Laboratorio de Biología Celular y Tisular, Universidad Autónoma de Aguascalientes, Av. Universidad 940, Aguascalientes, 20131, Aguascalientes, Mexico.
In Vitro Cell Dev Biol Anim. 2011 Dec;47(10):681-8. doi: 10.1007/s11626-011-9459-7. Epub 2011 Oct 20.
A new system for sustained release of growth factors, such as osteogenic protein 1 (OP-1) and transforming growth factor β1 (TGF-β1), intended to repair and promote dental tissue regeneration in rats was designed and characterized in this work. The release system was made with microparticles of sodium alginate, produced by ionic gelling dripping technique. The release profiles of OP-1 and TGF-β1 from biopolymer matrix were determined by high-performance liquid chromatography (HPLC), and with this purpose, an HPLC-UV method was developed. About 80% of each growth factor was released in the first 24 h, reaching almost 100% in 168 h. The system was tested during the tissue repair in rat molars in comparison with calcium hydroxide and both growth factors not encapsulated. The dentin sialoprotein (DSP) was used as a repair marker. It was detected by immunohistochemistry, after 14- and 28-d post-treatment. X (2) test (p ≤ 0.001) and Fisher exact test (p ≤ 0.05) were applied for assessment of the amount of immunostaining. The treatment with encapsulated OP-1 showed an increased DSP immunostaining after 14 d and did not find any significant difference with the immunostaining observed for calcium hydroxide treatment. Treatment with TGF-β1 did not show significant difference with calcium hydroxide. Treatment with both factors OP-1 and TGF-β1 showed higher DSP immunostaining in comparison with calcium hydroxide treatment. In conclusion, the combination of both growth factors encapsulated showed more DSP immunostaining in comparison with each one separated, either encapsulated or not.
本研究设计并表征了一种新的生长因子（如骨形成蛋白 1（OP-1）和转化生长因子 β1（TGF-β1））持续释放系统，旨在修复和促进大鼠牙组织再生。该释放系统由海藻酸钠微球制成，采用离子凝胶滴注技术制备。采用高效液相色谱法（HPLC）测定生物聚合物基质中 OP-1 和 TGF-β1 的释放曲线，并为此目的开发了 HPLC-UV 方法。约 80%的每种生长因子在最初的 24 小时内释放，在 168 小时内达到近 100%。该系统在大鼠磨牙组织修复中进行了测试，与氢氧化钙和未包裹的两种生长因子进行了比较。牙本质涎磷蛋白（DSP）被用作修复标志物。用免疫组织化学法在治疗后 14 天和 28 天检测到。X（2）检验（p≤0.001）和 Fisher 确切检验（p≤0.05）用于评估免疫染色的数量。在第 14 天，与氢氧化钙治疗相比，包裹 OP-1 的治疗显示出更高的 DSP 免疫染色量，且没有发现任何显著差异。TGF-β1 治疗与氢氧化钙无显著差异。与氢氧化钙治疗相比，同时包裹两种因子 OP-1 和 TGF-β1 的治疗显示出更高的 DSP 免疫染色量。总之，与单独包裹或不包裹的两种生长因子相比，同时包裹两种生长因子显示出更高的 DSP 免疫染色量。
