unrelated test model. Any previously evaluated test or control articles did not cause a response in the animals. Complete history of animal usage is
traceable in laboratory records. Animals used for previous evaluations are identified in
the report.
Experimental Procedure:
The fur was clipped from the area over the paravertebral muscles and the rabbits were sedated with acepromazine maleate (0.2 ml/kg dose). After the muscle area was wiped clean with 70% alcohol, each paravenehral muscle was infiltrated with 1% lidocaine to achieve local anesthesia on both sides of the vertebral
column.
Needles containing test article were inserted into the muscle at a 45 angle. A stylet was pliced in (he needle and the needle was withdrawn, leaving the sample in the paravertebral muscle. Six test article sections were implanted in the right paravenebral muscle of each rabbit. In the opposite muscle, four USP negative control strips were similarly impli-ited.
Rabbits were returned 10 their respective cages' following the procedure. Rabbits were observed daily for general health. Body weights were recorded prior to implantation and at termination.
LcontaiiiTSCACBI
35
BBAinKA World Leader in Tiilint Sirvieci for Iht Medical Diriec litdallr)
2261 Trcy RMB
Nonhwooa. OH 43619
Phone 41M6&945S
FAX
41M66-29M
AuttKXIUItai lor duplicttion ol ttila r*poi1, c*p( " whole, 13 naaivad ending NAmSA't tfflnn amifovil
Page 5 of 7
TU014-807
DuPont HLO 907-96 Mb No. 96T 1221100
At 1 week, the rabbits were weighed and euthanatized by an intravenous injection of a sodium pentobarbital based drug. The paravenebral muscles were dissected free and methodically cut to locate four of the test article sites and two of the USP negative control sites in each rabbit. Capsule formation or other evidence of irritation was scored using appropriate magnification and an auxiliary light source. Scores were assigned and recorded as
follows:
0 No capsule, no adverse reaction (other than minimal hemorrhage) 1 Up to 0.5 mm capsule or reaction area 2 0.6 to 1.0 mm capsule or reaction area 3 1.1 to 2.0 mm capsule or reaction area 4 > 2.0 mm capsule or reaction area
Average macroscopic scores for test implants were compared to average scores of the control sites. Calculations were rounded off to the nearest 0.1. A Reaction Index difference of 0.0 to 0.5 in scores (test minus control) was regarded as "not signify an," 0.6 to 1.0 as a "trace/ 1.1 to 2.0 as "slight," 2.1 to 3.0 as "moderate" and 2:3.1 as "marked." The requirements of the USP test were met if (he difference between test
and control score averages (macroscopic) was not greater than 1.0. The requirements of die test were not met if the difference between the test and control score for two (or more) implant sites exceeded 1 for any animal
implanted.
RESULTS
Clinical Observations: All animals appeared clinically normal throughout the duration of the study. Body weight data for individual rabbits were considered acceptable. Macroscopic Observations: The findings for the macroscopic evaluation are shown below.
Weight (kg)
Rabbit
Number
DayO
Day 7
Site
1
96487*
2.8
2.8
2
3
4
1
96476*
2.8
2.7
2
3
4
Test
0 0 0 0
0 0 0 0
USP Negative Control
0 0
0 0
Average:
0.0
0.0
Previous use history traceable in laboratory records.
Macroscopically, there was no difference between the reactions of the test and control materials. This resulted in a macroscopic reaction of "not significant" tissue contact irritation.
Results and conclusions apply only to the test article tested. No funher evaluation of these results is made by NAmSA. Any extrapolation of these data to other samples is the responsibility of the sponsor.
ompany Sanitized. Does not contain TSCA CBl
36
N A m S A V'srU Uadir in Tfilwi Smites for Ilii MHUcsl Dtflci Iniuitrf
2261 Tricy Rud
Nonhwood. OH 43619
Phon* 416C-9455
FAX
1Be.29SJ
AuthorlulKm Iw duplication ol ihli report eicapi In -ton, is r--rvd pTiding NAmSA'i irttun gppn>l
Page 6 of 7
gqn^agiySas^aad. Does aot contain TSCACq
hi 7 of 7
N A i l i X A World Leader in Testing Servicrs far Ihe Mediral Devicr Industry
DuPont HLO 907-96
2261 Tracy Road Northwood, OH 43619 Phone 419*666*9455 FAX 419*666*2954
LAB NO. P.O. NO.
96T 12212 00 LGX 592565
E. I. du Font de Nemours & Co. HASKELL LABORATORY
ELKTON RD. P.O. BOX 50
NEWARK, DE
19714^)050
ATTN: DR. SUSAN A. MACKENZIE
ID NO. H-22074
CYTOTOXICITY - AGAROSE OVERLAY, SOLID
Test Article: H-22074 (test item)
Test Article Description: Sheet 1 cm2 piece
Procedure:
A monolaycr of L-929 mouse fibroblast cells was grown to confluency and overlaid with Minimum Essential Medium supplemented with serum, antibiotics, neutral red, and 2X agarose. The test article, a 0.5 cm x C.5 cm piece of 725871H as a positive control, and a 1.0 cm length piece of UP-1 as a negative control were placed on the solidified overlay surface. Following incubation for 24 hours, the culture was macroscopically examined for evidence of cell dccolorization to determine the zone of cell lysis. Any decolorized zone present was examined microscopically to confirm cell lysis.
Score
Nontoxic (N) Toxic (T)
Observations
Normal cell morphology in proximity to test sample.
Cellular death and degeneration associated with the area beneath me test sample and possibly extended beyond the perimeter of the test sample. Where a zone of lysis was observed, the distance from the edge of the sample to the edge of the zone was measured and reported in millimeters (mm).
^ San.Si.ed. Does not contain TSCACB1
l.s
Completed'?-^-^C>
^
Approved tBarnfftfelpI
/
- 38 -
MG030-110 Page 1 of 2
Aulhuii/alii'in lor rjup!ii::itni "I Inr. rfporl. i-xcrpi in whole, is roservyri ponding NAmSA i written ;ipproval
NAuiSSA ^^| f.ro World Leader in Testing Service for ihe Medical Device Industry
DuPont HLO 907-96
226 ITracy Road Northwood, OH 43619 PhO- a 419*666*9455 e.fi^ 419*666*2954
LAB NO. 96T 12212 00
CYTOTOXICITY - AGAROSE OVERLAY, ?OUD
Results:
Teat/Control Articles
Score
?oneon.y-is (mm)
Test Article
N
Negative Control: UP-1
N
NAmSA Positive Control: 72587 IH
T
Conclusion: Comments:
Results and conclusions apply only to the test article tested. No further evaluation of these results is nude by NAmSA. Any extrapolation of these data to other samples is the responsibility of the sponsor.
The above test article was nontoxic for L-929 mouse fibroblast cells under the above described test parameters.
Placed me "tealest' side of test article down on agarose. All raw data pertaining to this study and a copy of the final report are to be retained in designated NAmSA archive files.
Test Article Received: 9-4-96 Date Prepared: 9-10-96
Date Terminated: 9-11-96
swan saniti2ed- ^s not contain TSCA A
h ^ n la*
Completed
Approved_________(See puff. I)
- 39 -
Page 2 of 2
Aulhon^dlio.i for dup'ic.ilion ';! 'his fBpurl. BXC"|'I in whole, is reserved ponding NAmSA's written approval
Confidential TU013.ROO
DuPont HLO 907-96
Lab No.
96T 12212 00
P.O. No. LGX 592565
STUDY TITLE: USP INTRACUTANEOUS TOXICITY STUDY IN THE RABBIT
(Extracts)
TEST ARTICLE:
H-22074 (test item)
IDENTIFICATION NO.:
H-22074
SPONSOR:
DR. SUSAN A. MACKENZIE E. I. du Font de Nemours & Co. HASKELL LABORATORY
- 40 -
^W^gg^ ELKTON RD. P.O. BOX 50
NEWARK. DE
'
^e Does Rot contain TS( 1 of
N A m S A WorU UaSir in riiUni Siniw for Ihl Mliical tl,,Ul InSuitr,
'
2261 Truy Roia
Northwood. OH 43619
''""
FAX
l9<66-955 419<fl649M
Authortzjtkxi lor dupBcjIion al fflU nyM. ia|H In whola, 1( ruenrod penOng NAmSA's writton gpprovBl.
TU013-800
DuPont HLO 907-96
Lab No. 96T 12212 00
TABLE OF CONTENTS
SUMMARY .................................................. INTRODUCTION ............................................... MATERIALS ................................................. METHODS ................................................... RESULTS .................................................... CONCLUSION .................................. ............. RECORD STORAGE .............................................
TABLE
I. USP INTRACUTANEOUS TOXICFTY OBSERVATIONS ..................
PWNTO^T
3 4 4 4 6 6 6
7
)oes not cor';ia?n TSCA "Bi
- Al -
NAmSA Worltf Uaitt in TfHtils Smiw for lln MfUcsttmici Is^utin
2281 Tflcy RfKd
Nolttliood. OH 43619
"''
FAX
<lfl6644SS 41066a8S4
Auttonutkxi (or duplication d (hi* nnoH, ttesCH ti eAote. a feaamtl pvnttny NAmSA'* wnH*n ppnwil
Page 2 of 7
TU013-800
DuPont HLO 907-96
Lab No. 96T 12212 00
SUMMARY
The test article, H-22074 (test item), H-22074, was extracted in 0.9% sodium chloride USP solution and cottonseed oil, NF. These extracts were evaluated for inuacutaneous toxicity in accordance with the guidelines of the current USP.
A 0.2 ml dose of the appropriate test article extract was injected by the intiacuianeous route into five separate sites on the right side of the back of each rabbit. Similarly, the corresponding blank vehicle was injected on the left side of the back of each rabbit. Two rabbits were used for each pair of extracts. Observations for erythema and edema were conducted at 24, 48, and 72 hours after inuacutaneous injection.
Under the conditions of this study, there was no evidence of significant irritation or toxicity from the extracts injected intracutaneously into rabbits. Each test article extract met the USP requirements.
Study and Supervisory Personnel:
Donald P. Barda, LAT Karen D. Cobum. ALAT Barbara M. Monroe Tina M. Cox. BS, LAT
Approved by:
) ll'UL'vLy Ufi-LtM fcjL La^
1}
IT
Timothy Jansen, BLS
Section Manager, Testing Services
CNO-qio
Date Completed
3, Gc-~'a ",':>' c"''":';' "'""'rv. /r/las
NAinSA ,h_^l v^H_r
42
-_______________2?_61 _Trj_cy _Ro_ad _
Northwood. OH 4361 B
Warm Uadir in TllliHS SirfU,! for Iki MiMcal Dttlct IH^IIIIIJ
""" 'Ax
<'fr666-9455 'M66.?95<
AuthoduUon lor aupllcalion of thu rpo<1. >c*(X m wtioM. a levmsa ponding NAfflSA'* wriinn pprovil
Page 3 of 7
TUO 13-800
DuPont HLO 907-96 Lab No. 96T 12212 00
INTRODUCTION
The test article identified below was extracted and the extracts were evaluated for biocompatibility a accordance with the guidelines of the current USP. The purpose of the study was to detnrmine whether leachables extracted from the material would cause local dermal irritant or toxic effects following injection into rabbit skin. The test article was received on September 4, 1996. The animals were injected on September 10, 1996, and the observations were concluded on September 13, 1996.
MATERIALS
The sample provided by the sponsor was identified and handled as follows:
Test Article:
H-22074 (test item)
Identification No:
H-22074
Storage Conditions:
Room temperature
Vehicles: Preparation:
0.9% sodium chloride USP solution (SC) cottonseed oil, NF (CSO)
A ratio of 120 caf'.W ml (surface area of test article to volume of vehicle) was used for each preparation. The test article was extracted in SC aad CSO at 121 C for 1 hour. The extraction vehicles without test article were
similarly prepared to serve as control blanks.
Condition of Extracts:
SC: CSO:
Test clear with test article particulates clear with test article particulates
Control clear clear
Sample Disposition:
Any remaining sample was uiscarded. METHODS
Test System:
Species:
Breed:
Source: Sex:
Body Weight Range:
Age: Acclimation Period:
Number of Animals:
Identification Method:
Rabbit fOryctolagus cuniculus)
New Zealand White Mohican Valley Rabbitry No particular gender was prescribed for this test No particular body weight range was prescribed for this test No particular age was prescribed for this test Minimum 2 days Two per pair of extracts
Ear tag or tattoo
Justification of Test System:
The intraculaneous injection test in rabbits is specified in the current USP and has been used historically to evaluate biomaierial extracts.
Animal Management:
Husbandry:
Conditions conformed to Standard Operating Procedures which are based on the "Guide for the Care and Use of Laboratory Animals.*
Food:
PROLAB* High Fiber Rabbit Diet was provided daily. gSSfflBpa"S?anitized. Does not contain TSCA CBI
43
KAiBiSA Wartd Uadtr in TflliHI Stnicii /or (Ac MiMcal Drier Infinity
'
226' TiacyRcjd
rolhooa. OH 43619
Pl.on* FAX
41M66.MS5
41668.28S4
Aulhonuten lu llupiieflion of ftu rapon. eictfl In whole. iMivd pfdmg NAmSA'i wnun pprovtl
Page 4 of 7
TU013-800
DuPont HLO 907-96
Lib No. 96T 12212 00
Water: Contaminants:
Freely available, municipal (Toledo, OH) water was delivered through an an^We
watering system.
Reasonably expected contaminants in feed or water suppliesdid not have the potential to influence the outcome of this test.
Housing: Environmental:
Animals were individually housed in stainless steel suspended cages identified by a card indicating the lab number, animal number, test code, sex, and date dosed.
The room temperature was monitored daily. The temperature range for the rabbit was 64-74-P.
The room humidity was monitored daily. The humidity range for the rabbit was 30-70%.
The light cycle was controlled using an automatic tinier (12 hours light, 12 hours dark).
Facility:
NAmSA is an AAALAC accredited facility.
Personnel: Selection:
Associates involved were appropriately qualified and trained.
Only healthy, thin-skinned animals free of mrfhanical irritation or trauma that could interfere with the test were selected.
Experimentfj Procedure:
Prior to treatment, each rabbit was clipped free of fur from the back and bom sides of the spinal column to yield a sufficient injection area. Two rabbits were prepared per pair of extracts. A 0.2 ml dose of test article extract was injected intncutaneously into five separate sites on the right side of the back of each rabbit; 0.2 ml of the control blank was injected into five separate sites on the corresponding left aide of me back. Injections were spaced approximately 2 cm apart. The animals were returned to their respective cages following the
procedure.
Observations for erythema and edema were conducted at 24. 48, and 72 hours after injection. Reactions were scored on a 0 to 4 basis. Any reaction at the injection sites was also noted. The reactions were evaluated
according to the following subjective rating scale:
ERYTHEMA (ER)
0 Nociythcma 1 Very slight (barely perceptible) 2 Well-defined (pink) 3 Moderate to severe (red) 4 Severe (beet redness) to slight eschar
formation (injuries in depth)
EDEMA (ED)
0 No edema 1 Very slight (barely perceptible) 2 Slight (edges of area well-defined by
definite swelling) 3 Moderate (raised approximately 1 mm) 4 Severe (raised more than 1 mm, may
extend beyond the area of exposure)
The cumulative average erythema and edema score for each test extract and corresponding control blank was calculated. For each extract, a difference in average scores (test minus control blank) of 1.0 or less was considered to be acceptable. A difference of 0.6 to 1.0 indicated a slight, but acceptable, reaction. A difference > 1.0 was considered to be unacceptable. Additionally, the average score for each test extract and blank was calculated for each interval. Any adverse reaction noted in the test extract was compared to the
corresponding blank.
t^notcontainTSCACBf
- 44 -
N A m S A WarU Ltdir in TeHtnf Stntrit far lln Mitical Dntci Munry
2201 Trac Rird
Northwood. OH 43619
Phon* FAX
<1'-<55 41M6640&4
Authortutlan XH dupNuiion d am upon, mpl h iholi. 1 r--iv*d [wndmg NAmSA't wrtcn tgptwil
Page 5 of 7
TU013-800 Results of (cores for individual nbbitt i
DuPont HLO 907-96 Lib No. 96T 12212 00
RESULTS ,
in Table 1. The findings are summarized below:
Extract SC CSO
Avenge TcjU-j)AvcraeControl" DiRerence A*"**'"**'*
0.0
0.0
0.0
--fu---s--ed-a
1.6
1.7
0.0
Paced
the difference in average scores was c
scores did not exceed 1.0 at any obser
eatable. The diRerence between test aod control average i interval.
Results and conclusions apply only to the test article tested. No further evaluation of these results is made by NAmSA. Any extrapolation of these data to other samples is the respoosibuity of the sponsor.
WNqyaQN
Under the conditions of this study, there was no evidence of significant irritation or toxidty from the extracts injected intracutaneously into rabbits. Each test article extract met the USP requirements.
RECORD STORAGE
All raw dar.. .leitainiog to this study and a copy of the final report are to be retained in designated NAmSA
archive files.
Company Saniteed. Does not wnSaiiinTSCACBl
- 45 -
N A m S A WfrU UUn to TutJrf Sr*(c*i /r (* Mci 0r<cf ln4llrj
2261 Tncy Riw)
Noithwood. OH 43818
Phan* 41B4W44JS
FAX
4i<8fr2W4
Aunonaaan lor iHt^-f^r- d Wt coot. fttUfA tn ho0. b waived pandhg HAmSA'l wMn tflpro--l
Page 6 of 7
TU013-800
DuPont HLO 907-96
Lab No. 96T 12212 00 TABLE I USP INTRACUTANEOUS TOXICrTY OBSERVATIONS
RABBIT NUMBER 97215 97216
97215 97216
EXTRACT
SC SC CSO
cso
1TEST EXTRACT
Scoriol laiemi (Houn)
24
48
72
ER
ED ER ED
ER EO
0
0
0
0
0
0
0
0
0
0
0
0
2
2
2
1
1
1
2
2
2
2
1
ER = Erythema
ED = F^r""
SC = 0.9ft sodium chloride USP solution CSO = cottonseed oil, NF
CONTROL BLANK
ScuriBi tanml (Boon)
24
48
72
ER
ED ER ED ER EO
0
0
0
0
0
0
0
0
0
0
0
0
2
2
2
1
1
2
2
2
2
1
2
1
OTOy Sanitteetf. Does not contain TSCA CBI
46
NAmM
WorU L**J<r In Tmliii Sinleti forth* MtttealDfflct Infsury
2261 Ticy Road
Nonhwood. OH 43819
Ptiore 41Befr94S5
FAX
41-68&2a54
AuauMteaflon tof duptoffllon al VMs rapon. ixcepi m to--, l* r--wvd p--KUngNAmSA'i wrtfn ppimit.
Page 7 of 7
Confidential TU014-807
DuPonfc HLO 907-96
Lab No.
96T 12212 00
P.O. No. LOX 592965
STUDY TITLE: LSP MUSCLE IMPLANTATION STUDY IN THE RABBIT
(1WEEK) TEST ARTICLE:
H-22074 (tect item)
IPENTmCATIONNO.:
H-22074
SPONSOR:
DR. SUSAN A. MACKENZIE
i
E. I. du Font de Nemoun & Co.
I
HASKELL LABORATORY
ELKTON RD. P.O. BOX 50
' ^bmpaffl<Ysaniiize<jD. oes not contain TSCA CB I
- 47 -
Page lof7
NAuiSA WirUUf^rtTuUlvSi^Ufl / (* MM( Dulci Mmlrj
2M1TaeyRa*d Hontwnod. OH 43eiB
""" <.\9m9w
"* 'w'W
Aiiftwullan In dupklllan al H* iwail. aiiil h tioli, i* nMnad pOTlftg NAniSA'i rtMn iipnwd.
TU014-W7
SUMMARY . . . INTRODUCTION
MATERIALS ..
METHODS . . . .
RESULTS......
CONCLUSION . . . RECORD STORAGE
TABLE OP CONTENTS
DuPonfc HLO 907-96 UbNo. 96T 12212 00
48 -
N A m S A WwU UUlt If, TitUni Sinlctt fw (** UtMnl DivUi imsitrj AuftaiteiNai hi dupk*tlan al ik npon. nawi m ilioc. M i
Company Sanitized. Does not contain TSCA CBS
221TneyRaU
Naianmod. OH 4381B
Phon* 4M985
FAX
1MM4
ng NAniSAI udMn pplOMI.
TU014-807
DuPont HLO 907-98
LA No. MT 12212 00
SUMMARY
The tot iticte, H-22074 (let Item), H-22074. wn implied In nude tissue of the rtbbit. T-- auedi ws evaluated for evidence of irritation or toxicity in eeontaace with (he (uUdiDM of the cuneal VSf.
ImfriKtt umples ud USP ae|ive control (trip
implanted nd were thca euduiutizad 1 week tater.
CT----rfnil HMMIMMIpL-lly
iloadBd
idleiidctefiUzedbyi
ruiirif HIIIIIT T iim nrliml inil itr Implmi ll- i.e
Under the condition* of this study, the nucnMcopic n.ictiao mi aot ispAVntt i icoa|Mml(otheUSP
negative comiol implant material. The impluued tet article met ae US? i
Study and Supervisory rffVfWW'
Christina M. ToUe*
Jaun A. Kantea Paul A. BeU, HT(ASCP) Michael A. Safioa. AAS (HT) Tina M. Cox, BS, LAT
Approved by:
7 ^ ^ Timothy Janita^BlJ
Section Manner,Testinig Services
Dafte -Col ^ --id ^
&9 M^i^ianS.1 /nun
- 49 -
WsfU Lufir In Tllll*t Sirricii ffftln UtMcil Dflici MSMI Auto--on to dikiHonalM
Company sanitized.
Does TOBtCOfl^"TSCAGgi
Pte3of7~^
t
i
iMITncyRiMd
NWHKiod. OH Mr
Pnon* iinam8
inane NAmSA't ----> wont.
TU014.W7
DuPont HLO 907-96
Lab No. 96T 12212 00
INTROnUCTION
The test uticle identified below w-- evaluated for biocompitibility in accordance with the (uidelinea of the
current USP. The puipose of the study w-- to evaluate the potential for a local irritant or toxic response to
material implanted in direct contact with onude time. The lest article waa received on September 4,1996. The animals were implanted on September 6.1996, and muscles were explained on September13.1996.
MATERIALS
The sample provided by the sponsor was identified and handled as follows:
Test Article:
H-22074 (test itsm)
Identification No:
H-22074
Storage Conditions: Control Article:
Preparation:
Room temperature
USP plastic 1 mm diameter neytivc control reference strips purchased by NAmSA from the offices of the <JS Pharmacopeial Convention and cut into 10 mm sections.
A ^nnmmm of six sections of the test article per nbbit, each approximately
1 mm x 10 mm, were loaded into 16 gauge needles. For each rabbit, a
minimum offourUSP strips were loaded into the same size needles as used for the test article. Test and control articles were sterilized by steam.
S.!'To>le Disposition:
Any remaining sample was discarded.
METHODS
Test System:
Species:
Breed: Source: Sex: Body Weight Range: Age: Acclimation Period:
Number of Animals: Identification Method:
Rabbit fOrvctolamis cuniculm)
New Zealand White Mohican Valley Rabbitry No oartimdar gender was prescribed for this test 2.9 kg to 3.0 kg at implantation No particular age was prescribed for this test Minimum 2 days
Two
Ear tag or tattoo
Justification of Test System:
The nbbit is sugge.ieu qs an appropriate animal model for evaluating polymer materials by the current USP testing guidelines. The muscle tissue has been used historically because me response to implanted material is easily graded and compared to a known negative control material.
- 50 -
M i i i S B J i WarU Usisr t TiiUKt Sinlcis far In HfttcslOntf Mulrf AuttKxtt--on tot dupliollon ij Oir nport. UMpI m o"0-- *'
""^f&Va Company Sanity. Does nog
2f1 Tr*cy Rud Northwod, OH 3<B
Phon* 41frS!
FAX
418fr2M4
ntfna NAmSA'i own pprevl.
TU014-807
DuPont HLO 907-96 Lib No. 96T 12212 00
Animal Manflgtmcnt:
Husbandly:
Conditions conformed to Standard Openting Procedure* which are bawd on the "Guide for the Care and Use of Laboratory Animals."
rBOuOuu4:.
PROLAB* High Fiber Rabbit Diet was provideddaily.
Water: Contaminants:
Freely available, municipal (Toledo. OH) water waa delivered through an aulomMic watering system.
Reasonably expected contaminants in feed or water supplies did not have die potential to influence the outcome of this test.
Housing:
Animals were individually housed in stainless steel suspendedcages identified by a end jiytic^ting the lab number, animal number, test code, sex, and date implanted.
Environmental:
The room temperature was monitored daily. The temperature range for the rabbit was 64-74-F.
The room humidity was monitored daily. The humidity nmge for the rabbit was 30-70.
The light cycle was controlled using an automatic timer (12 hqura light, 12 hours dark).
Facility:
NAmSA is an AAALAC accredited facility.
Personnel:
Associates involved were appropriately qualified and trained.
Selection:
Healthy animals were selected. To reduce the number of animals used for testing, and to comply with the directives of the NAmSA IACUC, rabbits on this study may have been used previously in an unrelated test model. Any previously evaluated test or control articles did not cause a response in the animals. Complete history of animal usage is
traceable in laboratory records. Animals used for previous evaluations are identified in
the report.
)E3q>erimertalProcedure:
The fur was clipped from the area over the paravertebral muscles and the rabbits were sedated with
acepromazine maleate (0.2 ml/kg dose). After the muscle area was wiped clean with 709 alcohol, each paravenebral muscle was infiltrated with 1% Udocaine to achieve local anesthesia on bom sides of the vertebral
column.
Needles containing teat article were inserted into the muscle at a 45 angle. A stylet was placed in die needle and the needle was withdrawn, leaving the sample in the paravenebral muscle. Six test article sections were implanted in the right paravenebral muscle of each rabbit. In the opposite muscle, four USP negative control strips were similarly implanted.
Rabbits were returned to their respective .-ages following the procedure. Rabbits were observed daily for general health. Body weights were recorded prior to implantation and at termination.
51 -
MinSA
WfrM LsUlt in Tllllsi Srric
/T llii M(ff D"(c liduilrj
tWmastUm let 6Wtsn d 8X3 repsrt. sow In oW, r r
22B1 Titcy Rna Northwooa. OH WIB
Phon* 4lfra4ii
FAX
4tfr6M4854
ng NAmSA'l ortlfn ppw).
iTSCAcfel
Page 5 of 7
TU014-807
DuPont HLO 907-96 Lab No. 96T 12212 00
At I week, the nbbits were weighed and euthanatized by in intraVenoul injection of a (odium peniobaibital bued drag. The paraveitebral muscles were rfiimrtfd free and methodically cut to locate four of the ten article sites and two of the USP negative control sitei in each rabbit. Capsule formation or other evidence of irritation
w scored using appropriate magnification and an auxiliary light source. Scores were assignedand recorded as
luuows:
0 No capsule, no advene reaction (other than minimal hemorrhage) 1 Up to 0.5 mm capsule or reaction area 2 0.6 to 1.0 mm capsule or reaction area
3 I.I to 2.0 mm capsule or reaction area
4 > 2.0 mm capsule or reaction area
Average macroscopic scores for test implants were compared to average scores of the control sites. Calculations were rounded off to die nearest 0.1. A Reaction Index difference of 0.0 to 0.5 in scores (test minus control) was regarded as "not significant,' 0.6 to 1.0 as a "trace," 1.1 to 2.0 as "slight." 2.1 to 3.0 as "moderate" and 23.1 as marked." The requirements of the USP test were met if the difference between test and control score averages (macroscopic) was not greater than 1.0. The requirements of the test were not met if the difference between the test and control score for two (or more) implant sites exceeded 1 for any animal
implanted.
.
RESULT
Clinical Observations: AU animals appeared clinically nonnal throughout the duradon of die study. Body weight data for individual rabbits were considered acceptable.
Macroscopic Observationa: The findings for the macroscopic evaluation are shown triow.
Weight (kg)
Rabbit
Number
DayO
Day?
Site
1
96517*
3.0
2.9
2
3
4
1
96477*
2.9
2.8
2
3
4
Test
0 0 0 0
0 0 0 0
USP Negative Control
0 0
0 0
Average:
0.0
0.0
Previous use history traceable in laboratory records.
Macroscopically, there was no difference between the reactions of the test and control materials. This resulted in a macroscopic reaction of "not significant* tissue contact irritation.
Results and conclusions apply only to the test article tested. No further evaluation of these results is made by NAmSA. Any extrapolation of these data to other samples is the responsibility of the sponsor.
- 52
M m S j K WerU Lasiti is r(f0ff Sinlesi far tin Ulttesi Dulct lUsilrj AlAortmion tor ducallan d Wi report, icepi In hoM. u r
Company Sanitized. Does not contain TSCA CBJI
Page 6 of 7
2211 Trcy ROM
Nonhoaod. OH 4381B
PhiMM 41MBM455
FAX
4(M8S4
no tUmSA'* rffien ippiwil.
TU014-807
CONCLUSION
DuPont HLO 907-96 Labr'v 96T 12212 00
Under the condiiions of this study, the macroscopic reaction was not significant as compared to the USF negative control implan' material. The implanted test article met the USP requirements.
RECORD STORAGE
All raw data pertaining to this study and a copy o(' the final report are to be retained in designated NAmSA
archive files.
Company Sanitized. Does not contain TSCA Cl
^A*
-53-
World l.tadtr in Tfilini Services for the Medical Device Industry
?26' Tracy Head
iOf'^wood OH 41611
Phone 4l-S61>-')4SS
PAX
.11.6M..?95t
Authonzalio^ tor dupdcalion of this repori. eiceDi m whotfl. fs reaarved pending NAmSA's wrilten approvBl
Page 7 of 7