角质形成细胞在与透明质酸和壳聚糖混合的聚(3-羟基丁酸酯)和聚(4-羟基丁酸酯)多孔膜上的生长。

Growth of keratinocytes on porous films of poly(3-hydroxybutyrate) and poly(4-hydroxybutyrate) blended with hyaluronic acid and chitosan.

作者信息

Peschel Gundela, Dahse Hans-Martin, Konrad Anke, Wieland Gerhard Dieter, Mueller Peter-Juergen, Martin David P, Roth Martin

机构信息

Leibniz Institute for Natural Product Research and Infection Biology, Hans Knoell Institute, Beutenbergstr.11a, 07745 Jena, Germany.

出版信息

J Biomed Mater Res A. 2008 Jun 15;85(4):1072-81. doi: 10.1002/jbm.a.31666.

Abstract

The objective of this study was to develop novel absorbable films suitable for use as a tissue-engineering scaffold for keratinocytes as a therapy for replacement of damaged skin. Poly(4-hydroxybutyrate) (P(4HB)) and poly (3-hydroxybutyrate) (P(3HB)) were blended with small amounts of the polysaccharides hyaluronic acid (HA), chitosan (CH), pectin and alginic acid, and were solution cast to produce porous films. The resulting composites had favorable mechanical properties, and these films were compared with two commercially available implantable films made of poly(L-lactide-co-D,L-lactide) (PLA copolymer) and HA benzyl ester. Tensile testing demonstrated that a high level of flexibility of P(4HB) was retained in the P(4HB)-polysaccharide composite films, whereas the P(3HB) film and its polysaccharide composites were stiffer and more brittle. The proliferation kinetics of adherent HaCaT keratinocytes on the films was examined in vitro. The porous surface of the P(4HB) and P(3HB) films blended with HA or CH promoted the growth of keratinocytes significantly. The order of maximum cell numbers on these films was P(4HB)/HA > P(4HB)/CH > P(3HB)/HA > P(3HB)/CH > P(3HB)/pectin > P(3HB)/alginic acid. Scanning electron microscopy and confocal laser scanning microscopy revealed differences in cell growth. Cells formed clusters on P(3HB) and its composites, while the cells grew as a confluent layer on P(4HB) and its composites. HaCaT cells formed large numbers of filaments only on P(4HB) films, indicating the excellent biocompatibility of this material. For the nonporous PHB films, the proliferation rate of cells was found to increase with decreasing hydrophobicity in the order: P(4HB) > P(3HB)/P(4HB) blend > P(3HB).

摘要

本研究的目的是开发新型可吸收薄膜,用作角质形成细胞的组织工程支架,以治疗受损皮肤的替代。将聚(4-羟基丁酸酯)(P(4HB))和聚(3-羟基丁酸酯)(P(3HB))与少量的多糖透明质酸(HA)、壳聚糖(CH)、果胶和海藻酸混合,并通过溶液浇铸制备多孔薄膜。所得复合材料具有良好的机械性能,并将这些薄膜与两种市售的由聚(L-丙交酯-co-D,L-丙交酯)(PLA共聚物)和HA苄酯制成的可植入薄膜进行比较。拉伸测试表明,P(4HB)-多糖复合薄膜保留了P(4HB)的高柔韧性,而P(3HB)薄膜及其多糖复合材料则更硬且更脆。在体外研究了贴壁的HaCaT角质形成细胞在薄膜上的增殖动力学。与HA或CH混合的P(4HB)和P(3HB)薄膜的多孔表面显著促进了角质形成细胞的生长。这些薄膜上最大细胞数的顺序为P(4HB)/HA > P(4HB)/CH > P(3HB)/HA > P(3HB)/CH > P(3HB)/果胶 > P(3HB)/海藻酸。扫描电子显微镜和共聚焦激光扫描显微镜揭示了细胞生长的差异。细胞在P(3HB)及其复合材料上形成簇,而细胞在P(4HB)及其复合材料上以汇合层生长。HaCaT细胞仅在P(4HB)薄膜上形成大量细丝,表明该材料具有优异的生物相容性。对于无孔的PHB薄膜,发现细胞的增殖速率随着疏水性的降低而增加,顺序为:P(4HB) > P(3HB)/P(4HB)共混物 > P(3HB)。

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