Acellular tendon or ligament collagenous fiber material and preparation method thereof
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A technology of collagen fibers and collagen fiber bundles, which is applied in the field of natural biological materials - decellularized tendon/ligament collagen fiber materials and its preparation, to achieve the effect of rich materials, good biomechanical properties, and low cost
Active Publication Date: 2012-12-12
陈雄生 +2
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Problems solved by technology
How to prepare it into a substitute or tissue engineering scaffold with the structural characteristics of tendon and ligament ECM has not been satisfactorily resolved
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Embodiment 1
[0023] Example 1: Preparation of bovine tendon decellularized collagen fibers or fiber bundles
[0024] 1) Take fresh bovine tendon, remove the outer membrane and synovial tissue on the surface of the tendon band.
[0025] 2) Put the tendons in a vacuum freeze-dryer (VirTis BENCHTOP, USA) at -56°C for 12 hours.
[0026] 3) Loosen the low-temperature dried tendon structure by extrusion and hammering.
[0027] 4) The loose tendon is further combed to separate tendon collagen fibers or collagen fiber bundles.
[0028] 5) Decellularize the isolated collagen fibers or fiber bundles:
[0029] Use 10mM hypotonic tris buffer + serine protease inhibitor (0.05% phenylmethylsulfonyl fluoride ethanol solvent, 35ml / L 50ml ethanol + 25mgPMSF) for 96h, and change the medium every 24h. Depending on the thickness of the fiber bundles, a small amount of detergent (such as TritonX-100, butyl triphosphate, etc.) and trypsin can be added as appropriate during the decellularization process. All...
Embodiment 2
[0031] Example 2: Preparation of bovine ligament decellularized collagen fibers or fiber bundles
[0032] Replace bovine tendon with bovine ligament, and all the other are the same as in embodiment 1.
Embodiment 3
[0033] Example 3: Histological examination of bovine tendon decellularized collagen fibers or fiber bundles
[0034] Collagen fibers or fiber bundles before and after decellularization were taken for paraffin embedding, sectioned (5 μm), and HE stained for observation. Observe the nuclear components (hematoxylin staining), collagen arrangement (eosin staining), figure 2 In A, collagen fiber bundles before decellularization; in B, collagen fibers or fiber bundles after decellularization, the cell components are completely removed.
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Abstract
The invention relates to the field of biological fiber materials. The conventional tendon and ligament substituent mainly comprises autograft, allograft or heteroplastic graft and synthetic material graft which all have unavoidable defects. The invention aims to provide a natural biological material with excellent biomechanical properties and biocompatibility, namely an acellular tendon / ligament collagenous fiber material, and a preparation method thereof. In the preparation method, a mechanical structural unit and a cell attachment structure which tendon / ligament collagenous fibers or fiber bundles originally have are reserved; the tendon / ligament fibers are processed by adopting a low-temperature drying method, and the separated fibers reserve certain mechanical strength; and the collagenous fibers or fiber bundles are selected by a physical loosening and carding method and then acellular treatment is performed. The tendon / ligament acellular collagenous fiber / fiber bundle material of the invention has wide source, large quantity and low price and not only can be used for ligament and tendon reconstruction and rehabilitation, but also can be widely applied to other biomedical fields because the tendon / ligament acellular collagenous fiber / fiber bundle material has properties of the fiber material.
Description
technical field [0001] The invention relates to the field of biological fiber materials, in particular to a new natural biological material-decellularized tendon / ligament collagen fiber material and a preparation method thereof. Background technique [0002] Current tendon and ligament substitutes mainly include autografts, allografts or xenografts, and synthetic material grafts, but all have their unavoidable shortcomings. Although autologous ligament transplantation is the most effective method of repair and reconstruction, it takes a long time to recover and the functional recovery is not satisfactory, so it cannot be widely used in clinic. The strong antigenicity of allogeneic or heterogeneous tendon is its biggest shortcoming. Artificial synthetic materials such as carbon fiber, polyester materials and other non-degradable materials can provide sufficient mechanical strength in the short term, but they cannot be regenerated, and long-term use is easy to cause wear and ...
Claims
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