CN2794471Y - Tissue engineering bone transplant containing innervation - Google Patents
Tissue engineering bone transplant containing innervation Download PDFInfo
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- CN2794471Y CN2794471Y CNU2005200571216U CN200520057121U CN2794471Y CN 2794471 Y CN2794471 Y CN 2794471Y CN U2005200571216 U CNU2005200571216 U CN U2005200571216U CN 200520057121 U CN200520057121 U CN 200520057121U CN 2794471 Y CN2794471 Y CN 2794471Y
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- innervation
- carrier bracket
- organizational project
- bone graft
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Abstract
The utility model belongs to the technical field of constructing artificial organs by a tissue engineering method for biomedicine engineering and relates to a tissue engineering bone transplant containing innervation and a constructing method thereof. The tissue engineering bone transplant containing innervation of the utility model comprises a carrier bracket and a seed cells, wherein the seed cell is attached on the carrier bracket, forming a compound body with a bone tissue three-dimensional structure and biologic activity. The carrier bracket is biologic degraded material containing the innervation characterized in that sensory nerve beams are implanted in the carrier bracket. The seed cell is a marrow base stem cell which is taken from self-body marrow and is separated and increased to the cell with the density of 10< 6 > to 10< 7 > piece /ml. The tissue engineering bone transplant containing innervation of the utility model can promote nerve generation and increase the osteogenesis function of the tissue engineering bone transplant. The utility model has the advantages of simple operation, high repair speed and good repair effect on repairing bone coloboma.
Description
Technical field
This utility model belongs in the biomedical engineering with Method of Tissue Engineering and makes up the artificial organ technical field, relates to a kind of innerv organizational project bone graft and construction method thereof of containing.
Background technology
In recent years, along with the development of tissue engineering technique, the report that research and development have reparation, keep or improve the biological substitution thing of damaged tissue function increases year by year, and the research report of related organization engineering bone graft is quite a lot.At present to the organizational project bone graft development and development and use become the focus of large segmental bone defect repairing and treating.
The organizational project bone graft of research and development at present, mostly be to utilize seed cell and the damaged osseous tissue of sentencing the reconstruction disappearance of biodegradable material compound implant internal skeleton, also have some to study and made up the organizational project bone graft of vascularization, the effect of repairing bone defect is better than the organizational project bone graft that the former does not have vascularization.But the organizational project bone graft of vascularization also reaches the effect of ideal repairing bone defect far away.
Only have the adequate blood supply and perfect innerv organizational project bone graft is only the most close with normal bone tissues, also being to meet bionics principle most, can guarantee that tissue-engineered bone survives in vivo and can obtain the effect of best repairing bone defect, is more satisfactory solution.Promoting to have a lot of achievements aspect the organizational project bone graft vascularization research at present, but aspect the neuralization structure, also having no talent both at home and abroad at present and explore.
The utility model content
The purpose of this utility model is to provide a kind of innerv organizational project bone graft that contains, can impel neuralization, increase the ossification of organizational project bone graft, have advantage simple to operate, that reparation speed is fast, repairing effect is good for the bone defect repair.
Described in the utility modelly contain innerv organizational project bone graft, comprise carrier bracket and seed cell, seed cell is attached on the carrier bracket, formation has osseous tissue three dimensional structure and bioactive complex, described carrier bracket is for containing innerv Biodegradable material, and described innervation is to be implanted into the sensory nerve bundle at carrier bracket; Described seed cell is a bone marrow stroma stem cell.
Described bone marrow stroma stem cell is taken at autologous bone marrow, is 10 through separating, being expanded to cell density
6-10
7Individual/ml.
Further, described innervation is to unite the implantation motor tract in carrier bracket.
Further, go back the implantable intravascular bundle.
In the such scheme, described Biodegradable material is the natural inorganic salt, is preferably the β tricalcium phosphate, i.e. β-TCP.
Described in the utility modelly contain innerv organizational project bone graft and can be used for preparing the functional organization through engineering approaches bone graft of repairing large segmental bone defect.
This utility model is by the skeletonization effect of different neuralization method for reconstructing in the assessment organizational project bone graft body, established a kind of skeletonization effect and contained innerv organizational project bone graft preferably, and provide how to make up the feasible program that large volume organizational project bone graft survives in vivo and plays a role rapidly.Neural is extensive and conclusive to osseous tissue growth, formation and metabolic influence, and adequate blood supply and perfect innervation are the deciding factors that guarantees that engineered diaphysis internal memory is lived.The composite tissue engineering bone graft that has innervation and vascular system meets the bionics viewpoint, is ideal solution.The utlity model has advantage simple to operate, that reparation speed is fast, repairing effect is good.
Description of drawings
Fig. 1 is the structural representation that contains the organizational project bone graft of sensory nerve domination described in the utility model;
Fig. 2 is for feeling and the structural representation of the organizational project bone graft of nervus motorius domination described in the utility model containing;
Fig. 3 is the structural representation of while implantable intravascular in the described organizational project bone graft of Fig. 1;
Fig. 4 is the structural representation of while implantable intravascular in the described organizational project bone graft of Fig. 2.
The specific embodiment
Described in the utility modelly contain innerv organizational project bone graft, as shown in Figure 1, comprise carrier bracket 1 and seed cell 2, seed cell 2 is attached on the carrier bracket 1, formation has osseous tissue three dimensional structure and bioactive complex, described carrier bracket 1 is for containing innerv Biodegradable material, and described innervation is to be implanted into sensory nerve bundle 3 at carrier bracket; Described seed cell 2 is bone marrow stroma stem cell (BMSc).
Described bone marrow stroma stem cell is taken at autologous bone marrow, is 10 through separating, being expanded to cell density
6-10
7Individual/ml.
As shown in Figure 2, described innervation is to unite to implant sensory nerve bundle 3 and motor tract 4 in carrier bracket 1.
As shown in Figure 3 and Figure 4, go back implantable intravascular bundle 5 in the described organizational project bone graft complex.
Described Biodegradable material is the natural inorganic salt, is preferably the β tricalcium phosphate, i.e. β-TCP.
Fig. 1 to Fig. 4 is a microcosmic sketch map of the present invention, and its size is not represented actual size.
The construction method that contains innerv organizational project bone graft described in the utility model, with bone marrow stroma stem cell as seed cell, with the Biodegradable material is carrier bracket, implants the sensory nerve bundle, is built into the organizational project bone graft by Method of Tissue Engineering; Described bone marrow stroma stem cell is taken at autologous bone marrow, is 10 through separating, being expanded to cell density
6-10
7Individual/ml.
Further, described innervation is to unite the implantation motor tract in carrier bracket.
Further, go back the implantable intravascular bundle.
Described Biodegradable material is the natural inorganic salt, is preferably the β tricalcium phosphate, i.e. β-TCP.
Further, implantable intravascular bundle in the implantable neural bundle.
Experimental design of the present utility model utilizes the method for microsurgery, respectively sensory nerve bundle, motor tract, arterial blood tube bank and sensation, motor tract is united implantation and cultivates and induce the bone marrow stroma stem cell of differentiation (cell density of implantation is 10 through amplification in vitro
6-10
7/ ml) and the biomaterial of absorbable and degradable (in the complex of β-TCP), implant the damaged place of animal body internal skeleton, impel tissue engineered bone graft tissue generation neuralization, observe the skeletonization effect after the short neuralization of distinct methods takes place.
Laboratory animal: 30 of healthy new zealand white rabbits, available from Nanfang Medical Univ (former No.1 Military Medical Univ.) animal center, 6 monthly ages, body weight 2-3kg.
Experiment grouping: A group: organizational project bone graft matched group (BMSc+ β-TCP); B group: simple sensory nerve bundle implantation group (BMSc+ β-TCP+ sensory nerve bundle is implanted) C group: simple vascular bundle implantation group (BMSc+ β-TCP+ vascular bundle is implanted); D group: simple motor tract implantation group (BMSc+ β-TCP+ motor tract is implanted); E group: sensation, motor tract implantation group simultaneously (BMSc+ β-TCP+ sensation, motor tract are implanted).The sensory nerve of implanting is a saphenous nerve, and nervus motorius is a rami musculares nervi femoralis, and the vascular bundle of implantation is a femoral artery.Other gets 2 rabbits and does blank group.
The concrete structure divided three steps:
1, cell culture
The new zealand white rabbit of growing up adopts speed sleep new, ketamine and the intramuscular anesthesia of atropine mixed liquor.With No. 16 medullo-puncture needle punctures, extract red bone marrow 5ml behind the bilateral ilium place preserved skin, sneak into the DMEM complete medium, the anticoagulant of 500u/ml heparin solution, the centrifugal 15min of 1000r/min abandons supernatant behind the mixing, crosses 100 mesh filter screens, adds culture medium numeration back inoculation.37 ℃, 5%CO
2Incubator is cultivated, and later half amount was changed liquid in 3 days, and full dose was changed liquid 1 time in later 2～3 days, treats that cell merges into behind the monolayer with 0.25% trypsinization, and numeration (contains 10% hyclone, 50ug/ml ascorbic acid, 10 with the DMEM conditioned medium
-8The mol/L dexamethasone, 10
-3The mol/L sodium) cultivation of going down to posterity.Changed liquid 1 time in conventional 3 days.Cultivated for 2 weeks with 10
6-10
7The density of individual/ml with β-TCP that BMSc inoculates organizational project bone graft group, simple sensory nerve bundle implantation group and simple motor tract implantation group last (material specification is φ 12mm * 15mm), 37 ℃, 5%CO
2Incubator was cultivated 4-6 hour, and conditioned medium adds complete cladding material along the culture dish edge, continued to cultivate the damaged place of implantable bone after 72 hours.
2, the preparation of animal model
The long otch of inboard arc before the hind leg of left side cuts skin, subcutaneous tissue and deep fascia, separates entering along the sharp property in rectus femoris and vastus medialis gap.4 moulding good hole general steel plates are placed in the outside before femur, and drive screw is fixed.Go in ring and peel off femur stage casing periosteum, between steel plate the 2nd, 3 holes,, excise the periosteum of this section correspondence, cause scarce property bone of section and the periosteum of 1.5cm damaged with the sawed-off femur of scroll saw.The organizational project bone graft is organized in the damaged place of bone and only embeds β-TCP+ through inductive BMSc complex.The simple damaged model preparation method of sensory nerve bundle implantation group bone is the same, in femoral triangle, separate and cut the saphenous nerve that sends by femoral nerve, free enough length, sharp property cut-out saphenous nerve is implanted the saphenous nerve bundle in the side channel of the BMSc complex of β-TCP+ through inducing differentiation; Simple motor tract implantation group is separated the flesh that cuts the domination vastus medialis that is sent by femoral nerve and is propped up, and a free segment length is cut off, and this nerve tract is implanted in the side channel of cell material complex.Sensation, motor tract implantation group are simultaneously separated the flesh Zhi Yiqi that cuts the domination vastus medialis that saphenous nerve and femoral nerve send and are implanted in the side channel of cell material complex.Sensory nerve bundle and vascular bundle implantation group simultaneously are that saphenous nerve and femoral artery are implanted in the side channel of cell material complex together.
3, detection method
(1) gross examination of skeletal muscle: the ordinary circumstance of tightly observing the new zealand white rabbit postoperative.4,8,12 weeks of postoperative respectively put to death 2 animals and draw materials, the surface variation of tissues observed engineering bone graft, the formation of interior crust, bone defect repair situation and surrounding tissue reaction.
(2) radiological examination: A, B, C, D, each treated animal of E (are thrown range from 1m in all taking the photograph left side femur normotopia X-ray sheet in 4,8,12 weeks of postoperative, throw according to condition 46KV, 50Ma, time of exposure 0.14s), blank is organized in 12,18 weeks of postoperative and is taken the photograph sheet, observes the bone defect healing situation.All photos are according to [Yang CY such as Yang, Simmons DJ, Lozano R.Thehealing of grafts combining freeze-dried and demineralized allogeneic bone in rabbits.ClinOrthop.1994,298:289] standards of grading carry out the radiology scoring.The resistance of bone defect x line is penetrated density and is measured by computer image analysis.Density is penetrated in resistance, and to penetrate density with the resistance of same photo equal area cortical bone be 100, and equal area photo background color is 0 to measure relative value, and the resistance of same photo is penetrated density and got averages that piece shadow density is penetrated in all visible resistances.
4, result
(1) gross examination of skeletal muscle has 2 rabbits that wound infection takes place, the equal first phase healing of all the other rabbit wounds.See that the wound of infection has purulent secretion 4 weeks when drawing materials, amount is few, is the inflammatory granulation tissue around the material, and a routine pus filling is few, and another routine pus filling is more, does not have obvious callus formation.A group postoperative 4 all material surfaces and defective region have a small amount of callus formation; 8 when week material surface and bone defective region callus increase, wherein 1 rabbit bridge part heals with the osteotomy end; Postoperative 12 all materials are most of to be wrapped up by callus.B group postoperative 4 all material surfaces and the visible more amount callus formation of defective region; 8 when week material surface and defective region callus showed increased, wherein 4 rabbit bridge parts heal with the osteotomy end; Postoperative 12 all bone defective regions are the callus parcel.C group postoperative 4 all material surfaces and bone defective region see that callus formation is more; Callus formation showed increased during 8 weeks wherein has the healing of 3 bridge parts and osteotomy end; Postoperative 12 all materials see that the bone defective region is that callus connects.D group postoperative during 4 weeks material surface and bone defective region see that a small amount of callus formation is arranged; Postoperative 8 all material surfaces and bone defective region callus increase, wherein 1 rabbit bridge part and osteotomy end healing; Postoperative 12 all materials are most of to be that callus is wrapped up.E group postoperative 4 all material surfaces and bone defective region see that more callus formation is arranged, and matter is hard; Postoperative 8 all material surfaces and defective region callus showed increased, wherein 3 rabbit bridge parts and osteotomy end healing; The damaged place of postoperative 12 all bones is the filling of callus institute.It is the same substantially to observe the damaged size of bone when blank 12 weeks of group postoperative and 24 all putting to death, the sclerosis of two epiphysis, and the medullary cavity sealing, fibrous tissue filling pulp cavity does not have obvious growth of spur.
(2) X-ray film is observed
Begin to blur around A group postoperative 4 all bone defective regions and the material, be a little cotton-shaped resistance projection; Postoperative 8 during week around the material and the bone defective region be patch shape resistance projection, the material visible absorbance, tissue-engineered bone skeletonization amount increases, and fogs with two-section osseous part gap, wherein one is connected healing with host bone; Implanting tissue through engineering approaches bone and bridge part are linked to be big lamellar resistance projection during 12 weeks, and shape becomes very irregular after the visible material degraded and absorbed, and the attenuating of the resistance projection of material, and density unevenness is even in it.
Occur bluring around B group postoperative 4 all bone defective regions and the material, point-like and cotton-shaped resistance projection are arranged; See that material reaches the bone defective region on every side and is bigger lamellar resistance projection in 8 weeks of postoperative, the material visible absorbance has 4 bridge parts to be connected healing with the two-section epiphysis; Postoperative 12 during week the bone defective region see big lamellar resistance projection, increase in density, material obviously absorbs the back and shows unclear.
Fog around C group postoperative 4 all bone defective regions and the material, cotton-shaped resistance projection is arranged; See that material reaches the bone defective region on every side and is bigger patch shape resistance projection in 8 weeks of postoperative, shape becomes irregular after the material visible absorbance, has 3 bridge parts to be connected healing with the two-section epiphysis; Postoperative 12 during week the bone defective region see big lamellar high density resistance projection, it is unclear to have developed after the absorbed.
Begin to fog around D group postoperative 4 all bone defective regions and the material, be a little cotton-shaped and small lamellar resistance projection; Postoperative reaches the bone defective region and is patch shape resistance projection around the material during 8 weeks, visible material absorbs, and tissue-engineered bone and two-section osseous part gap fog, and wherein a bridge part is connected healing with host bone; Implanting tissue through engineering approaches bone and bridge part are linked to be big lamellar resistance projection during 12 weeks, and big of bone defective region is fuzzy, and visible material obviously absorbs back resistance projection and lowers, and it is inhomogeneous that density becomes.
E group postoperative sees around the material during 4 weeks and the bone defective region fogs, and cotton-shaped resistance projection is arranged; The bone defective region is big patch shape resistance projection during 8 weeks, and material visible absorbance, shape are irregular, has 3 rabbits to see that the existing bone of organizational project bone graft and two-section epiphysis is connected; Postoperative 12 all bone defective regions are big lamellar resistance projection, and the edge smoother is neat, and absorbed is a lot.
Blank 12 weeks of group postoperative and 24 all bone defective regions still are radiolucent shadow, epiphysis sclerosis, fuzzy.
(3) X-ray film iconography appraisal result
Table 1 bone defective region iconography score value (x ± s, n=6, scoring) Roentgenographic scores at bone defects (x ± s, n=6, scores)
| Grouping groups | Time (week) time (w) | ||
| 4 | 8 | 12 | |
| A B C D E | 5.333±0.517 8.333±0.816 5.500±0.836 8.767±0.081 10.166±0.753 | 7.166±0.408 11.333±0.516 8.167±0.307 11.964±0.344 13.167±0.752 | 9.083±0.376 12.500±0.894 10.083±0.801 12.333±0.516 14.167±0.752 |
Table 2 bone defective region bone density value (x ± s, n=6, relative value) Values of callus density at bone defects (x ± s, n=6, relative values)
| Grouping groups | Time (week) time (w) | ||
| 4 | 8 | 12 | |
| A B C D E | 43.501±2.725 58.663±2.541 44.578±2.948 57.572±2.164 60.783±1.509 | 54.638±1.265 62.375±0.992 54.203±1.556 63.071±0.866 65.406±0.783 | 71.683±2.101 84.582±1.017 73.585±1.975 83.127±1.744 86.738±1.144 |
X-ray film iconography appraisal result:
Can find out from table 1, B, D, the scoring of E group normotopia photo bone defect repair are all learned scoring than the irradiation image of A, C group in 4,8,12 weeks and are excellent (P＜0.05), C group and A organize the also no significant difference (P all＞0.05) of comparing, but compare mutually between B group, D group and E group, no significant difference between D group and B group, E group, and the E group is better than the B group.
Density measurement is penetrated in positive bit slice bone defect resistance:
Can find out that from table 2 B, D, E organize 4,8, the damaged resistance of the bone in 12 weeks is penetrated density relative value and all is better than than A, C group (P all＜0.05), and C group and A group are compared also no significant difference (P all＞0.05), but B, D, E compare between organizing mutually, no significant difference between D group and B group, E group, and the E group is better than the B group.
5, conclusion
(1) in the organizational project bone graft, implant the sensory nerve bundle after, its repairing bone defect effect all is significantly improved than simple organizational project bone graft and the repairing effect of implanting motor tract in the organizational project bone graft;
The skeletonization effect of (2) feel, motor tract being united the implantation group is better than sensory nerve bundle implantation group;
(3) the skeletonization effect of vascular bundle implantation group is better than simple organizational project bone graft.
This description of test: the method for utilizing the sensory nerve bundle to implant can improve the ossification of organizational project bone graft, does not observe this effect and implant the motor tract group.If implant sensation, motor tract simultaneously, can improve the ossification of organizational project bone graft effectively.If the method for taking vascular bundle to implant again can more effectively improve the ossification of organizational project bone graft.
Claims (6)
1, contains innerv organizational project bone graft, comprise carrier bracket (1) and seed cell (2), seed cell (2) is attached on the carrier bracket (1), formation has osseous tissue three dimensional structure and bioactive complex, it is characterized in that: described carrier bracket (1) is for containing innerv Biodegradable material, and described innervation is to be implanted into sensory nerve bundle (3) at carrier bracket; Described seed cell (2) is a bone marrow stroma stem cell.
2, according to claim 1ly contain innerv organizational project bone graft, it is characterized in that: described innervation is to unite to implant motor tract (4) in carrier bracket (1).
3, according to claim 1 and 2ly contain innerv organizational project bone graft, it is characterized in that: go back implantable intravascular bundle (5).
4, according to claim 1ly contain innerv organizational project bone graft, it is characterized in that: described Biodegradable material is the natural inorganic salt.
5, according to claim 4ly contain innerv organizational project bone graft, it is characterized in that: described natural inorganic salt is β tricalcium phosphate, i.e. β-TCP.
6, according to claim 1ly contain innerv organizational project bone graft, it is characterized in that: described bone marrow stroma stem cell is taken at autologous bone marrow, is 10 through separating, being expanded to cell density
6-10
7Individual/ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU2005200571216U CN2794471Y (en) | 2005-04-18 | 2005-04-18 | Tissue engineering bone transplant containing innervation |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU2005200571216U CN2794471Y (en) | 2005-04-18 | 2005-04-18 | Tissue engineering bone transplant containing innervation |
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| CN2794471Y true CN2794471Y (en) | 2006-07-12 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112528727A (en) * | 2015-05-28 | 2021-03-19 | 阿克松根股份公司 | Quantitative structural determination of nerve grafts |
-
2005
- 2005-04-18 CN CNU2005200571216U patent/CN2794471Y/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112528727A (en) * | 2015-05-28 | 2021-03-19 | 阿克松根股份公司 | Quantitative structural determination of nerve grafts |
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| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20060712 Termination date: 20140418 |