The Technology Of Autologous MSCs/Chondro-Gide^? Engineered Cartilage Regeneration Preliminary Exploration Of Clinical Application Standardization

Background and ObjectiveBackground:Articular cartilage defects are one of the most common disorders in clinic. Although a variety of procedures are being employed to repair cartilage damage,methods that result in consistent durable repair tissue are not yet available.Tissue engineering,a prospective technology,has been successfully applied in abroad,such as Verigen's MACI technology in Germany,FBA's Hyalograft technology in Italy,Ars's CaRes in USA etc.At the same time,the standards of such technology and product are produced parallelly.Objective:To track and collect actively relevant standards of tissue engineering cartilage regeneration technology,then summarize these information to get received standards and guidelines.On the base of our previous research results,to establish our own autologous MSCs/Chondro-Gide^? engineered cartilage regeneration standardization, targetting to clinical application.To verify the availability of the standardization by the way of animal experiments.To realize normalization of tissue engineering cartilage regeneration in clinic through the development of related industry standards.To promote research level of engineering cartilage regeneration,the standardization of technology and the process of industrialization.Methods:1.Establishing the standard of tissue-engineered cartilage regeneration.(1) Research and refer to tissue engineered cartilage repair-related standards of the American Society of Testing Materials.(2) Research and refer to relevant provisions of'biological material and medical equipment biological evaluation criteria",enacted by the International Organization for Standardization.(3) Research and refer to relevant provisions of the national Drug Administration Medical Device Quality Supervision and Inspection and the Chinese Center for Drug testing.(4) Research and refer to relevant standards of the Association of the International Cartilage Repair.2.Establishing the Standardization of the Autologous MSCs/Chondro-Gide^? engineered cartilage regeneration in clinic.(1) Refer to the first part of the study.(2) Refer to the standardization of the autologous matrix-induced chondrogenesis technology.(3) Refer to the technology of culturing autologous serum MSCs in vitro.3.Repair cartilage defect in animal experiment through the way in accordance with the standardization of the Autologous MSCs/Chondro-Gide^? engineered cartilage regeneration technology.(1) Establish the cartilage defect model of goat's femur trochlea.Prepare the bone marrow and circulation blood which are enough to repair 4 cm² defects.(2) Acquire goat blood and bone marrow by standard process.Then extract autologous serum from blood and culture MSCs in vitro.(3) Establish the cartilage defect model of goat's femur trochlea.Repair goat cartilage defects by standardization process of the autologous MSCs/Chondro-ide^? engineered cartilage regeneration technology.(4) The third generation MSCs cell suspension(2ml/knee) injected in goats knee cartilage defect which has been covered by Autologous MSCs/Chondro-Gide^? engineered cartilage.After 4 week,preliminary evaluate integration and fixation between repair areas of articular cartilage defect and normal areas by general observation, imageology,arthroscopy and histology.Wakitani scores were counted to evaluate the repairing effect.Results:1.The establishment of the tissue engineering cartilage repair standardization.(1) Standardized tissue-engineered cartilage repair terminology.(2) Established a standardized tissue-engineered cartilage repair animal models.(3) Assessed the security of tissue engineering products.(4) Established collection of standardized cartilage seeds cells. (5) Established transport approach and related installation manuals after cartilage cells collected.2.The establishment of the standardization of the Autologous MSCs/Chondro-Gide^? engineered cartilage regeneration in clinic.(1) Established the standardization of preoperative assessment.(2) Established the standardization of the acquisition and cultivating of cartilage seeds cells in vitro.(3) Established the standardization process of the technology of autologous MSCs/Chondro-Gide^? engineered cartilage regeneration.(4) Established the standardization of post-operative rehabilitation.3.Repair cartilage defect in animal experiment through the way in accordance with the standardization of the Autologous MSCs/Chondro-Gide^? engineered cartilage regeneration technology.(1) Volume of bone marrow and circulation blood which are enough to repair 4cm² defects was 7.36 ml and 100 ml respectively.(2) Adequate cartilage seed cells can aquried according to the standardization of proliferating process.(3) Collagen gel fixed Chondro-Gide^? film is available.MRI and Arthroscopy illustrated implant was fixed well,the surface of cartilage defect repair is flat and the repair area integrated well with the surrounding tissue.And no synovium hyperplasy and fluidification were observed in the joint(4) After 4 week,the surface of cartilage defect repair is flat,and the color tend to integrate well with the surrounding tissue in experience group.Histology illustrated implant and surrounding implant tissue integrated well,naive cartilage cells evenly distributed beneath the repair surface and inflammatory response and immune rejection reaction were not observed in experience group.Histology of AMIC group and vacant group illustrated fibrous tissue cover area of cartilage defect.4w Wakitani scores in experience group,AMIC group and vacant group are 4.63±1.18,8.70±0.82,11.35±1.86 respectively.8w Wakitani scores are 3.95±1.02,7.06±1.13,8.25±1.38 respectively.Conclusion:1.At present a international common sense in cartilage tissue engineering,include its terminology,collection and transport of seed cell,animal models and assessing security of products and technology,is unified.There is no uniform standardization in clinical application of tissue-engineered cartilage regeneration.This study track,collect relevant information and summarize a common standards,which set up a foundation to apply this technology in clinic.2.On the base of the common stands,a clincal standardization of Autologous MSCs/Chondro-Gide^? engineered cartilage regeneration technology was set up,which norms collecting and culturing of MSCs and clinical procedure.Engineered cartilage can be implanted in a minimally invasive,easily operable and secure way in accordance with this clincal standardization.3.According to the standardized processes,the technology can be easily completed in arthroscopic.Implants fixed firmly without falling out.After 4 week,The cartilage defect repair surface formation,The engineered cartilage tend to integrate well with the surrounding tissue.Histological observation:Restoration Area and normal tissue integration. Beneath the surface there are more naive cartilage cells which evenly distributed,no inflammatory response and immune rejection reaction.4.Cartilage defect could repaired in animal experiment under arthroscopy through the way in accordance with the standardization of the Autologous MSCs/Chondro-Gide^? engineered cartilage regeneration technology.Cartilage defect could repaired well which were confirmed by MRI,arthroscopy and histology.4w,8w upgraded Wakitani scores illustrated the repair effect of experience group surpass that of AMICgroup and vacant group.