U.S. patent number 3,810,473 [Application Number 05/312,210] was granted by the patent office on 1974-05-14 for liquid-laid, non-woven, fibrous collagen derived surgical web having hemostatic and wound sealing properties.
This patent grant is currently assigned to Avicon, Inc.. Invention is credited to Orlando A. Battista, Carmine Cirolla, Mamerto M. Cruz, Jr., LaVerne C. Tressler.
United States Patent |
3,810,473 |
Cruz, Jr. , et al. |
May 14, 1974 |
**Please see images for:
( Certificate of Correction ) ** |
LIQUID-LAID, NON-WOVEN, FIBROUS COLLAGEN DERIVED SURGICAL WEB
HAVING HEMOSTATIC AND WOUND SEALING PROPERTIES
Abstract
Liquid-laid, non-woven, fibrous web having hemostatic and
adhesive properties sufficient to seal a wound formed of fibers of
ionizable, water-insoluble, partial acid salts of collagen, the web
having certain specified absorbency and porosity characteristics.
In order to prevent excessive swelling of the fibers with resulting
excessive hornification or densification upon drying, it is
essential that mixtures of water-miscible organic liquids such as
ethanol with specified minor porportions of water be utilized as
the slurrying media.
Inventors: |
Cruz, Jr.; Mamerto M.
(Pennington, NJ), Battista; Orlando A. (Fort Worth, TX),
Tressler; LaVerne C. (Trenton, NJ), Cirolla; Carmine
(Morrisville, PA) |
Assignee: |
Avicon, Inc. (Ft. Worth,
TX)
|
Family
ID: |
23210381 |
Appl.
No.: |
05/312,210 |
Filed: |
December 4, 1972 |
Current U.S.
Class: |
606/213;
106/157.3; 128/DIG.8; 106/156.1; 606/229 |
Current CPC
Class: |
A61F
13/0223 (20130101); A61F 13/0289 (20130101); A61F
13/00034 (20130101); A61L 15/325 (20130101); A61L
15/58 (20130101); A61L 15/58 (20130101); C08L
89/06 (20130101); A61F 2013/15821 (20130101); A61F
2013/00472 (20130101); A61F 2013/00744 (20130101); A61L
2400/04 (20130101); A61F 13/8405 (20130101); Y10S
128/08 (20130101); A61F 2013/00221 (20130101) |
Current International
Class: |
A61F
13/00 (20060101); A61L 15/16 (20060101); A61L
15/58 (20060101); A61L 15/32 (20060101); A61F
13/15 (20060101); A61b 017/04 (); A61b
017/08 () |
Field of
Search: |
;128/334R,335,DIG.8
;106/161 |
References Cited
[Referenced By]
U.S. Patent Documents
Other References
Peacock, E.E. Jr. - Annals of Surgery, Vol. 161, Feb. 1965,
pp.238-247, "Use of . . . Collagen Sponges . . . ".
|
Primary Examiner: Medbery; Aldrich F.
Claims
What is claimed is:
1. A hemostatic adhesive dressing for severed biological surfaces
comprising a liquid-laid, non-woven web having hemostatic-adhesive
properties comprising hemostatic-adhesive fibers of ionizable,
water-insoluble, partial acid salts of collagen, the web having an
absorbency of a mixture of 90 volumes of ethanol and 10 volumes of
water of between about 100 percent and about 300 percent by weight
and a porosity of from about 4 to about 15 seconds per 6.45 sq. cm.
based upon a web having a basis weight of about 175.5 lbs. per
3,000 sq.ft., the fibers of the partial acid salt of collagen
containing from about 50% to about 90% of the theoretical
stoichiometric amount of ionizable acid.
2. A dressing as defined in claim 1 wherein the web has a Stiffness
Factor of from about 15.degree. to about 100.degree. based upon a
web having a basis weight of about 175.5 lbs. per 3,000 sq.ft.
3. A dressing as defined in claim 1 wherein the fibers are partial
hydrogen chloride salts of collagen.
4. A dressing as defined in claim 2 wherein the fibers are partial
hydrogen chloride salts of collagen containing from 60 percent to
85 percent of the theoretical stoichiometric amount of hydrogen
chloride.
5. A dressing as defined in claim 1 wherein the fibers have a fiber
length distribution by the McNett Classifier such that about
45 to 55% are retained on a 20 mesh screen,
20 to 25% are retained on a 35 mesh screen,
3 to 6% are retained on a 65 mesh screen,
0.5 to 1.5% are retained on a 150 mesh screen, and not more than
30% pass a 150 mesh screen.
6. A dressing as defined in claim 2 wherein the fibers are partial
hydrogen chloride salts of collagen containing from 60% to 85% of
the theoretical stoichiometric amount of hydrogen chloride and the
fibers have a fiber length distribution by the McNett Classifier
such that about 48% are retained on a 20 mesh screen, about 22% are
retained on a 35 mesh screen, about 3.6% are retained on a 65 mesh
screen, about 0.7% are retained on a 150 mesh screen and the
balance passes a 150 mesh screen.
7. The method of forming a hemostatic adhesive liquid-laid,
non-woven web dressing for severed biological surfaces from
hemostatic-adhesive fibers of ionizable, water-insoluble, partial
acid salts of collagen including the steps of slurrying the fibers
in a mixture of a water-miscible organic liquid and water, sheeting
the fibers to form a web and drying the web, the fibers of the
partial acid salt of collagen containing from about 50% to about
90% of the theoretical stoichiometric amount of ionizable acid, the
mixture of organic liquid and water consisting of from about 95
volumes of organic liquid and about 5 volumes of water to about 80
volumes of organic liquid and 20 volumes of water.
8. The method as defined in claim 7 wherein the fibers have a fiber
length distribution by the McNett Classifier such that about
45 to 55% are retained on a 20 mesh screen,
20 to 25% are retained on a 35 mesh screen,
3 to 6% are retained on a 65 mesh screen,
0.5 to 1.5% are retained on a 150 mesh screen, and
not more than 30% pass a 150 mesh screen.
9. The method as defined in claim 7 wherein the organic liquid is
ethanol.
10. The method as defined in claim 7 wherein the mixture consists
of from about 95 volumes of ethanol and about 5 volumes of water to
about 80 volumes of ethanol and about 20 volumes of water and the
web is freeze dried.
11. The method as defined in claim 7 wherein the mixture consists
of from about 95 volumes of ethanol and about 5 volumes of water to
about 85 volumes of ethanol and about 15 volumes of water and the
web is vacuum dried.
12. The method as defined in claim 7 wherein the fibers of the
partial acid salt of collagen contain from 60% to 85% of the
theoretical stoichiometric amount of hydrogen chloride, the mixture
consists of 90 volumes of ethanol and 10 volumes of water, the
fibers have a fiber length distribution by the McNett Classifier
such that about 48% are retained on a 20 mesh screen, about 22% are
retained on a 35 mesh screen, about 3.6% are retained on a 65 mesh
screen, about 0.7% are retained on a 150 mesh screen and the
balance passes a 150 mesh screen and the web is freeze dried.
13. The method of joining severed biological surfaces in a living
warm blooded animal which comprises placing between and in contact
with the severed surfaces a liquid-laid, non-woven fibrous web of
hemostatic-adhesive fibers of ionizable, water-insoluble, partial
acid salts of collagen, allowing the web to become wetted with
blood, pressing the web into contact with the severed surfaces
until hemostasis has been effected and then releasing the pressure,
the web having the property of combining with blood to effect
hemostasis and to form a mass with the blood that is self-adherent
to the severed surfaces and thereby seal the wound, the web having
an absorbency of a mixture of 90 volumes of ethanol and 10 volumes
of water of between about 100% and 300% by weight and having a
porosity of from about 4 to about 15 seconds per 6.45 sq.cm. based
upon a web having a basis weight of about 175.5 lbs. per 3,000
sq.ft., the fibers of the partial acid salt of collagen containing
from about 50% to about 90% of the theoretical stoichiometric
amount of ionizable acid.
Description
This invention relates to liquid-laid, non-woven sheets or webs
particularly suited for medical and surgical purposes consisting
essentially of finely-divided fibers derived from collagen.
It has been known that collagen in various treated and prepared
forms is useful in medical and surgical procedures and in the
treatment of wounds. Collagen in certain forms has hemostatic
properties when used as a wound dressing and has a low level of
anti-genicity. In the copending application of Orlando A. Battista,
Mamerto M. Cruz, Jr., and Merritt R. Hait, Ser. No. 76,638, filed
Sept. 29, 1970 and now U.S. Pat. No. 3,742,955 there is described a
fluffy, finely-divided fibrous collagen product derived from
natural collagen which when wet with blood has hemostatic
properties and a unique adhesive property which is sufficient to
join together severed biological surfaces. This form of collagen
demonstrates an unexpected and entirely unique adhesive property
when wet with blood in live warm blooded animals and in many
instances can actually be used to adhere severed tissues without
the use of sutures. The disclosure of this above identified
application is herein incorporated and made a part of the present
disclosure.
As indicated in the above entitled application, the
hemostat-adhesive material is in a fluffy, finely-divided fibrous
form and the form for use in this invention consists essentially of
a water-insoluble, ionizable, partial salt of collagen, the fibrous
mass having a density of not more than about 8 pounds per cubic
foot, preferably the bulk density is between 1.5 and 6.0 pounds per
cubic foot. The mass when combined with blood in a wound forms a
mass that is self-adherent to the tissue surfaces and will seal the
wound without the use of sutures. The partial salt of collagen
consists essentially of an ionizable, water-insoluble, partial acid
salt of collagen consisting from about 50 percent to about 90
percent of the theoretical stoichiometric amount of the ionizable
acid. The fluffy, finely-divided fibers for use in the present
invention may be prepared as described in the aforementioned
application.
Because of the low density and fluffiness of the product of the
aforementioned application, it is necessary to transfer the product
to the wound by the use of forceps or manually with rubber gloves.
In such handling procedures fibers become dislodged from the bulk
being handled and portions will adhere to the forceps or rubber
gloves.
In copending application Ser. No. 76,638, it is stated that the
fluffy, finely divided fibrous collagen product may be converted
into non-woven webs by a wet method such as commonly employed in
forming papers. It is also stated that in such method the fibrous
collagen is slurried in a water-miscible organic liquid of the type
used in producing the fibrous collagen.
In the conventional production of water-laid fibrous sheets or webs
such as papers, naturally occurring cellulose fibers such as wood
pulp and cotton linters are mechanically hydrated as by beating and
mixing in a paper mill beater in the presence of a large excess of
water. In forming a pulp slurry by a beating operation, the fibers
become hydrated and exhibit a microscopic and submicroscopic
peeling of individual fibrils along the surface of the fibers and
at the end of the fiber bundles. The slurry is then passed to a
suitable screen to lay down the fibers as a sheet or mat. The
physical properties of the sheet, such as strength, tear and burst
are dependent to a large extent on the hydration of the fibers and
an interlocking of the hydrated fibers and of the fibrillae on the
fibers and the fiber-to-fiber bonding which develops upon
drying.
Because of the greater sensitivity to water of the fibers of the
fluffy, finely divided partial salt of collagen having the
hemostatic and unique adhesive characteristics as described in the
afore-mentioned application, water-laid fibrous sheets or webs
formed from such fibers are harsh and boardy and parchment-like in
structure with complete loss of hemostatic and adhesive properties
thereby rendering such sheets unsatisfactory for surgical
purposes.
The principal purpose of the present invention is to provide a
liquid-laid, non-woven web which retains the hemostatic-adhesive
properties of the fluffy, fibrous partial salt of collagen.
A further purpose of the invention is to provide a method of
forming the liquid-laid, non-woven sheet or web of the fluffy,
finely-divided fibrous collagen derived product which is flexible,
non-flaking, and has sufficient strength to permit its application
to a wound without separation into individual fibers.
A further purpose of the present invention is to provide a method
of forming a liquid-laid, non-woven fibrous web of the
finely-divided fibrous collagen derived product which retains the
hemostatic and unique adhesive properties of a finely-divided
fibrous mass as described in the aforementioned application.
Further objects and advantages of the present invention will become
apparent to those skilled in the art from the following description
of the method and product.
The present invention is based upon the discovery that non-woven
webs or sheets which retain the hemostatic and unique adhesive
properties of the fluffy, finely-divided fibers of the partial salt
of collagen can be formed by slurrying the fibers in a mixture of a
water-miscible organic liquid and water in certain relative
proportions and depositing the fibers from such slurry.
In the drawings:
FIG. 1 is a graph illustrating the freeness of the fibers when
slurried in ethanol and ethanol-water mixtures of varying
composition; and,
FIGS. 2 through 7 are graphs illustrating the variations in the
physical properties of the liquid-laid, non-woven sheets or webs
prepared from slurries of the fibers in ethanol and ethanol-water
mixtures of varying composition.
In order to avoid the production of products having a
parchment-like structure and to provide the web or sheet with
requisite characteristics, it is essential that the slurrying
liquid consists of a mixture having a composition, by volume, in
the range of from 95 percent organic liquid and 5 percent water to
about 85 percent organic liquid and 15 percent water. Where the
amount of water is less than about 5 volumes of water to 95 volumes
of the organic liquid, the finished product is unsatisfactory in
that it has no dry web strength and is extremely flaky; that is,
upon handling, fibers readily separate from the sheet. At least
this minimum amount of water must be present so as to provide a
sufficient degree of bonding between the fibers to form a coherent
and handleable sheet. In those instances where the sheet is vacuum
dried the amount of water preferably should not exceed about 15
volumes of water to 85 volumes of organic liquid, preferably 95
volumes of organic liquid and 5 volumes of water. If the amount of
water exceeds this ratio, the fibers become so firmly bonded
together that there is a loss in absorbency and flexibility and a
marked loss of the hemostatic and adhesive properties of the sheet.
Where the sheet is to be freeze dried the liquid may consist of up
to about 20 volumes of water to 80 volumes of organic liquid,
preferably 90 volumes of organic liquid and 10 volumes of water.
Here again, where the amount of water exceeds this value there is a
loss in the desired properties.
The water-miscible organic liquid consists of low molecular weight
alcohols, ketones and the like such as, for example, methanol,
ethanol, isopropanol, amyl alcohol, methylethyl ketone, acetone,
and mixtures of these organic liquids. It is possible to increase
the amount of water providing that a salt which hydrates is present
such as sodium sulfate and calcium chloride. The use of the organic
liquids other than ethanol is feasible, however, where the product
is intended for surgical uses it is essential that the product be
freed of such solvent.
After depositing or sheeting the fibers upon a suitable collecting
screen excess liquid may be separated as by pressing and the
application of vacuum. The product is then freeze dried or vacuum
dried to preserve the absorbency, flexibility and
hemostatic-adhesive properties. Freeze drying is preferred because
it provides softer, more absorbent and more flexible products that
exhibit no noticeable decrease in the hemostatic-adhesive
properties of the fibrous material. One of the unique
characteristics of these liquid-laid non-woven sheets or webs is
that upon heat sterilization as by heating at 120.degree.C. for
from 20 to 30 hours there is an improvement in the tear strength of
the sheet as well as in the hemostatic properties.
The fibrous collagen product can be prepared from any undenatured
collagen in the natural state or delimed edible forms of collagen
including, for example, hide, gut, tendon, cartilage or other high
fibrous collagen source material preferably chopped up for easier
handling. The collagen is preferably in a wet and never-dried state
or, if dried, drying has been effected under conditions which
minimize denaturation. Satisfactory raw materials for the collagen
include, for example, fresh cowhides and calfhides, salted down
cowhides, wet moosehide, pigskins, sheepskins and goatskins as
conventionally used for making leather. Special technical hide
collagen prepared from hide splits and possessing a minimal reduced
bacteria count is also satisfactory. The preferred raw material,
because of availability is corium derived from never-dried cowhide
or technical grades of collagen prepared from cowhide and other
animal hides.
The wet collagen source material such as hide is diced or chopped
into small fragments of from one-fourth to one-half inch sizes in a
cutting or grinding mill, such as, for example, an Urschel Mill.
Alternatively, these fragments may be mixed with crushed ice and
then passed through the Urschel Mill with cutting heads of smaller
dimension to fiberize the collagen into a coarse fibrous
product.
If swelling or hydration of the collagen fibers is not controlled
during the subsequent treatment wherein the collagen is subject to
mechanical shredding or opening in a liquid medium, excessive
hornification or densification will occur when the material is
dried down thereby effectively preventing the satisfactory
deaggregation of the collagen fibrils during the final mechanical
treatment. The initial swelling of the collagen fibers in the wet
state affords many more sites for hornification that is desirable
thus resulting in a dense, non-absorbent material. When
hornification and densification occurs, the product will not have
the required hemostatic and adhesive characteristics.
The wet or moist collagen source material is mechanically dispersed
or slurried in an aqueous liquid which controls the swelling of the
fibers. The aqueous liquid comprises water and a water-miscible
organic liquid such as low molecular weight alcohols, ketones, and
the like, such as, for example, methanol, ethanol, isopropanol,
methylethyl ketone, acetone and the like in a weight range of
between about 90 percent of the organic liquid and 10 percent water
and about 50 percent organic liquid and 50 percent water. Where the
proportion of water is too high, the collagen fibers swell to such
a great extent that they lose their fiber identity with an
attendant densification during the subsequent drying step. When
this occurs, it becomes commercially unfeasible to subsequently
deaggregate or fluff the fibrous product and attain the bulk
density requirements necessary for the present invention. Although
such product will possess some hemostatic properties, it does not
possess the desired adhesion to severed biological surfaces and
will not provide the required mechanical properties of the
collagen-blood matrix between the severed surfaces to seal the
wound.
The bulk of the liquid is drained from the mass and the fibrous
collagen slurried and washed with a water-miscible organic liquid
such as the alcohol and again the bulk of the liquid is separated
from the partially swollen wet collagen material. Preferably, the
collagen material is slurried in the organic liquid to reduce the
water content to a minimum. In general, the use of three slurrying
steps with the organic liquid will reduce the amount of water
present to about 1 percent. The organic liquid is removed as by
centrifugation and final drying. Drying may be effected either by
oven drying or vacuum drying as at, for example, 40.degree.C. under
a 29 inch vacuum for about 16 hours. In general, this vacuum drying
will reduce the volatile content to under 1 percent.
The partial acid salt of collagen is formed by incorporating the
required amount of an ionizable acid in the aqueous liquid wherein
the collagen is dispersed or slurried. The amount of acid
incorporated in the aqueous liquid is such as to provide the
product with a bound acid content of from about 50 percent to 90
percent preferably about 60 percent to 85 percent of the
theoretical stoichiometric bound acid content. After the acid has
reacted with the dispersed collagen, the reaction mass is subjected
to slurrying and washing with the water-miscible organic liquid and
the collagen salt processed as above described.
Before the final deaggregation into constituent fibers or fluffing
operation to produce the product having the required bulk density,
the fibrous material is preferably conditioned to contain about 8
percent to 15 percent volatiles. This conditioning may be readily
effected by allowing the product to remain at normal atmospheric
temperatures and humidities (for example, 70.degree. -
75.degree.F., 40% - 60% R.H.) for from about 8 to 24 hours. The
final fiber deaggregation or fluffing operation is necessary to
provide the requisite bulk density. This operation is an "opening"
operation whereby the diced material is converted into bundles of
individual fibers. In forming the product of the present invention,
the final fiber deaggregation or fluffing operation does not
separate all of the dried bundles into ultimate individual fibrils
but the product does contain finer fiber bundles (smaller in
diameter) as compared to the coarser fiber bundles obtained at the
end of the drying and conditioning operations. This deaggregation
or fluffing may be effected by apparatus such as a hammer mill type
comminution mill such as a Fitz Mill.
Alternatively, the wet collagen source material is merely diced or
chopped into small fragments as described hereinbefore and then
introduced into and mixed in a water-miscible organic liquid such
as ethanol or isopropanol. Mixing is continued for about 1 hour so
as to permit thorough penetration of the organic liquid into the
small fragments. The bulk of the liquid is then separated as by
draining or centrifuging and the recovered fragments again
introduced into and mixed in the organic liquid for about 1 hour.
Again, the bulk of the liquid is separated and the procedure
repeated. At the end of this period, the liquid is centrifuged from
the mixture and the wet fragments dried as by oven drying or vacuum
drying. The resulting product, after conditioning as described
above, is then subjected to a fiberizing and deaggregation or
fluffing operation. To produce a partial ionizable salt of
collagen, the desired amount of acid may be mixed with the organic
liquid in any one of the above described steps, preferably in the
second step. In such instances, the time of treatment with the
organic liquid containing the acid should be prolonged to permit
the required reaction between the acid and the collagen. Obviously,
the time periods may be reduced by operating under pressure.
In order for the fibrous product to exhibit the desired
hemostatic-adhesive properties and required physical properties the
starting fibers of the partial salt of collagen should possess
effective lengths within certain ranges. The effective fiber
lengths may be determined in accordance with TAPPI Standard Method
T233 su-64 utilizing a McNett Classifier having four tanks provided
with a 20 mesh screen (openings 840 microns), a 35 mesh screen
(openings 500 microns), a 65 mesh screen (openings 230 microns) and
a 150 mesh screen (openings 100 microns), respectively.
Because of the excessive swelling of the fibers in water of the
partial salts of collagen, these fibers are unsatisfactory for use
in a fiber classifier. Accordingly, the partial salt fibers are
converted to collagen fibers while preventing excessive swelling of
the partial salt fibers. Such conversion may be effected as by
slurrying the partial salt fibers at a solids concentration of
about 1 percent for about 30 minutes in a mixture of 90 volumes
ethanol and 10 volumes water adjusted to and maintained at a pH of
10.5 by addition of ammonium hydroxide solution. The fibers are
recovered by the use of a nylon filter fabric in a Buchner funnel
and using a water asperator. The recovered fibers are slurried at a
1 percent solids concentration in a mixture of 90 volumes ethanol
and 10 volumes water for about 30 minutes to wash out soluble salt
and the fibers recovered by the same type filtration. The recovered
fibers are then given two additional washes using 100 percent
ethanol. The finally recovered alcohol-wet fibers should not be
allowed to dry but are slurried in the required volume of water at
the recommended solids content for the specific classifier.
When ten gram samples of partial salt fibers satisfactory for the
purposes of the present invention are subjected to testing by the
use of the McNett Classifier the samples exhibit a fiber length
distribution of about
45 to 55 percent retained on a 20 mesh screen (840 microns)
20 to 25 percent retained on a 35 mesh screen (500 microns)
3 to 6 percent retained on a 65 mesh screen (230 microns)
0.5 to 1.5 percent retained on a 150 mesh screen (100 microns)
30 percent maximum passing a 150 mesh screen.
Where the proportion of long fibers is too high the partial salt
fibers upon slurrying in the ethanol-water mixture, the fibers
floculate and agglomerate into ropey masses. Where the proportion
of short fibers is too high, there is a loss in the adhesive
properties of the liquid-laid sheets.
A measure of the fluffiness and a rough indication of a
satisfactory fiber length distribution of the fibers is bulk
density.
The bulk density is measured by adding the fibrous collagen
products as initially fluffed to a 100 ml. graduate cylinder
without any compression step and determining the weight of the
added 100 mls. of the product.
In forming the partial salt of collagen, hydrochloric acid is the
preferred acid and is used in the examples which follow merely
because it is relatively inexpensive and allows ready flexibility
and ease of control. Other ionizable acids, both inorganic and
ionizable organic acids, such as, for example, sulfuric acid,
hydrobromic acid, phosphoric acid, cyanoacetic acid, acetic acid,
citric acid and lactic acid are satisfactory. Sulfuric acid, for
example, is satisfactory, but control of the action is difficult.
Citric acid may be substituted for hydrochloric acid with about
equal results. "Ease of control" has reference to the ability to
arrest the swelling and hydrolysis of the collagen fibers so as to
prevent the rapid degradation of the material to a water-soluble
product.
In the examples which follow, the fluffy, finely-divided fibrous
collagen product was derived from wet or green bovine corium. The
collagen product was a water-insoluble, ionizable, partial hydrogen
chloride salt of collagen containing approximately 84% of the
theoretical stoichiometric bound acid content. The product was
deaggregated or fluffed by passing it through a Fitz Mill (Model
DA50-6-5634) operated at 6,250 rpm. equipped with a No. 4 screen
having openings of 0.243 inch. It was then subjected to a second
pass using a special slotted sceen having openings 0.062 inch
.times. 0.5 inch with the slots at an angle of 30.degree. to the
sides of the screen.
The fiber length distribution of the fibrous partial salt of
collagen determined by the use of a McNett Classifier was about
48 percent retained on a 20 mesh screen
22 percent retained on a 35 mesh screen
3.6 percent retained on a 65 mesh screen
0.7 percent retained on a 150 mesh screen
25.7 percent passing a 150 mesh screen.
The bulk density of the fluffed, fibrous partial salt of collagen
was 2.0 - 2.5 pounds per cubic foot.
Upon disintegrating a sample of the fluffy, finely-divided fibrous
material in water at a solids concentration of 0.5 percent by
weight by subjecting the mixture to the action of a Waring Blendor
at high speed for 30 minutes, a stable dispersion was formed having
a pH of 3.20.
In the production of the liquid-laid, non-woven webs of the present
invention, conventional apparatus such as used in the papermaking
industry and in the production of non-woven webs may be used. The
finely-divided, fibrous collagen product is slurried in the
water-miscible organic liquid-water mixture by the use of a
suitable mixing device such as a beater wherein the beater is used
solely as a mixing device since the fibrous product does not
require hydration or fibrillation. The slurry or furnish may
contain from about 0.1 percent to about 3 percent of the fibrous
product, preferably about 0.5 percent. The slurry or furnish is
then passed to a suitable collecting screen where the fibrous
product is sheeted or deposited. Fourdrinier, cylinder vat,
Rotoformer and other sheet forming devices are satisfactory. After
removing the wet-laid sheet or web from the collecting screen,
excess liquid may be removed by passing the sheet between press
rolls and then drying the sheet.
The degree of swelling or hydration of the fibers in ethanol and
ethanol-water mixtures is illustrated by a freeness test using a
Canadian Standard Freeness Tester. The method of testing was in
accord with TAPPI Standard Test Method T227 m-58. In this series of
tests, slurries containing 0.3% fibers were formed in ethanol and
ethanol-water mixtures, as set forth in Table 1, by adding the
fibers to the liquid and manually agitating with a wide blade
spatula for 5 minutes. 1,000 mls of the slurry were then poured
into the cylinder of the Freeness Tester and measuring the volume
of liquid discharged from the side discharge tube. This volume in
mls. is reported in Table 1 as CSF No. and the data illustrated in
FIG. 1. This test illustrates that as the proportion of water in
the slurrying liquid increases, the swelling of the fibers
increases and, accordingly, the freeness decreases.
The variations in absorbency, tear strength, burst factor, tensile
strength, stiffness as well as other properties of the
hemostatic-adhesive non-woven webs of the present invention may be
illustrated by reference to the preparation of handsheets utilizing
liquids varying in relative proportions of ethanol and water. In
the preparation of these handsheets a modified 8 inch .times. 8
inch Williams handsheet mold was utilized. The normal wire mesh
screen at the bottom of the handsheet mold was covered with a
polypropylene filter fabric consisting of a 2/2 twill weave
structure, the fabric having a porosity of 85-90 CFM (Chicopee
Polypropylene 6016800 fabric) the edges of the fabric being sealed
to the wire mesh.
In each instance the ball valve located at the bottom of the
hand-sheet mold's water leg was closed and the specified liquid,
about 4,000 mls., poured through the polypropylene filter cloth to
bring the level of the liquid to just cover the polypropylene
filter cloth. For the preparation of hand-sheets of approximately 1
mm. thick (approximately 175.5 lbs. per 3,000 sq.ft.), 9.0 gms. of
the fluffy, fibrous material was added to 3,000 mls. of the liquid,
forming a slurry of about 0.37% by weight of the fibers, and the
slurry gently agitated for approximately 5 minutes. The slurry was
then poured into the handsheet mold and the slurry agitated by
lowering and raising a perforated plunger 3 times. The ball valve
was then opened to allow most of the liquid to drain by hydrostatic
pressure usually requiring from about 20 to 30 seconds. About 1 to
2 inches of liquid was allowed to remain over the polypropylene
filter fabric and thin areas or voids in the sheet on the filter
fabric were filled in with suspended fibers by gently agitating the
slurry with a spatula and moving fibers to the thin areas or voids.
When the sheet appeared to be quite uniform, the ball valve was
opened completely and all liquid allowed to drain.
The mold was then opened and the formed wet non-woven web covered
with a plain weave polypropylene filter fabric having a porosity of
about 90 CFM (Chicopee Polypropylene 6970500 fabric). Dry blotting
paper was sandwiched between very fine Dacron cloths and then
placed over the polypropylene fabric and gently pressed to absorb
some of the liquid. This procedure with the sandwiched blotting
paper was repeated a second time. A similar sandwich of dry
blotting paper and Dacron cloth was then placed over the
polypropylene fabric and the forming plate with the sandwiched wet
laid web removed from the mold and gently placed upside down with
the sandwiched blotting paper being placed on a sheet of polyester
film. The forming plate was then lifted and the wet laid web gently
peeled from the polypropylene filter fabric covering the forming
plate wire mesh. The exposed side of the removed wet laid web was
then covered with a polypropylene filter fabric (Chicopee
Polypropylene 697050 fabric) and a sandwiched blotting paper
between Dacron cloth placed over the fabric. The assembly was then
gently rolled with a printer's rubber roller to remove additional
liquid and this procedure repeated by rolling at 90.degree. to the
direction of the first rolling. A dry sheet of sandwiched blotting
paper was then applied after removal of the wetted sandwiched
blotting paper and the assembly turned over and a dry sheet of
sandwiched blotting paper placed on the upper side and the rolling
procedure was repeated. The sandwiched blotting paper and
polypropylene filter fabrics were then removed and the wet laid web
placed between polyester films and both sides gently rolled with a
printer's roller. The wet-laid webs between polyester films were
then stored in polyethylene bags until sufficient numbers of the
sheets were prepared to subject them to a drying step using either
a photographic dryer, freeze dryer, Noble and Wood hot plate or
vacuum oven.
Freeze drying was effected in a Repp Freeze Drier, Model 40, with
an initial shelf temperature of about -40.degree.C., a vacuum of 50
microns, heating cycle to 38.degree. C. over a period of 2 hours
and a condenser temperature of about -60.degree. C. Vacuum drying
was effected in a conventional vacuum oven at a vacuum of about 30
inches of mercury and a temperature of 35.degree. to 40.degree.C.
overnight.
Handsheets prepared as above described and dried by freeze drying
and vacuum drying were subjected to various physical testing after
heat sterilization by heating in an air oven at 120.degree. C. for
28 hours. Samples of the hand-sheets when dispersed at 0.5 % solids
in water in a Waring Blendor at high speed for 30 minutes formed
dispersions having a pH of 3.22 which was substantially identical
to the pH of dispersions formed from the original fibers.
The absorbency of freeze dried and vacuum dried products formed
from the slurries in different compositions was determined by the
use of a mixture of 90 volumes of ethanol and 10 volumes of water.
The central portions (1 1/2 inch in diameter) of plastic screw tops
of 4 oz. jars were removed leaving a sufficient annular flange to
cover the top edge of the jars. Samples of the various non-woven
sheets in the form of circular discs 2 inches in diameter were
placed on the top edges of the jars and the cut screw tops placed
on the jars to secure the circular discs on the jar tops. The
liquid was allowed to drip through a distance of one-half inch on
to the center of the sheet drop by drop allowing each drop to be
absorbed by the sheet before the succeeding drop was added. This
was allowed to continue until a drop of liquid dripped from the
bottom of the sheet into the jar. The flow of liquid was then
arrested. The percentage of absorbence was calculated from the
weight of the original dry sample and the weight of the wetted
sample, the data, the average for two samples, being presented in
Table 1. The data are plotted in FIG. 2.
The tensile strength of the various sheets was measured by the use
of a Thwing-Albert electrohydraulic tensile tester Model No. 37-4
at a 5.5 Kg and 31.8 Kg loading. In accordance with TAPPI Standard
Test Method T220 m-60, the test was applied to six strips of each
of the sheets cut 15 mm. wide and the results reported in Table 1
are the average breaking load in Kg for the 15 mm strips. The data
are plotted in FIG. 3.
Samples of the sheets were subjected to a tear test in accordance
with TAPPI Standard Test Method T220 m-60 using an Elmendorf Tear
Tester. The Tear Factor (average of two sheets) was calculated from
the average force in grams to tear a single sheet. The Tear Factor
as reported in Table 1 is equivalent to the number of square
decimeters of the sheet, the weight of which, if applied to a
single sheet would cause a tear in the sheet to progress. The Tear
Factor is plotted in FIG. 4.
Samples of the sheets were also subjected to a bursting test in
accordance with TAPPI Standard Test Method T403 ts- 63 using a
Mullen Burst Tester. The Burst Factor was calculated from the
pressure in psi required to burst the samples. The values reported
in Table 1 and plotted in FIG. 5 are the average values for two
samples.
The stiffness of the sheets was measured by maintaining samples
wrapped on a glass cylinder for 24 hours at room temperature
(73.degree.F) and a relative humidity of about 50 percent and then
releasing the samples and allowing them to flatten under their own
weight. The samples were cut into 3 in. .times. 7 in. strips,
wrapped around a glass tube 2.25 in. in diameter and held in place
by scotch tape. At the end of the 24 hour-period the tube was
placed on a horizontal surface with the mid portion of the sample
sheet in contact with the horizontal surface. The tape was then cut
and the free ends of the samples allowed to separate from the glass
tube and unfold or uncurl under their own weight. The amount of
uncurling was determined by measuring the angle between the
horizontal surface and the uncurled ends of the samples. The angle
in degrees is termed the Stiffness Factor, which is the average for
four sheets, both ends, and is reported in Table 1 and plotted in
FIG. 6.
The porosities of the sheets were determined by the use of a Gurley
Densometer (Closed-Top Model) in accordance with TAPPI Standard
Test Method T460 os-68. This test measures the air resistance of
the sheets and is reported in Table 1 as the average time in
seconds required to displace 100 ml. of air through an area of 6.45
sq.cm. of the paper. The reported values are averages for two to
six tests. These data are plotted in FIG. 7 and reported in Table
1. ##SPC1##
The data in Table 1 and FIGS. 2 through 7 illustrate that more than
about 15% water by volume in the ethanol-water mixtures used in
forming the slurries or furnishes for the preparation of the
liquid-laid, vacuum dried sheets causes excessive swelling of the
fibers and the sheets begin to exhibit hornification or
densification. Similarly, in the case of the freeze dried sheets,
where the ethanol-water mixtures contain more than about 20 percent
water by volume the sheets begin to exhibit hornification or
densification. This is particularly evidenced by the leveling off
of the absorbency (FIG. 2). Tear Factor (FIG. 4) and Stiffness
Factor (FIG. 6) and by the rapid rise in the tensile break load
(FIG. 3), Burst Factor (FIG. 5) and porosity (FIG. 7) of the
sheets.
Similar properties result when the other water-miscible organic
liquids are substituted for the ethanol. As indicated hereinbefore,
where the product is intended for surgical procedures it is
essential that such organic liquid and/or salt used in the
slurrying liquid must be removed.
The fibrous collagen products prepared in the above examples were
employed in surgical test procedures designed to provide the
efficacy of the material both as a hemostat and adhesive for
severed biological surfaces in a warm blooded animal when wet with
blood. "Severed" biological surfaces for the purposes of this
invention includes cut, sliced, ripped, abraded, torn, punctured,
burned, and tissue severed by any means or method whereby a fresh
biological surface is present. Biological surfaces will include
tissue, cartrilage, vessels, bone or other normal organic parts of
the warm blooded animal which may require mending or joining.
The in vivo surgical procedure was carried out on anaesthetized
mongrel dogs. The liver of the dog was exposed and a centimeter
square lesion was created by sharp excision of 1 mm. thick cortical
tissue to yield a uniformly bleeding surface. Swatches were cut
from samples of the various hand-sheets prepared as described
hereinbefore. The investigators were provided with swatches without
knowledge of the history of the samples so that all testing was
"blind." Each swatch was placed over the uniformly bleeding lesion
and pressure applied for 60 seconds, after which pressure was
removed. In the case of satisfactory samples, hemostasis was
effected within this time period. After 5 to 10 minutes, removal of
the excess outer marginal portions of the swatch was attempted by
pulling off such portions. Subsequent to rating the swatch, it was
forcibly removed and a uniformly bleeding lesion reestablished and
another swatch placed over the lesion as described and the swatch
rated.
The investigators evaluated the samples and rated them upon their
hemostatic efficacy, degree of adhesiveness to the bleeding surface
and delamination property. The rating was on an arbitrary scale of
0 to 5, 0 indicating that the sample exhibited no hemostatic
property, no adhesiveness to the wound surface and that the sample
could not be delaminated; that is, excess marginal portions could
not be removed without overcoming the adhesiveness and thereby
permitting resumption of bleeding. The delamination is highly
desirable so that in an internal surgical procedure, no more of the
material be allowed to remain than is necessary to effect
hemostasis and seal the wound and excess material be removed. A
surgical cotton gauze pad used as control was ineffective in
producing hemostasis, was not adhesive to the wound and, of course,
the entire pad could be lifted from the wound. Upon this basis it
received a 0 rating and was unsatisfactory. A rating of 1
indicating poor and unsatisfactory properties. A rating of 2
indicating a fair functioning but unsatisfactory. A rating of 3
indicating a good functional action and the acceptability of the
material is questionable. A rating of 4 indicating a very good
functional action and satisfactory. A rating of 5 indicating an
excellent functional action and satisfactory.
The investigators' ratings of the samples based upon the
evaluations and based upon the foregoing considerations are set
forth in Table 2. In addition, the investigators also observed the
handling characteristics of the samples. These characteristics
included a consideration of the physical properties such as
cohesiveness and flakiness of the sheet, the friability, the
stiffness and the ability to cut swatches of the required size from
the sheet with scissors to form a clean cut. The investigators
comments with respect to these characteristics are set forth in the
Table 2. ##SPC2##
Although the sheets prepared from the 100% ethanol slurries
received high in vivo ratings, based upon hemostatic efficacy,
adhesiveness and delamination, the sheets are not deemed
satisfactory from a handling viewpoint. Because of the loose,
crumbly and friable characteristics of these sheets, the products
may be said to be delicate and can not withstand normal shipping
and handling. Upon handling, the sheets exhibit excessive flaking
and loss of fibers. Such sheets can not withstand slight
abrasion.
Where the sheets are too stiff and are too harsh and boardy, they
have a low hemostatic efficacy, low adhesive quality and cannot be
delaminated without opening the wound. Furthermore, such sheets do
not readily conform to irregular surfaces encountered in a wound
and, hence, are unsatisfactory.
From the foregoing discussion and the in vivo evaluation, it is
clear that severed biological surfaces may be joined and a wound
sealed without the use of sutures by the application of the sheets
or webs of this invention. In sealing the wound, the web when
wetted with blood combines with the blood to form a mass that is
self-adherent to the wound tissue to seal the wound. It is only
necessary to apply pressure to the web for a short period
sufficient only until hemostasis has occured after which pressure
may be removed.
Based upon the evaluations of the various handsheets, products
satisfactory for the purposes of the present invention have
physical properties within the following ranges:
Absorbency of a mixture of 90 volumes of ethanol and 10 volumes of
water from about 100 % to about 300 % by weight;
Tensile strength in Kg. per 15 mm. strip from about 1 to about
4;
Tear Factor from about 5 to about 50;
Burst Factor from about 1.2 to about 6;
Stiffness Factor from about 15.degree. to about 100.degree.;
and,
Porosity in seconds to pass 100 ml. of air per 6.45 sq.in. from
about 4 to about 15.
The absorbency and porosity values and Stiffness Factor are
probably the simplest and most readily determinable properties.
These properties considered together for sheets or webs of a basis
weight of approximately 175.5 pounds per 3,000 square feet are not
only a fair indication of the ability of the sheet to withstand
normal and ordinary shipping and handling without flaking, but also
a fair indication of the hemostatic-adhesive efficacy.
Freeze dried products prepared from slurries of the fibers in 90
volumes of ethanol and 10 volumes of water and vacuum dried
products prepared from slurries of the fibers in 95 volumes of
ethanol and 5 volumes of water have closely similar physical
properties and from the in vivo evaluations are about equivalents
and the preferred products.
* * * * *