多孔壳聚糖膜的制备及皮下植入组织学观察

Preparation of porous chitosan membrane and observation of its subcutaneously implanted histology

目的 分析交联剂戊二醛对多孔壳聚糖膜孔隙率与孔径的影响，并探讨该多孔膜作为皮下植入式葡萄糖传感器保护膜的可行性。方法 通过致孔剂法利用硅胶和不同量的戊二醛制成多孔壳聚糖膜，并采用密度法和切片法分析其孔隙率与孔径等结构参数，扫描电子显微镜观察其表面形态。将无交联的多孔膜植入到9只SD大鼠皮下，第7、17、45天取材切片染色，观察组织形态学变化，定量分析多孔膜内外胶原沉积与血管增生密度。结果 戊二醛能提高多孔壳聚糖膜的孔隙率，5％交联度时孔隙率最大达76．2％／73．0％（密度法／切片法），但同时降低了多孔膜孔径。使用孔径最大的不交联多孔膜（38．5斗m）皮下植入，膜内胶原沉积含量由第7天的6．74％增加到第45天的22．5％，而Masson染色测量的膜内增生血管密度由O．37％增加到2．56％，与HE染色测量结果一致（由0．11％增加到1．65％）。第45天时膜外胶原沉积含量38．3％，是膜内的1．7倍。结论多孔壳聚糖膜能降低材料．组织界面纤维成分的致密性，增加血管增生，具有良好的组织相容胜，有望应用于植入式葡萄糖传感器。

Objective To analyze the impact of glutaraldehyde crosslinker on porosity and pore size of porous ehitosan membrane （PCSM） and to explore the feasibility of PCSM as the proiective membrane of glucose sensor. Methods The PCSM was prepared by particle leaching method using silica gel and different amounts of glutaraldehyde. Density method and slice method were used to analyze the structural parameters of PCSM such as porosity and pore size and its surface morphology was observed with scanning electron microscope （SCM）. Thereafter, the uncrosslinked porous membranes were subeutaneously transplanted into 9 SD rats and extracted to observe their histomorphological changes by section staining at the 7th, 17th and 45th day after transplantation. Quantitative analysis was then performed on collagen deposition inside and outside of the porous membranes and vascular proliferation density. Results The glutaraldehyde could increase the PCSM porosity that could reach the maximum of 76.2%/73.0% with 5% degree of crosslinking （density method vs slice method）, but it could also decrease the pore size of porous membrane. The uncrosslinked porous membranes with the largest pore size （38.5 μm） were used and subcutaneously transplanted, which resulted in an increase of intramembranous collagen deposition content from 6.74% at the 7th day to 22.5% at the 45th day. Moreover, the intramembranous proliferated capillary density was increased from 0.37% to 2.56% using Masson staining, which was consistent with the measured outcomes using HE staining （ranged from 0.11% to 1.65% ）. At the 45th day, the content of collagen deposition outside of the membrane was 38.3% that was 1.7 times of the intramembranous collagen deposition content. Conclusions PSCM may decrease the compactness fibrous composition of material-tissue interfacebut increase vascular proliferation, with favorable histocompatibility. Thus, PCSM appears promising inapplication in transplanted glucose sensor.