BACKGROUND:Freeze-dried bone has strong immunogenicity due to insufficient removal of xenoantigen.Deproteinized bone and completely-decalcified bone have weak antigenicity,but the fomer has no osteoinductive property,and the latter has poor biomechanical property,so both of tem are limited in clinical application.OBJECTIVE:To observe the change of rabbit peripheral blood T lymphocyte subsets after transplantation of tissue engineered bone constituted by partially-decalcified freeze-dried bone scaffold and the histological changes of transplanted tissue.DESIGN,TIME AND SETTlNG:Randomized grouping,controlled animal observation.Performed in the State Key Laboratory of Biotherapy(I.E.Department of Stem cells and Tissue Engineering),Huaxi Hospital,Sichuan University between June 2006 and June 2007.MATERlALS:Tissue-engineered bone was in vitro constructed using osteoblasts.Which were derived from rabbit periosteum and used as seeding cells,and xenogeneic cancellous bone,which were antigen self-digested,partially-decalcified freeze-dried bone.METHODS:Forty-eight rabbits were randomly divided into the following 4 groups,with 12 rabbits in each group:partially-decalcified freeze-dried bone group(partially-decalcified bone group),tissue engineered bone group,autogenous bone group.And allogeneic bone group.Partially-decalcified freeze-dried bone,tissue engineered bone,autogenous bone,and allogeneic bone were respectively implanted into the 1 cm segmental defect in rabbit radius in above-mentioned groups.MAINOUTCOME MEASURES:Prior to and 1,2,and 4 weeks after implantation,the change of rabbit peripheraI blood T lymphocyte subsets were examined by flow cytometry；At 2,4,8,and 12 weeks after implantation,osteogenesis of the 4 materials was examined by routine histological examination.RESULTS:①In the partially-decalcifled bone group,peripheral blood CD4+and CD8+1r lymphocytes were significantly increased at 1 and 2weeks afterimplantationthan priortoimplantation(P<0.05).At 4 weeks after implantation.CD4+T lymphocytes were increased,but not significantly,compared with prior to implantation(P>0.05).In the autogenous bone group,CD4’and CD8+T lymphocytes were increased,but not significantly(P>0.05).In the allogeneic bone group,CD4’and CD8+T lymphocytes were significantly increased at weeks 1,2,and 4 after implantation than prior to implantation and the synchroale phase in the other groups(P<0.05).②inthetissue engineeredbonegroup,at week 2 after implantation,osteoblasts and chondroblasts were visible in the material porous,in addition,a new mixed tissue containing bone and cartilage formed and surrounded by osteoclasts,and partial rack was destroved and absorbed.At week 4,newly formed bone had turned into woven bone.At week 8.Lamellar bone was foand.And partially-decalcified freeze-dried bone was completely degraded and absorbed.At week 12,the implant had been completely substituted by lamellar bone,and medullary cavity was recanalized.CONCLUSION:Tissue-engineered bone constituted by taking partially-decalcified freeze-dried bone as scaflfold led to an increase in peripheral blood T lymphocytes,but which did not influence its good repair capabmtv of bone defects.

0.05),hints that the tenocyte's function was not disturbed. The DNA index of cells of GDBM group was 0.96 and 2.1% higher than the control group,indicating that the tenocytes grow and proliferate faster when being combine cultured on GDBM. Conclusion GDBM show good biocompatibility combined with tenocytes and they are promising extracellular matrix scaffold for cell transplantation in tendon tissue engineering.

Objective To observe the clinical effect of acupuncture combined with language training on subcortical aphasia.Methods 32 cases in treatment group and 26 cases in control group were given routine medication,in addition,treatment goup accepted acupuncture and language training.Results After 4 weeks of treatment,patients' language function improved significantly in treatment group compared with that of control(P<0.05).The total effective rate was 87.5% in treatment group,while it was 61.5% in control group(P<0.05).Conclusion Acupuncture combined with language training is effective on subcortical aphasia.

ObjectiveTo study the variant differentiated phase of the human bone marrow stromal cell (MSC) during induced adipogenesis course by morphological observation.MethodsAfter proliferated in vitro, MSCs isolated from bone marrow of the female adults volunteers were cultured with the adipogenetic inducers for 6—30 days. Morphological changes of the cells were observed everyday under Olympus contrast phase microscopy, and some specimens were stained with Oil-red-O.ResultsMSC showed long spindle shape before inducing, and then changed gradually to oval or round shape and the figure enlarged simultaneously. There were specifically morphological changes with lipid droplet emergence under plasmalemma and confluence gradually to lipid drop during differentiated into the phase of immature and mature adipocyte.ConclusionIt is easy to detect the MSC differentiated into the phase of immature and mature adipocyte with the specific morphological change of lipid droplet, while in the phase of preadipocyte and adipoblast, there is no special morphological change, and it may need some specific markers to detect.

Objective To design a modular two-direction scalpel handle with easy assembly and disassembly to solve the problems of common handle in function singleness,directivity,assembling and disassembling,pick-up and etc.Methods A scalpel handle module involving in a single-groove two-direction tailstock,multifunctional handle and etc was designed according to international standards,which consisted of more than 10 kinds of instruments for orthopedic surgery and etc such as two-direction scalpel handle with easy assembly and disassembly,bent wrench,probe introducer,needle-knife remover and measuring tools.Simulation experiment,clinical trial and control test were carried out to verify the efficacy of the handle module.Results It's proved that the handle module gained advantages in safety,convenience,prevention of sharp instrument injury,decrease of human errors and etc.Conclusion The handle module behaves well in modularity,integration,multifunction,two-direction adaptability,easy assembly and disassembly,safety,storage and carrying,high costperformance ratio and etc,meets the requirements of Joint Commission on Accreditation of Healthcare Organization,and is suitable for military and civilian uses.

BACKGROUND:Hypertrophic differentiation of chondrocytes is the sign of starting endochondral ossification, and it is also an essential step in endochondral ossification, which is a cascade reaction and difficult to be blocked once started. The end result is the formation of bone structure. RNA interference is a post-transcriptional gene silencing. Relevant studies have shown that the use of RNA interference to block the expression of core binding factorα1 (Cbfα1) can effectively inhibit the formation of heterotopic ossification. OBJECTIVE:To use RNA intereference technology to suppress Cbfα1 expression so as to achieve the purpose of blocking the hypertrophic diferentiation of chondrocytes. METHODs: We constructed an adenovirus containing siRNA against Cbfα1 (Ad-Cbfα1-siRNA). Retinoic acid and interleukin-1α were used to induce hypertrophic differetiation of chondrocytes, and then Ad-Cbfα1-siRNA was utilized to inhibit the hypertrophic differentiation of chondrocytes. Immunohistochemistry method was used to analyze the expression of Cbfα1. RESULTS AND CONCLUSION:After induction with retinoic acid and interleukin-1α, the chondrocytes in the negative control virus group appeared to have hypertrophy and the expression of Cbfα1 was positive. In the Ad-Cbα1-siRNA group, the expression of Cbfα1 was negative. These findings suggest that the inhibition of Cbfα1 by RNA interference can be a powerful way to prevent the hypertrophic differentiation of chondrocytes .

This study was aimed to evaluate the cellular compatibility of the small intestinal submucosal(e) (SIS). Prepared by use of pig jejunum. SIS were cocultured with human embryonic periosteal osteoblasts (HEPOB), human embryonic skin fibroblasts (HESFB) and rabbit renal vascular endothelial cells (RRVEC) respectively. The cell growth, attachment, cell cycle, cell apoptosis rate were detected to evaluate the cellular compatibility of SIS. The three kinds of cells attached onto SIS and grew well. SIS accelerated the growth of RRVEC. No effects of SIS were detected on cell cycle and cell apoptosis rate in the three kinds of cells. SIS has good cellular compatibility without cytotoxicity. The porous structure of SIS is suited for the growth of HEPOB, HESFB and RRVEC in three dimensions in the scaffold. SIS is a good bio-derived material of tissue engineering.
Animals ; Cell Differentiation ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Coculture Techniques ; Extracellular Matrix ; physiology ; Histocompatibility ; Intestinal Mucosa ; cytology ; Jejunum ; cytology ; Osteoblasts ; cytology ; physiology ; Swine ; Tissue Engineering

This paper is aimed to investigate the feasibility of applying the small intestine submucosa (SIS) as the scaffold in constructing tissue engineering cartilage in vitro. We obtained SIS from the small intestine of specific pathogen-free pigs. Then we isolated tunica submucosa layer from the mucosal, muscular, and serosal layers by gentle mechanic abrasion. The SIS was made acellular by combination of detergent and enzyme digestion. The chondrocytes were seeded onto the SIS and were cultured for 3 weeks. The cell growth, attachment and distribution were detected by histochemical stain, immunohistochemical stain and scan electron microscope. The chondrocytes could adhere and grow well on the matrix surface, and synthesize a large of the GAG and type U collagen. However, the chondrocytes grew only on the surface andsuperficial layer of the scaffold, they did not move into the inner part of the scaffold. It could be concluded that SIS has good cellular compatibility without cytotoxicity and provides temporary substrate to which these anchorage-dependent cells can adhere, and stimulate the chondrocytes anchored on the scaffold to proliferate and keep differentiated phenotype. Further study will be needed to promote the ability of chondrocyte chemotaxis in order to distribute the chondrocytes into the whole scaffold uniformly.
Animals ; Cell Adhesion ; Cell Culture Techniques ; Cell Proliferation ; Chondrocytes ; cytology ; Chondrogenesis ; physiology ; Intestinal Mucosa ; cytology ; Intestine, Small ; cytology ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

Experiments have been performed to investigate why the biomechanical strength of repaired tendons is lower than that of the normal tendon when the engineered tendons are implanted in vivo to replace the tendon defects. We seeded the primary culture tendon cells derived from Roman chickens' digital flexor tendons on the degradable polyglycolic acid meshes to construct tissue-engineered tendons. The flexor tendon defects (0.5 cm-0.8 cm) excised in second digit bilaterally in 20 Roman chickens, had been repaired with the constructed tissue-engineered tendons. The samples of repaired tendons were collected at 2, 4, 6 and 8 weeks after operation. Tests for scaffold weight, hydroxyproline content, and mechanical strength of the samples were performed. We found that from 2 weeks to 8 weeks afteroperation, the weight of the scaffolds decreased significantly, almost disappearing at 8 weeks; the hydroxyproline content determining the total collagen content increased gradually without significance; mechanically, both energy at break and tensile strength showed a tendency of drastic decrease at first 4 weeks afteroperation and a gradual increase afterwards, but the tensile strength at 8 weeks afteroperation was only 23% of that of the normal tendon. We conclude that the lower biomechanical strength of repaired tendons is owing to the serious mismatch between scaffold degradation and collagen synthesis.
Achilles Tendon ; injuries ; Animals ; Biocompatible Materials ; metabolism ; Bioprosthesis ; Cell Culture Techniques ; Chickens ; Collagen ; metabolism ; Female ; Prosthesis Implantation ; Tendon Injuries ; surgery ; Tendons ; cytology ; Tensile Strength ; Tissue Engineering ; methods

This is a study on the histologic pattern and mechanical properties of tissue-engineered tendon implanted for treatment of tendon defects. Tendons were resected from Roman chickens. Tendon cells were isolated from the tendons and cultured in vitro. The 2nd-4th passages of tendon cells were seeded on the degradable polyglycolic acid mesh to form cell-scaffold composites, which were further cultured for 7-10 days to construct tissue-engineered tendons. The tendon defects, 0.5 cm-0.8 cm in length, were made in the second digit flexor tendon bilaterally in 20 Roman chickens and then bridged with the constructed tissue-engineered tendons. At 2 weeks, 4 weeks, 6 weeks, and 8 weeks post-operation, the samples of regenerated tendons were collected for gross examination, histologic staining and biomechanical test. After implantation of the tissue-engineered tendons, the wounds healed well. The gross appearance, the cells and collagen fibers arrangement of the regenerated tendons were similar to those of natural tendons, but there were relatively not many closely packed collagen fiber bundles organized in parallel with the tendons ("remodel"), so the maximum tensile force increased slowly and its value was 15.40+/-10.63 N at 8 weeks after surgery, reaching only 23% of that of natural tendon. The maximum strain was 22.49%+/-10.21% at 8 weeks, being 10% higher than that of natural tendons. Polyglycolic acid scaffolds are degraded in vivo so rapidly that the regenerated tendons lose the normal biomechanical stimulus and then are unable to be remodeled. As a result, the mechanical strength of regenerated tendons is much lower than that of natural tendons. These results suggest that the normal biomechanical stimulus may be an important factor for the regenerated tendons to remodel.
Animals ; Animals, Newborn ; Biomechanical Phenomena ; methods ; Cell Separation ; Cells, Cultured ; Chickens ; Female ; Implants, Experimental ; Tendon Injuries ; surgery ; Tendons ; cytology ; physiology ; surgery ; Tensile Strength ; Tissue Engineering ; methods