Tanzi M C, Farè S, Petrini P, Tanini A, Piscitelli E, Zecchi Orlandini S, Brandi M L
Bioengineering Department, Politecnico di Milano, Milano - Italy.
J Appl Biomater Biomech. 2003 Jan-Apr;1(1):58-66.
This work reports preliminary results on the development of biointegrable scaffolds, composed of biostable 3D polymer matrices and bioabsorbable inorganic salts, to be used for cell anchorage in bone regeneration. Three crosslinked polyurethane foams (PUFs), prepared by one-step bulk polymerisation from a polyether-polyol mixture, polymeric MDI and water as expanding agent, were tested for their ability to promote adhesion and growth of bone-derived cells. The open porosity of these foams ranged from 16 to 31% with an average pore size of 470 /600 microm, compressive strength (at 10% ε ) of 0.28/0.38 MPa and elastic moduli of 4.88/6.61 MPa. The human osteosarcoma line Saos-2, and primary cultures of normal human articular chondrocytes and bone marrow-derived (HBM) stromal cells were used for in vitro cytocompatibility tests. For cell adhesion and proliferation analysis, DNA synthesis was evaluated by 3 H-thymidine uptake. Osteoblastic differentiation of Saos-2 adherent cells was determined by measuring the enzymatic activity of alkaline phosphatase (ALP). All cell types were able to adhere to all tested PUFs and to synthesize DNA. At 48 hr culture, HBM stromal cells showed the maximal rate of adhesion with the highest rate of proliferation onto PUFs with the largest pore size, whereas both chondrocytes and Saos-2 appeared to adhere preferentially onto foams exhibiting the highest percentage of open porosity. Up to 8 days in culture Saos-2 cells were able to proliferate into all PUFs, with a time-dependent increase of DNA synthesis and ALP activity. At SEM, the morphology of cells adherent to PUF pores was spread with cytoplasmatic extroflessions, indicating a good metabolic activation. These results demonstrate a good cytocompatibility of the proposed 3D matrices, suggesting that their use in the preparation of composite scaffolds is worth further investigation. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 58-66)ABSTRACT: This work reports preliminary results on the development of biointegrable scaffolds, composed of biostable 3D polymer matrices and bioabsorbable inorganic salts, to be used for cell anchorage in bone regeneration. Three crosslinked polyurethane foams (PUFs), prepared by one-step bulk polymerisation from a polyether-polyol mixture, polymeric MDI and water as expanding agent, were tested for their ability to promote adhesion and growth of bone-derived cells. The open porosity of these foams ranged from 16 to 31% with an average pore size of 470 /600 microm, compressive strength (at 10% ε ) of 0.28/0.38 MPa and elastic moduli of 4.88/6.61 MPa. The human osteosarcoma line Saos-2, and primary cultures of normal human articular chondrocytes and bone marrow-derived (HBM) stromal cells were used for in vitro cytocompatibility tests. For cell adhesion and proliferation analysis, DNA synthesis was evaluated by 3 H-thymidine uptake. Osteoblastic differentiation of Saos-2 adherent cells was determined by measuring the enzymatic activity of alkaline phosphatase (ALP). All cell types were able to adhere to all tested PUFs and to synthesize DNA. At 48 hr culture, HBM stromal cells showed the maximal rate of adhesion with the highest rate of proliferation onto PUFs with the largest pore size, whereas both chondrocytes and Saos-2 appeared to adhere preferentially onto foams exhibiting the highest percentage of open porosity. Up to 8 days in culture Saos-2 cells were able to proliferate into all PUFs, with a time-dependent increase of DNA synthesis and ALP activity. At SEM, the morphology of cells adherent to PUF pores was spread with cytoplasmatic extroflessions, indicating a good metabolic activation. These results demonstrate a good cytocompatibility of the proposed 3D matrices, suggesting that their use in the preparation of composite scaffolds is worth further investigation. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 58-66).
本研究报告了用于骨再生中细胞锚定的生物可整合支架的初步开发成果,该支架由生物稳定的三维聚合物基质和生物可吸收无机盐组成。通过一步本体聚合由聚醚多元醇混合物、聚合MDI和水作为发泡剂制备了三种交联聚氨酯泡沫(PUF),测试了它们促进骨源细胞粘附和生长的能力。这些泡沫的开孔率在16%至31%之间,平均孔径为470/600微米,压缩强度(10%应变时)为0.28/0.38兆帕,弹性模量为4.88/6.61兆帕。人骨肉瘤细胞系Saos-2、正常人关节软骨细胞和骨髓来源(HBM)基质细胞的原代培养物用于体外细胞相容性测试。对于细胞粘附和增殖分析,通过3H-胸苷摄取评估DNA合成。通过测量碱性磷酸酶(ALP)的酶活性来确定Saos-2贴壁细胞的成骨分化。所有细胞类型都能够粘附到所有测试的PUF上并合成DNA。在培养48小时时,HBM基质细胞在孔径最大的PUF上显示出最大的粘附率和最高的增殖率,而软骨细胞和Saos-2似乎优先粘附到开孔率最高的泡沫上。在培养长达8天的时间里,Saos-2细胞能够在所有PUF中增殖,DNA合成和ALP活性随时间增加。在扫描电子显微镜下,粘附在PUF孔上的细胞形态呈扩散状,有细胞质外突,表明代谢活性良好。这些结果证明了所提出的三维基质具有良好的细胞相容性,表明它们在复合支架制备中的应用值得进一步研究。(《应用生物材料与生物力学杂志》2003年;1:58-66)摘要:本研究报告了用于骨再生中细胞锚定的生物可整合支架的初步开发成果,该支架由生物稳定的三维聚合物基质和生物可吸收无机盐组成。通过一步本体聚合由聚醚多元醇混合物、聚合MDI和水作为发泡剂制备了三种交联聚氨酯泡沫(PUF),测试了它们促进骨源细胞粘附和生长的能力。这些泡沫的开孔率在16%至31%之间,平均孔径为470/600微米,压缩强度(10%应变时)为0.28/0.38兆帕,弹性模量为4.88/6.61兆帕。人骨肉瘤细胞系Saos-2、正常人关节软骨细胞和骨髓来源(HBM)基质细胞的原代培养物用于体外细胞相容性测试。对于细胞粘附和增殖分析,通过3H-胸苷摄取评估DNA合成。通过测量碱性磷酸酶(ALP)的酶活性来确定Saos-2贴壁细胞的成骨分化。所有细胞类型都能够粘附到所有测试的PUF上并合成DNA。在培养48小时时,HBM基质细胞在孔径最大的PUF上显示出最大的粘附率和最高的增殖率,而软骨细胞和Saos-2似乎优先粘附到开孔率最高的泡沫上。在培养长达8天的时间里,Saos-2细胞能够在所有PUF中增殖,DNA合成和ALP活性随时间增加。在扫描电子显微镜下,粘附在PUF孔上的细胞形态呈扩散状,有细胞质外突,表明代谢活性良好。这些结果证明了所提出的三维基质具有良好的细胞相容性,表明它们在复合支架制备中的应用值得进一步研究。(《应用生物材料与生物力学杂志》2003年;1:58-66)