甲状旁腺激素(1-34)、多聚赖氨酸应用于口腔种植体表面促成骨作用研究

 目的 研究甲状旁腺激素(parathyroid hormone, PTH)(1-34)、多聚赖氨酸应用于种植体表面的促成骨作用。方法 将PTH(1-34)和多聚赖氨酸通过层层自组装的方式涂层到种植体表面。按照涂层制备方法和PTH(1-34)浓度将种植体分为4个组: PLL载体组(A组)、10 μg组(B组)、100 μg组(C组)和空白对照组(D组)。使用ELISA法探索该涂层的体外药物释放规律;将小鼠胚胎成骨前体细胞MC3T3-E1接种于涂层后的种植体表面,使用MTT法、碱性磷酸酶半定量、PCR等方法,检测该涂层在体外条件下对成骨细胞生物学行为的影响。结果 通过层层自组装技术于钛种植体表面制成的PLL-PTH(1-34)聚电解质多层膜涂层在初始阶段表现出明显的释放过程,随后释放程度趋于平缓,第14天时仍可检出PTH(1-34)。MTT检测:第5 天时C组OD值(1.738±0.026)与第1天(1.663±0.045)相比有明显升高(P=0.0219)。碱性磷酸酶半定量:第14天时,C组(3.377±0.336)U/gprot明显高于A组(2.537±0.083)U/gprot、B组(2.843±0.060)U/gprot和D组(2.063±0.061)U/gprot,差异均有统计学意义(P<0.05)。PCR检测:ALP基因于第7天时各组表达水平最高,且C组明显高于A、B、D三组,差异均有统计学意义(P<0.05);OCN基因于第14天时各组表达水平最高,且C组明显高于A、B、D三组,差异均有统计学意义(P<0.05)。结论 在钛种植体表面可以通过层层自组装技术制成PLL-PTH(1-34)聚电解质多层膜涂层。通过该技术可以实现PTH(1-34)较长时间的局部释放,从而促进其表面成骨细胞的增殖和分化。

A study on osteogenesis promoted by PTH (1-34) and polylysine coating on the surface of implant

Objective To promote osteogenesis, parathyroid hormone which can promote bone formation was used to process surface of implant material.Methods Parathyroid hormone PTH (1-34) and polylysine were coated to the surface of the implant material through layer by layer self-assemble technique. ELISA was used to explore the in vitro pharmacokinetics regularity of the coating. The mouse embryos osteogenesis precursor cells MC3T3 E1 were seeded on the coated implant material surface. MTT, alkaline phosphatase semi-quantitative and PCR Methods were used to detect the in vitro influence of osteoblast biology behavior.Results PLL - PTH (1-34) polyelectrolyte multilayer membrane coating made by layer by layer self-assemble technique showed significant release process in the initial phase, then the release leveled off, and PTH (1-34) could be detected until 14 d. MTT: OD value of group C (1.738±0.026) on the 5 d elevated compared with 1 d (1.663±0.045) (P=0.0219). Alkaline phosphatase semi-quantitative: activity of alkaline phosphatase in group C (3.377±0.336) U/gprot was higher than in group A (2.537±0.083) U/gprot, B (2.843±0.060) U/gprot and D(2.063±0.061) U/gprot; the difference was statistically significant (P<0.05). PCR: expression level of ALP in four groups were highest on 7 d, and in group C was higher than in groups A, B and D (the difference was statistically significant, P<0.05); expression level of OCN in four groups were highest on 14 d, and in group C was higher than in groups A, B and D (the difference was statistically significant, P<0.05).Conclusions PLL - PTH (1-34) polyelectrolyte multilayer membrane can be made on implant material surface through layer by layer self-assemble technique. PTH (1-34) can release for a long time (14 d) by this technique, so as to promote proliferation and differentiation of the osteoblasts seeded on the surface.