| 【Background】Maxillofacial soft tissue defects due to maxillofacial trauma, oncologic resectivesurgery, facial burn and other variety of reasons causes a direct impact on a number ofphysiological functions like chewing, swallowing, breathing, speech and esthetics. Oral andmaxillofacial surgeons face major challenges on restoration of functions and improvepatients quality of life and needs to master basic and microvascular reconstructiontechniques. Major clinical reconstructive methods includes various flaps from differentpart of body, synthetic materials, fat transplantation from body adipose stem celltransplantation, etc. Autologous fat stem cells can increase the effect of fat transplantation,but there are some limitations for the application of body fat stem cells, such as cultured in vitro with autologous stem cell cycle is long, the part of the patient’s own stem cells islimited or unwilling to accept liposuction. As reported in the literature, allogeneic stemcell has the characteristics of low immunogenicity, and has been used in some clinical cases.【Objective】To compare the difference in survival of allogeneic fat stem cells and autologous fatstem, and to provide an experimental basis for clinical implementation of allogeneic fat stemcell transplantation.【Methods】1. Establishment of animal models: Rabbit ear artery end automatically, vein bifurcationof the ear artery and the inner ear edge vein at the junction of the mid-point of a circle with aradius of15cm. Separated from the skin and periosteum, periosteal stripping was required inaccordance with the experimental groups, with18#syringe2ml complex was transplantedto this site.2. Identification and culture of rabbits fat stem cells: under sterile conditions,approximately8ml of fat removed from the back of the rabbit, by using digestioncentrifugation and cultured adipose stem cells at37°C,5%CO2incubator. HE staining,the streaming detection and confirmation of pre-transplant fat stem cells is done.3. To transplanted the composites into the ear of the rabbits: In accordance with theexperimental groups,we put ADSCs, AG and PRF into the ear of the rabbits.4. Experimental group:30healthy New Zealand rabbits,were divided into5groups:GroupA:AG;GroupB:AG+PRF;GroupC:AG+ADSCs(adipose-derivedstem cells, fatstem cells);Group D: AG+ADSCs+PRF Group E: AG+PRF+allograft ADSCs.5. In vivo transplantation of the injectable fat particles complex: AG, autologous,allogeneic ADSCs prf grouped by experimental transplantation to rabbit ears correspondingparts.6. Morphological and immunological observations of complex after transplantation:general appearance, HE staining, rabbit ears light transmission experiments, caliper, ultrasound measurement of survival to observe the morphological changes. HE staining,CD4/CD8lymphocyte subsets, IL-2, IL-4observation immunological changes.7. Data collection and statistical analysis. Results were statistically analyzed usingSPSS18.0software, each group data were analyzed by ANOVA single factor analysis ofvariance, and comparison among the four groups of survival differences were made.【Results】1. Adipose stem cells isolated and cultured from the back of the rabbits differentiatedto the third generation, short spindle cells covering the bottom. On HE staining, cellswere spindle or polygonal, on flow analysis cells were CD29positive (87.39±2.85)%,CD31negative,(1.37±0.61)%. Osteogenic and adipogenic induction showed osteogenicand adipogenic differentiation potential of stem cells respectively.2. Morphological and immunological aspects comparison of composite transplantationin rabbits,In terms of morphology, the5packet after transplantation, there was no liquefactionnecrosis, infection, post-transplant rabbits general condition showed no obviousabnormalities under inspection. B ultrasound results, used to measure and calculate thesurvival rate of each group showed no significant difference (P>0.05) among group D andE but there is a significant difference with the other groups (P <0.05),Three months aftertransplantation fat survival rate of A~E were54.39±7.72,63.72±3.93,65.39±2.48,72.62±4.72,72.56±4.26,six months after transplantation fat survival rate of A~E were38.73±9.81,61.29±1.61,62.34±1.65,73.94±2.73,69.83±5.12; Under HE staining, adiposetissue structure were clear and complete, no significant liquefaction necrosis and infection.Histologically, D, E two groups no significant differences, but there are significantdifferences with the other groups. In immunology, by slicing HE staining in each group haveno significant liquefaction necrosis, and no significant immune rejection; found bymeasuring the CD4/CD8ratio in peripheral blood of rabbits at different time points, groupsD and E were significantly different from other groups(P>0.05). No significant immunerejection is detected, in the plasma of rabbits IL-2, IL-4levels change at different time points, and found that D group E group was no significant difference (P>0.05), and thus nosignificant immune rejection.【Conclusion】1. Adipose stem cells isolated and cultured from rabbit back has a strong proliferation,and differentiation capacity.2. AG PRF composite allogeneic ADSCs AG PRF composite autologous ADSCs twogroups after transplantation could improve the survival rate of the fat, and the fat graftsurvival after critical role.3. AG PRF composite allogeneic ADSCs transplantation has no significant immunerejection and thus can provide important experimental basis for transplantation of allogeneicfat in clinical applications. |
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