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Objective To explored the bio-compatibility and cartilage regeneration of the rabbits genipin cross-linked decellularized scaffold, to provide experimental and theoretical support for the clinical application of genipin cross-linked decellularized scaffold. Methods Detergent-enzyme method was used to prepare decellularized tracheal scaffolds. Cellular content of native trachea and decellularized trachea were compared by 4', 6-diamidino-2-phenylindole(DAPI) staining. Masson trichrome staining was used to compare the histological structure of the progenitor tube, decellularized trachea, and genipin cross-linked decellularized trachea. Nine adult New Zealand white rabbits were randomly divided into autologous tracheal transplantation group (negative control group), allogeneic tracheal transplantation group (positive control group), and genipin cross-linked decellularized tracheal transplantation group (experimental group). Autologous bone marrow mesenchymal stem cells were implanted on the surface of trachea in each group. The blood cells and type II collagen were detected to compare the inflammatory response and chondrocyte regeneration after tracheal orthotopic transplantation in the three groups. Results After DAPI staining and light microscope observation (×200), the cell content of the acellular 7-cycle trachea [(143.0 ± 71.1) cells/field] was significantly lower than that of the native trachea [(853.5 ± 149.6) cells/ field], and the difference was statistically significant (P<0.001). Masson's trichrome staining showed that the tissue structure of genipin cross -linked decellularized trachea was more complete. Blood cell analysis and type II collagen test results showed that genipin cross-linked decellularized trachea transplanted with bone marrow mesenchymal stem cells after transplantation in situ has little rejection and can be converted into chondrocytes by the action of related growth factors in vivo. Conclusions Genipin cross-linked decellularized tracheal scaffold combined with stem cell transplantation can successfully construct a tracheal in situ replacement model. This study provides a strong support for the research of tissue engineering trachea.
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@#Objective Toinvestigatetheeffectofhydroxyapatite(HA)ontheosteogenicdifferentiationpotencyof adipose-derivedmesenchymalstemcells(ADSCs).Methods ADSCswereisolatedandpurifiedfromC57BL/6mice.Toxic effectsofhydroxyapatiteonADSCsweredetectedbycellproliferationassay.Alkalinephosphatase(ALP)assaywasusedto detecttheeffectsofdifferentconcentrationsofHAonosteogenicdifferentiationofADSCs.RelativemRNAexpressionlevels ofosteogenicgenes(BGLAP, ALP, COL1A1, OPNandRunx2)weremeasuredbyRT-PCR.Results Thelowconcentration ofHA(≤20mg/L)showedlesseffectonproliferationofADSCs.WiththeincreaseofHAconcentration,thecellproliferation decreased.Theco-cultureof20mg/LHAwithADSCssignificantlyincreasedtheALPactivity,andpromotedtheexpression ofosteoblast-relatedgenes(P＜0.01).Conclusion Hydroxyapatitehavetheabilitytoinducetheosteogenicdifferentiation ofADSCs,whichprovideatheoreticalbasisforcombinatingHAandADSCsintoanewboneengineeringscaffold.
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ObjectiveTo investigate the effect of IκB kinase (IKK-β) on migration and proliferation of bone marrow mesenchymal stem cells(BMSCs) from patients with systemic lupus erythematosus (SLE).MethodsHuman bone marrow aspirates were collected from iliac of six donors and six SLE patients and cultured in vitro.Migration of BMSCs were observed by wound healing and transwell migration assays.Proliferation of BMSCs was quantified by cell counting kit-8 assay.Total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expression of IKK-β at transcription level.The expression of IKK-β and phospho-IKK-β(p-IKK-β) protein were determined by Western blotting analysis.Statistical analysis was conducted with or Mann-Whitney rank test.Results① The migration rate of BMSCs from SLE patients(5.2±3.8)‰ were significantly reduced as compared with normal controls (7.0±2.9)‰(P＜0.05 ).The proliferation of BMSCs of SLE patients (0.21±0.49)was lower than that from healthy controls ( 1.00±0.35 )(P＜0.05 ).②) No difference in IKK-β3 mRNA expression between SLE ( 1.9± 1.4) subjects and normal controls (1.9±2.4) (P＞0.05).IKK-β protein expression of BMSCs from SLE patients (1.41 ±0.19) increased significantly compared with healthy controls (0.93±1.24) (P＜0.05).③ Inhibitor of IKK-β caused a significant increase in cell migration (3.3±1.6)‰ and proliferation (1.13±0.26) of BMSCs from SLE patients compared with untreated cells (2.3±1.1)‰ and (0.81±0.17),respectively (P＜0.05).ConclusionMigration and proliferation of BMSCs are significantly decreased in SLE patients.IKK-β may be involved in migration and proliferation of BMSCs.