Abstract: Objective To assess the feasibility of transferring major histocompatibility complex (MHC) gene into the thymus to mitigate xenograft rejection. Methods By molecular cloning technique, we extracted and proliferated the-H-2K d gene from donor mice (MHC class Ⅰ gene of Balb/c mice) and constructed the expression vector plasmid of pCI-H-2K d. Twenty SD rats were selected as receptors, and by using random number table, they were divided into the experimental group and the control group with equal number of rats in each group. By ultrasoundguided puncture and lipofection method, the pCI-H-2Kd was injected into thymus of SD rats in the experimental group and meanwhile, empty vector plasmid of pCIneo was injected into thymus of SD rats in the control group. Subsequently, we transplanted the donor mice myocardium xenografts into the receptor rats, and observed the xenograft rejection in both the two groups. Results The survival time of the xenotransplanted myocardium in the experimental group was significantly longer than that in the control group (14.61±2.98 d vs. 6.40±1.58 d, t=-7.619,Plt;0.05). Microtome section of transplanted myocardium in the control group showed a relatively large amount of lymphocyte infiltration and necrosis occurred to most part of the transplanted myocardium, while microtome section of experiment group showed no lymphocyte infiltration and most of the cells of the transplanted myocardium were still alive. After mixed lymphocyte culture, the reaction of receptors to donor cells in the experiment group was obviously lower than that in the control group (t=4.758, P=0.000).After the count by flow cytometer, the xenoMHC molecules were expressed in the receptors’ thymus with a transfection efficiency of 60.7%. Conclusion Our findings suggest that xenograft rejection can be mitigated substantially by donor’s MHC gene transferring into receptor’s thymus. This may provide theoretical and experimental evidence for inducing xenotransplantation tolerance.
Objective Surface modification of nitinol (NiTi) shape memory alloy is an available method to prevent nickel ion release and coating with titanium-niobium (TiNb) alloy will not affect the superelasticity and shape memory of NiTi. To evaluate the bone histocompatibil ity of NiTi shape memory alloy implants coated by TiNb in vivo. Methods NiTi memory alloy columns which were 4 mm in diameter and 12 mm in length were coated with Ti (Ti-coating group) and TiNb alloy (TiNb-coating group) respectively by magnetron sputtering technique. And NiTi group were not coated on the surface. Fifteen mongrel dogs were divided into 3 groups randomly with 5 dogs in each group. NiTi, Ti-coating and TiNb-coating columns were implanted into the lateral femoral cortex of each group, respectively. There were 10 columns embedded in eachdog’s femur whose distance was 1.0 cm to 1.5 cm from each other. The materials were obtained 12 months after operation. After X-ray photography, only those columns which were perpendicular to the cortex of the femur shaft were selected for subsequent analysis. Push-out tests were performed to attain the maximum shear strength (the number of specimens of TiNi group, Ticoating group, and TiNb-coating group were 12, 10, and 14, respectively). Undecalcified sections were used for histological observation and the calculation of osseointegration rate (the number of specimens of TiNi group, Ti-coating group, and TiNb-coating group were 8, 5, and 10, respectively). Results The maximum shear strength of Ti-coating group (95.10 ± 10.03) MPa, and TiNb-coating group (91.20 ± 15.42) MPa were significantly higher than that of NiTi group (71.60 ± 14.24) MPa (P lt; 0.01). Gimesa staining showed that no obvious macrophage and inflammation cell was observed in 3 groups. The osseointegration rates of NiTi group, Ti-coating group, and TiNb-coating group were (21.30% ± 0.23%), (32.50% ± 0.31%), and (38.60% ± 0.58%), respectively; there were significant differences among 3 groups (P lt; 0.01). Conclusion The implants of 3 groups all have good bone histocompatabil ity. But the osseointegration rate and the shear strength in the Ti-coating group and the TiNb-coating group were better than those in the NiTi group, the TiNb-coating group is the best among them.
The biocompatible hydrogel was fabricated under suitable conditions with natural dextran and polyethylene glycol (PEG) as the reaction materials. The oligomer (Dex-AI) was firstly synthesized with dextran and allylisocyanate (AI). This Dex-AI was then reacted with poly (ethyleneglycoldiacrylate) (PEGDA) under the mass ratio of 4∶6 to get hydrogel (DP) with the maximum water absorption of 810%. This hydrogel was grafted onto the surface of medical catheter via diphenyl ketone treatment under ultraviolet (UV) initiator. The surface contact angle became lower from (97 ± 6.1)° to (25 ± 4.2)° after the catheter surface was grafted with hydrogel DP, which suggests that the catheter possesses super hydrophilicity with hydrogel grafting. The in vivo evaluation after they were implanted into ICR rats subcutaneously verified that this catheter had less serious inflammation and possessed better histocompatibility comparing with the untreated medical catheter. Therefore, it could be concluded that hydrogel grafting is a good technology for patients to reduce inflammation due to catheter implantation, esp. for the case of retention in body for a relative long time.
ObjectiveTo investigate the in vivo degradation and histocompatibility of modified chitosan based on conductive composite nerve conduit, so as to provide a new scaffold material for the construction of tissue engineered nerve.MethodsThe nano polypyrrole (PPy) was synthesized by microemulsion polymerization, blended with chitosan, and then formed conduit by injecting the mixed solution into a customized conduit formation model. After freeze-drying and deacidification, the nano PPy/chitosan composite conduit (CP conduit) was prepared. Then the CP conduits with different acetyl degree were resulted undergoing varying acetylation for 30, 60, and 90 minutes (CAP1, CAP2, CAP3 conduits). Fourier infrared absorption spectrum and scanning electron microscopy (SEM) were used to identify the conduits. And the conductivity was measured by four-probe conductometer. The above conduits were implanted after the subcutaneous fascial tunnels were made symmetrically on both sides of the back of 30 female Sprague Dawley rats. At 2, 4, 6, 8, 10, and 12 weeks after operation, the morphology, the microstructure, and the degradation rate were observed and measured to assess the in vivo degradation of conduits. HE staining and anti-macrophage immunofluorescence staining were performed to observe the histocompatibility in vivo.ResultsThe characteristic peaks of the amide Ⅱ band around 1 562 cm−1 appeared after being acetylated, indicating that the acetylation modification of chitosan was successful. There was no significant difference in conductivity between conduits (P>0.05). SEM observation showed that the surfaces of the conduits in all groups were similar with relatively smooth surface and compact structure. After the conduits were implanted into the rats, with the extension of time, all conduits were collapsed, especially on the CAP3 conduit. All conduits had different degrees of mass loss, and the higher the degree of acetylation, the greater the mass change (P<0.05). SEM observation showed that there were more pores at 12 weeks after implantation, and the pores showed an increasing trend as the degree of acetylation increased. Histological observation showed that there were more macrophages and lymphocytes infiltration in each group at the early stage. With the extension of implantation time, lymphocytes decreased, fibroblasts increased, and collagen fibers proliferated significantly. ConclusionThe modified chitosan basedon conductive composite nerve conduit made of nano-PPy/chitosan composite with different acetylation degrees has good biocompatibility, conductivity, and biodegradability correlated with acetylation degree in vivo, which provide a new scaffold material for the construction of tissue engineered nerve.