Transplant for scleroplasty (versions)

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to ophthalmosurgery, and in particular to scleroplasty. Transplant for scleroplasty has polymeric base, covered with porous layer of the same polymer. As polymer base, transplant includes layer, made from porous stretched polytetrafluoroethylene, which has nodular-fibrillar structure. As porous layer, it includes layer of porous polytetrafluoroethylene, which has volume fraction of void space 15-40%, specific surface of void space 0.25-0.55 mcm2/mcm3, average distance between voids in volume 25-30 mcm and average chord volume 8-25 mcm, with the total width of transplant constituting 0.15-0.35 mm (first version). Transplant for scleroplasty can also include porous layer of polymer, whose surface is processed to add compatibility with sclera tissue. Transplant surface is processed by application of allogenic dermal fibroblasts of 3-5 passages of culturing, with the total width of transplant being 0.15-0.35 mm (second version).

EFFECT: chemically and biologically inert transplant, which ensures effect of invasion of sclera tissues, is obtained.

2 cl

 

The invention relates to medicine, namely to ophthalmology, and in particular to scleroplasty.

In modern plastic ophthalmic surgery problem grafting remains quite relevant. The use of different grafts will undoubtedly increase the efficiency of many reconstructive surgeries. However, it is still relevant and was not fully resolved the issue of optimal, which meets modern requirements of ophthalmic surgery trasplantation material for scleroplasty, especially considering the major histocompatibility in each case [Zharov CENTURIES BIOPLAST - Department of procurement, preservation and production of plastic materials / V. Zharov, ER Tochilov. - Moscow, 2003. - 105 C.].

There are many indications for plastic surgery on the sclera, including such as extensive wounds with tissue defect, breaks sclera, ulcers of the cornea and sclera, severe burns of the sclera, degeneration and thinning of the cornea after depertment etc. [Som E.E. Scleroplasty / E.E. Kgs. - Saint Petersburg: Saint Petersburg Pediatric medical Institute, 1995. - 144 S.]. In many cases, the refusal scleroplasty can lead to loss of the eye.

As the plastic material for the scleroplasty used the auto - and allogenic materials, such as a private wide fascia bed is a, suspension Autorama with Autocraft, allogeneic Dura mater, the suspension of the placenta, AllUnion, a mixture of dry plasma and thrombin in a solution of novocaine [Vasilkova AS Autoandrophilia in the treatment and prevention of myopia and retinal dystrophy: author. Diss. Kida. the honey. Sciences. - M., 1989], however, used for this purpose various biological tissues (allesklar, canned Dura membrane, the amnion, the auto - and allegrad) as a result of engraftment are in the process of pathological changes in the sclera of the recipient [H.E. Trutta, L.D. Andreeva, etc. // Vestnik of ophthalmology. - 1999. No. 5. - P.8-10]. Transplanted allografts are substituted its own connective tissue of the recipient, resulting in not formed a complex sclera-implant" and the disease process continues. Postoperative inflammatory reaction occurs in the form of canonita and granulomatous inflammation [Wiswe I., GrungreiffJ., Schlotte H.W., Bansche I., Fortschr. Ophthalmol. - 1991. - Bd 88, No. 6. - S.881-884]. All this limits the scleroplasty using auto - or allotransplantation.

Famous transplant for scleroplasty, representing the allograft sclera, stitched threads biocompatible suture material with the formation of a mesh structure with a cell size of 1.0×1.0 mm [RU No. 2140242, IPC A61F 9/007, 1999].THE quality of Biosafety the CSOs suture material was used capron, polyester fiber or polyester. The graft is used for the treatment of rapidly progressive myopia; presents the results sclerocephalus effect after 6 months.

However, transplantation allostery is expressed inflammatory reaction and rough capsulorraphy [L.D. Andreeva, H.E. Trutta etc. // Vestnik of ophthalmology.- 1999. No. 3. - P.15-18]. Further, in 1.5-2 months after transplantation begins lysis of donor sclera and replacement with newly formed collagen recipient, as a result, sclerocephalus effect leveled [H.E. Trotta and others, 1999; Whitmore W.G., Harnson W., Curtin B.J. Ophthalmic Surg. - 1990. - Vol.21, No.5.- R-330].

Also known graft for scleroplasty, made in the form of a grid with a cell size of 1×1 mm of nylon or polyester fibers, stitched collagen fiber (suture "collagen 5,0") in increments of 1 mm [RU No. 2157159, IPC A61F 9/01, 2000]. The specified graft was also used for the treatment of progressive myopia.

The disadvantage of the graft, and graft on RU # 2140242, is the lack of stability of the used polymer fibers to biodegradation, as the cellular and non-cellular, as well as the disintegration and fragmentation of the material on a separate thread.

The closest to the essential features is the transplant for when ceroplastic, represents a polymer base, which is applied to the porous layer of the same or another polymer, the surface of which is treated to make it compatible with the tissue of the sclera [RU No. 2160123, IPC A61L 27/56, 2000]. The graft is made of a porous silicone (item 3 in the formula), the base and layer with pores can be performed at the same time (2 formulas); more porous layer that implantation should be addressed to the sclera, has pores, made in the form of perpendicular or inclined to the surface of the channels (4 formulas), located regular way (claim 7 of the formula). Layer with pores processed low-temperature plasma discharge (section 8 formula), which causes the formation in the surface layer of the graft transition structures containing polymer chain and fragments thereof (claim 1 of the formula). Transitional structure creates a permanent hydrophilicity of the working side of the graft, which creates favorable conditions for the germination of living tissue into the pores to increase thereby the mechanical strength of the connection of the graft with the sclera" (p.5, abs description of the invention).

However, in RU # 2160123 not provide specific examples of the manufacture of the claimed graft and there are no experimental data on scleroplasty. Production of graft on RU # 2160123 difficult; silicone polymers do not have sufficient chemical and biological ine what testu; the effectiveness of the transition hydrophilic layer has not been proven experimentally.

The technical result, which is aimed by the invention, is to get a transplant for a scleroplasty, chemically and biologically inert and provides the effect of germination tissue of the sclera.

This result is achieved in that the graft to scleroplasty, having a polymer base, which is applied to the porous layer of the same polymer as the polymer base includes a layer made of stretched porous polytetrafluoroethylene having a nodular-fibrillar structure, and as a porous layer includes a layer of porous polytetrafluoroethylene having a volume fraction of the space of voids 15-40%, specific surface space void of 0.25-0.55 m2/μm3the average distance between voids in the volume of 25-50 microns and an average chord volume 8-25 μm in total thickness graft of 0.15-0.35 mm (the first option).

This result is achieved by the fact that the transplant for scleroplasty having a porous polymer layer, the surface of which is treated to make it compatible with the tissue of the sclera, as the porous layer includes a layer of porous polytetrafluoroethylene having a volume fraction of the space of voids 15-40%, specific surface area simple is the Christianity of voids 0.25 to 0.55 m 2/μm3the average distance between voids in the volume of 25-50 microns and an average chord volume 8-25 μm, the surface of which is processed by the application of allogeneic dermal fibroblasts 3-5 passages cultivation with a total thickness graft of 0.15-0.35 mm (the second option)

Porous stretched polytetrafluoroethylene (PTFE) nodular-fibrillar structure is obtained by extraction of raw calendered film of PTFE emulsion, for example of the brand f-4D, as described in EN No. 2203685. The layer of polymer base graft includes 2-4 layer porous stretched film of this structure, the total thickness of this layer is 0.05 to 0.15 mm

Porous PTFE constituting the porous layer obtained from a suspension of PTFE, such as stamps, f-4, as described in EN No. 2270642. The thickness of the layer of porous PTFE in the graft is 0,10-0,30 mm

Porous PTFE on RU # 2270642 was used for the manufacture of the implant to strengthen the cornea in the form of a convexo-concave lens with a minimum thickness of 0.4 mm, which was introduced in corneal intralamellar pocket formed during operation. 4 months connective tissue and isolated blood vessels were grown in the space of voids between the elements of polymer; there were no cases of exclusion or postoperative complications.

However, for scleroplasty is required transplantati thicker than 0.35 mm; if the material RU # 2270642 cut plate of such thickness (or thickness of 0.15-0.20 mm), the transplant after surgery will not withstand voltage, and before the connective tissue will grow into the graft, it will be invalid.

Porous PTFE was obtained as follows.

Suspension of PTFE according to GOST 10007-80 in powder form termostatic at a temperature of 340-390°C and additionally crushed in the crusher. Powder scatter, selecting a fraction of 0.25 to 0.60 mm, from selected fractions pressed the workpiece at a pressure of 10-40 MPa, the workpiece is sintered at a temperature of 340-390°C for 5-6 hours. The graft is specified thickness receive machining sintered billets.

Volumetric parameters of the proposed transplant is determined by well-known methods [Panteleev VG PAMM HP Inorganic materials, 1986, vol. 22, No. 12, s-1951; Avtandil, and other System stereometry in the study of the pathological process. M, Medicine, 1981; Cherniavsky the HP Stereology in metallography. M, Metallurgy, 1977].

According to the definition of the structure of the porous PTFE layer has a volume fraction of the space of voids 15-40%, specific surface space void of 0.25-0.55 m2/μm3the average distance between voids in the volume of 25-50 microns and an average chord volume 8-25 μm,

For more material on the first paragraph of the formula preparation of p is ristoro PTFE, the obtained suspension method, combined overlay with 2-4 layers of biaxially-oriented film, and such a package was thermophysical between heated plates at a temperature of (330-360°C. and pressure (0,003-0,008) MPa.

To obtain material for the second claim were used dermal fibroblasts isolated from human skin. Source selection cells were the fragments of the eyelids, ears or face, taken in the result of cosmetic surgery. Fibroblasts were isolated by the method of spontaneous migration in the process of cultivation of these fragments in culture medium DMEM with addition of 10% (by volume) fetal cow serum. For sowing taken fibroblasts cultivated not later than 3-5th passages. It is known that fibroblasts are not immunocompetent cells, and populations of fibroblasts lacking immunocompetent cells; therefore, the use of allogeneic fibroblasts cannot have a negative impact on the status of the transplant recipient.

Sowing was carried out as follows.

All variants of the experiment were performed in 6 replicates, i.e. the cells of each variant were seeded in 6 wells of 96-hole Board. Suspension dermal fibroblasts were sown at a concentration of 2000 cells/well. Before seeding the cells in the wells VA is Ianto 2 and 3, laid disks with a diameter of 16 mm, cut from the porous PTFE thickness of 0.15-0.35 mm (pre-incubated in 10% fetal serum of cows).

After 1 hour after seeding cells on the achievement of their attachment to the substrate in the wells of option 3 (the disks are pre-incubated in nutrient medium) standard culture medium was replaced with conditioned medium. On the 5th day of cultivation, the contents of each well were photographed. Then the method colorimetry was performed determining the number (i.e., the degree of proliferation) of cells grown in each of the variants of the experience.

The morphological state of the cultured cells was assessed visually under an inverted microscope. The presence and condition of the cells on the membranes after 5 days of cultivation were evaluated by scanning electron microscopy.

Data colorimetric studies have shown that when seeding cells at a dose of 2000 cells/well after 5 days of cultivation in option 3 (the conditioned medium after incubation in her diaphragm disks) increased 18000 ((±500) cells, and in the control variant (standard culturing conditions) - 20000 (±650) cells. Obtained differences can be neglected and, consequently, it is possible to draw a conclusion about the absence of toxic effects of membrane material of the porous polytetrafluoroethylene on stump the dummy fibroblasts.

Morphological state (morphology of individual cells and the state of the monolayer of cells, formed during 5 days of cultivation) also indicates the absence of toxic effects of membranes on the cell.

Scanning electron microscopy of membranes after 5 days of cultivation on these cells showed their presence that can serve as evidence of the ability of cells to grow on this substrate.

The test of the proposed transplant was carried out on rabbits of the Chinchilla breed weight of 3.5-3.7 kg Test graft on the first claim was held on 15 animals; testing of the graft on the second claim was conducted in 3 animals.

The intervention was carried out under intravenous anesthesia. For local anesthesia was used instillation of alcaine (Alcon, USA). Area operations served as the upper outer quadrant of the eyeball. 1.5 mm from the limbus was performed incision of the conjunctiva with the underlying tenon's capsule length 5 mm Spatula formed a tunnel in the tenon space, then placed the graft oval size h mm, cut from the material of the first variant (15 animals) and ismaterial second option (3 animals), and the material was placed on the sclera porous party in the first case and the side with fibroblasts in the second case. The wound NAC who was Adivasi 2 knotted suture silk 8-0. Under the conjunctiva was injected with 0.05 g of Cefazolin and 0.002 g of dexamethasone.

After surgery for 5 days in the operated eye was buried 0,3% aq ciprofloxacin hydrochloride (cipromed; Promed, India).

The first two days after surgery instillation of antibiotic conducted 4 times a day, the rest of the year 2 times a day.

On the first day after surgery was noted mild conjunctival injection directly in the intervention area. Further, the injection took place entirely. Abnormal discharge from the conjunctival cavity were observed in any of the cases. For the whole period of observation of experimental animals was noted to have signs of damage to the transplant, rejection or migration.

1. The transplant for scleroplasty, having a polymer base, which is applied to the porous layer of the same polymer, characterized in that as the polymer base includes a layer made of stretched porous polytetrafluoroethylene having a nodular-fibrillar structure, and as a porous layer includes a layer of porous polytetrafluoroethylene having a volume fraction of the space of voids 15-40%, specific surface space void of 0.25-0.55 m2/μm3the average distance between voids in the volume of 25-50 microns and an average chord volume 8-25 μm, the total thickness is of transplantati 0,15-0,35 mm

2. The transplant for scleroplasty having a porous polymer layer, the surface of which is treated to make it compatible with the tissue of the sclera, characterized in that the porous layer includes a layer of porous polytetrafluoroethylene having a volume fraction of the space of voids 15-40%, specific surface space void of 0.25-0.55 m2/μm3the average distance between voids in the volume of 25-50 microns and an average chord volume 8-25 μm, the surface of which is processed by the application of allogeneic dermal fibroblasts 3-5 passages of cultivation, when the total thickness of the graft 0,15-0,35 mm



 

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