CN100464791C

CN100464791C - Method for preparing biological filler for injectable soft tissue

Info

Publication number: CN100464791C
Application number: CNB2005100440055A
Authority: CN
Inventors: 曹成波; 王勇; 邹玉萍; 吕荣晖
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2005-07-11
Filing date: 2005-07-11
Publication date: 2009-03-04
Anticipated expiration: 2025-07-11
Also published as: CN1718249A

Abstract

A process for preparing the biologic filler able to be injected in soft tissue for repairing it or beautifying skin includes such steps as providing the fresh skin of piglet, removing subcutaneous fat, immersing in defatting liquid for defatting, defatting again, alkali treating, removing alkali, cutting for layering, softening by enzyme liquid, cross-linking, water washing, bleaching, cutting to become particles, packing and Co-60 irradiation.

Description

A kind of preparation method of syringeability soft tissue biological filler

Technical field

The present invention relates to medical science shaping and beauty technical field, is a kind of injectivity soft tissue biological filler and preparation method thereof.

Background technology

In orthopedic clinical operation, often need to the congenital development deficiency of some position soft tissue of body surface or the day after tomorrow depressed deformity fill reparation, the filling reparation of implant is the necessary means of treatment.Ideal implant should satisfy the requirement of the following aspects: (1) at first is safety, and promptly You Yi histocompatibility does not cause toxic reaction, inflammatory reaction, foreign body reaction, allergy, no antigen, non-carcinogenesis; (2) certain intensity is arranged, physics, chemical stability are good, corrosion-resistant, do not produce the leachable of noxious substance, absorb fewly, are not shifted; (3) simple to operate, easily moulding, non magnetic, easy sterilization, easy storage, cheap and easy to get.

Collagen class biological filler mainly contains two classes in the market.The one, be the product of the commodity " Zyderm " and " Zyplast " by name of representative with the facial esthetic surgery center of U.S. Ba Feite (Parfitt Facial Cosmetic Surgery Center), be the pure collagen protein that extracts purification from Corii Bovis seu Bubali, form through the glutaraldehyde chemical crosslinking.This product needs the continuous several times injection because degraded is too fast, and filling effect is not good, and bounce-back rate height, does not sell at home as yet at present; The 2nd, with the commodity of the U.S. " Meronized Alloderm by name ^TM" being the micronize people's of representative acellular dermal matrix; this product has intactly kept the morphosis and the constituent of hypodermal cell epimatrix owing to removed the cell component that causes host's rejection, and mechanical property is good; antigenicity is low; can augmenting soft tissue, stimulate the collagen hypertrophy, but this material has certain absorption; and also raw material is people's cadaver skin; it is very limited to originate, and costs an arm and a leg, and does not obtain general application.

Summary of the invention

The object of the invention is to overcome above-mentioned the deficiencies in the prior art, and a kind of safety non-toxic is provided, and immunogenicity is low, and absorbability is little, easily for human body tolerance and can stimulate the collagen hypertrophy, injectivity soft tissue biological filler cheap and easy to get and preparation method thereof.

The present invention realizes in the following manner:

The little Corii Sus domestica of selecting newly to slaughter is a raw material; remove the foreign body on the fur; the subcutaneous tissue of pruning; handle, cut open layer, enzyme bating, glutaraldehyde cross-linking, bleaching processing, micronize processing, packing, cobalt 60 radiation treatment through defat, alkali liquor; the percentage ratio of materials is to be benchmark with Corii Sus domestica weight in its preparation method, and concrete steps are as follows:

1. defat: get fresh little Corii Sus domestica, remove the foreign body on the skin, the subcutaneous fat of pruning immerses degreaser, water 150～250%, and soda 1.5～2.0%, 37～38 ℃ of temperature are rotated 1.5～2.0h, washing;

2. secondary degreasing: water 200～300%, penetrating agent 0.3～0.5%, soda 1.5～3.0%, 37～38 ℃ of temperature are rotated 2～3h, control water;

3. alkali treatment: water 80～150%, akali sulphide 2～4%, pulverized limestone 2～6%, auxiliary liming agent 1～3%, rotate 2～3h, add pulverized limestone 2～5% again, rotate 0.5～2.0h, add water to 250～300%, rotated 30 ℃～38 ℃ of temperature, total time 10～20h 10 minutes in 50 minutes at interval, get rid of alkali liquor, it is sliding to be washed to Pi Bufa;

4. dealkalize: water 200～300%, ammonium chloride is regulated pH value 6～9, and degreasing agent 1～3%, rotates 2～5h, washing by 30 ℃～38 ℃;

5. cut open layer: cut open layer, deburring by thickness 0.3～0.4mm;

6. enzyme bating: water 200～300%, 30～38 ℃ of temperature, SDS0.2～2% is regulated pH value to 6～8 with ammonium sulfate, adds pancreatin 0.1～0.5%, rotates 0.5～2h, adds 1398 proteinase-10 .1～0.3% again, rotates 3～5h, puts clean waste liquid, washing;

7. crosslinked: water 100～300%, glutaraldehyde 2～5%, anhydrous sodium sulfate 5～10%, sodium bicarbonate 0.2～0.8%, after room temperature rotated 2～5h down, pH value 4～6 added sodium bicarbonate 3～5% again, 30 ℃～38 ℃ of temperature, be rotated further 2～5h, pH value 6～9 is washed to neutrality;

8. bleaching: water 100～400%, hydrogen peroxide 1.0～5.0%, room temperature stir bleaching 2～5h down, and it is pure white to look skin graft, washing;

9. the skin graft after bleaching cuts into microgranular, the length of side 0.3～0.5mm, and every 3g microgranule adds PBS liquid 5～10ml, and PVC is packaged into 1 bag, cobalt 60 irradiation, room temperature is preserved down.

Adopt prepared material of the present invention and preparation method thereof to have the following advantages:

1. material source is extensive, and is cheap and easy to get, and can the bark fetching multilamellar be material, greatly reduces cost, is easy to suitability for industrialized production;

2. to derive from zoodermic native protein macromolecule as host material, safety non-toxic is the human body tolerance easily;

3. material is handled through glutaraldehyde, antigenicity significantly reduce and degraded and absorbed few;

4. use this material and can realize the permanent reparation of soft tissues such as skin, not bounce-back.

The specific embodiment:

Preparing packing material of the present invention can carry out in the rustless steel rotary drum, and concrete grammar and step are as follows:

1. defat: get fresh little Corii Sus domestica of butchering, remove the foreign body on the skin, the subcutaneous fat of pruning immerses degreaser, water 150%, and soda 2%, 37～38 ℃ of temperature were rotated 90 minutes, washing;

2. secondary degreasing: water 230%, penetrating agent 0.3%, soda 1.5%, 37～38 ℃ of temperature were rotated 120 minutes, control water;

3. alkali treatment: water 80%, akali sulphide 2.1%, pulverized limestone 1.5%, auxiliary liming agent 1% acts on 120 minutes, add pulverized limestone 2% again, changeed 30 minutes, add water to 250～300%, rotated 10 minutes in 50 minutes at interval, 30 ℃ of temperature, total time, 15h, venting alkali liquor, it is sliding to be washed to Pi Bufa;

4. dealkalize: water 250%, ammonium chloride is regulated pH value to 8.4, and 1%, 30 ℃ of degreasing agent rotated 120 minutes, washing, control water;

5. cut open layer, deburring by thickness 0.3～0.4mm;

6. enzyme bating: 200%, 38 ℃ in water, SDS0.4%, ammonium sulfate is regulated pH value to 7.4, adds pancreatin 0.3%, rotates 0.5h, adds 1398 proteinase-10 .3% again, rotates 3.5h, puts clean waste liquid, washing;

7. crosslinked: water 100%, glutaraldehyde 3.75%, anhydrous sodium sulfate 10%, after sodium bicarbonate 0.8%, room temperature rotated 2h down, pH value 4.5 added 5%, 30 ℃ of sodium bicarbonate again, is rotated further 5h, pH value 8.3, its pH value 7.0～7.3 is surveyed in washing;

8. bleaching: water 200%, hydrogen peroxide 5.0%, room temperature stir bleaching 5h down, and it is pure white to look skin graft, washing;

9. it is microgranular that the skin graft after bleaching adopts physical method to cut into, the length of side 0.3～0.5mm, and every 3g microgranule adds PBS and preserves liquid to 5ml, and PVC is packaged into 1 bag, cobalt 60 irradiation 15-20kgy, room temperature is preservation down.

Claims

1. the preparation method of an injectivity soft tissue biological filler; it is characterized in that the little Corii Sus domestica that it selects newly to slaughter is a raw material; remove the foreign body on the fur; the subcutaneous tissue of pruning; handle, cut open layer, enzyme bating, glutaraldehyde cross-linking, bleaching processing, micronize processing, packing and cobalt 60 radiation treatment through defat, alkali liquor; the percentage ratio of materials is to be benchmark with Corii Sus domestica weight in its preparation method, and concrete steps are as follows:

(1) defat: get fresh little Corii Sus domestica, remove the foreign body on the skin, the subcutaneous fat of pruning immerses degreaser, water 150～250%, and soda 1.5～2.0%, 37～38 ℃ of temperature are rotated 1.5～2.0h, washing;

(2) secondary degreasing: water 200～300%, penetrating agent 0.3～0.5%, soda 1.5～3.0%, 37～38 ℃ of temperature are rotated 2～3h, control water;

(3) alkali treatment: water 80～150%, akali sulphide 2～4%, pulverized limestone 2～6%, auxiliary liming agent 1～3%, rotate 2～3h, add pulverized limestone 2～5% again, rotate 0.5～2.0h, add water to 250～300%, rotated 30 ℃～38 ℃ of temperature, total time 10～20h 10 minutes in 50 minutes at interval, get rid of alkali liquor, it is sliding to be washed to Pi Bufa;

(4) dealkalize: water 200～300%, ammonium chloride is regulated pH value 6～9, and degreasing agent 1～3%, rotates 2～5h, washing by 30 ℃～38 ℃;

(5) cut open layer: cut open layer, deburring by thickness 0.3～0.4mm;

(6) enzyme bating: water 200～300%, 30～38 ℃ of temperature, SDS0.2～2% is regulated pH value to 6～8 with ammonium sulfate, adds pancreatin 0.1～0.5%, rotates 0.5～2h, adds 1398 proteinase-10 .1～0.3% again, rotates 3～5h, puts clean waste liquid, washing;

(7) crosslinked: water 100～300%, glutaraldehyde 2～5%, anhydrous sodium sulfate 5～10%, sodium bicarbonate 0.2～0.8%, after room temperature rotated 2～5h down, pH value 4～6 added sodium bicarbonate 3～5% again, 30 ℃～38 ℃ of temperature, be rotated further 2～5h, pH value 6～9 is washed to neutrality;

(8) bleaching: water 100～400%, hydrogen peroxide 1.0～5.0%, room temperature stir bleaching 2～5h down, and it is pure white to look skin graft, washing;

(9) skin graft after the bleaching cut into microgranular, the length of side 0.3～0.5mm, every 3g microgranule adds PBS and preserves liquid 5～10ml, PVC is packaged into 1 bag, cobalt 60 irradiation, room temperature is preserved down.