| Part one:Preparation of the compound electrospun poly(ε-caprolactone)/gelatin fibrous scaffoldsObjective To study the preparation of the compound electrospun poly (e-caprolactone)/gelatin fibrous scaffolds.Methods Two solutions of gelatin and poly(ε-caprolactone)(mass ratio of 1:1) were mixed together under magnetic stirring to form a homogenous solution,than using electrospinning technology to obtain different porosities electrospun fibrous membrane. we using Scanning electron microscopy to observe the morphological characteristics, measureing porosity and fiber diameter. Through immersed in methylacrylic acid anhydride modified gelatin in order to bonding together by crosslinking reaction, under APS provided free radicals and TEMED as catalyster. MC3T3-E1 cells were added to compound scaffold for co-culture, than cell toxicity were carried out to examine the biocompatibility of the compound scaffold by CCK-8 kit.Results The electrospun poly(ε-caprolactone)/gelatin fibrous had smooth surface, homogeneous distribution and high porosity. One of the membrane pore rate is about 75%, the other is about 55%. The fiber diameter of the larger pore size membrane with an average of (2.7673±0.2238) μm, while the small pore size membrane with an average of (2.2514±0.2988) μm. Compare with simplex electrospun fibrous membrane, the compound electrospun poly(ε-caprolactone)/gelatin fibrous membrane scaffolds have better hydrophilicity and mechanical capacity. Provides a good foundation for cell adhesion and engraftment. After co-culture with MC3T3-E1 cells for seven days, Cell counting Kit-8 (CCK-8) was used to analyze the cell compatibility. The result of CCK-8 assay show that the cell viability of test group (1.9393±0.4174)had no difference compared with the control group(2.4574± 0.318).Conclusion The electrospun poly(ε-caprolactone)/gelatin fibrous membrane had smooth surface, homogeneous distribution and high porosity. Compound electrospun poly(ε-caprolactone)/gelatin scaffolds can be built through the crosslink reaction of modified gelatin with cytocompatibility.Part two:Experimental study of compound electrospun poly(ε-caprolactone)/gelatin fibrous scaffolds for mandibular bone defect regeneration in rabbit.Objective To study the biocompatibility and the effect of mandibular bone defects reconstruction of the compound electrospun poly(ε-caprolactone)/gelatin fibrous scaffolds in rabbit.Methods Created the size was 10 mm (long) x 8 mm (wide) x 5 mm (thickness) unilateral mandibular bone defect in New Zealand white rabbit. Then we planted the conpounded scaffold into experimental group bone defect, the blank group left untreated. Rabbit mandibular bone were harvested and put into 4% neutral paraformaldehyde solution for one day at 12 weeks postoperatively. Regeneration bone tissue was subjected to microcomputed tomographic(micro-CT) examination, in order to have tha information about neogenesis bone mineral density and neogenesis bone constituent ratio. After decalcified by Ethylenediaminetetraacetic acid disodium salt for 3 month, regular embedding, sectioning, the histologic section were stained with hematoxylin-eosin (HE) and Masson.Results The experimental group showed higher neonatal bone mineral density (BMD,0.220±0.253) and Bone Volume/Tissue Volume (BV/TV,11.941±3.046) than the control group(BMD,0.179±0.348; BV/TV,5.809±2.433)(P<0.05) by micro-CT examination. Histological study indicated that there was no fiber connective tissue invaded into bone defect areas in experimental group. Scaffolds had been partly degraded and regenerated bone with lamellar structure, also a lot of blood vessel formation was founded around the new bone.Conclusion This compound electrospun gelatin/poly(ε-caprolactone) fibrous scaffolds have a good biocompatibility, can effectively maintain space for bone regeneration and promote new bone formation and neovascularization. |
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