JPH05502178A - Hollow visceral prosthesis and implantation method - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Hollow visceral prosthesis and implantation method For products and techniques used to repair or restore the walls of excised organs or other body parts. related. More particularly, the present invention relates to transplantation or end-to-end transplantation of such organs or body parts. Restoration of part of the organ wall or relates to an implantable prosthesis that can be integrated into the surrounding tissue for the replacement of whole organ walls. .

State of the art: Numerous diseases or diseases requiring replacement of hollow internal organs or partial restoration of organ walls. has symptoms. A more common of these diseases or conditions is unresponsive stricture of the esophagus. Esophageal cancer, diabetes mellitus, with rates reported as 1% of all malignancies and 4% of gastrointestinal malignancies Neurogenic bladder, a non-responsive neurogenic cause of voluminous retention, present in some 83% of patients. Urine with ureteral or urethral obstruction as a result of sexual bladder, trauma, congenital defect or tumor formation Duct obstruction: Bladder tumor formation that accounts for 2.5% of all cancers.

There are many other diseases or conditions in which organ or tissue replacement or restoration would be invaluable, but The technology to do so is lacking. Examples of these are: New stoma opening and retrieval devices often cause significant pain, discomfort and inconvenience. There are various diseases that occur, such as cancer or idiopathic mucosal ulcerative colitis. For example, the trachea There are situations in which the trachea must be removed, and removal of more than 8 cm of the trachea requires Preventing end-to-end anastomotic repair. Currently, there are no reliable tracheal replacements.

Said diseases and symptoms may be caused by other hollow visceral organs instead of diseases, such as in the case of graft reconstruction. It was often treated by surgical replacement or replacement of tissue from other organs. example For example, the esophagus may be replaced by a replacement portion of the colon or, more commonly, by an esophagogastroresection. available. The lower ureter is treated by partial to total cystectomy along with ureteroileal anastomosis. It will be done. In addition, reservoir ileal fistula formation and mucosal rectal incision are performed along with ileoanal anastomosis. The use of ablation has been utilized after body cavity resection; A nearby operation was developed.

All of these operations require long surgical procedures and result in a significant number of postoperative complications. In some cases external collection equipment is required. In addition, some of these operations may be satisfactory. There is a certain obvious aesthetic effect that cannot be helped.

In addition to the use of donor tissue or organ substitutes to treat these conditions, polymer - and metals have been used alone or in combination for esophageal or lower ureteral replacement. Many A number of materials have been suggested for use as prostheses for tracheal, esophageal and ureteral replacement.

For example, E. F. Barbman, “Experiments on a part of the esophagus using a plastic tube. ``Replacement'', Volume 135, Ann's Sargieli - March 11952, No. 3 pp. 37-43 (E, F, Bergman, in″The Experiment dental Replacements of Portions of the Esophagus by a Plastic Tube", 135 A nnalsof Surgery, March 1952. pp, 337- 43) used methacrylate or polyethylene tubes as an alternative after esophageal surgery. Suggested the use of Bing. Furthermore, Battersby et al. ``Esophageal Replacement with a Tube'', A, M, A, Arches of Surgery-11 954, pp. 400-09 (Battersby, et at, “Eso Phageal Replacement with Plastic Tube s”, A, M, A, Archives of Surgery.

+954. pp, 400-09 (about polyethylene and nylon lamination) ``Placement of a Portion of the Canine Esophagus with a Composite Prosthesis and an Ornament'' ”, Volume 64, Journal of Thoracic and Cardiovascular Surgery C.Surgery, December 1972, pp. 892-96 (Barnes, et al,, “Replacement of Portion of Ca nineEsophagus with Composite Prosthe sis and Greater Omentum,”64 Jr., of 96 (disclosing the use of polyurethane) Tsukushima et al. Seven-year follow-up study after replacement of the esophagus with an engineered esophagus, Volume 93, Surgery Department January 11983, pp. 70-77 (Fukushima, et al., '5even-Year Follow-up 5tudy After th eReplacement of the Esophagus a n　Artificial　Esophagus　1nthe　Dog,”　9 3 SurgerY, January, +983. pp, 70-77 (discloses silicone rubber tubing with external Dacron tube) Light. E. Michelson et al. ”, Volume 41, Journal of Thoracic and Cardiovascular Surgery Tracheal Replacement Polymers Tested in Surg. 1961, June 748-58. A number of devices are discussed and discussed. Gore-Tex” ) has been used for ureteral replacement, but with limited success. S., Birdy et al., “New synthesis Substance, Ureteral Replacement with Core Tex” Volume 128, Journal of Uroro G. July 1982, pp. 171-75 (S. Vardy, et al. “Ureteral, Replacement with a New 5 ynthetic Material: Gore EX,” 128 Jr. of Llrolo (By, July, 1982.pp, 171-175 ).

If a synthetic prosthetic device is used for replacement or restoration, the prosthesis! and surrounding organizations Anastomotic leakage, infection, intraluminal or extracorporeal prosthesis after prosthesis suture line Several significant problems were observed, including protrusion of the tube, appearance of hollow tubes, and migration of the prosthesis. It was noticed. These problems are a result of the type of prosthesis used and the technique of implantation. occurs.

For example, suture leakage usually involves improper or insufficient sutures and viscous closure of the prosthetic tube. The result is the specific method employed to contact the membrane. Infection occurs at the anastomosis site. resulting from leakage or failure of aseptic technique during prosthesis implantation.

However, migration and suture arytenoids are a result of the unique properties of the epithelium. The epithelium is The epithelium continues to grow along the stroma until it comes into contact with other epithelium, and the epithelium is essentially nothing other than a mucous membrane. Nothing can be permanently recovered. In the case of non-biodegradable prostheses, the mucous membrane It begins to move inside the endothelial organ, but the blood vessels are coated with certain polymers, such as Silas glue. The mucous membrane cannot quickly replenish its blood supply because it cannot penetrate the silastic membrane. Instead, the mucous membrane falls off toward the suture line. Next, the arytenoids of the suture line are formed. Jiru. The mucous membrane then grows outside the prosthesis and drains its blood from the connective tissue around the prosthesis. Receive supply. This results in protrusion of the prosthesis into the lumen or formation of a hollow tube. .

Some of the problems that arise with implantation of synthetic matrix materials are due to the application of collagen to the synthetic matrix. Overcame. Fiber protein, the main extracellular structural protein of connective tissue and bone , collagen forms a structural continuum by binding cells together to form a weave. to be accomplished. Y. Shimizu et al., “Research on Collagen Copolymers and Synthetic Polymers” Research (First Report), Volume 5, Biomad Met Deb Art & O. 1977, pp. 49-66 (Y, Shimizu, et al, “5 tudieson Copolymers of ColCo11a and a 5ynthetic Polymer (First Rep-ortY, 58ioma1. Med, Dev, Art and Orq, 1 977, pp. 49-66), “On Composites of Collagen and Synthetic Polymers” ``Research (Second Report)'', Volume 6, Biomad Met Deb Art and ・Oga, 1978, pp. 375-391 and “Collagen and Synthetic Polymers” Research on Composite Materials', Proceedings of the Second Meet April 1979 ("5tucly on Cr+mpo site of ColCo11a and 5ynthetic Polym er,”Proceedings of the 5th Meeting g of + SAO, April 1979) discovered that collagen and synthetic A copolymer of substances is responsible for the formation and growth of natural connective tissue promoted by collagen. The results suggest that it is highly useful for use as a prosthetic biomaterial. non-absorbable fiber Collagen prosthetic implants using fibrous reinforcement include Artandi et al., U.S. Pat. No. 3,272,204, which can be used in surgical procedures. Collagen-coated silicone rubber is disclosed in U.S. Patent No. 3.955.012 by Saikimura et al. Disclosed in the specification.

In traditional synthetic matrix-collagen copolymers, collagen is attached to the epithelium throughout the matrix. It has been shown to promote growth. Collagen is eventually absorbed by the body and converted into synthetic radicals. Leaving a layer of epithelial tissue located on top of the epithelium. Epithelialization on the stroma is caused by these devices. However, serious problems still remained. For example, the growing epithelial layer lacks angiogenesis. growth is eventually restricted. It was also found that epithelialization does not occur completely on the stroma. Ta. Additionally, growth cannot be controlled and excessive connective tissue growth can cause narrowing of the ducts. . Ineffective methods of collagen application also cause graft leakage.

A problem with implanting synthetic prosthetic devices such as those described above is the interaction between the prosthesis and the body's connective tissues. promotes primary contact and promotes epithelial growth within the lumen of the prosthesis or organ, while vascular Providing a prosthesis that facilitates formation and anchors connective tissue with synthetic pores This will be overcome.

Summary of the invention The present invention addresses the difficulties encountered with conventional synthetic prosthetic forms in transferring tissue or internal organs of the body. Provides unique substances for planting. That is, by the controlled application of collagen to synthetic matrices, In addition, the present invention utilizes regulated growth of connective tissue for ultimate incorporation of the device into surrounding body tissues. Promote longevity.

The present invention provides that the pores of the matrix are occluded with collagen, thereby preventing fluid and bacterial entry. Collagen or porous synthetic polymer base applied to allow matrix retention It's also about quality. This impermeability is achieved by the present invention for reconstruction or replacement of the ureteral, respiratory tract or gastrointestinal tract. This is particularly important when used in pipes.

Synthetic substrates are polyethylene to polyurethane, polypropylene, silicone rubber or From any tissue compatible and relatively inert synthetic polymer such as Teflon Usually formed. The pores formed therein may have uniform or irregular shape and size. Ru. However, irregularity in porosity increases the strength of the epithelial bond to the stroma and only It also regulates the amount of granulation tissue (initial connective tissue) produced. like tantalum Porous synthetic metallic materials may also be used in the present invention. “Synthesis” as used here refers to the desired When in its desired form, it is not natural and must be specially processed for the purposes of this invention. Substances of natural origin, metals, or polymers.

Controlled growth of connective tissue into the pores of the matrix ensures optimal incorporation of the invention into the body. is important above. That is, the rate at which collagen is absorbed by the body and the resulting The formation of connective tissue on the substrate influences the rate of epithelialization on the substrate, which in turn influences the rate of final incorporation of the device. It also affects the degree. For example, in previous collagen-coated implants, insufficient If connective tissue ingrowth occurs from the stroma to the lumen, it should be Directed epithelial growth was observed to cease. This is the blood supply in connective tissue This is due to a lack of epithelium, which causes the epithelium to separate and fall off toward the suture line. epithelium is endogenous If the connective tissue does not grow over the long connective tissue, it will continue to grow and fill the lumen of the internal organs. This causes stenosis. Controlled application of collagen to the stroma results in a lack of connective tissue within the stroma. Improves insufficient or excessive ingrowth and regulates optimal growth of connective tissue.

The increase and control of connective tissue ingrowth into the stroma and subsequent epithelialization are caused by the coating on the stroma. This may be promoted by the addition of fibroblasts to the collagen. Fibroblasts are Fibrous cells found abundantly in connective tissue; collagen in connective tissue It is the source. When added to collagen, fibroblasts bind collagen to the matrix. interact with and produce their own collagen, thus increasing the amount of connective tissue on the matrix. Promote ingrowth to ensure complete integration of the graft into the surrounding tissue.

Collagen coating is exposed to radiation, ultraviolet light, glutaraldehyde or polyglyceride. Any of several means including role polyglycidyl ether (PPE) It may be crosslinked with Cross-linking increases the density and strength of collagen, and expand slightly within the matrix pores to strengthen collagen anchoring to the matrix. Melt. Does cross-linking increase the density and strength of collagen coatings on matrix pores? , crosslinking may be unnecessary or undesirable in some applications.

In addition to the porous synthetic polymer used as the matrix, elasticity can be added to the implant if desired. Alternatively, other synthetic materials may be used as substrates to provide prosthetic devices. i.e. some applications a certain amount of elasticity is desired. 11" Z, L) Implant expansion and contraction filling and prosthetic implants for the bladder required during evacuation. Typical bullets that may be used Examples of the material include silicone rubber (Silastic 6) or polyurethane.

The best efficacy of the graft is to combine the mucosal epithelium from the surrounding viscera or tissue with the intraluminal surface of the graft. Achieved by bonding. The epithelial lining of many organs is It is important for Noh. For example, the mucosal lining of the trachea moves debris from the lungs Contains fimbriae. Grafts of mucosal epithelium from the trachea or nasal sinuses are grafts directed into the lumen. It may also be attached to the surface of the body or prosthesis. With angiogenesis and epithelialization within the synthetic matrix , the endoluminal graft ultimately performs its function.

The incorporation materials of the present invention are suitable for a variety of medical or surgical applications. for example, Incorporation materials can be used to repair or repair holes in internal organs caused by accidental trauma or necessary surgical removal. are manufactured in batches to restore or dilate stenotic lumens.

As an example for this application, 9% of infants placed in front of a ventilator will have subglottic stenosis. Rub. This often progresses to the point where the entire glottis becomes obstructed. the current, There are no prosthetic devices available to widen the narrowed glottis, and only costal cartilage can be used. must be done. This maneuver usually involves a long tracheostomy and lane cut in the glottis. It takes. The use of a patch that rapidly epithelializes and integrates throughout can further improve subglottic stenosis. Reduce morbidity in severe cases and improve recovery in these patients.

On the other hand, incorporation materials may be used for transplantation into the trachea, esophagus or intestine where end-to-end anastomosis is not possible. It may also be formed into a tube. Most esophagectomy is performed for cancer of the esophagus or the cardiooesophageal junction. performed in many cancers. Esophagectomy is also required for some benign lesions and chemical burns. release The use of radiation therapy or chemotherapy, either individually or in combination, is recommended for the treatment of esophageal cancer. The intermittent need for surgical resection of the esophagus could not be resolved. saliva and food Nonsurgical palliative use of endoluminal esophageal tubes to preserve the lumen for passage is usually It was found to be insufficient as a palliative measure for esophageal cancer and inferior to surgical resection. . Endoluminal tubes are sometimes used as a last resort or not used at all.

Palliative esophagectomy and wide resection for esophageal cancer are associated with the use of chemotherapy and radiation. Without this, most patients have a very short survival time of less than 2 years from the time of diagnosis. It will be limited. Despite a limited survival prognosis, many patients Major surgical procedures involving surgical invasion of both the thorax and abdomen are required for the production of explants. It will be provided.

Currently, the maximum amount of trachea that can be successfully resected is 6-8 ann due to tension. neville (Nevile) prosthesis (polyester-coated Dacron soy on either end) Medical grade silicone copolymer tubing (with rings) is sometimes used. However, this does not allow for intraluminal connective tissue ingrowth or epithelial ingrowth, and does not cause protrusion or Or, fistulas are often formed. Most of these major surgical procedures It will develop a high morbidity rate in older or debilitated patients. Artificial segment of esophagus According to use, surgical procedures can be performed on the thorax and possibly the neck without the need for abdominal procedures. It can be limited. Resolution of abdominal procedures is through the gastrointestinal tract in most patients. Immediately restores nutritional intake after surgery. It also solves the problem of intestinal obstruction after flank incision. , shortening the postoperative hospital stay for these patients with a very limited prognosis. Shrink.

In another embodiment, the invention is used as a prosthetic replacement for a whole organ or organ system. Ru. For example, bladder replacement is required after cystectomy for bladder cancer. Reported cases of bladder cancer There are 37,000 to 40,000 cases per year, of which about 25% ultimately Cystectomy required. In addition, nerves related to cervical or rectal cancer or atrophic bladder Obstruction and removal of pelvic contents (selective organ removal) may require ureteral diversion or cystectomy. There's a lot to do. Current methods for cystectomy and urinary diversion usually involve the ileum or Utilizes the intestinal conduit from the colon. These procedures carry high complication rates and Requires the use of external instruments and places a urological handicap on the patient. It becomes.

A number of different repair methods have been proposed and tested in recent decades, although few Which methods fail due to infection, inflammation, rejection, protrusion, obstruction, leakage or fistula formation or was found to be insufficient.

The only method with limited success has been to leave part of the bladder intact and to This method provides a source for the formation of more transitional cell epithelium.

The present invention overcomes the consequences associated with previous prosthetic devices by being able to integrate into surrounding tissues; provides a functional device for applications such as

FIG. 1 is a partially cutaway perspective view of a porous synthetic substrate.

FIG. 2 is a cross-sectional view of a substrate showing an open-celled arrangement of pores: Figure 3 is a cross-sectional view of the matrix in contact with collagen; Figure 4 shows connective tissue ingrowth and tubes. In a cross-sectional view of an implanted prosthetic device showing the epithelial layer on the intraluminal and abluminal surfaces. Figure 5 is a cross-sectional view of the prosthetic device showing the host graft epithelium on the endoluminal surface. be; Figure 6 is a perspective view of an embodiment of the invention comprising a prosthetic punch; Figure 7 is a hollow tube. FIG. 8 is a perspective view of an embodiment of the invention comprising a bladder prosthesis; FIG. FIG.

Detailed description of the aspects shown The present invention can be applied to either collagen or a mixture of collagen and fibroblasts. Usually includes a porous matrix material that is touched. A layer of autograft or tissue cultured mucosal epithelium is applied to one surface of the substance, i.e. the intraluminal surface, to promote the growth of mucosal epithelium . The prosthetic material of the present invention can be used to implant patches, prosthetic tubes or prosthetic organs. It takes any form, including. The basic materials and prosthetic shapes are detailed below. The quality matrix can be any tissue-compatible porous synthetic material, including metallic materials and organic polymers. It may be a substance. Inert in the body, such as polyethylene or polyurethane. Organic polymers are preferred. In a preferred embodiment, the substrate is a polyethylene material. , and then a substantial number of irregularly shaped open-celled pores are formed. . An example of such a material is Porex Medica. l　Co, ) [Fairburn, Georgia (Fairburn, Ga.)] Medpor, manufactured by Medpor. As shown in Figure 1, this type The pores 22 in the synthetic material 20 are irregular in shape and size, and are Zushi is not limited to straight roads either. The diameter of the aperture is small enough to allow objects to pass through it. As usually measured in the diameter of Ru. For purposes of this invention, pore sizes of about 100 to 360 microns are preferred.

The thickness of the matrix affects the rate and amount of connective tissue ingrowth. That is, the substrate is thicker. As the size increases, the depth of the pores into which the connective tissue must grow increases. I wanted to The thickness of the substrate is approximately 0.25 millimeters (mm) to approximately 1.50 cm. It is within the range of Tor (Ro).

If a thin sheet of this porous synthetic polymer is cross-sectioned as shown in Figure 2, It can be seen that the holes 24 form a curved and irregular path. A substantial number of holes are opened. The air release bubbles 25 indicate that there are openings in the pores on both surfaces of the substrate. means. However, closed cell 26, i.e. openings on only one surface There may be several pores with . Irregular porosity in the matrix is Enhances the attachment of collagen or collagen and fibroblasts to the tissue. In addition, Ingrowth of connective tissue into irregularly shaped pores increases the strength and adhesion of the tissue to the matrix. The collagen is attached to the matrix and non-liquid to ensure the incorporation of the prosthesis into living tissue. Either method fills the pores with collagen sufficiently to create a permeable seal. The matrix is then brought into contact with the collagen. As shown in Figure 3, collagen 28 substantially fills the pores of matrix 30 and deposits a thin layer of collagen 32 on one surface of the matrix. Limit layers.

The prosthetic material of the present invention can control the ingrowth of vascularized connective tissue. can control the ingrowth of the epithelium. This is the collagen applied to the matrix. By varying the thickness and/or density (i.e. the concentration of collagen in solution) as well as by varying the thickness of the substrate. Therefore, collagen or The thinner the collagen fibroblast coating, the faster connective tissue ingrowth I can smell it. Conversely, if the collagen coating is thicker and/or more concentrated, In some cases, connective tissue growth slows down. The thickness of collagen is about 0.025 to about 1. It's oomm.

The faster the connective tissue ingrowth, the faster the epithelial growth. At the suture site Greater connective tissue ingrowth and epithelial growth are thinner and denser at that location. facilitated by a thinner collagen layer or thinner matrix. On the contrary, there is excess inside Luminal narrowing caused by connective tissue growth results in a thicker, denser collagen layer or more Avoided in thick substrates. This allows epithelialization to continue. collage The absorption rate and subsequent connective tissue ingrowth of collagen or collagen and Adjustment may be made by varying the depth to which the fibroblasts are applied in the pores of the matrix. Ko Collagen absorption is also regulated by cross-linking of collagen as described below.

FIG. 4 shows connective tissue in which collagen 36 grows into pores 40 of matrix 42. 38 will be replaced. Connective tissue angiogenesis 44 occurs as epithelium 46 grows. maintain the viability of the organization.

Any type of non-antigenic collagen may be used to coat the matrix. Like Alternatively, atelocollagen (Koken, Tokyo, Japan) may be used. Atelocholla Gen is an enzyme-solubilized collagen that lacks the strongest antigenic determinants, resulting in a It has almost no antigenic activity. Atelocollagen is biodegradable and has significant It exhibits little inflammatory response and is particularly suitable for sealing pores in matrices due to its viscosity. There is.

Purchased from Koken Co., Ltd. (161 Japan, 3-5-18 Shimoai, Shinjuku-ku, Tokyo). Telocollagen is kept in its dry form in a refrigerator at about 40°F. dried aterocola -gen in an appropriately sized glass vial with a screw cap, usually 20 mm. So 0. OI M Dissolved in CHsCOOH (glacial acetic acid).

As mentioned above, the rate of collagen absorption varies with the density of collagen applied to the matrix. It is adjusted by increasing the amount of water. Density as used in this disclosure refers to collagen in solution. related to the concentration of molecules. The concentration of collagen can vary from 1-5%.

For example, a 5% solution of collagen contains 1 gram of solubilized collagen at 0.10 M C Prepared by adding 9 milliliters (-) of HICOOHl. The solution is It is maintained for about 24 hours to completely dissolve the collagen. The solution is 2 during this period. Stir ~3 times. The collagen is then stored in a refrigerator. Collagen or dissolved Once understood, the solution should not be used within 4 days after preparation unless there are signs of contamination. It can be used. The concentration of collagen in the solution depends on the desired use of the device. determined accordingly. Therefore, if faster dissolution of collagen is required, lower percentage of collagen is used. The slower connective tissue ingrowth is required, the more A high collagen percentage is used.

One method of contacting atelocollagen with a matrix is as described in the Examples below. obtained by Example A- Metobore (Bole, Kuss, Fufu) with a pore size of approximately 100 to 300 micrometers Using scissors, cut a 2cm (on) x 4an piece of It was cut to approximately the size of a patch. When the batch was weighed, it was 3.14 g. . An equal amount of 14 g of prepared 5% solubilized collagen was weighed. collagen spa It was applied onto the methobore with a chula to cover the top surface of the methoboa. coated methobo The sample was immediately immersed in a 5% NaCf solution for 1 hour. Collagen is cloudy and began to form random non-parallel fibrils of collagen.

On the other hand, the coated Methobore pieces were prepared using phosphate buffer (PB) immediately after coating the Methobore pieces. S) May be immersed in 100-. This operation allows collagen to form natural or parallel fibrils. Form a rill.

Collagen attachment to matrix Collagen contact with irregular porous matrix penetrates into the intricate cavities of the pores. for collagen to enhance anchoring of collagen to the matrix. This machine As a result of mechanical anchoring, no further bonding between collagen and matrix is required. be. Furthermore, atelocollagen is added to fibroblasts in culture as shown in Example C below. When mixed with cells, the fibroblast culture medium usually contains the gelatinous material atelocollagen. raw and further liquefied in consistency. Atelocollagen to substrate in liquefied state The contact between cells and fibroblasts is due to the ability of atelocollagen to penetrate into the irregular pores of the matrix. increases the force so that no further bonding is required.

Collagen crosslinking 2.5-5% Atelocollagen is usually very soluble and does not naturally dissolve in liquid environments. not maintain oneself. Reduces the solubility of atelocollagen and before it is absorbed in the body To make it last longer, it may be crosslinked. Crosslinking is collagen poly Increases the density of collagen by inducing bonding between mer chains. Is it the degree of crosslinking? The higher it is, the denser and less biodegradable the collagen is. For this reason, The absorption rate of collagen depends on the concentration of collagen in the coating, and the Recognize that it depends on the amount of crosslinking and the porosity or porosity of the pores in the substrate. Can be done.

Crosslinking is a technique well known in the art and can be achieved by radiation, ultraviolet light, or glutaraldehyde. This can be done in many ways, including using different chemicals.

Method for inducing crosslinking by the use of glutaraldehyde Hari, T., Cheno and M.E. Nimni, “Mechanism of protein cross-linking by glutaraldehyde I: modeling. "Reaction with Compounds", Volume 10, Connective Tissue Research, Volume 187-9 9 pages, ■982 CD, T.

Cheung and M.E., Nimn1. “Mechanism of Cro sslinking of Proteins by Glutaraldehy der:Reaction with Model Compounds,” lOConnect.

Ti5s, Res, 187-99 (1982)) and D.T., Cheno and M., E. Nimni, “Mechanism of protein cross-linking by glutaraldehyde ■. : Reaction with monomeric and polymeric collagen, “Volume 10, Connective T. Tsu Research, pp. 201-16, 1982, and among them: Contents are incorporated herein for reference. The method of inducing crosslinking by the use of radiation is RJ, Davidson and Tori.

R. Cooper, “Effects of UV irradiation on acid-soluble collagen”, Vol. 105, Biochemical Journal, pp. 965-69, 1967 (R, J, pa vidson and D, R, Cooper, Go he Effect of U ltraviolet Irradiation on Ac1d-5olub le ColCo11a, “105 Bioch However” Yu, 965-69 (1967)) D. R. Cooper and R. J. Davidson, “Soluble CO “Effect of UV irradiation on Lagen”, Volume 97, Biochemical Journal , pp. 139-47, 1965 and T. Myata, T. Sonde, A.L., Lupi. N. and H. Stenzel, “On natural and telopeptide-deficient collagen. Acta of ultraviolet irradiation, pp. 672-680, 1971 (T, entry 1i ya) ta, T, 5onde.

A, L, Rubin, and K, H, 5tenze1. 'Effect s of Ultraviolet 1rradiation on Nati ve and Te1opeptide-poor ColCo11a,” The contents of 229 Biochi are incorporated herein for reference.

Examples of induced crosslinks are:

Example B: Coating the substrate to be measured for 2anX4an with collagen (as described in previous Example A) and precipitated the coated substrate patch by immersing it in a 5% NaCl solution. . 100 ml of glutaraldehyde crosslinking solution is mixed with 50 ml of MeOH and 1 ( ,050- was prepared by adding . Transfer the solution onto a magnetic stirring plate for 15 minutes. 0-Ersin Meyer flasks. The pH of the solution is IN NaOHO , 5yd was added to adjust it to 11.5. 25% glutaraldehyde 40mg Add Little to the solution and bring the pH to 12 by adding IN NaOHO, Irn1. It was adjusted. The solution was stirred slowly on the plate for 1 hour.

Immediately after sedimentation with NaCl in collagen, the batch was added to the glutarium in a capped vial. Immediately placed in aldehyde solution. The patch was completely submerged in the solution.

The vials were placed in a shaking water bath at 25-26°C for 2 hours to form a high degree of crosslinking. (It takes less time if fast dissolution of collagen is required).

After 2 hours, remove the batch from the glutaraldehyde solution and add sodium azide. (NaNs) containing 100% of phosphate buffer (phosphate buffer is potassium phosphate). Mu (KH, PO4) 0.272 g/12. Sodium phosphate (NaJPO -) 1.136 g/l and sodium chloride (NaC1) 8.474 g/l Add j7 to 250 mj' of distilled water and then fill to obtain a solution of 1000 mj'. Prepared by adding sufficient amount of distilled water. This is added to sodium azide (NaNf ) Add I g/l).

Batch in PBS and sodium azide in a shaking water bath at 25-26 °C for 5-7 days. I paused. The solution should be administered twice a day for the first 3 days, then once a day without walking. changed. After performing this solution exchange operation, it may have remained in the collagen. Unreacted glutaraldehyde was removed.

At the end of this period, the batch is kept in a saline solution containing sodium azide and used for surgical implantation. It was kept in the refrigerator for up to 24 hours.

Approximately 24 hours before implantation, the batch was removed from the sodium azide-warm water bath. next It was placed in a sterile container containing sterile saline. Brine the first 12 minutes using aseptic technique. I changed the time 5-6 times. The final exchange included 1 g of cefadil solution/100 O + t + j'. It was in bacteriostatic saline. The batch was stored in the refrigerator during this time.

At this point, the device manufactured by the above procedure is considered ready for implantation and is It is handled using techniques.

fibroblast The matrix can be contacted with atelocollagen alone or with atelocollagen as described above. and fibroblasts may be applied to the substrate.

Fibroblasts are ring-shaped dense fibrous elastic tissue-like fibrillar tissues and entire membranes. type and is a source of collagen fibrils. tissue cultured fibroblasts Atelocollagen is added and then brought into contact with the porous matrix. Fibroblasts are that They produce their own collagen, causing atelocollagen to contract and become more dense.

This shrinkage and increased density causes the Kolaken-fibroblast coating to further penetrate into the pores of the matrix. No crosslinking is necessary for strong adhesion. In addition, cross-linking damages viable fibroblasts or kill them. Intraviable cells formed by fibroblasts within the stroma The contents increase the ingrowth of connective tissue. This viable membrane within the matrix also contains cross-linked attenuators. Locolage> Transports nutrients to the substrate faster. Transport of nutrients is essential for connective tissue growth. is important and therefore ensures better survival of mucosal or tissue culture grafts. Ru.

One method of contacting the porous matrix with the collagen and fibroblast mixture is as described below. Illustrated in the examples.

The dog's oral mucosa was collected, washed with 1 liter (1) of phosphate buffered saline (PBS), and then Once removed, the cells were cut into small explants of approximately 2an x 2an. These explants are 25 cu. Place it on the upper seam of the lasco and add 10% tallow serum (FBS) containing Dulbecco's modified 9mi+ of sexual Eagle's medium (DMEM) was supplied.

Dulbecco's Modified Eagle Medium is available from Gibco, Grand Island, New York. Tissue culture medium purchased from Gibco Labs. The medium is 1000 yd of distilled deionized water, DMEM powder (#430-1600, Gibko Kata) Log) 10 grams (g), sodium bicarbonate 3; 7g, j! -Glutamine 0. 3g5N-2-hydroxyethylbiverazine-N'-2-ethanesulfonic acid (H EPES) 4,76 g, amphotericin BZ5■/10.5- and penicillin It is prepared by mixing together streptomycin and streptomycin. pH of mixture Increase to 7.3 with IMNaOH. Filter the mixture using aseptic technique. Then Utah Cow fetuses purchased from Hyclone Labs in Logan Add 100 - of baby serum (FBS). Fetal bovine serum is blood extracted from fetal bovine blood. It provides a nutrient medium for cell growth.

Place these flasks so that only the culture medium or the seams of the flasks are located (= slightly tilted 3 Incubate at 7° Celsius (C). Fibroblasts grow from the explant and It begins to move downward into SC. After 3 weeks when the fibroblasts have sufficiently proliferated, culture the cells. Transfer to a culture dish and grow in DME medium for 7 days.

fibroblast coating A 2anX3an patch of Metopore 1 porous synthetic matrix is applied by pipette. 1. Cover with 25% atelocollagen l-. The coated batch was heated at 37°C for 1 Leave for 6-24 hours to sole the pores in the substrate.

Then about 4X10' fibroblasts containing 1,25% atelocollagen 11711 were added. Pipette onto each batch. The coated batch was then treated with DM containing IO% FBS. Put it in E5rnl. As fibroblasts proliferate, they develop their own core. After 62 weeks of starting lagen production, the batch is tested for leakage. This is a medium Place the batch on a ring on a plate. DMEMo, 5E Little on batch Transfer the DMEM from the batch to a container and observe the batch for 5 minutes. Check the output put out

If any leakage is observed, add an additional 1.25% collagen and fibroblasts. Add 1d to each batch as above. Tests for leaks can be batch or normed. Do this every two weeks until the

Many cell types of epithelial and endothelial origin contain H-like extracellular matrix such as collagen. When placed on Trix, cell proliferation, cell differentiation and gene expression are promoted. Proliferates in such a culture environment. Such proliferation and differentiation functions are present in human liver cancer and human liver cancer. -Ing sarcoma, aorta, pulmonary artery, umbilical vessel, bovine vascular endothelium from aorta, adrenal cortex , using bovine capillary endothelium from the brain cortex and corpus luteum, and human upper respiratory epithelium from nasal polyps. It has been proven that Such cells have resisted previous successful attempts in culture. but However, when placed in serum-supplemented cultures on collagen, they remain firmly They attach, migrate, and proliferate to form flat, nonoverlapping, closely juxtaposed epithelial layers.

Functional characteristics such as mucus secretion, ciliary migration, expression of tissue-specific antigens and other similar characteristics signs are proven. Typically, such cultures have a Similar to its in vivo counterpart.

The use of autografts of epithelial tissue, such as oral mucosal epithelium, improves the physiology of the prosthesis in the short term. By making it more functional, the prosthetic device can be more easily integrated into the host tissue. It is clear that it can be done. “Autologous” as used here refers to internal organs or surrounding areas of the host. The present invention relates to donor tissue extracted from m-tissue. Tissue may be removed from similar individuals. , in which case surgical treatment is performed. Figure 5 shows collagen 54 and fibroblasts 5 6 shows the placement of a graft epithelium 50 on a substrate 52 coated with 6.

Autograft epithelium can be directly or in vitro cultured with collagen or collagen fibers. Applied to blast coated substrates. These two operations are performed in Example E and Example F. are shown.

Example E Place the collagen-coated matrix, ready for implantation, into the surgical bowl.

The matrix is then precoagulated with blood taken from the patient. Oral or respiratory mucosa 1/ 2. Collect an an-wide piece from the patient. Place the piece on the luminal surface of the stroma with absorbable sutures. suture to. The luminal surface is coated with synthetic materials such as fibrings to protect the graft. It may also be coated with a degradable substance. The matrix is sutured in place in the area to be repaired. transplant operation Afterwards, the device is moisturized with Dulbecco's tissue medium.

Example F: Medbore (Bolex, Fairburn, Georgia) punch at O.1% They were immersed in brine containing fazil for about 48 hours, and the residual ethanol that had been stored was removed. Remove alcohol or sodium azide.

They were then pre-equilibrated in Dulbecco's modified Eagle tissue medium for approximately 24 hours. let

The surgically removed dog oral cavity or ureteral tissue was washed with phosphate buffered saline (PBS). , dermatome centrated to cut the uppermost part of the dermis and epithelium at 500 μm. Remove with a slicer. Discard the lower dermis. Next, the epithelium-dermis is about 1-2 Cut into 2 small explants. Place these explants on the collagen-coated side of the batch. on the side that will be the intraluminal surface (approximately 5 grafts/aI).Then the batch is Place in a tissue culture dish and add tissue culture medium [keratinocyte growth medium (KGM), C1onetics Corp., San Diego, Calif. Luniaization] Supply 5 yam. In the case of ureteric skin, KGM is KGM90% lO Dulbecco's modified Eagle medium (DMEM) with a ratio of 10% DME containing % FBS Mix with. The batch is incubated at 37° for 14 days. At that point, the batch will be ported It is well epithelialized.

When the prosthesis is implanted, the collagen or collagen-fibroblast layer becomes infected with bacteria or It prevents contamination from penetrating into the matrix and at the same time removes connective tissue (granulation tissue) from surrounding tissue. grown in a substrate. As shown in Figure 4, collagen, collagen/fiber By the time the blastema layer has been biodegraded by the invading connective tissue, the pores 40 are filled with granulation tissue 38 (undeveloped). filled with a sterile layer of mature connective tissue). This granulation tissue is essential to the epithelium in order to survive. and provides the necessary blood supply to the epithelium as it grows inward from the suture site. collagen Fibroblasts also osmotically transport nutrients from surrounding tissues to keep the epithelium alive and growing. conduct.

prosthetic shape The basic material of prosthesis is a fairly large number of devices for reconstruction of ulcerated areas or for replacement of whole organs. can be formed into - In embodiments, the material is a compound as shown in FIG. ie, a single layer of collagen-coated matrix. This specific person a is an organization or is used to restore damaged areas in tissues or organs where end-to-end anastomosis is not possible. It is used for The batch is shaped or sized as indicated by the area being restored. You can also make it from scratch. For example, FIG. 6 is partially covered with a film of collagen 62, For this purpose, a round piece of porous matrix 6° is shown ready for implantation.

In another embodiment, the tube 64 as shown in FIG. 7 is a porous synthetic substrate. formed from and contacted with collagen 66 or collagen and fibroblasts. Can be done. An autograft epithelial layer may also be added. The tube thus formed is Is it possible to restore the trachea and esophagus where end-to-end anastomosis has become impossible due to excessive tissue removal? Or it can be used to change the length of the intestinal tract.

In yet another embodiment, the base material is used to replace a natural organ, such as the bladder. It may also be formed into a prosthesis. FIG. 8 is formed from a porous synthetic substrate 70 shown partially cut away. The bladder 68 shown in FIG. In this embodiment, the analogy shown in FIG. For example, it is desirable to attach an autologous layer of bladder mucosal tissue 76 to the intraluminal surface. patient's The ureter 72 and urethra 74 were sutured to the prosthesis.

Examples of operating techniques and results of device integration related to various aspects of device implantation. Examples are described below.

Example ■: Tracheal or glottic patch graft Make a longitudinal incision through the patient's trachea or glottis. A batch with the same vertical length as the defect select. The defect width is determined by the size of the area to be patched. 5% collage 25% collagen, 2.5% colacine/fibroblasts and 2.5% colacine 2 x 3 punches to grow coatings of fibroblasts and oral mucosal tissue cultures was transplanted. The mucosa is cut away from the cartilage to form a two-pronged flange of tissue.

The mucosal flange is sutured to the luminal surface of the batch with a continuous preabsorbable suture. mucosal suture After tying, the cartilage is placed in the groove or under the flange and intermittent non-absorbable sutures and cotton retraction sutures are placed. Fix it with.

There was no evidence of air leakage or infection. The first patch of 5% collagen is displaced and inside. It showed no growth. The edges of the 5% collagen batch show ingrowth. However, there was no epithelial growth from the edge. 2.5% colacine/fibroblasts The batch showed tissue growth in the middle of the batch. 2.5% colaseen/fibroblasts A batch of cystic oral mucosa showed tissue growth in the middle of the batch. 2.5% Colacy A batch of fibroblast oral mucosa was isolated from tissue culture based on some evidence of mucosal survival. It also showed endophytic growth. Channel-shaped patches can occur without mechanical fixation or connective tissue ingrowth showed that.

Tracheal tube implants up to 7 lengths made of 5% colacine-coated tubing are recommended for dogs. showed complete connective tissue ingrowth in the stromal and intraluminal connective tissue layers. Some substrate The narrowing in the center of the lumen resulted from excessive connective tissue growth.

Intraepithelial growth from the suture line occurred within 2 CI11 from each end. Free oral mucosal transfer One tube with an explant showed complete epithelialization, 7 an tube long.

Example ■・ A 5% collagen-coated tube up to a length of 50 mm was used to remove an intestinal segment in a dog. ported to the client. The intestinal segment was removed. Mucosa-submucosal tissue and serous fluid-muscular layer? I took it out. The mucosa was sutured into the lumen of the graft with a pre-continuous absorbable suture. Serum- The muscle layer was sutured to the outside of the implant with a continuous suture of non-absorbable material. of submitted intestines One end was closed and the other end was exposed for colonic fistula formation. Wrap the transplant in netting , the omentum was sutured in place around the tube. There were no leaks or infections. All Chu The tube formed a luminal layer of connective tissue 4 weeks after surgery. 60 day total, 0.0 from each end. Some tubes showed evidence of mucosal migration from the suture line toward the center of the device at a distance of 6 mm. (normal intestine shows only 0.8 mm of epithelial growth). removed intestine Mucosal and fluid production and high bacterial counts were present.

Example■ 5% Collagen, 25% Collagen, 2.5% Colashene/Fibroblasts and Collar A large tube that grows a coating with autologous ureteral epithelium or oral epithelial tissue cultured on A batch of 2×4an size was implanted into the bladder of a dog. The bladder was exposed. batch and A defect of the same size was created in the bladder. The mucosa-submucosa is removed from the serous-muscular layer. , 2-formed a flange. The mucosal layer is cultured in batches with a continuous suture of absorbable material ( (luminal) surface was sutured. The serous-muscular layer is sutured to the outside of the batch with absorbable interlocking sutures. did. The batch was covered with mesh and sutured in place. There was no evidence of infection or urine leakage . The pigeon batch showed connective tissue ingrowth around the edges. z5% collagen/ Fibroblast/culture batches have connective tissue ingrowth and tissue culture cell growth in the center of the batch. showed some evidence of its existence.

summary Something comes up.

June 8, 1992 Cheers

Claims (11)

[Claims] 1. A prosthetic device suitable for implantation in living tissue of the body, the device having an irregular shape a thin synthetic substrate having pores and defining an opposing surface; and a colloid in contact with at least a portion of said surface of at least one of said thin synthetic substrates; -gen substance; A prosthetic device consisting of. 2. The prosthesis of claim 1 further comprising fibroblasts in contact with the collagenous material. Device. 3. 3. The prosthetic device of claim 2, wherein the collagen is atelocollagen. 4. The collagen material is in solution, where the concentration of collagen is 1.5% The prosthetic device of claim 1, wherein the prosthetic device ranges from 5.0% collagen to 5.0% collagen. 5. Claim 1, wherein the thickness of the collagen material is in the range of 0.025 to 1.0 mm. The prosthetic device described. 6. A thin synthetic matrix of sufficient thickness so that the collagen material substantially occludes the pores. 2. The prosthetic device of claim 1, wherein the prosthetic device is in contact. 7. The thin synthetic substrate is 0.25 mm to 1.5 cm thick. 2. The prosthetic device of claim 1. 8. A thin synthetic matrix has a first intraluminal surface, and autologous epithelial tissue covers the first intraluminal surface. 2. The prosthetic device of claim 1, wherein the prosthetic device has collagen material attached thereto. 9. A method of manufacturing a transparent prosthetic device for implantation into living tissue of the body, the method comprising: providing a synthetic substrate having opposing surfaces and irregularly shaped pores; and A small amount of collagen material is added to the synthetic matrix to seal the pores with collagen material. applying on at least one of said surfaces. 10. Further processing of the synthetic matrix to effect collagen cross-linking in the collagen material 10. The synthetic substrate according to claim 9, wherein the synthetic substrate is soaked to remove residual substances. the method of. 11. Further contact of the synthetic matrix with cultured viable fibroblasts following application of collagen material 10. The method according to claim 9, wherein