Feasibility Study Of Seeding Fresh Bone Marrow Aspirate Directly On Scaffolds To Construct Tissue Engineering Ligament

Object: seed cells combined with scaffold materials is the basic method toconstruct tissue engineered ligament, but separated from the seed cells in vitro,cultured, passaged step is time-consuming, laborious, and prolong cycle of tissueengineering ligament used in clinical, which undoubtedly increase hospitalizationcycle and the cost of treatment. we refer to the literature, rarely mentioned on theapplication of fresh bone marrow aspirate directly being planted on scaffolds to buildtissue engineering ligament, but there were several reports about the application offresh bone marrow aspirate being injected directly into the intra-articular anteriorcruciate ligament (ACL) injury partial to repair ligament injury, so we assume thatfresh bone marrow aspirate seeded directly on the scaffolds can also promote the seedcell adhesion, proliferation, differentiation and secretion of extracellular matrix(ECM), and the effect of non-inferiority isolated and cultured of BMSCs plantingbracket effect in the past, through the evaluation of the two cell cultivation methods toexplore the feasibility of application of fresh bone marrow aspirate planted directly onscaffolds tissue engineering ligament, and trying to find a new, more easy and theexact effect of cell seeding method, as well as evaluating the feasibility of servingfibroin fiber/small intestinal submucosa (SIS) complexes as the ligament tissueengineering scaffolds.Method: constructing fibroin fiber/small intestinal submucosa (SIS) composite scaffold and evaluation materials biomechanical properties; extract fresh bonemarrow2ml of SD rats were divided into A, B two groups: A fresh bone marrowaspirate1ml planted directly on the the aforementioned scaffolds for bone marrowseeding group; B group: extract isolated and purified bone marrow by whole bonemarrow in vitro culture to the third generation BMSCs planted on the same scaffoldfor cell seeding group; another scaffold not compound any ingredient as a controlgroup Group C, detection of the three materials in vitro biocompatibility, adhesionand proliferation activity, type Ⅰ, type Ⅲ collagen secretion, and then through thestandardization of surgical reconstruction of the anterior cruciate ligament in therabbit transection material into animals,4weeks after drawn vivo biocompatibilityevaluation materials.Results: the biomechanical properties of the composite scaffold than normal wereno significant differences in the anterior cruciate ligament; marrow seeding group wassignificantly higher than the cell seeding cells were incubated for4h cell adhesiondensity; proliferation of the bone marrow seeding group at each time point weresignificantly higher than those of cell seeding; the marrow seeding group and cellseeding group type Ⅰ, Ⅲ collagen secretion dose was significantly higher than thatof the control group, but the collagen secretion of bone marrow seeding group andcell seeding group between the two groups no significant difference; composite cellscaffold implantation in vivo not cause fatal immune rejection and severeinflammatory reaction, no significant ligament regeneration and vascularization.Conclusion: the adhesion, proliferation and extracellular matrix secretion of cellson the material can be observed visibly via fresh bone marrow aspirate seeded directlyon scaffolds, and are biocompatible in vivo short-term and biomechanical propertiesof materials to meet the requirements of normal human general motion.