To evaluate the biocompatibility of poly(lactic acid/glycolic acid/asparagic acid-co-polyethylene glycol)(PLGA-ASP-PEG )tri-block copolymer in vitro,L929 fibroblast was co-cultured with the copolymer for cytotoxicity,hemolysis and pyrogen tests.And,compared with PLGA,the adhesiveness rate of the copolymer was calculated.The experimental results show that the toxicity gradation of the material was 0-1; L929 fibroblasts had a good cell morphology and proliferated rapidly on the surface of the material;hemolysis ratio was 3.08%;there was no pyrogen reaction.The adhesiveness of PLGA-ASP-PEG was better than that of the PLGA's(P<0.05).The results confirm that the PLGA-ASP-PEG has a good biocompatibility.
To experimentally evaluate the ectopic osteogenetic capacity of synthesized BMP2-derived peptide P24 combined with poly lactic-co-glycolic acid (PLGA), Wistar rats were di- vided into two groups: group A, in which BMP2-derived peptide P24/PLGA complex was implanted, and group B which received simple PLGA implant. The complex was respectively implanted into the back muscles of rats. Samples were taken the 1st, 4th, 8th, and the 12th week after the implantation. Their bone formation was detected by X-ray examination, and tissue response was histologically ob- served. Western blotting was used for the detection of the expression of collagen Ⅰ (Col-Ⅰ) and osteopontin (OPN). There was acute inflammation in the tissue around both types of implants at early stage. The cartilage was found around implant areas 4 weeks after the implantation of BMP2-derived peptide p24/PLGA complex, 8 weeks after the implantation, osteoblasts were found, and 12 weeks after the implantation, typical trabecular bone structure was observed. In group B, after 12 weeks, no osteoblasts were found. It is concluded that PLGA is an ideal scaffold material for bone tissue engi- neering. BMP2-derived peptide can start endochondral ossification and is more effective in inducing ectopic osteogenesis.
In this study, the bioactivity of a novel BMP2-derived oligopeptide P24 was investigated by using the model of rabbit femoral defect after loaded in the biodegradable poly (lactic acid / glycolic acid / asparagic acid-co-polyethylene glycol) (PLGA-[ASP-PEG]). A 1.5-cm unilateral segmental bone defect was created in the left femoral diaphysis in each of the 30 new zealand white rabbits. The defects of 18 legs filled with BMP2-derived peptide P24 combined with PLGA-[ASP-PEG] scaffold serves as the experimental group, and the defects in the rest 12 rabbits filled with (PLGA-[ASP-PEG]) without P24 as control group. The bone-repairing capability in the target region of the two group was grossly, radiologically, histopathologically and biomechanically evaluated 4, 8 and 12 weeks after the operation. Our results showed that in each group, primary healing of incision was achieved in the two groups. Radiographically, in experimental group, defects were filled with induced callus within 8 weeks, and a cortical bone-like structure was observed in some animals at the 12th week. According to the standardized stage of bone defect repair, 9 (64.28%) achieved grade-4 healing. In contrast, little bone formation was seen in the defects even 12 weeks after the operation, and 5 (62.50%) had grade 0 healing in this group. Histologically, tissue engineering material was mostly absorbed and cartilage was found around implants in the experimental group at the 4th week; 8 weeks after operation, the engineering material was completely absorbed, and formation of woven bone was observed and typical trabecular bone structure could be seen. In control group, 8 weeks after operation, the defect was filled with fibrous tissues, and no bone-like structure was observed. Statistical analysis showed very significant difference in biomechanical indicators between the two groups (P<0.05). It is concluded that new oligopeptide P24 can induce excellent bone regeneration and promote bone repair.
A new biomimetic bone tissue engineering scaffold material, nano-HA/ PLGA-(PEG-A_ SP)_n composite, was synthesized by a biologically inspired self-assembling approach. A novel biodegradable PLGA-(PEG-A_ SP)_n copolymer with pendant amine functional groups and enhanced hydrophilicity was synthesized by bulk ring-opening copolymerization by DL-lactide(DLLA) and glycolide(GA) with Aspartic acid (A_ SP)-Polyethylene glycol(PEG) alt-prepolymer. A Three-dimensional, porous scaffold of the PLGA-(PEG-A_ SP)_n copolymer was fabricated by a solvent casting, particulate leaching process. The scaffold was then incubated in modified simulated body fluid(mSBF). Growth of HA nanocrystals on the inner pore surfaces of the porous scaffold is confirmed by calcium ion binding analyses, SEM, mass increase measurements and quantification of phosphate content within scaffolds. SEM analysis demonstrated the nucleation and growth of a continuous bonelike, low crystalline carbonated HA nanocrystals on the inner pore surfaces of the PLGA-(PEG-A_ SP)_n scaffolds. The amount of calcium binding, total mass and the mass of phosphate on experimental PLGA-(PEG-A_ SP)_n scaffolds at different incubation times in mSBF was significantly greater than that of control PLGA scaffolds. This nano-HA/PLGA-(PEG-A_ SP)_n composite shows some features of natural bone both in main composition and hierarchical microstructure. The A_ SP-PEG alt-prepolymer modified PLGA copolymer provide a controllable high surface density and distribution of anionic functional groups which would enhance nucleation and growth of bonelike mineral following exposure to mSBF. This biomimetic treatment provides a simple method for surface functionalization and subsequent mineral nucleation and self-assembling on biodegradable polymer scaffolds for tissue engineering.
